S uc u al Analysis o Nanopa icles
by Small Angle X- ay Sca e ing
Disse a ion
Zu E langung des akademischem G ades eines
Dok o s de Na u wissenscha en (D . e . Na .)
in Fach Chemie de Fakul ¨a ¨u Biologie, Chemie und Geowissenscha en
de Uni e si ¨a Bay eu h
o geleg on
Ch is ophe N. Roche e, M. Sc.
gebo en in Bo deaux, F ank eich
Bay eu h, Donne s ag den 17. No embe 2011
Die o liegende A bei wu de in de Zei on No embe 2005 bis No embe 2009 am
Leh s uhl ¨u Physikalische Chemie I de Uni e si ¨a Bay eu h un e Be euung on He n
P o . D . Ma hias Ballau ange e ig .
Volls ¨andige Abd uck de on de Fakul ¨a ¨u Biologie, Chemie und Geowissenscha en
de Uni e si ¨a Bay eu h genehmig en Disse a ion zu E langung des akademischen G ades
eines Dok o s de Na u wissenscha en (D . e . na .).
Disse a ion einge eich am: 17.11.2011
Zulassung du ch die P ¨u ungskommission: 15.12.2011
Wissenschla liches Kolloquiums: 14.05.2012
Am ie ende Dekan:
P o . D . Bea e Lohne
P ¨u ungsausschuss:
P o . D . M. Ballau (E s gu ach e )
P o . D . S . F¨o s e (Zweigu ach e )
P o . D . A. M¨ulle (Vo si z)
P o . D . B. Webe
2
This wo k is dedica ed o Jean de Be ie , my G and a he .
3
4

Con en s
1 In oduc ion 8
2 Theo y o SAXS 10
2.1 Basiso SAXS heo y.............................. 10
2.2 Sca e ing unc ion o monodispe se pa icles . . . . . . . . . . . . . . . . . 12
2.2.1 Sphe ical pa icles . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.2.2 Disk-like pa icles . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.2.3 Local luc ua ions............................ 13
2.3 Polydispe si y.................................. 13
2.4 In e pa icle in e ac ions . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
2.4.1 Agg ega ion............................... 14
2.4.2 Polyme Re e ence In e ac ion Si e Model - PRISM . . . . . . . . . 15
2.5 Con as Va ia ion ............................... 15
3 Gene ali ies 17
3.1 Calci ica ion................................... 17
3.2 Polybu adiene.................................. 21
3.3 Polye hylene................................... 22
4 S udy o he ea ly s age o calci ica ion 26
4.1 In es iga ion o he ea ly s age . . . . . . . . . . . . . . . . . . . . . . . . . 26
4.1.1 Ma e ials ................................ 27
4.1.2 Expe imen al .............................. 28
4.2 Resul s and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
4.2.1 Nuclea ion................................ 29
4.2.2 S uc u al e ec o Fe uin-A on o he CPPs . . . . . . . . . . . . . 33
5 Polybu adiene 41
5.1 Expe imen al .................................. 41
5.2 Theo e icalmodeling.............................. 42
5.3 Resul s and Discussions . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
5.3.1 C yo-TEM................................ 43
5
5.3.2 SAXS .................................. 44
5.4 Conclusion.................................... 46
6 Polye hylene 48
6.1 Expe imen al .................................. 48
6.2 Resul s and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
6.2.1 C yo-TEM................................ 49
6.2.2 SAXS .................................. 50
6.3 Va ia ion o annealing empe a u e . . . . . . . . . . . . . . . . . . . . . . 53
7 Expe imen al 58
7.1 SAXS ...................................... 58
7.1.1 K a ky-Compac -Came a . . . . . . . . . . . . . . . . . . . . . . . . 58
7.1.2 ID02................................... 58
7.2 Dynamic Ligh Sca e ing . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
7.3 Densi ome y .................................. 59
8 Summa y 61
8.1 Calci ica ion................................... 61
8.2 Polybu adiene.................................. 62
8.3 Polye hylene................................... 63
9 Zusammen assung 64
9.1 Calci izie ung .................................. 64
9.2 Polybu adien .................................. 65
9.3 Polye hylen ................................... 66
Appendices 67
A Theo y o SAXS 68
A.1 E ec o polydispe si y............................. 68
A.2 C++ p og ams.................................. 69
A.2.1 o m ac o o homogenous sphe ical polydispe se pa icles . . . . . 69
A.2.2 s uc u e ac o o he agg ega ion o sphe ical pa icles . . . . . . . 71
A.3 Modi ied hambu ge model . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
B Addi ional expe imen al in o ma ion 76
B.1 TEM pic u es o calcium phospha e complexes . . . . . . . . . . . . . . . . 76
B.1.1 Wi hou addi ion o Fe uin-A . . . . . . . . . . . . . . . . . . . . . 77
B.1.2 Wi h 30 µMo Fe uin-A........................ 78
B.2 WAXS signal o calcium phospha e complexes . . . . . . . . . . . . . . . . 79
6
B.3 Syn hesis o he polye hylene nanopa icles . . . . . . . . . . . . . . . . . . 80
B.4 In luence o he annealing p ocess on N
V.................... 81
B.5 Con as se ieso PL78............................. 82
B.5.1 O iginal sys em - PL78 . . . . . . . . . . . . . . . . . . . . . . . . . 82
B.5.2 Sys em annealed a 90◦C........................ 83
B.5.3 Sys em annealed a 105◦C ....................... 84
B.5.4 Sys em annealed a 115◦C ....................... 85
B.6 Syn hesiso sPB ................................ 86
B.7 X- ay di ac ion o BK280 . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
B.8 DLSo BK280.................................. 88
B.9 Models in es iga ed o he SAXS-CV o BK280 . . . . . . . . . . . . . . . 89
7
Chap e 1
In oduc ion
Du ing he las decades, Small Angle X- ay Sca e ing (SAXS) became a powe ull ech-
nique in colloidal science o he de e mina ion o shape, size and in e nal s uc u es o
nanoscale objec s in he size ange o ew nanome e s [1] up o abou 300 nm [2].
Calci ied pa icles ha e been ex ensi ely s udied wi hin he las decades [3–5]. Thei
o ma ions a e o impo ance o clinical easons and a e in es iga ed om he second
pa o las cen u y [6–8]. Amo phous calcium phospha e o ma ion has been he subjec
o nume ous in es iga ions in he pas conce ning hei s uc u e [9,10], s abili y [9] and
ans o ma ion in o o he c ys alline phases [11]. Howe e , he e olu ion o hese com-
plexes emains poo ly unde s ood.
Few yea s ago, he o ma ion o calcium ca bona e p ecu so pa icles ha e been suc-
cess ully p esen ed by Time-Resol ed-SAXS (TR-SAXS) [12] and i has been shown ha
a double hyd ophilic block-copolyme is esponsible o he slow down o he agg ega ion
o hese pa icles [13]. The au ho s o e e ence [12,13] demons a ed ha he o ma ion
o calci ied objec s can be moni o ed and in es iga ed as well by he help o TR-SAXS ex-
pe imen s. La ely, he in e ac ion be ween p o eins and calcium phospha e pa icles has
been disco e ed [14,15] bu no s udy in es iga ed he ea lies s age o calcium-phospha e
o ma ion up o ha ime [16,17].
The i s pa o his hesis is o in es iga e he o ma ion a he ea lies s age o cal-
cium phospha e pa icles and o s udy he e ec o a p o ein called Fe uin-A on o he
calci ica ion p ocess. TR-SAXS by using a Synch o on sou ce was used o explo e his
o ma ion.
Polybu adiene is one o he mos syn hesized polyme sys em which is ound o in-
s ances in y es [18,19]. Since he Second Wo ld Wa , new syn heses in bulk o hese sys-
8
2.5 Con as Va ia ion Theo y o SAXS
Ndeno es he e he o al numbe o pa icles. g( ) is he pa icle pai -co ela ion unc ion
and desc ibes he p obabili y o ha ing wo pa icles sepa a ed by a dis ance 2 om each
o he . By aking in o accoun he size ξo he clus e , he pai -co ela ion unc ion is
ende ed as:
g( ) = A
d−d e− /ξ (2.19)
whe e Ais a cons an . Using he ela ion in equa ion 2.18 and pe o ming a Fou ie
ans o m esul o:
S(q) = 1 + S0
[1 + q2ξ2](d −1)/2
sin[(d −1) an−1(qξ)]
(d −1)qξ (2.20)
wi h S0de ines as:
S0=C(d −1)Γ(d −1)ξd (2.21)
whe e
Γ(x) =
∞
Z
0
x−1e− d (2.22)
and C is a cons an . Since
lim
x→0
sin( an−1(x))
x= 1 (2.23)
S(0)=1+S0de ines he numbe o pa icles pe agg ega e.
2.4.2 Polyme Re e ence In e ac ion Si e Model - PRISM
The PRISM heo y is de i ed om he Re e ence In e ac ion Si e Model (RISM) de el-
opped by Chandle and co-wo ke s [40–44]. In his pa , only he gene al aspec behind
he RISM will be desc ibe.
The PRISM bases on liquid s a e in eg al equa ion heo y which was o iginally de el-
opped o a omic and small molecule luids [45]. Gi en an in e pa icle po en ial, he
heo y p o ides an in e pa icle pai co ela ion unc ion. This pai co ela ion unc ion
is connec ed wi h he s uc u e ac o S(q) measu ed in SAXS by Fou ie ans o m. In
he pa icula case o his s udy, he PRISM desc ibes he equilib ium s uc u e and p op-
e ies o polyme s in bulk solu ion. The main app oxima ion he eby is ha all sides o
a pa icle a e equi alen . Thus he pa icle in e ac ion p oblem is educe o a simple
p oblem. Mo e in o ma ion abou he PRISM heo y and applica ion could be ound in
e e ences [45,46].
2.5 Con as Va ia ion
The sca e ed in ensi y I(q) o an ensemble o N pa icles o olume VPin a olume V is
gi en by:
15

2.5 Con as Va ia ion Theo y o SAXS
I(q) = N
V(∆ρ)2V2
PP(q)S(q) (2.24)
wi h ∆ρ=ρs-ρmwhe e ρsand ρmde ine he mean elec on densi y o he sys em and he
medium. Acco ding o equa ion 2.24, a ying he elec on densi y o he medium ρm
lead o di e en esul ing in ensi ies. Figu e 2.3 displays a schema ic iew o he con as
a ia ion echnique: changing he elec on densi y o he sol en co esponds o a change
o he colo o he backg ound. The e a e wo ex eme cases: in ini e con as and ze o
con as . A a con as o ±∞ (case a and e in igu e 2.3), only in o ma ion abou he
o e all size and shape could be ob ained. When he elec on densi y o he sol en is equal
o one he mean elec on densi y o he pa icle, hen only he de i a ions, due o di e en
in e nal con as , a e isible. Measu ing he same sys em wi h di e en con as lead o
a comple e in es iga ion o he s uc u e o he e ogenous sys ems. In p axis, con as
a ia ion is no o en used in con en ional SAXS due o he di icul y o change only he
elec on con as ρmo he medium wi hou modi ying he s uc u e o he in es iga ed
sys em in he same ime.
Figu e 2.3: Schema ic ep esen a ion o he con as a ia ion. Di e en elec on densi ies
o he sol en help o de ec subs uc u es o he sys em. This me hod pe mi s hen o
s udy he o al s uc u e and he di e en phases p esen in he analyzed pa icles in
de ail. ∆ρde ines he elec on con as o he s udied sys em ela i e o he medium.
16
Chap e 3
Gene ali ies
3.1 Calci ica ion
Calci ica ion is he p ocess in which he mine al calcium builds up in so issue, caus-
ing i o ha den. The i s appea ance o biomine aliza ion in His o y is documen ed in
he P ecamb ian in e eb a e Cloudina [47], a shelly ube-like ossil. In o de o con ol
his mine aliza ion, po en inhibi o s o spon aneous calci ica ion mus exis . The mul i-
plici y o he exis ing phases o calcium phospha e complexes (see able 3.1) is as well an
impo an pa ame e in he complexi y o he chemical equilib ium engaged. Ve eb a es,
including Human, con ain hese phases which a e ound mos ly e eywhe e in he body
(bones, ligamen s and e en muscles) bu a di e en concen a ions. The mechanism o
egula ion o calcium and phospha e concen a ion in he Ve eb a es is eally a om
being comple ely unde s ood.
Phase Ac onym Empi ical o mula Ksp
Amo phous calcium phospha e ACP Ca3(PO4)2·xH2O -
Dicalcium phospha e dihyd a e DCPD CaHPO4·2H2O 1.87 ×10−7M2
Dicalcium phospha e anhyd ous DCPA CaHPO49.2 ×10−7M2
β- icalcium phospha e TCP Ca3(PO4)29.2 ×10−29 M5
Oc acalcium phospha e OCP Ca8H2(PO4)6·5H2O 2.5 ×10−99 M8
Hyd oxyapa i e HAP Ca10(PO4)6(OH)25.5 ×10−118 M9
Table 3.1: Di e en phases o calcium phospha e complexes and hei espec i e solubili y
p oduc s Ksp.Muni is mol/L. Da a ob ained om e e ence [48].
The unde s anding o he o ma ion o calcium phospha e complexes is ha dly in es i-
ga ed especially since he middle o las cen u y. In 1967, Wal on and co-au ho s demon-
s a ed ha Hyd oxyapa i e and Oc acalcium phospha e may no be he ini ial phase o
calcium phospha e complexes [49]. The e is no doub abou he na u e o p ecipi a ion o
hese ions a ea ly s age o calci ica ion bu his ini ial p oduc is s ill no p ecisely known
and depends mainly on pa ame e s such as empe a u e, pH o sol en . I is now gene ally
17
3.1 Calci ica ion Gene ali ies
ecognized ha he i s compound o med om soluble sal s is a me as able p ecu so
phase [48]. This p ecu so phase, also called Amo phous Calcium Phospha e (ACP), has
been widely s udied by changing he calcium o phospha e a io [50], hei ini ial mola
p oduc [6] o he expe imen al empe a u e [51]. Kine ic s udies o he s uc u a ion o
calcium phospha e complexes has been pe o med ecen ly [52]. These au ho s mixed Di-
calcium Phospha e DiHyd a e (DCPD) wi h Calcium Oxide (CaO) and de ec ed he o -
ma ion o nanoHyd oxyApa i e (nano-HA) pa icles wi hin ew hou s by he combina ion
o X-Ray Di ac ion (XRD) and Di e en ial Scanning Calo ime y (DSC) measu emen s.
The s uc u a ion o he ini ial calcium phospha e complexes (DCPD) is e y simila o
ACP acco ding o he p esen s udy (see page 29 in chap e Calci ica ion a ea ly s age).
Vascula calci ica ion (VC), ha is deposi ion o calcium phospha e mine al in ca dio-
ascula issues including a e ies, hea al es and cad iac muscles, is o en encoun e ed
in he de eloppmen o a he oscle o ic in imal lesions and is a common consequence o
aging [53]. VC is posi i ely co ela ed wi h inc eased isk o myoca dial in a c ion and o
dissec ion a e angioplas y [54]. In 2000, Jono and cowo ke s demons a ed ha di e en
le els o phospha e egula es human smoo h muscle cell calci ica ion h ough a sodium-
dependen phospha e anspo e -sensi i e mechanism and implica e his mechanism in
he de eloppmen o ec opic calci ica ion in i o [55].
Kine ically and s uc u ally speaking, he o ma ion o he p ecu so pa icles o cal-
cium phospha es complexes leading o he p ecipi a ion o he ions is unclea . A huge
numbe o scien i ic a icles a e p oposing Mon e-Ca lo simula ions in o de o be e in-
e p e such a beha iou (see o ins ance e e ences [56, 57]). Au ho s o e e ence [57]
demons a ed ha he solu ion may be di ided in o h ee egimes: he i s one p esen s
in idual monome s in solu ion, in he second egime, small clus e s o monome s a e o m-
ing and in he las case, la ge pa icles a e o med.
Howe e , he calci ica ion p ocess is a much mo e complica ed and i is e iden ha
addi ional molecules play a signi ican ole in he o ma ion o ee h o bones [58]. The
o ma ion o calcium phospha e complexes has been in es iga ed by using unc ional-
ized mac omolecules as empla es. Fo ins ance, Hol and cowo ke s [59] used β-casein
phosphopep ides as s ablising agen and demons a ed ha he pep ide co e ed nanopa -
icles o calcium phospha e. Enlow e al. [60] c ea ed an o ganized ne wo k o 20 nm
diame e calcium phospha e nanosphe es by he help o copolyme s, Li [61] showed ha
β-cyclodex in is he only mac omolecule o his amily which is able o s abilize he
amo phous phase o ACP. In 2002, Combes and Rey s udied he g ow h o calcium phos-
pha e complexes in p esence o BSA p o eins and p oposed a schema ical ep esen a ion
o c ys alline OCP co e ed by an adso bed laye o BSA o p e en u he g ow h o he
nanopa icles [62]. The long way o unde s and he complexi y o calci ica ion is a om
being achie ed.
18
3.1 Calci ica ion Gene ali ies
Figu e 3.1: Model o he h ee domains o Fe uin-A. D1 and D2 emain o he Cys a in
Supe amily while D3 has a s uc u al homolgy wi h an inse ion domain. Pic u e aken
om e e ence [63].
Since some yea s, he p ope ies o a pa icula p o ein a e o ema kable in e es in
he ield o mine aliza ion: α2-HS-glycop o ein (ahsg) also called Fe uin-A (see s uc u al
model o he p o ein in igu e 3.1). The name α2-HS-glycop o ein e e s o he ac ha
his p o ein mig a es wi h he α2 ac ion o se um p o eins upon adi ional cellulose
ace a e pape based elec opho esis. H and S eminds o He emans [65] and Schmid [66],
Figu e 3.2: Hypo he ical model o a calcip o ein pa icle (CPP) consis ing o agg ega ed
calcium-phospha e-Fe uin complexes. Figu e aken om e e ence [64].
19
3.1 Calci ica ion Gene ali ies
Figu e 3.3: Radiological analysis o 9-mon hs-old mice. The mouse in he igh is gene i-
cally de icien in Fe uin-A p oduc ion while he one on he le is a wild ype mouse. The
lack o Fe uin-A is cha ac e ized by a s ong calci ica ion in ex acelula space. Pic u e
aken om e e ence [72].
he co-disco e s o his p o ein in humans [67]. The name Fe uin is coming om he la in
wo d e us and has been gi en o his p o ein o i s abundance in e al cal se um [68].
Finally, his glycop o ein is called Fe uin-A a e he ecen disco e y o a second Fe-
uin, Fe uin-B [69]. The abundance o Fe uin-A in bone sugges s ha he glycop o ein
may ha e a ole in bone o ma ion o emodeling. In 1996, Schinke and cowo ke s [70]
we e he i s o sugges a possible ole o inhibi o in mine aliza ion o his p o ein.
In he ollowing yea s, i has been shown ha Fe uin-A ac s as a sys emic inhibi o o
calci ica ion [15,71] and ha Fe uin-A is esponsible o he o ma ion o a Fe uin-mine al
complex called calcip o ein pa icle (CPP [64]). An hypo he ical model o CPP is dis-
played in igu e 3.2. A ecen wo k [72] showed ha mice gene ically modi ied no o
syn hesize Fe uin-A displayed a e 4 mon hs a se e e sys emic calci ica ion pheno ype
(see igu e 3.3).
The i s pa o his hesis will p esen a s udy o he in luence o Fe uin-A on o he
ea lies s age o calci ica ion by he combina ion o ime- esol ed SAXS measu emen s,
dynamic ligh sca e ing and ansmission elec on mic oscopy.
20

3.2 Polybu adiene Gene ali ies
3.2 Polybu adiene
Depending on he s uc u al a ia ions o he componen s in he polyme ic ma e ials,
he chemical s uc u es o polyme s is subjec o change. Polybu adiene (PB) consis s o
h ee isome ic uni s: cis-1,4, ans-1,4 and 1,2- inyl. Fu he mo e, he 1,2- inyl s uc u e
has h ee possible sequence a angemen s along he backbone chain: iso ac ic-, syndio-
ac ic and a ac ic-1,2 uni s [73,74]. O he ca alys sys ems ha can p oduce syndio ac ic
polyme s wi h con ollable cons i u ion and con igu a ion ha e been used and some wo ks
on syn hesis, he mal beha io , c ys alliza ion, s uc u e and mo phology o syndio ac ic
polybu adiene (sPB) ha e been published [75–79].
sPB is a he moplas ic elas ome o indus ial in e es due o i s p ope ies o bo h
plas ics and ubbe s. This polyme may be ound in “packaging b ea hing” i ems o
ui s, ege ables and sea ood because o i s highe ca bon oxide gases and oxygen pe me-
abili y and be e esis ance agains we ing and slippage, in molding applica ion such as
molded bo les, adhesi e, oil pain , pho osensi i e esin, plas ic ma e ials, i e eadings
in adhesi es o in oo wea s o ins ance [80, 80, 81]. The adjec i e syndio ac ic means
ha c ys alline sPB has a s e eo egula s uc u e in which he side-chain inyl g oups
a e loca ed al e na i ely on he opposi e sides in ela ion o he polyme ic main chain.
Figu e 3.4 p esen s a schema ical ep esen a ion o sPBD.
Figu e 3.4: Schema ical ep esen a ion o syndio ac ic 1,2-poly(1,3-bu adiene). We no e
a egula al e na ion o he CH-CH2bond behind and be o e he plane o he acyl chain.
sPB was i s syn hesized by Na a [82] in 1955 and i s s uc u e was de e mined one
yea a e by he help o X- ay di ac ion [83]: o ho hombic packing (Pacm, a=10.98 ˚
A,
b=6.60 ˚
Aand c=5.14 ˚
A). Such polyme s samples a e now syn hesized wi h a pu i y o 1,2
con en o 97% (see e. g. e e ence [84]). Up o now, li le a en ion has been paid o he
s uc u a ion o nanopa icles o sPB [74,85,86].
Using an in-si u cobal ca alys [87], e y small semic ys alline nanopa icles o sPB
could be syn hesized (see syn hesis o hese nanopa icles in appendix page 86). These
pa icles ha e a diame e o he o de o 14 nm and do p esen a c ys allisa ion pe cen age
a ound 50% [20]. The second pa o his hesis is o p esen he s uc u al analysis o
hese sPB nanopa icles by using he combina ion o SAXS and c yo-TEM expe imen s.
21
3.3 Polye hylene Gene ali ies
3.3 Polye hylene
C ys alliza ion o polye hylene (PE) is among he classical subjec s o polyme sci-
ence. In 1957, Till [88], Fishe [89] and Kelle [90] independen ly demons a ed ha PE
c ys allizes by chain olding leading o a lamella s uc u e. In o de o ini ia e he spon-
aneous o ma ion o chain- olded lamellae, he c ys alliza ion empe a u e needs o be
high enough o pe mi he equisi e molecula mo ions.
In he special case o su ace nuclea ion, he la e al su ace ee ene gy σcould be
w i en as [91,92]:
σ=T∆h
Tm
a0
2
lb
lu
1
C∞
(3.1)
whe e ∆h is he hea o usion, Tm he mel ing poin , a0 he wid h o he chain, lb he
bond leng h, lu he C-C dis ance and C∞is an empi ical pa ame e de e mined o 6.7
o he special case o polye hylene [93]. Two di e en ypes o olds a e desc ibed in
he li e a u e [93]: sha p- old and igh - old. The sha p- old is de ined as an eme gen
chain ha execu es a ansi o an adjacen whe e i e-en e s he lamella o o igin. The
igh - old is desc ibed as one eme gen chain ha e-en e s wi hin he lamella o o igin
wi h a mimal a e se leng h wi h li le amo phous con ibu ion (See he schema ical
ep esen a ions in igu e 3.5).
Figu e 3.5: Schema ical ep esen a ion o sha p- and igh - olded chains o c ys alline
polyme s.
The kine ic o he g ow h o he c ys alli es along he la e al axis depends mainly o
he ep a ion in he mel . The ep a ion e e s o he de o ma ion o he chains so as o
c ea e he old i sel . This ep a ion highly depends on he iscosi y ηwi hin he amo -
phous phase o he polyme and hus on he molecula weigh o he polyme . Fo he
case o polyme s wi h high molecula weigh , he ep a ion may occu on di e en pa s
o one chain and, a e a c i ical pe iod o ime, he esul ed s uc u es may be emo ed
leading o a dec ease o he kine ic o he g ow h o he c is alli es. The eade may e e
o pa II o e e ence [93] o mo e de ails abou he ep a ion.
22
3.3 Polye hylene Gene ali ies
Solu ion-g own c ys als o PE usually ha e la e al dimensions o he o de o some
mic ome e s and a e di icul o handle. A low empe a u e, PE c ys als p ecipi a e on
cooling o o m pla ele s which ha e he appea ance o a lozenge [94]. Such a mo phology
could be explain as shown in igu e 3.6 [95–99]: The aces 100 disappea as hey g ow
as e han he 110 esul ing o a lozenge shape. In he li e a u e, he c ys alline hickness
o PE a ies be ween 10 and 20 nm and a co ela ion be ween he in e se o his hickness
and he empe a u e o he syn hesis has been ound [100].
Figu e 3.6: Schema ic g ow h o a nascen PE nanoc ys al. The aces 100 disappea as
hey g ow as e han he 110 (le ), esul ing o a lozenge shape ( igh ). The le e G
e e s o he g ow h along he ace.
Du ing he las decades, he mechanism o ec ys alliza ion o lamella PE c ys als
a e mel ing has been ex ensi ely s udied [101,102]. Howe e , he p ocess in which e-
c ys alliza ion occu s is no ye ully unde s ood. In 1975, Windle analyzed he annealing
o mul i-lamella PE c ys allized in solu ion by an X- ay di ac ion s udy and sugges ed
an asymme ic s ep-like hickening o he c ys alline phase [103]. Twen y yea s ago,
Sadle and Spells demons a ed by he combina ion o neu on sca e ing and in a ed
spec oscopy ha a localized solid-s a e ans o ma ion is in ol ed du ing hea ing close
o he mel ing empe a u e [104, 105]. In 1997, Ras ogi and cowo ke s showed ha PE
c ys alline lamellae a e doubling in size du ing annealing and explained i by s acking
o c ys alline adjacen lamellae [106]. In 2001, mic oscopic s udies [107] show ha he
annealing p ocess may occu a empe a u e sligh ly abo e he c ys alliza ion empe a-
u es and a below he mel ing empe a u es o he c ys als. Howe e , none o hese
s udies explain he mo phology o swiss-cheese-like shapes obse ed a e he annealing o
lozenge-like c ys alline PE pa icles [101]. Mos o he p e ious s udies so a ha e been
pe o med on PE bulk o by means o mac oscopic PE c ys als.
Up o he end o las cen u y, polyme la exes we e polyme ized exclusi ely by ee-
adical p ocesses [108,109]. Since he begining o he 21s cen u y, a new ou e o polyme -
23
3.3 Polye hylene Gene ali ies
iza ion o PE pa icles eme ged: ca aly ic polyme iza ion in aqueous dispe sion [30–32].
S able aqueous dispe sions o su ac an -s abilized polyme nanopa icles in he ange o
50 o 500 nm diame e a e ob ained. In 2007, Webe and cowo ke s [33] p esen ed a com-
ple e analysis o hese PE nanopa icles by using a combina ion o c yo-TEM and SAXS
expe imen s. These pa icles consis o a ema kably hin c ys alline single laye o PE
(6.5 nm) sandwiched be ween wo amo phous polyme shee s (“nano-hambu ge s”).
The modynamically, he o ma ion o he polyme c ys alli es had become o a s ong
in e es in he las ew yea s [110,111]. In 2009, S obl p oposed an hypo he ical scheme
(see igu e 3.7), depending exclusi ely on he empe a u e T and on he numbe o s uc-
u e uni pe c ys alline chain n, ha migh explain he o ma ion and ec ys alliza ion o
polyme c ys alline pa icles [34]: A p ecu so o ma ion o a mesomo phic inne s uc u e
(poin (1) in igu e 3.7) occu s and polyme chains ea ange hemsel es o quasi-s e ched
con o ma ions bu no close enough o each o he o o m a densi y equi alen o he c ys-
al i sel (be ween poin s (1) and (2)). Once he s e ched chains each a ce ain limi ,
he laye s hicken o o m he na i e c ys alline phase (poin (2)). The passage om he
c ys alliza ion line up o he ec ys alliza ion one is s ill ma e o deba e. In 2005, S obl
assumed ha no elonga ion o he c ys alline hickness occu s du ing he annealing while,
Figu e 3.7: The modynamic scheme o polyme c ys als p oposed in li e a u e [34]. n
de ines he numbe o s uc u e uni pe c ys alline chain, see ex o u he explana ions.
This scheme is s ill unde deba e especially he pa hway om poin 2 o 3.
24
4.2 Resul s and discussion Calci ica ion a ea ly s age
size a ound 100 nm. Due o he expe imen al p ocedu e, he ea lies DLS measu emen s
we e achie ed ca. 30 seconds a e he mixing p ocess. No change in he size o he
o med pa icles we e isible du ing he i s ou hou s a e he mixing p ocess. Thus,
DLS measu emen s do no pe mi o de ec he nuclea ion o he p ima y pa icles.
Figu e 4.4: E olu ion o he SAXS in ensi ies wi h ime in he abscence o Fe uin-A a
he e y ea ly s age. The g ow h in in ensi y is cha ac e is ic o he nuclea ion phase,
id es , he o ma ion o he i s p ecu so pa icles. The ime poin o measu emen is
epo ed in he legend.
Time- esol ed SAXS expe imen s by using a synch o on sou ce ha e been used in o de
o de ec he o ma ion o calcium-phospha e complexes. Figu e 4.4 p esen s he sca e ed
in ensi ies eco ded o he sample con aining no Fe uin-A a he e y ea ly calci ica ion
s age, du ing he i s 0.05 second. The e a e wo main poin s: a g ow h in in ensi y
a all q- alues which is cha ac e is ic o a g ow h o he numbe o pa icles. And he e
is absolu ely no change in he shape o he sca e ed in ensi ies, which means ha he
mo phology and he size o he pa icles do no e ol e. A q- alues below 0.15 nm−1, he
h ee ea lies sca e ed in ensi ies o e lap hemsel es. This is a ibu ed o he emaining
low in he capilla y (quasi s eady s a e no eached). This c ea es a e ac s in he signal.
No u he change in in ensi y we e seen o longe imes meaning ha he numbe o
pa icles pe olume s ays cons an . This shows he e y as kine ic o he nuclea ion o
calcium phospha e pa icles. As well, no oscilla ion o he sca e ed in ensi ies could be
seen which is ypical o a e y high polydispe si y in size o he sys em. The sca e ed
in ensi ies in igu e 4.4 we e ob ained by changing he ini ial so-called dead- ime, ha is
he ime be ween he end o he mixing p ocess and he ime co esponding o he i s
measu emen . The in ensi y I(q) is p opo ional o he numbe o pa icles pe olume N
V,
o he squa e o he elec on con as o he sys em ∆ρand o he squa e o he olume
o he pa icle VP(see equa ion 2.24 page 16). In he special case o nuclea ion, he
31

4.2 Resul s and discussion Calci ica ion a ea ly s age
size o he sys em and i s densi y a e cons an and he s uc u e ac o S(q) is equal o
uni y. The sca e ed in ensi y is hen di ec ly p opo ionnal o he numbe o pa icles
pe olume N
V. The ime-e olu ion o he sca e ed in ensi y a a ce ain q- alue pe mi s
hen o quali a i ely s udy he nuclea ion p ocess. A q- alues below 0.15 nm−1, a e ac s
appea and hey a e a ibu ed o he emaining low in he capilla y ( igu e 4.4). Da a a
q- alues highe han ca. 0.45 nm−1a e mo e subjec o noise and hese in ensi ies will be
la e on a ibu ed mainly o he densi y luc ua ion o he amo phous sys em. I esul s
ha he nuclea ion o his sys em could be s udied by ollowing he sca e ed in ensi ies
eco ded be ween 0.15 and 0.45 nm−1.
Figu e 4.5: Time-e olu ion o he in ensi ies collec ed a q=0.25 nm−1 o he ea lies
SAXS in ensi ies collec ed. The o ma ion o he p ima y pa icles ollows a kine ic o
i s o de . The kine ic cons an k is di ec ly p opo ionnal o he slope and is dec easing
wi h he amoun o Fe uin-A. Da a collec ed o he sys em wi hou added p o ein (blue)
is compa ed o he sys ems including 1 µM ( ed), 5 µM (black) and 15 µM (g een) o
Fe uin-A.
Conside ing as i s s ep he o ma ion o dicalcium phospha e dihyd a e (DCPD), and
since he mola a io used implies mo e calcium ions, he limi ing ion du ing he o ma-
ion o he p ecu so phase is HPO2−
4. Thus, he nuclea ion is di ec ly p opo ional o
he ini ial phospha e concen a ion and o he kine ic cons an ko he i s o de . SAXS
in ensi ies a q=0.25 nm−1ha e been ollowed o each sample a he e y ea ly s age o
calci ica ion du ing he i s 0.05 second and a e displayed in igu e 4.5. The cons an k
is di ec ly p opo ionnal o he slope o hese se s o da a. Wi hin he limi o e o , i
seems e iden ha Fe uin-A has an in luence om he e y begining o he nuclea ion p o-
cess: The lowe he kine ic cons an , he highe amoun o he glycop o ein (see igu e 4.5).
32
4.2 Resul s and discussion Calci ica ion a ea ly s age
Figu e 4.6: In luence o Fe uin-A on o he size o he CPPs. The DLS se up was he -
mos a ed a 37◦C and 1.8 mL o calcium solu ion (wi h he app op ia e concen a ion
o Fe uin-A) was inse ed in a glass cell. A solu ion o phospha e ions was as well he -
mos a ed a 37◦C and 0.2 mL o his solu ion we e inse ed in he cell. The solu ion was
hen mixed by hand and he DLS measu emen s s a ed ca. 30 seconds a e he mixing
p ocess. The inal concen a ion o calcium and phospha e ions was espec i ely 10 mM
and 6 mM. The sys em wi hou added p o ein (da k blue) is compa ed o he ones wi h
1µM ( ed), 5 µM (black), 15 µM (g een) and 30 µM (ligh blue) o Fe uin-A.
4.2.2 S uc u al e ec o Fe uin-A on o he CPPs
In his pa , a compa ison be ween he esul s ob ained in e e ence [17] and he da a
eco ded in p esence o di e en amoun s o Fe uin-A om 1 µM up o 15 µM, he la e
co esponding o a physiological concen a ion o he p o ein, will be ealized.
Size o CPPs in p esence o Fe uin-A
In his pa ag aph, he ionic concen a ion o calcium and phospha e a e he same han in
he wo k o Heiss [17]. The e ec o he o e all size o he CPPs ha e been in es iga ed
by using DLS measu emen s and is p esen ed in igu e 4.6 page 33. A d as ical change in
he size o he pa icles is obse ed. None o he expe imen s in ol ing 10 mM o Ca2+
and 6 mM o PO3−
4led o a mean hyd odynamic adius o less han 20 nm.
Table 4.1 p esen s he di e en mean alue o he hyd odynamic adii ob ained when
11.1 mM o calcium and 60 mM o phospha e ions a e mixed ( olume a io 9:1 o ge a
calcium- o-phospha e a io o 10/6) in p esence o di e en concen a ions o he glyco-
p o ein. The sizes epo ed in his able co espond o he ones once he i s equilibi um
s a e is eached, ca. 60 seconds a e he mixing p ocess. As expec ed by he wo k o
Heiss and cowo ke s [17], he e is a dec ease o he sizes o he o med pa icles wi h he
33
4.2 Resul s and discussion Calci ica ion a ea ly s age
inc ease o he concen a ion o Fe uin-A. S udies abou he size o he p ima y pa icles
is missing in li e a u e. Addi ional expe imen s by DLS ha e been in es iga ed in o de
o es ima e his size.
Single pa icles
Figu e 4.7: DLS measu emen s o CPPs ob ained by mixing 10 mM o Ca2+ wi h 6 mM
o PO3−
4. The in luence o he addi ion o 4 µM o Fe uin-A ( ed) is well highligh ed
compa ed o he sys em wi hou added p o ein (blue).
In his pa ag aph, lowe concen a ions o calcium and phospha e ions ha e been used
bu he calcium- o-phospha e- a io is kep a 10/6.
As p e iously shown in igu e 4.3 page 30, he weigh ac ion o calcium and phospha e
ions ha e a s ong impo ance on o he agg ega ion p ocess. Se e al a emp s o DLS
measu emen s we e ealized by using di e en amoun s o Fe uin-A. As an example, ig-
u e 4.7 p esen s he esul s o DLS measu emen s pe o med by mixing 10 mM o calcium
ions wi h 6 mM o phospha e ions wi hou any glycop o ein and wi h 4 µM o Fe uin-A.
[Fe uin-A] (µM) RH(nm)
0>1000
1 150
5 90
15 70
30 45
Table 4.1: Hyd odynamic adius o he pa icles o med depending on he concen a ion
o he glycop o ein. The mix was ealized a 37◦C by using solu ions o 11.1 mM o
calcium (1.8 mL) and 60 mM o phospha e (0.2 mL).
34
4.2 Resul s and discussion Calci ica ion a ea ly s age
A he excep ion o he sys em wi hou any addi ion o p o ein, all measu emen s in-
cluding 1 µM o Fe uinA lead o a mean hyd odynamic adius o ca. 10 nm a oom
empe a u e and a 37◦C. These esul s a e in ag eemen wi h TEM mic og aphs which
we e aken a oom empe a u e o he sys ems wi hou and wi h 30 µM o Fe uin-A
( igu e 4.8).
Figu e 4.8: TEM mic og aph o a sample ob ained by mixing 20 mM o Ca2+ wi h 12 mM
o HPO2−
4wi hou addi ion o Fe uin-A (le ) and in p esence o 15 µM o he glycop o ein
( igh ).
The le pa o igu e 4.8 p esen s a TEM mic og aph o he sys em wi hou any addi ion
o Fe uin-A. Agg ega es o sphe ical-like pa icles a e obse ed. The sphe ical subpa i-
cles ha e a size o he o de o ca. 10 nm and he size o he agg ega e is o he o de o
ca. 50 nanome e s. Some agg ega es o mo e han one mic ome e we e as well obse ed
on o he a eas o he g id (see in he appendix page 77). The igh pa o igu e 4.8
p esen s a TEM mic og aph o a sample ob ained by mixing 20 mM o Ca2+ and 12 mM
o HPO2−
4in p esence o 15 µM o he glycop o ein. The gene al obse a ion was he
p esence o single sphe ical-like pa icles wi h a mean size o he o de o 10 o 20 nm.
A e y low numbe o agg ega es o 2 o 3 sphe ical pa icles was ound. They we e sus-
pec ed o be o med du ing he blo ing o he excess solu ion by he help o a il e pape .
Quan i a i e analysis
SAXS expe imen s ha e been pe o med in o de o quan i a i ely s udy he agg ega ion
p ocess highligh ed by he help o DLS and TEM measu emen s. The op pa o igu e
4.9 p esen s he e olu ion o he sca e ed in ensi ies o he sample con aining no addi-
ion o Fe uin-A. A weak minimum a ound 0.3 nm−1and he abscence o oscilla ion in
hese in ensi ies a e cha ac e is ic o he high polydispe si y o he sys em. This is in
ag eemen wi h bo h DLS and TEM expe imen s. Sca e ing pa e ns olde han 0.26
35
4.2 Resul s and discussion Calci ica ion a ea ly s age
s did no p esen any change in he shape o in in ensi y (measu emen s ealized up o
145 seconds). This demons a es he e y as kine ic o mine aliza ion o calcium and
phospha e ions.
Figu e 4.9: E olu ion o he SAXS in ensi ies wi h ime in he abscence o Fe uin-A.
La e SAXS in ensi ies did no show any e olu ion o he shape o he in ensi y. This
unde lines he e y as kine ic o ea ly calci ica ion leading o he i s equilib ium phase
o calcium phospha e pa icles. Time poin o measu emen s a e: 0.035 s (blue), 0.255 s
( ed) and 1.355 s (g een) a e he mixing p ocess.
Measu emen s ealized by adding he glycop o ein ga e simila da a a q- alues bigge
han 0.3 nm−1. As an example, he igu e 4.10 p esen s he e olu ion o he SAXS in ensi-
ies eco ded o he sample con aining 15 µM o he glycop o ein, ha is a physiological
concen a ion. The main di e ence as compa ed o igu e 4.9 in he end o he sca e ed
in ensi ies is si ua ed a small q- alues. Howe e , i is o cou se necessa y o i he SAXS
in ensi ies in o de o ge quan i a i e piece o in o ma ion o he ole o Fe uin-A on o
he ea ly s age o calci ica ion.
The i ing p ocedu e was ealized by assuming he p ima y pa icles as homogenous
sphe es by using equa ion 2.6 page 12 as expec ed om he wo k o A. Heiss using TEM ex-
pe imen s [64]. The polydispe si y in size o he p ima y pa icles was aken in o accoun
by assuming a no malized Gaussian dis ibu ion. Due o he p esence o an amo phous
sys em (WAXS measu emen s in solu ion we e ealized and did no show any B agg peak,
see igu e B.2 page 79 in appendix), i is necessa y o add an addi ional e m o simula e
he he mal luc ua ions acco ding o he heo y o O ns ein-Ze nike (see e e ence [38]).
This has been aken in o accoun by he use o he equa ion 2.12 page 13. The up- u n
a small q- alues was quan i ied by he help o he s uc u e ac o (equa ion 2.23 page
36

4.2 Resul s and discussion Calci ica ion a ea ly s age
Figu e 4.10: E olu ion o he SAXS in ensi ies wi h ime in he abscence ( op) and
in p esence o 15 µM (bo om) o Fe uin-A. La e SAXS in ensi ies did no show any
e olu ion o he shape o he in ensi y. This demons a es he e y as kine ic o ea ly
calci ica ion leading o he i s equilib ium phase o calcium phospha e pa icles. Time
poin o measu emen s a e: 0.035 s (blue ci cles), 0.255 s ( ed squa es), 0.695 s (black
iangles) and 6.415 s (blue squa es) a e he mixing p ocess.
15). The elec on con as o he calcium phospha e complex and o he Fe uin-A a e
de e mined espec i ely o 536 and ca. 50 e/nm3. Thus, i is assumed ha he sca -
e ed in ensi ies is only due o he calcium-phospha e complex. The esul ing heo e ical
equa ion ha has been used in o de o i he expe imen al sca e ed in ensi ies I(q) is
hen:
I(q) = N
V(∆ρ)2V2
pS(q)[I0(q) + I luc(q)] (4.3)
Figu e 4.11 p esen s he sca e ed in ensi ies collec ed 0.89 s a e he mixing p ocess o
all samples s udied. No addi ional s uc u e ac o was needed in o de o i he da a o
he sample in ol ing 15 µM o Fe uin-A. Figu e 4.12 p esen he s uc u e ac o s S(q)
esul ing om he modelling o he SAXS da a eco ded 0.89 s a e he mixing p ocess.
The igu e 4.13 p esen s he e olu ion o he adius o he p ima y sphe ical pa icles Rpsp
o all s udied samples. A e y as kine ic is de ec ed o all s udied samples since wi hin
one second, he p ima y pa icles do no g ow anymo e. Howe e , his kine ic is oo as
o ge a quan i a i e analysis o he g ow h o he p ima y pa icles.
Quan i a i ely, he di e en adii o he p ima y pa icles a e in e es ing: he highe
he concen a ion o Fe uin-A, he bigge he p ima y pa icles. Such a esul could
no be de e mined jus by he help o DLS o mic oscopic measu emen s. The igu e
37
4.2 Resul s and discussion Calci ica ion a ea ly s age
Figu e 4.11: Expe imen al da a o measu ed samples and hei espec i e i s 0.89 second
a e he mixing p ocess. The poin s ep esen he expe imen al da a. Fo he sake o
cla i y, only one ou o wo poin s is ep oduced. The dashed lines exhibi he heo e ical
i s o polydispe se homogenous non in e ac ing sphe es, including he he mal luc ua-
ions and he ull lines e lec he comple e i when he s uc u e ac o is needed. The
g aphic shows he di e en concen a ions s udied: 0 µM, 1 µM, 5 µM and 15 µM o
Fe uin-A om bo om o op.
Figu e 4.12: E olu ion o he s uc u e ac o S(q) ob ained om he modelling o he
SAXS da a eco ded 0.89 s a e he mixing p ocess. The up- u n a small q- alues
cha ac e izes he agg ega ion o he nanopa icles. Mixes con ain 0 µM (blue), 1 µM
( ed), 5 µM (black) and 15 µM (g een) o Fe uin-A.
38
4.2 Resul s and discussion Calci ica ion a ea ly s age
Figu e 4.13: Time-e olu ion o he adius o he p ima y sphe ical pa icles o calcium
phospha e as a unc ion o he concen a ion o he p o ein. The p ima y sphe ical pa -
icles g ow wi h a e y as kine ic wi hin 1 s o all samples s udied. The dashed lines
a e guide lines o he e olu ion o he adius o he p ima y sphe ical pa icles. Da a
in ol ing a concen a ion o 1 µM o Fe uin-A a e no ep oduced o he sake o cla i y
and is in e media e o he one o 0 and 5 µM.
Figu e 4.14: Time-e olu ion o he numbe o p ima y sphe ical pa icles pe agg ega e
(S(0)+1). The main e ec o Fe uin-A is seen he e: inhibi ion o he agg ega ion. The
dashed lines a e guide lines o he e olu ion o he pa ame e s. Mixes con ain 0 µM
(blue), 1 µM ( ed), 5 µM (black) and 15 µM (g een) o Fe uin-A.
39
4.2 Resul s and discussion Calci ica ion a ea ly s age
4.14 p esen s he ime-e olu ion o he numbe o p ima y pa icles pe agg ega e. This
plo pe mi s o be e unde s and he ole o Fe uin-A on o he calci ica ion a ea ly
s age. As demons a ed by igu es 4.11 and 4.12, he e we e no need o simula e any
in e pa icle in e ac ions in o de o i he sys em in ol ing a physiological concen a ion
o he p o ein (15 µM). The esul s ob ained om he o he s udied concen a ions o
he glycop o ein show ha he lowe he amoun o Fe uin-A, he bigge he numbe o
pa icles pe agg ega e. This is he p oo ha α2-HS-glycop o ein/Fe uin-A is inhibi ing
he agg ega ion o he p ima y sphe ical pa icles o calcium phospha e and is s abilising
hese pa icles om he ea lies s age o mine aliza ion.
Acco ding o he esul s ound in his s udy and o p e ious wo k in li e a u e [17],
an hypo he ical model o he o ma ion o calcium phospha e pa icles induced by he
p esence o Fe uin-A could be desc ibed as ollow: Du ing he nuclea ion p ocess, Fe uin-
A has a weak e ec un il he p ima y pa icles g ow o ca. 10 nm in adius. Once he
nuclea ion p ocess is ended, Fe uin-A may co e he calcium-phospha e pa icles and ac
as a shield o p e en agg ega ion. Acco ding o Heiss [64], he glycop o ein in e ac s
wi h 6 di e en calcium ions (see igu e 4.15), which is p obably he way o he p o ein
o c ea e a laye o i sel on o he calci ied pa icles.
Figu e 4.15: Binding o he D1 domain o Fe uin-A on o he su ace o hyd oxyapa i e.
The posi i e cha ges o Ca2+ a e ma ked in blue and he nega i e phospha e cha ges a e
ep esen ed in ed. Figu e aken om e e ence [63].
40
5.4 Conclusion Polybu adiene
o he di e en phases (c ys alline, amo phous sPB and SDS) we e le ee. The bes
heo e ical in ensi ies we e achie ed wi h an elec on densi y o he c ys alline laye co -
esponding o a weigh densi y o 0.962 g/cm3. This alue is, acco ding o Na a [83], he
weigh densi y o c ys alline sPB. The elec on densi y o he su ac an ’s phase (SDS)
was o he same o de as he one de e mined o he polye hylene s udy (see nex chap e ).
This unde lines and con i ms he e aci y o he esul s ob ained. The numbe o polyme
chain pe pa icle has been as well de e mined by he help o equa ion 5.4 page 43 and
i esul ed o only wo polyme chains which is d as ically lowe han o he PE sys em
s udied in e e ence [33] whe e an a e age o 14 polyme chains we e cons i u ing each
nanopa icle.
47

Chap e 6
Polye hylene
6.1 Expe imen al
Fo his s udy (collabo a ion wi h D . Qiong Tong om he g oup o P o Mecking,
Kons anz), wo di e en PE sys ems PL39 and PL78 we e in es iga ed due o he limi ed
amoun o each sample. The samples PL39 and PL78 con ain 1.7 w % and 1.6 w % PE,
espec i ely. The su ac an sodium dodecyl sul a e (SDS) is needed o s abilize he PE
pa icles agains coagula ion. The weigh ac ions o SDS a e 0.87 w % o PL39 and 0.36
w % o PL78. Hence, he amoun o SDS in PL39 has been inc eased as compa ed o he
p e ious s udy [33]. The PE sys ems di e in he labile ligand o he ca alys used du ing
he syn hesis (TPPTS o PL39, NH2-PEG o PL78, see addi ional piece o in o ma ion
in he appendix page 80). The longe li e ime o he ca alys wi h he NH2-PEG ligand
esul s in he di e en amoun o SDS du ing syn hesis. This does no ha e a signi ican
in luence on ei he he molecula weigh o in molecula s uc u e o he ob ained PE
chains.
Annealed samples we e p oduced by placing a 5 mL glass bo le con aining he o iginal
sys em on o a me al box hea ed a he annealed empe a u e o 20 minu es. PE nanopa -
icles annealed o longe ime (up o 60 minu es) did no e eal any u he inc ease o
he hickness o he c ys al. The annealed sys ems we e sligh ly mo e concen a ed since
e apo a ion could no be comple ely a oided.
Dynamic Ligh Sca e ing measu emen s we e pe o med wi h an ALV/DLS/SLS-5000
compac goniome e sys em (ALV Lange ) equipped wi h a He-Ne lase (632.8 nm) and a
he mos a (Ro ilabo, ±0.1◦C) a 25◦C. Dilu ed samples (0.0013 w %) we e analyzed om
30 o 80◦. Acco ding o he S oke-Eins ein equa ion, he hyd odynamic adius o each
sample was de e mined espec i ely o 11.2 ±0.2 nm and 11.4 ±0.2 nm o he o iginal
and annealed sample. These ligh sca e ing expe imen s did no pe mi o de ec any
48
6.2 Resul s and discussion Polye hylene
di e ence be o e and a e he annealing p ocess. In o de o ha e a be e unde s anding
o he mo phology o he pa icles s udied, c yogenic ansmission elec on mic oscopy
expe imen s ha e been pe o med.
6.2 Resul s and discussion
6.2.1 C yo-TEM
Specimens o c yo-TEM expe imen s we e p epa ed a oom empe a u e as explained
on page 42. The concen a ion used he e was highe by a ac o o 10 leading o 0.3
w %. Figu e 6.1 p esen s ypical o e iews o he c yo-TEM mic og aphs eco ded o he
Figu e 6.1: Typical c yo-TEM mic og aphs o he o iginal PL39 sys em ( op) and o he
annealed PL39 sample (bo om). Inse s in each mic og aph p esen s a single pa icle in
which he di ec ion o he main axis is o h hogonal and pa allel o he di ec ion o he
elec on beam. The annealing p ocess was ca ied a 125◦C o 20 minu es. Fo bo h
mic og aphs, he weigh pe cen age s udied is 0.3 w %.
49
6.2 Resul s and discussion Polye hylene
o iginal sys em ( op) and o he annealed one (bo om). These pa icles a e well dispe sed
in he solu ion. Di e en mo phologies could be ema ked and his comes om he angle
be ween he main axis o he pa icle and he one o he elec on beam. The di e ence
o con as be ween one pa icle and ano he is ela ed o di e en angles be ween he
no mal o he pla ele s and he di ec ion o he elec on beam. This is mo e p ononced in
he op-inse whe e he main axis o a single pa icle is pa allel o he elec on beam. Such
pa icles appea as da k ods. A he opposi e, he bo om-inse p esen a single pa icle
in which he main axis is o hogonal o he di ec ion o he beam. These pa icles appea
as ligh e hexagons. This hexagonal shape ag ees well wi h he heo e ical g ow h o PE
nanopa icles ( igu e 3.6). The annealed nanopa icles (bo om mic og aph in igu e 6.1)
p esen he same mo phology as shown in he inse . Howe e , a change appea ed in he
dimensions ( adius and hickness) o he sys em. De i ed om he image analysis o 20
pa icles wi h hei no mal o ien ed pe pendicula o he elec on beam, he heigh o he
pla ele s was de e mined o 7 ±1 nm be o e and 13 ±2 nm a e annealing and he pla ele
adii dec eased om 14 ±4 nm o 9 ±2 nm espec i ely. One has o ake in o accoun
ha i is no possible o de ec he amo phous laye o hese PE nanopa icles because
i s elec on densi y is e y simila o he one o he su ounding medium (low densi y
amo phous ice, [33]; ligh g ay in he backg ound ep esen s he low densi y amo phous
ice). In o de o elucida e he shape and s uc u e o he PE pa icles in mo e de ails,
he sys ems ha e been in es iga ed using SAXS and con as a ia ion echnique.
6.2.2 SAXS
SAXS expe imen s we e pe o med a he ID02 beamline a ESRF, G enoble, F ance
and in Bay eu h by using a K a ky Compac Came a. Figu e 6.2 p esen s he sca e ed
in ensi ies o he PL39 PE nanopa icles be o e ( op) and a e (bo om) he annealing
p ocess. The olume ac ion o he con as agen a ies om 0% (blue poin s), 4% ( ed
poin s), 10% (black poin s) up o 14% (g een poin s), while he olume ac ion o he
nanopa icles dec ease: 2.5, 2.4, 2.3, 2.2 ol% o he o iginal sys em ( op) and 2.6, 2.5, 2.3
and 2.2 ol% o he annealed sample (bo om) espec i ely. The h ee lowe mos in en-
si ies a e di ided by ac o s o 10, 102and 103 o sake o cla i y. The he o e ical SAXS
in ensi ies ha e been modelled by using equa ions 5.1 o 5.3 and a e p esen ed in igu e
6.2. The solid lines ep esen he esul o he non-in e ac ing polydispe se disks. The
sho dashed lines (q<0.14 nm−1) a e ob ained using he PRISM in eg al heo y. PRISM
heo y is well known o accoun o in e ac ions o anis opic pa icles and su ace cha ges.
The di e ence be ween he dashed and he solid lines e lec he in e molecula in e ac-
ions be ween he nanopa icles. The modelling including he in e pa icle in e ac ion
was ealized by P i .-Doz. D . Ludge Ha nau.
50
6.2 Resul s and discussion Polye hylene
Figu e 6.2: No malized sca e ed in ensi ies o he PL39 PE sys em be o e ( op) and a e
(bo om) he annealing p ocedu e. All in ensi ies a e no malized by he olume ac ion
o he sample. The olume ac ion o he con as agen a ies om 0 (blue poin s), 0.04
( ed poin s), 0.10 (black poin s) up o 0.14 (g een poin s), while he olume ac ion o
he nanopa icles dec ease: 2.5, 2.4, 2.3, 2.2 ol% o he o iginal sys em ( op) and 2.6,
2.5, 2.3 and 2.2 ol% o he annealed sample (bo om) espec i ely. The h ee lowe mos
in ensi ies a e di ided by ac o s o 10, 102and 103 o sake o cla i y. The e ical g ay
dashed lines show he q- alue below which he s uc u e ac o is needed. Fo he sake
o cla i y, only one ou o 5 poin s is shown. The dashed lines ep esen s he esul o
he modeling o he SAXS da a assuming a dispe sion o non-in e ac ing polydispe se
pla ele s. The solid lines ep esen he sca e ing in ensi y calcula ed o a dispe sion o
in e ac ing polydispe se pla ele s.
51
6.2 Resul s and discussion Polye hylene
E iden ly, and as expec ed by he c yo-TEM mic og aphs ( igu e 6.1), he anneal-
ing p ocess changed he s uc u e o he o iginal sys em. The e a e mo e oscilla ions
in he sca e ed in ensi ies o he annealed sys em han in he one o he o iginal PE
nanopla ele s (see igu e 6.2). One can no e ha he s uc u e ac o S(q) in e ene only
a long dis ances, i.e. small magni udes o he sca e ing ec o q, below 0.14 nm−1. This
de ines he a e age dis ance dbe ween wo pa icles wi h he equa ion d= 2π/q= 45 nm.
This alue emains cons an be o e and a e he annealing p ocess. This demons a es
ha he numbe o pa icles pe olume is cons an , hus no usion o he pa icles occu
du ing he annealing p ocess. This is u he con i med by he ac ha I(q=0)/[φ(∆ρ)2]
is cons an o he o iginal and annealed sys em (see igu e B.4 in he appendix page 81).
The change o he mo phology o he nanopla ele s can di ec ly be seen by he SAXS
da a: he loca ion o he side maxima and minima shi o lowe q alues a e annealing.
Sys em O iginal Annealed
R [nm] 10.0 ±3.0 7.5 ±3.0
Lc[nm] 6.5 ±1.0 13.0 ±1.0
La[nm] 3.1 ±0.8 3.8 ±1.0
Table 6.1: Pa ame e s ob ained om adap ing equa ions 5.1 o 5.3 o he expe imen al
da a. Lcand Lade ine he o e all c ys alline and amo phous hickness o he pa icle.
Table 6.1 esumes he pa ame e s ob ained by adjus ing equa ions 5.1 o 5.3 o he
expe imen al da a o he o iginal and annealed sys em (see igu e 6.2). The hickness o
he c ys alline laye inc eases by a ac o o 2 ( om 6.5 o 13 nm espec i ely o he o ig-
inal and annealed sys em). The hickness o he amo phous laye emains nea ly cons an
( espec i ely 3.1 nm and 3.8 nm). Wi hin he limi o e o , he adius dec eases om 10
nm o 7.5 nm. By using a Gaussian polydispe si y du ing he i ing o he SAXS da a,
he a e age numbe o polyme chain nchains has been de e mined acco ding o equa ion
5.4 (ρc i=339 e.u./nm3and ρamo=302 e.u./nm3). This a e age numbe o polyme chains
was de e mined o nchains=8 be o e and a e he annealing p ocess.
The combina ion o he c yo-TEM expe imen s and o he SAXS con as a ia ion
da a pe mi o demons a e ha annealing he sample PL39 a 125◦C o 20 minu es leads
o a doubling o he c ys alline hickness (6.5 nm o 13 nm). In he li e a u e, wo scena ios
o he hickening p ocess a e p oposed. The i s one is an unlooping o he polyme
chains wi hin one c ys alline pla ele ( igu e 6.3 (a), e e ences [120,121]). Inc easing he
empe a u e leads o an inc ease o he chain mobili y and o coope a i e mo ion o he
monome uni s. This hypo hesis leads o a su ace o he c ys alline phase di ided by wo
as compa ed o he o iginal sys em and o he same numbe o pa icles. The second model
52

6.3 Va ia ion o annealing empe a u e Polye hylene
explains he hickening by he s acking o wo adjacen c ys alline laye s in o one o he
same su ace and double hickness ( igu e 6.3 (b) and e e ences [106,122]). Such a model
is alid o sys ems in which he c ys alline phases a e connec ed by amo phous egions
and lead o a numbe o pa icles di ided by wo. F om he i ing pa ame e s, he e is
a doubling o he hickness o he c ys alline phase. Mo eo e , he numbe o pa icles
pe olume emains cons an be o e and a e he annealing p ocess. This beha iou is
cha ac e is ic o he unlooping model p oposed by e e ences [120,121]. Thus model (b)
in igu e 6.3 is uled ou .
Figu e 6.3: 2D-model o he unlooping p ocess expec ed by some au ho s [106,120–122].
(a): The ull unlooping p ocess o one single c ys alline nanopa icle lead o a doubling o
he c ys alline hickness o he same pa icle. (b): The s ack o wo c ys alline pa icles
lead o one pa icle wi h a doubled c ys alline hickness.
6.3 Va ia ion o annealing empe a u e
The s udy o he e ec o he annealing p ocess has been p esen ed in he p e ious sec-
ion. Well-de ined nanopa icles o polye hylene ha e been subjec o a he mic ea men
o 20 minu es a 125◦C, ha is jus below he mel ing empe a u e Tm=128◦C acco ding
o DSC measu emen s. The las sec ion demons a ed ha , du ing he annealing p ocess,
no usion occu s and a simple un olding o he c ys alline chains is esponsible o he dou-
bling o he c ys alline hickness. The PE nanopa icles a e hen pa icula ly in e es ing
o be e unde s and he annealing p ocess o PE c ys alline phase. These expe imen s
ha e been pushed o wa d by a ying he annealing empe a u e o lowe empe a u es.
The annealing p ocess was ca ied a 90◦C, 105◦C and 115◦C o 20 minu es by using he
sample PL78 due o he lack o sample PL39. C yo-TEM expe imen s ha e been pe -
o med on o hese samples by using he same se up as he sys em PL39 and he g ow h
o he o e all hickness is highligh ed (see igu e 6.4). A e y b oad polydispe si y o e
he adius o he pa icles is obse ed as expec ed by he syn hesis (use o a ca alys
leading o a longe li e ime and s onge polydispe si y, see appendix). Thus, c yo-TEM
53
6.3 Va ia ion o annealing empe a u e Polye hylene
mic og aphs only pe mi o insu e ha he nanopa icles a e well-de ined.
Figu e 6.4: C yo-TEM mic og aphs o PL78 sample (o iginal and di e en annealed em-
pe a u es). The black ba ep esen s 50 nm.
Figu e 6.5: Two-dimensional schema ical ep esen a ion o pa ial unlooping o PE chains
wi hin a nanopa icle. A pa ial unlooping leads o a e y small dec ease o he adius R
o he pla ele , p obably a dec ease oo small o be de ec ed by SAXS expe imen s.
Howe e , i seems e iden ha he hickness o he c ys alline phase inc eases wi h
54
6.3 Va ia ion o annealing empe a u e Polye hylene
he annealing empe a u e as expec ed. Because o hese e y small dimensions and his
high polydispe si y, he esolu ion o he SAXS expe imen s do no allow o de e mine
p ecisely hese adii. In he p e ious sec ion, o he sys em PL39, he nanopa icles
we e ully annealed ( he hickness o he c ys alline phase ully doubled). Due o pa ial
annealing, i is impossible o de e mine he p ecise loca ion and amoun o unlooped PE
chains ( igu e 6.5). In combina ion wi h he gi en polydispe si y o adius Rand he
limi ed esolu ion o SAXS, he exac de e mina ion o he adius is impossible. Thus,
o his sys em, a adius o 6 nm was kep cons an du ing i ing ou ine. Ne e heless,
i is s ill kep in mind ha he numbe o pa icles pe olume is cons an and ha he
adius o each pa icle mus sligh ly dec ease wi h he annealing empe a u e.
SAXS measu emen s ha e been pe o med by he same app oach han o he PL39
sys em. Figu e 6.6 p esen s he expe imen al da a o he samples PL78 in wa e and hei
co esponding i s acco ding o equa ions 5.1, 5.2 and 5.3 including he in e pa icula
in e ac ions gi en by S(q). The di e ences be ween he measu ed sca e ing in ensi ies o
he o iginal sample PL39 (lowe symbol in he op g aphic o igu e 6.2) and he o iginal
sample PL78 (lowe symbols in igu e 6.6) is due o he di e en amoun o SDS added.
C ys alline hicknesses o 6.5 ±1 nm, 8.5 ±1.4 nm, 10.2 ±1.4 nm and 11.8 ±1.4 nm
esul ed o he o iginal sys em and he ones annealed a 90◦C, 105◦C and 115◦C espec-
i ely. The da a o he whole con as se ies a e displayed in he appendix page 82 o 85.
Figu e 6.6: Expe imen al da a and co esponding i s o he o iginal sample PL78 dis-
pe sed in pu e wa e (blue ci cles). The olume ac ions o he sys ems a e: 2.1 ol%,
1.6 ol%, 2.4 ol% and 2.6 ol% espec i ely o he o iginal sys em (bo om) and he
ones annealed a 90◦C ( ed squa es), 105◦C (black iangles) and 115◦C (g een ci cles).
55
6.3 Va ia ion o annealing empe a u e Polye hylene
Figu e 6.7: Expe imen al da a o he ecip ocal o he c ys alline hickness depending
on he annealing empe a u e. The da a wi h iangula and squa e symbols ha e been
ob ained om mic on c ys alls o iginally c ys allized a T=85◦C and T=95◦C espec i ely
[123]. The blue ci cles ep esen he da a o his s udy (nanoc ys als eely suspended).
Figu e 6.7 displays a plo o he annealing empe a u e agains he ecip ocal o he
c ys alline hickness Lc esul ed om he i s o he SAXS da a. As sugges ed by he
he modynamic equa ion 3.3 page 25, hese da a (blue ci cles) desc ibe a linea depen-
dency agains he ecip ocal o he c ys alline hickness. As a compa ision, expe imen al
da a o mic on c ys als on subs a e a e plo ed [123]. By ex apola ion, all da a lead
o a c ys aliza ion empe a u e o an in ini e c ys al o he o de o T∞
c=182◦C. This
beha iou may seem in iguing o he case o wo di e en sys ems, bu by ex apola ing
a an in ini e c ys al, he susb a e e ec and he SDS molecules can be neglec ed.
I is also in e es ing o compa e he esul s o he he modynamic scheme o polyme s
p oposed in li e a u e (see igu e 3.7 page 24). This leads o igu e 6.8 which p esen s he
ec ys alliza ion line ob ained om his s udy ( ed squa e), he hicknesses o c ys alline
lamella o PE collec ed om mel (blue ci cle [94]) and pos - hickened c ys alline PE
lamella as de e mined by elec on mic oscopy (g een illed ci cles [123]). The esul ing
ec ys alliza ion line de ines a linea ela ionship be ween he annealing empe a u e and
he ecip ocal o he c ys alline lamella laye . As expec ed by he co esponding he -
modynamic equa ions 3.2 and 3.3 page 25, he ex apola ion o he c ys alliza ion and
ec ys alliza ion lines a an in ini e c ys alline hickness lead o he same empe a u e
T∞
c≈182◦C. Acco ding o igu e 6.7, his T∞
cco espond o he mel ing empe a u e o
56
8.3 Polye hylene Summa y
8.3 Polye hylene
Finally, in con as o ecen li e a u e on bulk polye hylene (PE), his hesis in es iga ed
eely suspended nanopa icles o PE. As sugges ed by Webe and cowo ke s [33], he com-
bina ion o SAXS and c yo-TEM has been used o his s udy. The s uc u e o indi idual
PE nanoc ys als has been de e mined in de ail and an imp o ed model o he o m ac o
(SAXS) has been de elopped in close collabo a ion wi h P i .-Doz. D . Ludge Ha nau.
The second pa o his hesis mainly deals wi h he annealing o hese PE pa icles. Fo
he i s ime, i is shown ha he e ec o he annealing p ocess esul s in a doubling
o he c ys alline laye o he PE nanopa icles. This beha iou could be aced back o
he unlooping o he PE chains. In addi ion, a linea ela ionship be ween he ecip ocal
o he c ys alline laye and he annealing empe a u e has been expe imen ally d awn.
This line was p edic ed by he Gibbs-Thomson equa ion acco ding o he li e a u e [34].
This esul is impo an because i allows o con ol he c ys alline hickness and physical
p ope ies o he sys em, by he empe a u e.
Figu e 8.2 is a ske ch o he ob ained esul s on he PE nanoc ys als: The o iginal
semic ys alline PE nanopa icles ha e a e y hin c ys alline lamella. By annealing a
a empe a u e a below he mel , a hickening o he lamella occu s. This beha iou ends
up a he mel ing line.
Figu e 8.2: Schema ical esume o he expe imen s ealized on o he PE nanopa icles
annealed a di e en empe a u es. T is he empe a u e and n is he numbe o monome
uni s in he c ys alline hickness.
63

Chap e 9
Zusammen assung
Das Ziel diese A bei wa , du ch eine Kombina ion on Kleinwinkel ¨on gens euung
(SAXS), Elek onenmik oskopie (TEM und K yo-TEM) und dynamische Lich s euung
(DLS) Teilchen mi G ¨oßen im Nanome e be eich im De ail zu analysie en. Dazu wu -
den zwei Sys eme mi seh un e schiedliche Mo phologie und Zusammense zung un e -
such : Kugel ¨o mige Teilchen on Calciumphospha -P o ein-Komplexen und he e ogene
Scheibchen on Polye hylen/Polybu adien.
9.1 Calci izie ung
Die A bei en ¨ube Calciumphospha -P o ein-Komplexen be ass en sich mi dem Ein luss
des P o eins Fe uin-A, auch AHSG genann , au die Calci izie ung zu ¨uhen Zei punk en.
Dazu wu den Calcium- und Phospha ionen mi und ohne Fe uin-A in eine Pu e l¨osung
mi pH = 7,4 gemisch . In einem e s en Sch i wu den DLS-Messungen du chge ¨uh ,
um den Ein luss de absolu en Gewich san eile de Ca2+ und PO3−
4-Ionen besse zu e -
s ehen. Du ch diese Expe imen e wu de he ausge unden, dass ohne Zugabe on AHSG
die gebilde en Teilchen mi kleine em Gewich san eil diese Ionen kleine sind.
In wei e en Expe imen en wu de de Ein luss on Fe uin-A au den P ozess de Calci-
icie ung w¨ah end de e s en Minu e un e such . Die ¨uhe Bildung on Calciumphospha -
Komplexen konn e e olg eich mi TR-SAXS e olg we den. Dabei wu de eine schnelle
Nukleie ung on Nanopa ikeln inne halb eine Sekunde beobach e . Zum e s en Mal
konn e die Rolle des Glycop o eins Fe uin-A in de seh ¨uhen Phase de Calci izie ung
quali a i besch ieben we den: AHSG e hinde die Agg ega ion de Calciumphospha -
Teilchen. Fe uin-A spiel demnach eine wich ige Rolle im ¨o alen Se um in de Vo bildung
des Skele s on Wi bel ie en. Die Un e suchung ha e ne gezeig , dass eine physiolo-
gische Konzen a ion on 15 µM dieses Glycop o eins aus eich , um die Agg ega ion de
Calciumphospha -Teilchen olls ¨andig zu e hinde n.
In Abbildung 9.1 sind die E gebnisse zusammenge ass . Die TEM-Au nahmen au de
ech en Sei e zeigen das Sys em ohne P o ein (un en) und mi 15 µM AHSG (oben),
64
9.2 Polybu adien Zusammen assung
de physiologischen Konzen a ion. Im ¨uhen S adium de Calci icie ung zeig sich die
Nukleie ung de Calciumphospha -Teilchen. Wenn diese Teilchen eine bes imm e G ¨oße
e eich haben, wi k Fe uin-A, wenn es in de L¨osung o handen is , als Abschi mung
und umh¨ull die calci izie en Teilchen, um so eine wei e e Agg ega ion zu e hinde n.
Sys eme ohne P o ein ode mi eine ge ingen Konzen a ion on Fe uin-A weisen Agg e-
ga ion au .
Figu e 9.1: Vo geschlagenes Modell des Ein lusses on Fe uin-A ( o ) au die Bildung
on Calciumphospha -Teilchen (o ange). Die Bilde au de ech en Sei e zeigen TEM-
Au nahmen de Calciumphospha -Komplexen, die ohne (un en) und mi 15 µM Fe uin-A
(oben) gebilde wu den.
9.2 Polybu adien
Es wu den ei suspendie e Nanopa ikel aus syndio ak ischem Polybu adien un e such .
Mi els eine Kombina ion aus c yo-TEM und SAXS wu de gezeig , daß die Pa ikel eine
beme kenswe d¨unne k is alline Polyme lamelle besi zen. Ve schiedene Modelle ¨u ho-
mogene und he e ogene Nanopa ikel (mi zwei ode d ei e schiedenen S eul¨angendich en
inne halb eines Teilchens) wu den mi einande e glichen um eine op imale heo e ische
Besch eibung de expe imen ellen R¨on genkleinwinkels euda en zu e hal en. Das Vo -
liegen on amo phem und k is allinem Polybu adienbe eichen wu de mi els R¨on gendi -
ak ion gezeig . Die No wendigkei eine zus¨a zlichen SDS-Schich bei de Modellie ung
is au den SDS-¨
Ube schuß w¨ah end de Syn hese de Polybu adienpa ikel (Gewich an eil
ca. 1:1) zu e kl¨a en. Eine Besch eibung de heo e ischen R¨on genkleinwinkels euin ensi ¨a en
is ungen¨ugend wenn die Anwesenhei on SDS nich be ¨ucksich ig wi d. Die Bildung
on semik is allinen PE-Nanopa ikeln e m¨oglich eine neue Syn hese ou e zu Nanopoly-
me en mi in e essan en physikalisch-chemischen Eigenscha en wie sie in Halblei e n ode
pho o ol aische Komponen en ge unden we den.
65
9.3 Polye hylen Zusammen assung
9.3 Polye hylen
Im Gegensa z zu neue en Li e a u ¨ube Polye hylen (PE) im Bulk wu den in diese A -
bei ei suspendie e PE-Nanopa ikel un e such . Wie on Webe e al. [33] o geschla-
gen, wu de dazu eine Kombina ion aus SAXS und K yo-TEM e wende . Die S uk-
u on einzelnen k is allinen PE-Nano eilchen wu de im De ail besch ieben und ein
e besse es Model ¨u den Fo m ak o ¨u SAXS in enge Zusammena bei mi P i .-Doz.
D . Ludge Ha nau en wickel . De zwei e Teil de A bei besch¨a ig sich mi dem Tem-
pe n diese PE-Teilchen. Zum e s en Mal wu de gezeig , dass de Tempe ungsp ozess eine
Ve dopplung de k is allinen Schich dicken de PE-Nanopa ikel bewi k . Dieses Ve hal-
en kann au ein En al en de PE-Polyme ke en zu ¨uckge ¨uh we den. Zus¨a zlich wu de
expe imen ell eine linea e Beziehung zwischen de Rezip oken de k is allinen Schich dicke
und de Tempe empe a u he ges ell . Diese Linie wu de au Basis de Gibbs-Thomson-
Gleichung in de Li e a u [34] o ausgesag . Dieses wich ige E gebnis zeig , dass die
k is alline Dicke und physikalischen Eigenscha en des Sys ems du ch die Tempe a u
kon ollie we den kann.
Abbildung 9.2 zeig schema isch die E gebnisse ¨ube die PE-Nanok is alle: Die k is alline
Lamelle de u sp ¨unglich semik is allinen PE-Nanopa ikel is seh d¨unn. Du ch Tempe n
bei Tempe a u en wei un e halb de Schmelze inde eine Ve dickung de Lamelle s a .
Dieses Ve hal en ende an de Schmelzku e.
Figu e 9.2: Schema ische Zusammen assung de Expe imen e mi PE-Nanopa ikeln und
e schiedenen Tempe empe a u en.
66
Appendices
67
Appendix A
Theo y o SAXS
A.1 E ec o polydispe si y
E ec o he polydispe si y on o he sca e ing in ensi y o an ensemble o non-in e ac ing
sphe ical pa icles o mean adius R0=10 nm. Polydispe si ies used he e a e 5%, 10% and
20% in gaussian dis ibu ion (co esponding espec i ely o 0.5 ( ed squa e), 1.0 (black
iangle) and 2.0 nm (blue ci cles) o hei s anda d de ia ions σ). ∆ρis he e equals o
1 e.u.nm−3and N
Vis equal o uni y. The in ensi y a q=0 g ow wi h he polydispe si y.
A high polydispe si y, he i s peak is ha dly isible and hus piece o in o ma ion a e
los .
68

A.2 C++ p og ams Appendix
A.2 C++ p og ams
A.2.1 o m ac o o homogenous sphe ical polydispe se pa i-
cles
69
A.2 C++ p og ams Appendix
70
A.2 C++ p og ams Appendix
A.2.2 s uc u e ac o o he agg ega ion o sphe ical pa icles
71
A.3 Modi ied hambu ge model Appendix
A.3 Modi ied hambu ge model
72
B.2 WAXS signal o calcium phospha e complexes Appendix
B.2 WAXS signal o calcium phospha e complexes
The p e ious igu e p esen s WAXS signal o he calcium-phospha e complexes wi hou
(blue ull line) and in p esence o 15µM o Fe uin-A ( ed bold line). Due o he abscence
o c ys alline peak, he sys ems ha e been assumed as amo phous.
79

B.3 Syn hesis o he polye hylene nanopa icles Appendix
B.3 Syn hesis o he polye hylene nanopa icles
The syn hesis o he nanopa icles we e ealized in Kons anz, Ge many, by D . Qiong
Tong.
Dispe sion syn hesis was ca ied ou in a 300 mL s ainless s eel mechanically s i ed (1000
pm) p essu e eac o equipped wi h a hea ing/cooling jacke supplied by a he mos a
con olled by a he mocouple dipping in o he polyme iza ion mix u e. To a mix u e o
750 mg o SDS (Fluka, 98%) and 10 µmol o he Ni(II)-complex in a 250 mL Schlenk lask
was added 100 mL o dis illed and degassed wa e a oom empe a u e. The esul ing
homogenous solu ion was hen cannula- ans e eed o he a gon lushed eac o cooled
a 12◦C. The eac o was p essu ized o a cons an e hylene p essu e o 40 ba while
he empe a u e was adjus ed a 15◦C. A e 30 min eac ion ime, e hylene eeding was
in e up ed, he eac o was ca e ully dep essu ized, and he ob ained dispe sion was
il a ed h ough a plug o glass wool.
The le molecule is he ca alys used o he syn hesis o PL39. The igh molecule is he
ca alys used o he syn hesis o PL78.
80
B.4 In luence o he annealing p ocess on N
VAppendix
B.4 In luence o he annealing p ocess on N
V
No malized sca e ed in ensi ies o he PE sys ems be o e (blue) and a e ( ed) he
hea ing p ocedu e ba hing in he highes glucose solu ions. All in ensi ies a e no malized
by he olume ac ion o he sample: espec i ely 6.26 ol% and 6.96 ol% o he o iginal
and annealed pa icles. The olume ac ion o he added D+-glucose a e 14.25 ol% and
14.29 ol% espec i ely. The equali y o he sca e ed in ensi ies a low sca e ing ec o s
demons a e ha he numbe o pa icles pe olume is cons an be o e and a e he
annealing p ocess.
81
B.5 Con as se ies o PL78 Appendix
B.5 Con as se ies o PL78
B.5.1 O iginal sys em - PL78
Expe imen al con as a ia ion SAXS da a o he o iginal PL78 sys em and hei
espec i e heo e ical in ensi ies acco ding o equa ions 5.2 and 5.3 page 42. The olume
pe cen ages o he sys ems a e om bo om o op: 2.1 ol%, 1.9 ol%, 1.8 ol% and 1.7
ol% and hei espec i e olume pe cen ages o added glucose a e: 0.0 ol%, 4.4 ol%,
9.9 ol% and 15.0 ol%. The mean c ys alline hickness measu es he e 6.5 nm.
82
B.5 Con as se ies o PL78 Appendix
B.5.2 Sys em annealed a 90◦C
Expe imen al con as a ia ion SAXS da a o he PL78 sys em annealed a 90◦C and
hei espec i e heo e ical in ensi ies acco ding o equa ions 5.2 and 5.3 page 42. The
olume pe cen ages o he sys ems a e om bo om o op: 1.6 ol%, 1.5 ol%, 2.0 ol%
and 1.9 ol% and hei espec i e olume pe cen ages o added glucose a e: 0.0 ol%, 4.4
ol%, 9.9 ol% and 14.9 ol%. The mean c ys alline hickness measu es he e 8.5 nm.
83
B.5 Con as se ies o PL78 Appendix
B.5.3 Sys em annealed a 105◦C
Expe imen al con as a ia ion SAXS da a o he PL78 sys em annealed a 105◦C
and hei espec i e heo e ical in ensi ies acco ding o equa ions 5.2 and 5.3 page 42.
The olume pe cen ages o he sys ems a e om bo om o op: 2.4 ol%, 2.2 ol%, 2.1
ol% and 2.0 ol% and hei espec i e olume pe cen ages o added glucose a e: 0.0
ol%, 4.4 ol%, 9.9 ol% and 15.1 ol%. The mean c ys alline hickness measu es he e
10.2 nm.
84

B.5 Con as se ies o PL78 Appendix
B.5.4 Sys em annealed a 115◦C
Expe imen al con as a ia ion SAXS da a o he PL78 sys em annealed a 115◦C
and hei espec i e heo e ical in ensi ies acco ding o equa ions 5.2 and 5.3 page 42.
The olume pe cen ages o he sys ems a e om bo om o op: 2.6 ol%, 2.4 ol%, 2.3
ol% and 2.1 ol% and hei espec i e olume pe cen ages o added glucose a e: 0.0
ol%, 4.4 ol%, 9.9 ol% and 14.9 ol%. The mean c ys alline hickness measu es he e
11.8 nm.
85
B.6 Syn hesis o sPB Appendix
B.6 Syn hesis o sPB
The syn hesis o he nanopa icles we e ealized in Kons anz, Ge many, by D . B igi e
Ko hals.
A oluene solu ion (5 ml) o cobal (II) 2-e hylhexanoa e (1.60 mmol) was in oduced
unde a gon o a mechanically s i ed 10 ml p essu e glass eac o equipped wi h a hea -
ing/cooling jacke con olled by a empe a u e senso dipping in o he eac ion mix u e.
A e e apo a ing o oluene in acuum, 32.5 g o bu adiene we e condensed a -5◦C. An
e hanol solu ion (15 ml) o sodium bo ohyd ide (3.55 mmol) was added, a o ding a so-
lu ion o he p eca alys [Co(C8H13)(C4H6)]. An aqueous solu ion o su ac an (46.5 g
SDS/ 20 g pen anol and 400 g H2O) was hen ans e ed o he eac o by means o a
pump unde s i ing (1000 pm) and he empe a u e was se o 20◦C, a o ding a ans-
pa en bu adiene/p eca alys mic oemulsion. A oluene solu ion o ca bon disul ide (1.6
mmol, [CS2] / [Co] = 1) was pumped in o he eac o , which was hen apidly hea ed o
he desi ed empe a u e (T = 40◦C). A e 3h he eac ion was s opped by cooling and
eleasing esidual gas p essu e, a o ding a ligh b own anspa en la ex.
86
B.7 X- ay di ac ion o BK280 Appendix
B.7 X- ay di ac ion o BK280
X- ay di ac ion o he polybu adiene sys em BK280 (blue ull line) and o he glas
susb a e ( ed bold line).
X- ay di ac ion o he polybu adiene sys em BK280 alone. This da a we e ob ained
by sub ac ing he signal o he glas and o he amo phous pa . The hkl peak o he
c ys alline syndio ac ic 1,2-polybu adiene (Pacm, a=10.98 ˚
A, b=6.60 ˚
Aand c=5.14 ˚
A) a e
epo ed when isible.
87
B.8 DLS o BK280 Appendix
B.8 DLS o BK280
Figu e B.1: DLS da a o he polybu adiene sys em. The di usion coe icien was de e -
mined o 3.81×10−23 m2/s leading o an hyd odynamic adius o 6.4 nm. The sys em was
dilu ed in wa e a a concen a ion o 0.00013 w %.
88
LIST OF FIGURES Appendix
4.14 Time-e olu ion o he numbe o p ima y sphe ical pa icles pe agg ega e
(S(0)+1). The main e ec o Fe uin-A is seen he e: inhibi ion o he agg e-
ga ion. The dashed lines a e guide lines o he e olu ion o he pa ame e s.
Mixes con ain 0 µM (blue), 1 µM ( ed), 5 µM (black) and 15 µM (g een)
o Fe uin-A.................................... 39
4.15 Binding o he D1 domain o Fe uin-A on o he su ace o hyd oxyapa i e.
The posi i e cha ges o Ca2+ a e ma ked in blue and he nega i e phospha e
cha ges a e ep esen ed in ed. Figu e aken om e e ence [63]. . . . . . . 40
5.1 Hambu ge model used in his s udy. Two addi ional shee s o SDS ha e
o be aken in o accoun o model he s uc u e. L , L=Lc+Laand Lc
ep esen he hicknesses o he whole pa icle, o he polyme and o he
c ys alline phase espec i ely, while R is he adius o he disk. . . . . . . . 43
5.2 C yo-TEM image o he nanopa icle o polybu adiene. The concen a ion
was 0.03 w %. The scale ba o he main mic og aph ep esen s 50 nm
while he ones in he inse s ep esen 10 nm. . . . . . . . . . . . . . . . . . 44
5.3 No malized sca e ing in ensi ies o he polybu adiene nanopa icles. Fo
he sake o cla i y, he ou lowe mos sca e ed in ensi ies we e di ided by
ac o o 101, 102, 103and 104. F om bo om o op, he concen a ion o
glucose (and o he sample) in solu ion was 0.0 (1.19), 3.2 (1.23), 6.5 (1.24),
9.8 (1.24) and 17.3 (1.32) ol%. . . . . . . . . . . . . . . . . . . . . . . . . 45
5.4 Plo o he empe a u e as a unc ion o he ecip ocal o he c ys alline
hickness o syndio ac ic polybu adiene sys ems. The ed squa es a e ob-
ained om e e ence [119] and he blue ci cle ep esen s his s udy. The
bold ed and blue lines a e guide-lines o he ec ys alliza ion and c ys al-
liza ion lines espec i ely. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
6.1 Typical c yo-TEM mic og aphs o he o iginal PL39 sys em ( op) and o
he annealed PL39 sample (bo om). Inse s in each mic og aph p esen s
a single pa icle in which he di ec ion o he main axis is o h hogonal
and pa allel o he di ec ion o he elec on beam. The annealing p ocess
was ca ied a 125◦C o 20 minu es. Fo bo h mic og aphs, he weigh
pe cen age s udied is 0.3 w %. . . . . . . . . . . . . . . . . . . . . . . . . . 49
95

LIST OF FIGURES Appendix
6.2 No malized sca e ed in ensi ies o he PL39 PE sys em be o e ( op) and
a e (bo om) he annealing p ocedu e. All in ensi ies a e no malized by
he olume ac ion o he sample. The olume ac ion o he con as
agen a ies om 0 (blue poin s), 0.04 ( ed poin s), 0.10 (black poin s)
up o 0.14 (g een poin s), while he olume ac ion o he nanopa icles
dec ease: 2.5, 2.4, 2.3, 2.2 ol% o he o iginal sys em ( op) and 2.6, 2.5,
2.3 and 2.2 ol% o he annealed sample (bo om) espec i ely. The h ee
lowe mos in ensi ies a e di ided by ac o s o 10, 102and 103 o sake o
cla i y. The e ical g ay dashed lines show he q- alue below which he
s uc u e ac o is needed. Fo he sake o cla i y, only one ou o 5 poin s is
shown. The dashed lines ep esen s he esul o he modeling o he SAXS
da a assuming a dispe sion o non-in e ac ing polydispe se pla ele s. The
solid lines ep esen he sca e ing in ensi y calcula ed o a dispe sion o
in e ac ing polydispe se pla ele s. . . . . . . . . . . . . . . . . . . . . . . . 51
6.3 2D-model o he unlooping p ocess expec ed by some au ho s [106,120–122].
(a): The ull unlooping p ocess o one single c ys alline nanopa icle lead o
a doubling o he c ys alline hickness o he same pa icle. (b): The s ack
o wo c ys alline pa icles lead o one pa icle wi h a doubled c ys alline
hickness. .................................... 53
6.4 C yo-TEM mic og aphs o PL78 sample (o iginal and di e en annealed
empe a u es). The black ba ep esen s 50 nm. . . . . . . . . . . . . . . . 54
6.5 Two-dimensional schema ical ep esen a ion o pa ial unlooping o PE
chains wi hin a nanopa icle. A pa ial unlooping leads o a e y small
dec ease o he adius Ro he pla ele , p obably a dec ease oo small o
be de ec ed by SAXS expe imen s. . . . . . . . . . . . . . . . . . . . . . . 54
6.6 Expe imen al da a and co esponding i s o he o iginal sample PL78 dis-
pe sed in pu e wa e (blue ci cles). The olume ac ions o he sys ems
a e: 2.1 ol%, 1.6 ol%, 2.4 ol% and 2.6 ol% espec i ely o he o igi-
nal sys em (bo om) and he ones annealed a 90◦C ( ed squa es), 105◦C
(black iangles) and 115◦C (g een ci cles). . . . . . . . . . . . . . . . . . . 55
6.7 Expe imen al da a o he ecip ocal o he c ys alline hickness depend-
ing on he annealing empe a u e. The da a wi h iangula and squa e
symbols ha e been ob ained om mic on c ys alls o iginally c ys allized
a T=85◦C and T=95◦C espec i ely [123]. The blue ci cles ep esen he
da a o his s udy (nanoc ys als eely suspended). . . . . . . . . . . . . . . 56
96
LIST OF FIGURES Appendix
6.8 Expe imen al he modynamic scheme ob ained by he help o da a in e -
e ence [124] (blue ci cles) and om he p esen s udy ( ed squa es). The
blue, ed and black lines ep esen espec i ely he c ys alliza ion, ec ys-
alliza ion and mel ing lines o he he modynamic scheme p oposed in li -
e a u e [29,94]. The bold g een line is a guide line o he pos - hickened
expe imen al da a poin s [123]. . . . . . . . . . . . . . . . . . . . . . . . . 57
7.1 Schema ic ep esen a ion o he USAXS/SAXS/WAXS equipmen a he
ERSF in G enoble, F ance. Pic u e aken om h p://www.es .eu/Use s
AndScience/Expe imen s/So Ma e /ID02/BeamlineLayou on Jully, he
1s 2009...................................... 59
7.2 De e mina ion o he densi y o a calcium phospha e sys em in bu e so-
lu ion. F om he slope, ρis de e mined o 1.67 ±0.05 g/cm3. ....... 60
8.1 Hypo he ical model o he in luence o Fe uin-A ( ed) on o he o ma ion
o calcium phospha e complexes (o ange). Black and whi e images on
he igh p esen TEM mic og aphs o calcium-phospha e pa icles o med
wi hou (bo om) and in p esence o 15 µM o Fe uin-A ( op). . . . . . . . 62
8.2 Schema ical esume o he expe imen s ealized on o he PE nanopa icles
annealed a di e en empe a u es. T is he empe a u e and n is he
numbe o monome uni s in he c ys alline hickness. . . . . . . . . . . . . 63
9.1 Vo geschlagenes Modell des Ein lusses on Fe uin-A ( o ) au die Bildung
on Calciumphospha -Teilchen (o ange). Die Bilde au de ech en Sei e
zeigen TEM-Au nahmen de Calciumphospha -Komplexen, die ohne (un-
en) und mi 15 µM Fe uin-A (oben) gebilde wu den. . . . . . . . . . . . 65
9.2 Schema ische Zusammen assung de Expe imen e mi PE-Nanopa ikeln
und e schiedenen Tempe empe a u en. . . . . . . . . . . . . . . . . . . . 66
B.1 DLS da a o he polybu adiene sys em. The di usion coe icien was de e -
mined o 3.81×10−23 m2/s leading o an hyd odynamic adius o 6.4 nm.
The sys em was dilu ed in wa e a a concen a ion o 0.00013 w %. . . . . 88
97
LIST OF FIGURES Appendix
B.2 Theo e ical SAXS signal o he di e en s uc u al models. The in ensi ies
esul ing om models B and C do no lead o any d as ical change o he
SAXS in ensi ies while he model used in his s udy as well as he co e-
bishell model do lead o a s onge change in in ensi ies a small q- alues.
Fo he model used in his s udy, ”model PE/sPB”, he pa ame e s used
a e: R = 12.0 ±2.0 nm, Lc= 6.5 ±0.4 nm, La= 3.1 ±0.8 nm and Lsds
= 2.0 nm. Fo model B: R = 12.0 ±2.0 nm, Ra= R-2.0 nm (Radeno es
he adius o he amo phous phase alone), Rc= Ra-1
2La(Rcdeno es he
adius o he c ys alline phase alone) Lc= 6.5 ±0.4 nm, La= 3.1 ±0.8
nm and Lsds = 2.0 nm. Fo model C: R = 12.0 ±2.0 nm, Ra= R-2.0 nm
(Rpdeno es he adius o he polyme phase alone), Lc= 6.5 ±0.4 nm,
La= 3.1 ±0.8 nm and Lsds = 2.0 nm. Fo he co e bi-shell model: Rc
= 6.5 ±0.4 nm, ha= 3.1 ±0.8 nm and hsds = 2.0 nm. Fo all models
in his appendix, he elec on densi ies ha e he ollowing alues: 333.57
e.u.nm−3, 339 e.u.nm−3, 302 e.u.nm−3and 397 e.u.nm−3 espec i ely o
he sol en , c ys alline, amo phous and sds phase. . . . . . . . . . . . . . . 91
98
Acknowledgemen s
Fi s o all, I would like o exp ess my deepes g a i ude o P o . D . M. Ballau o
gi ing me he in e es ing subjec s ha I ha e been in es iga ing. I would like o hank
him o being e y help ul o he publica ions and his hesis.
I am e y g a e ul o P o . D . W. Jahnen-Dechen and P o . D . S. Mecking o
hei help ul discussions and o kindly p o iding samples du ing ou collabo a ions.
Many hanks a e gi en o D . Sabine Rosen eld o he c i ical eadings o my wo ks
and especially his hesis. I gi e my acknowlegemen o D . Alexande Heiss o all alu-
able discussions abou he calci ica ion p ojec and o P i .-Doz. D . Ludge Ha nau
o his ous anding con ibu ion o he simula ion wo k on in e pa icle in e ac ions o he
polye hylene nanopa icles. D . Ma kus D echsle is acknowledged o his g ea in es i-
ga ions by elec on mic oscopies on he di e en samples ha I s udied.
I am e y g a e ul o D . J´e ˆome C assous o his daily good mood and o ou alu-
able discussions a any ime. I would like o hank him as well o he e y in e es ing
p ojec ha we ha e been in es iga ing oge he .
I am g a e ul o all my colleagues who ha e cons uc ed a e y iendly a mosphe e o
wo king. I gi e my acknowledgemen s especially o Ad iana and Se gio Mihu o hei
help du ing he s udy o he polye hylene nanopa icles and o gi ing me some piece o
ad ice o he p og amming o he i ing p og ams espec i ely. I would like o hank,
D . Ka ja Henzle o ealizing he SAXS expe imen s o he i s polye hylene sample a
he ESRF, G enoble, F ance. A e y big hanks is gi en o D . Theyenche i Na ayanan
o p o iding beam ime a he Synch o on sou ce o all s udied sys ems. Qiong Tong
is g ea ly hanked o he syn hesis o he PE nanopa icles and o all impo an dis-
cussions o a be e unde s anding o hese objec s. I easu e my iendship wi h my
colleagues F ank Polze , Ch is ian Schneide , Michael Zeise , D . Yan Lu, D . Alexande
Wi emann, Mi iam Siebenb¨u ge and D . S eena h Bolise y.
I hank Elisabe h D¨ung elde o he bu eauc a ic wo k wi h a lo o pa ience and kind-
ness and Ka lheinz Lau enbach o his a ailibili y and echnical suppo .
My special hanks goes o my amily. My pa en s ha e encou aged me and shown hei
app ecia ions o my e e y p og ess in hese wo ks. I am e y g a e ul oo o my b o he s
o kindly gi ing me he oppo uni y o use unix se e s o simula ing SAXS heo e ical
in ensi ies and o hei encou agemen s.
99
Financial suppo by he Deu sche Fo schungsgemeinscha , SFB 481, Bay eu h, and
by he Ma ie Cu ie Resea ch T aining Ne wo k (POLYAMPHI) a e g a e ully acknowl-
edged.
100

Bibliog aphy
[1] Fajun Zhang, Maximilian W. A. Skoda, Rob M. J. Jacob, Richa d A. Ma in,
Ch is ophe M. Ma in, and F ank Sch eibe . P o ein in e ac ions s udied by saxs:
E ec o ionic s eng h and p o ein concen a ion o bsa in aqueous solu ions.
Jou nal o Physical Chemis y B, 111 (1):251–259, 2007.
[2] D. Pon oni, T. Na ayanan, and A. R. Rennie. T -saxs s udy o nuclea ion and
g ow h o sillica colloids. Langmui , 18:56–59, 2002.
[3] G. MacLennan and C. A. Bee e s. The c ys al s uc u e o dicalcium phospha e,
cahpo4. Ac a C ys , 8:579–583, 1955.
[4] C. A. Bee e s. The c ys al s uc u e o dicalcium phospha e dihyd a e, cahpo4.2h2o.
Ac a C ys , 11:273–277, 1958.
[5] K. Suda sanan and R. A. Young. S uc u e o s on ium hyd oxide phospha e,
s 5(po4)3oh. Ac a C ys , 28:3668–3670, 1972.
[6] J. D. Te mine and A. S. Posne . Calcium phospha e o ma ion in i o. A chi es
o Biochemis y and Biophysics, 140:307–317, 1970.
[7] J. L. Meye and E. D. Eanes. A he modynamic analysis o he amo phous o
c ys alline calcium phospha e ans o ma ion. Cacli ied Tissue Resea ch, 25:59–68,
1978.
[8] Hans E. Lundage -Madsen, Isabel Lopez-Vale o, Vic o ia Lopez-Ace edo, and
Roland Bois elle. The o ma ion p oduc o amo phous icalcium phospha e a
37¨ı¿1
2c. Jou nal o C ys al G ow h, 75:429–434, 1986.
[9] F. Be s, N. C. Blumen hal, and A. S. Posne . Bone mine aliza ion. Jou nal o
C ys al G ow h, 53:63–73, 1981.
[10] M. U. Nylen, E. D. Eanes, and J. D. Te mine. Molecula and ul as uc u al s udies
o non-c ys alline calcium phospha es. Calci ied Tissue Resea ch, 2:95, 1972.
[11] Lj. B ece ic and H. F¨ı¿1
2 edi-Milho e . P ecipi a ion o calcium phospha es om
elec oly e solu ions. Calci ied Tissue Resea ch, 10:82–90, 1972.
101
[12] J. Bolze, B. Peng, N. Dingenou s, P. Panine, T. Na ayanan, and M. Ballau . Fo -
ma ion and g ow h o amo phous colloidal caco3 p ecu so pa icles as de ec ed by
-saxs. Langmui , 18:8364–8369, 2002.
[13] J. Bolze, D. Pon oni, M. Ballau , T. Na ayanan, and H. C¨ı¿1
2l en. Time- esol ed
saxs s udy o he e ec o a double hyd ophilic block-copolyme on he o ma ion o
caco3 om supe sa u a ed sal solu ion. Jou nal o Colloid and In e ace Science,
277:84–94, 2004.
[14] Willi Jahnen-Dechen , And eas T indl, Jasmina Godo ac-Zimme mann, and We ne
M¨ı¿1
2lle -Es e l. Pos ansla ional p ocessing o human ahsg (human e uin) - e i-
dence o he p oduc ion o a phospho yla ed single-chain o m by hepa oma cells.
Eu opean Jou nal o Biochemis y, 226:59–69, 1994.
[15] Willi Jahnen-Dechen , C. Sch¨ı¿1
2 e , A. Heiss, and J. G ¨ı¿1
2 zinge . Sys emic inhi-
bi ion o spon aneous calci ica ion by he se um p o ein ahsg. Zei un ¨ı¿1
2 Ka di-
ologie, 90:47–56, 2001.
[16] A. Heiss, W. Jahnen-Dechen , H. Endo, and D. Schwahn. S uc u al dynamics o a
colloidal p o ein-mine al complex bes owning on calcium phospha e a high solubili y
in biological luids. Bioin e phases, 2(1):16–20, 2007.
[17] Alexande Heiss, Thomas Ecke , Anke A e z, Wal e Rich e ing, Wim an Do p,
Co a Sch¨ı¿1
2 e , and Willi Jahnen-Dechen . Hie a chical ole o e uin-a and acidic
se um p o eins in he o ma ion and s abiliza ion o calcium phospha e pa icles.
Jou nal o Biological Chemi s y, 283(21):14815–14825, 2008.
[18] J.-P. Lin, C.-Y. Chang, C.-H. Wu, and S.-M. Shih. The mal deg ada ion kine ics
o polybu adiene ubbe . Polyme Deg ada ion and S abili y, 53:295, 1996.
[19] P. T. Williams and S. Besle . Py olisis- he mog a ime ic analysis o y es and y e
componen s. Fuel, 74:1277, 1995.
[20] B. Ko hals, M. C. Mo an -Minana, M. Schmid, and S. Mecking. Func ionaliza ion
o polyme nanopa icles by hiol-ene addi ion. Mac omolecules, 43:8071, 2010.
[21] T. Yamamo o. Molecula dynamic o e e sible and i e e sible mel ing in chain-
olded c ys als o sho polye hylene-like polyme . Mac omolecules, 43:9384–9393,
2010.
[22] And zej Pawlak and And zej Galeski. Ca i a ion du ing ensile d awing o annealed
high densi y polye hylene. Polyme , 51:5771–5779, 2010.
[23] A. Galeski, Z. Ba czak, T. Kazmie czak, and M. Slou . Mo phology o unde o med
and de o med polye hylene lemella c ys als. Polyme , 51:5780–5787, 2010.
102
[24] Cou ney P. Ennis and Ral I. Kaise . Mechanis ical s udies on he elec on-induced
deg ada ion o polyme s: polye hylene, poly e a luo oe hylene, and polys y ene.
Physical Chemis y Chemical Physics, 12:14884–14901, 2010.
[25] S. C. Moyses. Low empe a u e annealing in low densi y polye hylene. saxs and dsc
s udies. Polyme Jou nal, 32:486–491, 2000.
[26] S¨ı¿1
2 e ine Humbe , Oli ie Lame, Jean-Ma c Chenal, Cy ille Rochas, and
G¨ı¿1
2 a d Vigie . Small s ain beha io o polye hylene: In si u saxs measu emen s.
Jou nal o Polyme Science, Pa B: Polyme Physics, 48:1535–1542, 2010.
[27] M. Kakiage, M Sekiya, T. Yamanobe, T. Komo o, S. Sasaki, S. Mu akami, and
H. Ueha a. In si u saxs analysis o ex ended-chain c ys alliza ion du ing mel -
d awing o ul a-high molecula weigh polye hylene. Polyme , 48:7385–7392, 2007.
[28] N. S ibeck, A. Almenda ez Cama illo, U. N¨ı¿1
2chel, P. B¨ı¿1
2secke, and R. K.
Baye . Ea ly o ien ed iso he mal c ys alliza ion o polye hylene s udied by high-
ime- esolu ion saxs/waxs. Analy ical and Bioanaly ical Chemis y, 387:649–661,
2007.
[29] G: S obl. A he modynamic scheme ea ing polyme c ys alliza ion and mel .
Eu opean Physical Jou nal E, 18:295–309, 2005.
[30] R. Soula, C. No a , A. Tomo , R. Spi z, J. Cla e ie, X. D ujon, J. Malinge, and
T. Saudemon . Ca aly ic polyme iza ion o e hylene in emulsion. Mac omolecules,
34:2022–2026, 2001.
[31] Flo ian M. Baue s and S e an Mecking. High molecula mass polye hylene aqueous
la exes by ca aly ic polyme iza ion. Angewand e Chemie In e na ional Edi ion,
40(16):3020–3022, 2001.
[32] Flo ian M. Baue s and S e an Mecking. W¨ı¿1
2ss ige la ices aus hochmolekula em
polye hylen du ch ka aly ische polyme isa ion. Angewand e Chemie, 113(16):3112–
3115, 2001.
[33] C. H. M. Webe , A. Chiche, G. K ausch, S. Rosen eld , M. Ballau , L. Ha nau,
I. G¨ı¿1
2 ke -Schne mann, Q. Tong, and S. Mecking. Single lamella nanopa icles o
polye hylene. Nano Le e s, 7 (7):2024–2029, 2007.
[34] G. S obl. Colloquium: Laws con olling c ys alliza ion and mel ing in bulk poly-
me s. Re iews o mode n physics, 81:1287–1300, 2009.
[35] P. Debye and A. M. Bueche. Sca e ing by an inhomogeneous solid. Jou nal o
Applied Physics, 20:518–525, 1949.
103
[36] And e Guinie . Small-Angle Sca e ing X- ays. John Wiley & Sons, 1955.
[37] G. Fou ne . Theo e ical and expe imen al s udy o he di usion o x- ays by dense
agg ega es o pa icles. Bulle in de la Soci¨ı¿1
2 ¨ı¿1
2F ancaise de Mine alogie e de
C is allog aphie, 74:37–17, 1951.
[38] H. E. S anley. In oduc ion o phase ansi ions and c i ical phenomena. Cla endon
P ess, 1971.
[39] B. B. Mandelb o . The F ac al Geome y o Na u e. F eeman, 1983.
[40] H. C. Ande sen and D. Chandle . Op imized clus e expansions o classical luids.
i. gene al heo y and a ia ional o mula ion o he mean sphe ical model and ha d
sphe e pe cus-ye ick equa ions. Jou nal o Chemical Physics, 57:1918, 1972.
[41] D. Chandle and H. C. Ande sen. Op imized clus e expansions o classical luids.
ii. he o y o molecula liquids. Jou nal o Chemical Physics, 57:1930, 1972.
[42] D. Chandle . De i a ion o an in eg al equa ion o pai co ela ion unc ions in
molecula luids. Jou nal o Chemical Physics, 59:2742, 1973.
[43] L. J. Lowden and D. Chandle . Solu ion o a new in e g al equa ion o pai co e-
la ion unc ions in molecula liquids. Jou nal o Chemical Physics, 59:6587, 1973.
[44] L. J. Lowden and D. Chandle . Theo y o in e molecula pai co ela ions o molec-
ula liquids. applica ions o he liquids ca bon e achlo ide, ca bon disul ide, ca bon
diselenide and benzene. Jou nal o Chemical Physics, 61:5228, 1974.
[45] K. S. Schweize and J. G. Cu o. In eg al-equa ion heo y o he s uc u e o polyme
mel s. Physical Re iew Le e s, 58:246, 1987.
[46] A. Ye hi aj, B. J. Sung, and F. Lado. In eg al equa ion heo y o wo-dimensional
polyme mel s. Jou nal o Chemical Physics, 122:94910, 2005.
[47] J. L. Ki sch ink and J. W. Hagado n. Biomine aliza ion. Wiley-VCH, Weinheim,
2000.
[48] Ma s S.-A. Johnsson and Geo ge H. Nancollas. The ole o b ushi e and oc acalcium
phospha e in apa i e o ma ion. C i ical Re iews in O al Biology and Medicine,
3:61–82, 1992.
[49] A. G. Wal on, W. J. Bodin, H. Fu edi, and A. Schwa z. Nuclea ion o calcium
phospha e om solu ion. Canadian Jou nal o Chemis y, 45:2695, 1967.
104
[123] I. G. Voig -Ma in. A quan i a i e elec on-mic oscopic s udiy o a linea polye hy-
lene ac ion c ys allized a di e en empe a u es. Jou nal o Polyme Science:
Polyme Physics Edi ion, 19:1769–1790, 1981.
[124] P. J. Ba ham, R. A. Chi e s, A. Kelle , J. Ma inez-Salaza , and S. J. O gan. The
supe cooling dependence o he ini ial old leng h o polye hylene c ys allized om
he mel : uni ica ion o mel and solu ion c ys alliza ion. Jou nal o Ma e ials
Science, 20:1625–1630, 1985.
[125] B. Wunde lich and G. Czo nyj. A s udy o equilib ium mel ing o polye hylene.
Mac omolecules, 10:906–913, 1977.
[126] H. S abinge and O. K a ky. A new echnique o he measu emen o he absolu e
in ensi y o x- ax small angle sca e ing. he mo ing sli me hod. Mak omolekula
Chemie, 179:1655, 1978.
[127] Nico Dingenou s and Ma hias Ballau . S uc u al in es iga ion o la exes by small
angle x- ay sca e ing: Measu emen and e alua ion o da a. Ac a Polyme ica,
49:178–183, 1998.
[128] P. B¨ı¿1
2secke. The high- lux beamline a he es . Re iew o Scien i ic Ins umen s,
63:438, 1992.
[129] Pe e Boesecke. Reduc ion o wo-dimensional small- and wide angle x- ay sca e ing
da a. Jou nal o Applied C ys allog aphy, 40:423–427, 2007.
111

Publica ions
•C. N. Roche e; S. Rosen eld ; A. Heiss; T. Na ayanan; M. Ballau ; W. Jahnen-
Dechen :
A shielding opology s abilizes he ea ly s age p o ein-mine al complex o Fe uin-A
and calcium-phospha e
ChemBioChem, 10(4), 735-740, 2009.
•C. N. Roche e; S. Rosen eld ; K. Henzle ; F. Polze ; M. Ballau ; Q. Tong; S.
Mecking; M. D echsle ; T. Na ayanan; L. Ha nau:
Annealing o single lamella nanopa icles o polye hylene
Mac omolecules, 44(12), 4845-4851, 2011.
112
Cu iculum Vi ae
P i a e:
Name: Ch is ophe Nicolas Roche e
Add ess: 5 ue de Sa oie
33600 Pessac
FRANCE
Ma i al s a us: Single
Schools:
1984-1988: Ecole p imai e de la Glaci`e e, M´e ignac, F ance
1988-1990: Ecole p imai e de Cap de Bos, Pessac, F ance
1990-1994: Coll`ege Ladonne, Pessac, F ance
1994-1995: Lyc´ee Pape Cl´emen , Pessac, F ance
1995-1998: Lyc´ee Sain e Ma ie G and Leb un, Bo deaux, F ance
Scien i ic s udies:
1998-2000: Ins i u e o Technology, Physical Measu emen s, Talence, F ance
2000: Resea ch assis an : ”S udies o he polu an e ec o aluminium
ions on sun lowe s”, TAMK, Tampe e, Finland - Supe iso :
P o . D . Ma jukka Dye
2000-2004: Mas e o Chemis y, Uni e si y o Bo deaux 1, Talence, F ance
2003: Resea ch assis an : ”Memb ane s uc u e and in e ac ions o
an ibio ic pep ides om Aus alian ee ogs”, School o
Chemis y, Uni e si y o Melbou ne, Vic o ia, Aus alia -
Thesis supe iso : P o . D . F ances Sepa o ic
2004: Resea ch assis an : ”S udy o he con inemen o anionic
pep ides on o ca ionic amphiphiles”, Eu opean Ins i u e o
Chemis y and Biology, Uni e si y o Bo deaux 1, F ance -
Thesis supe iso : P o . D . Reiko Oda
PhD:
Since No embe 2005: ”S uc u al Analysis o Nanopa icles by Small Angle
X- ay Sca e ing”, Bay eu h Cen e o Colloids and In e ace
Science, Uni e si y o Bay eu h, Ge many -
Thesis supe iso : P o . D . Ma hias Ballau
113
E kl¨a ung
Hie mi e kl¨a e ich, dass die A bei selbs ¨andig e ass und keine ande en als die on
mi angegebenen Quellen und Hil smi el benu z habe.
Fe ne e kl¨a e ich, dass die ande wei ig mi ode ohne E ol nich e such habe, diese
Disse a ion einzu eichen. Ich habe keine gleicha ige Dok o p ¨u ung an eine ande e
Hochschule endg¨ul ig nich bes anden.
Bay eu h,
Ch is ophe N. Roche e
114