biomedicines
A icle
E alua ion o Roo -End P epa a ion wi h Two
Di e en Endodon ic Mic osu ge y Ul asonic Tips
Paulo J. Palma 1,2,* , Joana A. Ma ques 1, Ma ga ida Casau 3, And éSan os 3,
F ancisco Ca amelo 4, Rui I. Falacho 5and João Miguel San os 1,2
1Ins i u e o Endodon ics, Facul y o Medicine, Uni e si y o Coimb a, 3000-075 Coimb a, Po ugal;
[email p o ec ed] (J.A.M.); [email p o ec ed] (J.M.S.)
2Cen e o Inno a ion and Resea ch in O al Sciences (CIROS), Facul y o Medicine, Uni e si y o Coimb a,
3000-075 Coimb a, Po ugal
3Den is y Depa men , Facul y o Medicine, Uni e si y o Coimb a, 3000-075 Coimb a, Po ugal;
[email p o ec ed] (M.C.); [email p o ec ed] (A.S.)
4Coimb a Ins i u e o Clinical and Biomedical Resea ch (iCBR), Labo a o y o Bios a is ics and Medical
In o ma ics (LBIM), Facul y o Medicine, Uni e si y o Coimb a, 3000-548 Coimb a, Po ugal;
[email p o ec ed]
5Ins i u e o O al Implan ology and P os hodon ics, Facul y o Medicine, Uni e si y o Coimb a,
3000-075 Coimb a, Po ugal; [email p o ec ed]
*Co espondence: [email p o ec ed]; Tel.: +351-239-249-151
Recei ed: 11 Augus 2020; Accep ed: 25 Sep embe 2020; Published: 28 Sep embe 2020


Abs ac :
The aim o his s udy is o compa e oo -end p epa a ion pe o med wi h wo di e en
ul asonic ips—CVDen us and NSK—and espec i e ime equi emen s. A e oo -end esec ion,
32 ee h we e andomly di ided in wo g oups, acco ding o he ul asonic ip used o oo -end
p epa a ion. P epa a ion ime was eco ded. Pho omic og aphs we e aken o assess he ollowing
pa ame e s: oo su ace mic oc acking, ma ginal in eg i y and p esence o deb is. One ul asonic
ip om each g oup was analyzed h ough scanning elec on mic oscopy be o e and a e oo -end
p epa a ion. The signi icance le el was se a
α
=0.05. Incidence o mic oc acks in bo h g oups
was 12.5%. Solely in acanal mic oc acking was ound, consis en ly posi ioned wi hin he wides
side o he emaining den ine. No s a is ically signi ican di e ences we e e i ied be ween bo h
expe imen al g oups ega ding ma ginal in eg i y (p=0.102) and p epa a ion ime (p=0.780),
whe eas s a is ical di e ences (p=0.003) we e ound conce ning he p esence o deb is ( he minimum
egis e ed sco e was mos ly e i ied in CVDen us g oup). NSK ips showed majo mo phological
changes, wi h ex ensi e su ace wea and no iceable loss o pa icles, which was no e i ied on
CVDen us ips. Ou indings sugges signi ican di e ences ega ding oo -end p epa a ion walls
quali y, wi h CVDen us ips showing be e esul s. Conce ning mic oc acking, as well as p epa a ion
ime and ma ginal in eg i y, bo h ul asonic ips showed simila esul s. Quali a i e analysis exposed
NSK ips majo mo phological changes and wea a e use, which was no e i ied on CVDen us ips.
Keywo ds:
apical su ge y; endodon ic mic osu ge y; oo -end p epa a ion; ul asonic; ul asonic ips
1. In oduc ion
Apical pe iodon i is comp ises he hos ’s esponse o pa hogenic mic oo ganisms colonizing
he oo canal sys em o he oo h [
1
,
2
]. The p incipal goal o con en ional endodon ic ea men
is p e en ion and/o elimina ion o apical pe iodon i is [
1
,
2
]. Today, he success a e o endodon ic
ea men s ands be ween 85% and 95%, being equen ly applied o ea i e e sible in lamma ion
o nec osis o he oo canal con en [
3
]. Endodon ic mic osu ge y is o en he las op ion when
nonsu gical e ea men ails, is un easible o unlikely o imp o e he ini ial endodon ic ea men [
4
].
Biomedicines 2020,8, 383; doi:10.3390/biomedicines8100383 www.mdpi.com/jou nal/biomedicines
Biomedicines 2020,8, 383 2 o 19
In pa icula , only su gical in e en ion may esol e cases in ol ing a pe sis en lesion wi h e iology
ela ed o complex canal ana omy, ex a- adicula in ec ion, o eign body eac ion ma e ial, and/o
cys ic issue [3].
Pe iapical su ge y acili a es comple e deb idemen o he oo canal and placemen o a oo -end
illing o ensu e adequa e apical sealing. The su gical app oach comp ises se e al sequen ial p ocedu es
in o de o ul ill he a o emen ioned goals: (a) pe iapical esec ion (apicoec omy), (b) p epa a ion o
he oo -end ca i y, and (c) sealing o he oo canal sys em by means o a bioac i e and biocompa ible
oo -end illing ma e ial placemen [4,5].
The ad en o no el diagnos ic ools, ins umen s, and ma e ials has g ea ly bene i ed endodon ic
su ge y [
5
,
6
]. Fo y- i e-deg ee oo -end esec ion be els, bu d i en e og ade p epa a ions and
amalgam o in e media e es o a i e ma e ials o oo -end illing we e o many yea s conside ed
he s a e-o - he-a wi h inconsis en success a es anging om 44.2% o 59% epo ed p io o he
in oduc ion o mic osu gical echniques [7,8].
The in oduc ion o he den al ope a i e mic oscope (DOM) in he ea ly 1990s led o a new e a in
mode n mic osu gical endodon ics [
9
]. Besides magni ica ion, con empo a y echniques inco po a e
he use o ul asonic ips, mic osu gical ins umen s and illing ma e ials which exhibi supe io
biocompa ibili y such as mine al ioxide agg ega e (MTA), Bioden ine [
7
,
10
–
13
], and p emixed
icalcium silica e pu y (To alFill FS pu y) [
14
]. This new mic osu gical app oach allowed o a
signi ican imp o emen in success a es, eaching le els abo e 91% [7,15].
The a ailable li e a u e highligh s he impo ance o an adequa e oo -end p epa a ion o a a o able
p ognosis, wi h i s quali y being di ec ly ela ed o he ea men success [
16
]. Roo -end p epa a ion
should be pa allel o he long axis o he oo , 3 mm deep, and cen e ed wi hin he oo in o de o
p ese e adequa e wall hickness and e ain a biocompa ible illing ma e ial [4,5,17].
Ul asonic ips p esen an al e na i e o he con en ional o a y bu s and show se e al ad an ages
when used o pe o m oo -end p epa a ion. In ac , he ad en o ul asonic ips esul ed in he
imp o emen o oo -end p epa a ion mainly due o he a ailabili y o ips wi h di e en shapes
and angula ions, which a e me iculously selec ed acco ding o he oo ea u es and loca ion [
18
].
Mo eo e , ul asonic ips ca y nume ous ad an ages including he possibili y o pe o ming a mo e
conse a i e os eo omy and o ob aining oo -end esec ion wi h minimal o inexis en be el angles [
19
],
hus educing he numbe o exposed den inal ubules and consequen ly he possibili y o apical
leakage [
20
]. Addi ionally, hese ips enable he emo al o is hmus issue p esen be ween wo canals
wi hin he same oo [
5
] and exhibi lowe isk o damaging he su ounding so issues du ing he
su gical p ocedu e [
7
]. Ul ima ely, ul asonic p epa a ion esul s in oo -end ca i ies ha a e smalle ,
cleane , and mo e e en i e, as well as mo e cen ally placed and aligned wi h he di ec ion o he
o iginal oo canal [
17
]. Howe e , he incidence o apical mic oc acks ollowing oo -end p epa a ion
wi h ul asonic ips has been epo ed [21–24].
Al hough no o mally es ablished, mic oc acks may inc ease he chance o apical leakage
and jeopa dize he o e all s eng h o he oo -end [
25
], wi h nega i e in luence in he long- e m
ou come o endodon ic mic osu ge y [
10
]. Op ical magni ica ion wi h o wi hou he use o dyes [
9
,
26
],
his ologicalsec ions[
26
], s e eomic oscopy[
27
], scanningelec onmic oscopy(SEM)[
28
], and luo escence
con ocal mic oscopy [
26
] a e commonly used me hods o de ec ing mic oc acks esul ing om
ul asound-ac i a ed oo p epa a ion. A ew ac o s ha e been iden i ied ha po en ially con ibu e o
he occu ence o mic oc acks—namely, he use o dehyd a ed ex ac ed ee h, absence o pe iodon al
ligamen , imp ope powe se ings o he ul asound uni , spu e -coa ing o specimens o SEM
examina ion, ime equi ed o oo -end p epa a ion, ini ial oo condi ion, and emaining den inal
hickness [
29
]. Addi ionally, he ype o coa ing o he ul asonic ips may play a signi ican ole in
mic oc ack de elopmen [30].
Recen ly some a emp s ha e been made o imp o e ul asonic ins umen s bo h in e ms o
usabili y, as well as pe o mance. New zi conium-coa ed and diamond-coa ed oo -end p epa a ion
ips ep esen a ele an issue in his ield [
31
]. Howe e new echnologies a ise, such as he chemical
Biomedicines 2020,8, 383 3 o 19
apo deposi ion which comp ises he o ma ion o a hick pu e diamond laye ha shall p oduce a
single s one co e ing he en i e ip’s su ace [32].
The aim o he p esen ex i o s udy is o compa e oo -end p epa a ion pe o med wi h wo
di e en ul asonic ips—chemical apo deposi ion CVDen us (CVDen us, S
ã
o Paulo, B azil) and
diamond-coa ed NSK (NSK, Tochigi, Japan)— ega ding oo su ace mic oc acking ( h oughou he
se en-day e alua ion pe iod), quali y o he oo -end ca i y ma gins, p esence o deb is, oo -end
p epa a ion ips’ wea , and espec i e ime equi emen s.
The null hypo hesis s a es he e a e no di e ences be ween bo h ul asonic ips ega ding he
e alua ed pa ame e s.
2. Ma e ials and Me hods
2.1. Specimen Selec ion
The p esen s udy has been app o ed by he E hical Commi ee o he Facul y o Medicine o he
Uni e si y o Coimb a (no i ica ion CE001/2013, 2 Feb ua y 2015) and ollowed he guidelines o he
Decla a ion o Helsinki. Fo y single- oo ed p emola s wi h ully de eloped apices, eshly ex ac ed
o o hodon ic easons, we e selec ed. Sample size calcula ion was based on a p e ious, unpublished,
pilo s udy using G* Powe (3.1.9.3 so wa e, Kiel, Ge many), conside ing a signi icance le el o
5% and a powe o 80%. All ee h we e imme sed in 1% sodium hypochlo i e (NaOCl, CanalP o,
Col ene/Whaleden AG, Al s a en, Swi ze land) o 15 min, immedia ely a e ex ac ion. A e wa d,
so issue and deb is we e emo ed om he ex e nal oo su aces wi h pe iodon al scale s.
The in eg i y o he oo s was assessed using DOM (Leica M300 Su gical mic oscope, Leica Mic osys ems,
We zla , Ge many) unde 16
×
magni ica ion. Tee h we e hen kep imme sed in 0.5% chlo amine T o
a pe iod o one o h ee weeks in an incuba o (Gallenkamp, London, Uni ed Kingdom) wi h con olled
empe a u e o 37 ◦C, o simula e he o al en i onmen clinical condi ions.
2.2. Specimen P epa a ion
Tee h we e deco ona ed using a high-speed conical unk diamond bu (In ini y, CVDen us, S
ã
o Paulo,
B azil) unde con inuous wa e sp ay. The wo king leng h was de e mined using a size 10 K- ile (a 0.5 mm
dis ance om he apex was conside ed as e e ence o wo king leng h de e mina ion). Roo canals we e
hen cleaned and mechanically p epa ed up o F2 (P oTape uni e sal, Den sply Maille e , Baillaigues,
Swi ze land) applying a c own-down echnique. The oo canals we e i iga ed wi h 1 mL o 1% NaOCl
be ween each ile usage, o aling a olume o i igan solu ion o 4 mL. When p epa a ion was comple ed,
a inal inse wi h 2 mL o 70% alcohol (Meda Pha ma, Lisboa, Po ugal) was pe o med, and oo canal
sys em was hen d ied using s e ile abso ben pape poin s (Zippe e Abso ben Pape Poin s Endo
Easy E icien , VDW; Munich, Ge many).
The single cone echnique was hen used o oo canal illing wi h calib a ed gu a-pe cha
P oTape poin s F2 (P oTape uni e sal, Den sply Maille e , Baillaigues, Swi ze land) and AH Plus
(Den sply, Kons anz, Ge many) as seale . Sec ion o he gu a-pe cha cone was pe o med a he cemen
enamel junc ion (CEJ) by using a hea ed ins umen and hen e ically condensed wi h Buchanan
Sys em B Plugge s (Syb onEndo, O ange, CA, USA).
Following ob u a ion, each oo h was numbe ed, and an X- ay image was aken o con i m he
quali y o ob u a ion. Finally, ee h we e posi ioned p io o he subsequen p ocedu es by placing he
oo ’s wo co onal hi ds in high— iscosi y silicone ma e ial (Col
è
ne Lab-Pu y, Col
è
ne/Whaleden
AG, Swi ze land).
All specimens we e s o ed in an incuba o (Gallenkamp, London, UK) a 37
◦
C and 98% humidi y
h oughou he expe imen al pe iod.
Biomedicines 2020,8, 383 4 o 19
2.3. Roo -End Resec ion
The sec ion le el was se a 3 mm om he apex, and all he oo s we e esec ed acco ding
o a 90-deg ee angle o hei longi udinal axis. Roo -end esec ion was pe o med using a H23LR
(Kome , Geb . B assele , Lemgo, Ge many) ca bide ungs en ope a i e bu and he sec ion su ace
was pos e io ly smoo hed wi h a H375R (Kome , Geb . B assele , Lemgo, Ge many) ca bide ungs en
inishing bu .
He ea e , he oo su aces we e checked by an examine , wi h a DOM (Leica M300 Su gical
mic oscope, Leica Mic osys ems, We zla , Ge many) unde 16
×
magni ica ion, o assess he p esence
o mic oc acks. Pho omic og aphs o he cu ing sec ion we e aken wi h a s e eomic oscope
(objec i e HR Plan Apo 1X WD 54—Nikon SMZ 1500, Tokyo, Japan) be o e and a e me hylene
blue dye 1% (Canal blue, Den splySi ona, Kons anz, Ge many), which was applied di ec ly on he
su ace du ing 5 min, and ollowed by insing wi h abundan wa e o 1 min, in o de o imp o e
mic oc acks isualiza ion.
2.4. Roo -End P epa a ion
Thi y- wo ee h ha did no p esen any mic oc acks o ac u es we e s a i ied by ans e sal
oo shape and su ace a ea, and andomly di ided in wo g oups (s a i ied andom sampling me hod),
acco ding o he ul asonic ip used in oo -end p epa a ion:
•
G oup 1 (n =16): ul asonic chemical apo deposi ion ip TOF-L (CVDen us, S
ã
o Paulo,
B azil)—lo numbe E7009;
•
G oup 2 (n =16): ul asonic diamond-coa ed ip E32D (NSK, Tochigi, Japan)—lo numbe Z217211.
Roo -end p epa a ion was pe o med using he ma ching ul asound uni and ollowing he
manu ac u e s’ ecommenda ions ega ding in ensi y, namely 30% in ensi y o powe when using
CVDen us Ul asonic Sys em (CVDen us; S
ã
o Paulo, B azil) and Endo mode le el 6 o in ensi y
wi h Va ios 970 (NSK iPiezo engine, Tochigi, Japan), unde con inuous saline solu ion i iga ion.
Roo -end p epa a ion was pe o med applying in e mi en and minimal p essu e, wi h in-and-ou
mo ion un il an apical ca i y 3 mm deep om he esec ed su ace was achie ed, ollowed by
ci cum e en ial mo emen s o comple e he en i e p epa a ion. Specimens we e kep in he silicone
blocks and main ained hyd a ed h oughou he p ocedu es. Each ip was used on a maximum
o eigh oo s and eplaced in case o ip ac u e. This p ocedu e was accomplished by a single
ope a o , using a DOM (Leica M300 Su gical mic oscope, Leica Mic osys ems, We zla , Ge many)
unde 16
×
magni ica ion. The oo -end ca i y was conside ed inished when he ope a o deemed o
ha e ob ained a isibly deb is- ee p epa a ion. All p epa a ions we e class I (acco ding o Black’s
classi ica ion). P epa a ions we e eco ded using a ideo came a and ime was measu ed using a ideo
playback so wa e in o de o ge a mo e p ecise measu emen , coun ing solely he ac ual ime o
ip- oo -end con ac .
Pho omic og aphs we e aken ollowing p epa a ion o each oo be o e and immedia ely a e
(T
PO
) applying me hylene blue dye 1% (Canal blue, Den splySi ona, Kons anz, Ge many) as p e iously
desc ibed, as well as 24 h (T24H) and se en days (T7D) a e oo -end p epa a ion.
2.5. Da a Analysis
The p eope a i e and he pos ope a i e pho omic og aphs we e coded and e alua ed by wo
blinded ope a o s. The examine s assessed he ollowing c i e ia, h ough pho omic og aphs analysis
unde 20×and 40×magni ica ion:
•
The numbe , ype and loca ion (in ela ion o den inal walls) o oo su ace mic oc acking
(Table 1a);
•The quali y o oo -end ca i y ma gins p oduced by he ul asonic ips (Table 1b).
Biomedicines 2020,8, 383 5 o 19
Table 1.
Classi ica ions adap ed o e alua e (
a
) oo su ace mic oc acking, (
b
) quali y o oo -end
ca i y ma gins, and (c) p esence o deb is wi hin he apical ca i y, ollowing oo -end p epa a ion.
(a)
Roo -End Su ace Mic oc acking.
Adap ed Classi ica ion om Rainwa e e al. [24] and De B uyne and De Moo [33].
Type Loca ion
Designa ion Comple e Incomple e Na owe Wide
In acanal Ex a-canal In a-den inal
Desc ip ion
F om he
oo canal o
he oo
su ace
O igina ing
om he oo
canal and
adia ing in o
he den ine
O igina ing
om he
oo
su ace
adia ing
in o he
den ine
Con ined o he den ine
Loca ed a
he na owe
side o he
emaining
den ine
su ace
Loca ed a
he wide
side o he
emaining
den ine
su ace
(b) Ma ginal In eg i y
Adap ed Classi ica ion om Taschie i e al. [31].
Sco e 0 1 2 3
Desc ip ion
Ideal
p epa a ion,
wi h no
de ec s
A single isible de ec
p oduced by he con ac
be ween he angle o he ip
and he ca i y ma gin
Chipped, agged ca i y
ma gin
Chipped, agged ca i y
ma gin plus some de ec s
due o he ips bouncing
o he oo du ing
oo -end p epa a ion
(c) P esence o Deb is
Adap ed Classi ica ion om Khabbaz e al. [34].
Sco e 0 1 2 3 4
Desc ip ion Clean walls Deb is on 1 wall Deb is on 2
walls
Deb is on
3 walls Deb is on 4 walls
Addi ionally, di ec s e eomic oscope isualiza ion unde 40
×
magni ica ion allowed o he
e alua ion o he p esence o deb is (den inal and/o gu a-pe cha emnan s)—Table 1c.
The sco es and numbe o c acks we e assessed independen ly by wo examine s, and in case o
disag eemen bo h examine s discussed un il a consensus was eached.
In addi ion, one ul asonic ip om each expe imen al g oup was andomly selec ed and analyzed
h ough scanning elec on mic oscopy (SEM) be o e and a e oo -end p epa a ion, wi h he pu pose
o e alua ing ip wea due o use.
2.6. S a is ical Analysis
S a is ical analysis was ca ied ou using he comme cially a ailable IBM SPSS .24 so wa e (Chicago,
IL, USA) o assess he di e ences be ween he expe imen al g oups. In o de o e alua e he incidence o
mic oc acks be o e and a e (T
PO
, T
24H
, T
7D
) oo -end p epa a ion, as well as c acking ype and loca ion,
esul s ob ained o each g oup we e analyzed h ough desc ip i e s a is ics. The Mann—Whi ney es
was pe o med o e alua e he di e ences ega ding mic oc acks, ma ginal in eg i y (quali y o apical
ca i y ma gins) and p esence o deb is be ween g oups. Conce ning he ime equi ed o oo -end
p epa a ion, he no mali y o da a dis ibu ion es ing was ca ied ou using he Shapi o–Wilk es .
The Mann—Whi ney es was used o de ec signi ican di e ences be ween he g oups as da a did no
ollow he no mal dis ibu ion. The signi icance le el was se a α=0.05.
3. Resul s
3.1. Roo -End Su ace Mic oc acking—Numbe , Type, and Loca ion
Table 2shows he esul s o he wo s udy g oups ega ding he numbe , ype and loca ion o c acks.
No isible c acks we e de ec ed a e oo -end esec ion, independen ly o he ip ype. Rega dless o he
imepoin (T
PO
, T
24H
, T
7D
) ollowing oo -end p epa a ion, in acanal oo mic oc acking was obse ed

Biomedicines 2020,8, 383 6 o 19
in wo samples o each expe imen al g oup. The e o e, an incidence o 12.5% was eco ded conce ning
he occu ence o ac u es in bo h g oups. No p opaga ion o ac u es, no he appea ance o new
ones, was e i ied h oughou he comple e se en-day pe iod o e alua ion.
Mo eo e , he maximum
numbe o mic oc acks eco ded o he same sample was one. No ex a-canal, in a-den ine, o comple e
mic oc acks we e ound. Rega ding loca ion, oo su ace mic oc acking was consis en ly posi ioned
wi hin he wides side o he emaining den ine su ace, hus egis e ing a equency o 100% o
he “wide ” pa o he oo loca ion. Figu e 1(CVDen us g oup) and Figu e 2(NSK g oup) display
ep esen a i e images o oo su ace mic oc acking o bo h expe imen al g oups.
Table 2.
Resul s o he wo expe imen al g oups ega ding he numbe , ype, and loca ion o mic oc acks.
Immedia ely a e
Roo -End Resec ion TPO T24H T7D
CVDen us NSK CVDen us NSK CVDen us NSK CVDen us NSK
Numbe 0 0 2 2 2 2 2 2
Type
In acanal 0 0 2 2 2 2 2 2
Ex a-canal 0 0 0 0 0 0 0 0
In a-den inal
0 0 0 0 0 0 0 0
Comple e 0 0 0 0 0 0 0 0
Loca ion Na owe 0 0 0 0 0 0 0 0
Wide 0 0 2 2 2 2 2 2
Biomedicines 2020, 8, x FOR PEER REVIEW 6 o 18
o e alua ion. Mo eo e , he maximum numbe o mic oc acks eco ded o he same sample was
one. No ex a-canal, in a-den ine, o comple e mic oc acks we e ound. Rega ding loca ion, oo
su ace mic oc acking was consis en ly posi ioned wi hin he wides side o he emaining den ine
su ace, hus egis e ing a equency o 100% o he “wide ” pa o he oo loca ion. Figu e 1
(CVDen us g oup) and Figu e 2 (NSK g oup) display ep esen a i e images o oo su ace
mic oc acking o bo h expe imen al g oups.
Table 2. Resul s o he wo expe imen al g oups ega ding he numbe , ype, and loca ion o
mic oc acks.
Immedia ely a e
Roo -End Resec ion T
PO
T
24H
T
7D
CVDen us NSK CVDen us NSK CVDen us NSK CVDen us NSK
Numbe 0 0 2 2 2 2 2 2
Type
In acanal 0 0 2 2 2 2 2 2
Ex a-
canal 0 0 0 0 0 0 0 0
In a-
den inal 0 0 0 0 0 0 0 0
Comple e 0 0 0 0 0 0 0 0
Loca ion Na owe 0 0 0 0 0 0 0 0
Wide 0 0 2 2 2 2 2 2
Figu e 1. Pho omic og aphs o one sample om CVDen us g oup wi h no isible mic oc ack, unde
20× magni ica ion: (a) immedia ely a e oo -end esec ion; (b) immedia ely a e oo -end
p epa a ion—T
PO
; (c) 24 h a e oo -end p epa a ion—T
24H
; (d) se en days a e oo end
p epa a ion—T
7D
.
Figu e 1.
Pho omic og aphs o one sample om CVDen us g oup wi h no isible mic oc ack,
unde 20
×
magni ica ion: (
a
) immedia ely a e oo -end esec ion; (
b
) immedia ely a e oo -end
p epa a ion—T
PO
; (
c
) 24 h a e oo -end p epa a ion—T
24H
; (
d
) se en days a e oo end
p epa a ion—T7D.
Biomedicines 2020,8, 383 7 o 19
Biomedicines 2020, 8, x FOR PEER REVIEW 7 o 18
Figu e 2. Pho omic og aphs o one sample om NSK g oup p esen ing one in acanal mic oc ack
(a ow) loca ed wi hin he “wide ” pa o he emaining den ine walls, unde 20× magni ica ion: (a)
immedia ely a e oo -end esec ion; (b) immedia ely a e oo -end p epa a ion—TPO; (c) 24 h a e
oo -end p epa a ion—T24H; (d) se en days a e oo -end p epa a ion—T7D.
3.2. Ma ginal In eg i y (Roo -End Ca i y Ma gins)
Rega ding ma ginal in eg i y (Table 3a), he maximum alue o “3” was ound in one oo om
NSK g oup, o aling 6.2% o he samples om he e e ed expe imen al g oup. The minimum
egis e ed sco e was “0” mos ly e i ied in specimens om CVDen us g oup (25% o he samples
om he g oup). The sco e “1” was he one wi h highes incidence in CVDen us g oup wi h a
pe cen age o 62.5%, whe eas in NSK g oup he alue wi h he highes incidence was “2” (43.8%). No
s a is ically signi ican di e ences (U = 84.00; Z = −1.783; P = 0.102) we e e i ied be ween bo h
CVDen us and NSK g oups ega ding ma ginal in eg i y (Figu e 3a).
Table 3. F equencies and pe cen ages ob ained in he wo expe imen al g oups ega ding (a) ma ginal
in eg i y and (b) p esence o deb is.
CVDen us NSK CVDen us NSK
(a) Ma ginal in eg i y *
0 4 3 25.0 18.8
1 10 5 62.5 31.2
2 2 7 12.5 43.8
3 0 1 − 6.2
(b) P esence o deb is **
0 9 2 56.2 12.5
1 6 6 37.5 37.5
2 1 7 6.2 43.8
3 0 1 − 6.2
4 0 0 − −
* N = 32; Mann—Whi ney es ; P = 0.102, ** N = 32; Mann—Whi ney es ; P = 0.003.
Figu e 2.
Pho omic og aphs o one sample om NSK g oup p esen ing one in acanal mic oc ack
(a ow) loca ed wi hin he “wide ” pa o he emaining den ine walls, unde 20
×
magni ica ion:
(
a
) immedia ely a e oo -end esec ion; (
b
) immedia ely a e oo -end p epa a ion—T
PO
; (
c
) 24 h a e
oo -end p epa a ion—T24H; (d) se en days a e oo -end p epa a ion—T7D.
3.2. Ma ginal In eg i y (Roo -End Ca i y Ma gins)
Rega ding ma ginal in eg i y (Table 3a), he maximum alue o “3” was ound in one oo om
NSK g oup, o aling 6.2% o he samples om he e e ed expe imen al g oup. The minimum
egis e ed sco e was “0” mos ly e i ied in specimens om CVDen us g oup (25% o he samples om
he g oup). The sco e “1” was he one wi h highes incidence in CVDen us g oup wi h a pe cen age o
62.5%, whe eas in NSK g oup he alue wi h he highes incidence was “2” (43.8%). No s a is ically
signi ican di e ences (U =84.00; Z =
−
1.783; p=0.102) we e e i ied be ween bo h CVDen us and
NSK g oups ega ding ma ginal in eg i y (Figu e 3a).
Biomedicines 2020,8, 383 8 o 19
Table 3.
F equencies and pe cen ages ob ained in he wo expe imen al g oups ega ding (
a
) ma ginal
in eg i y and (b) p esence o deb is.
F equency Pe cen ages (%)
CVDen us NSK CVDen us NSK
(a) Ma ginal in eg i y *
0 4 3 25.0 18.8
1 10 5 62.5 31.2
2 2 7 12.5 43.8
3 0 1 −6.2
(b) P esence o deb is **
0 9 2 56.2 12.5
1 6 6 37.5 37.5
2 1 7 6.2 43.8
3 0 1 −6.2
4 0 0 − −
* N =32; Mann—Whi ney es ; p=0.102, ** N =32; Mann—Whi ney es ; p=0.003.
Biomedicines 2020, 8, x FOR PEER REVIEW 8 o 18
Figu e 3. Sco e dis ibu ion wi hin he es ed g oups ega ding (a) ma ginal in eg i y (Mann—
Whi ney es ; P = 0.102) and (b) p esence o deb is. (Mann—Whi ney es ; P = 0.003).
3.3. P esence o Deb is (Walls Quali y)
Conce ning he p esence o deb is ollowing oo -end p epa a ion (Table 3b), he maximum
alue o “3” was ound in one sample om NSK g oup (6.2% o he samples om he e e ed
expe imen al g oup). The minimum egis e ed sco e was “0” mos ly e i ied in specimens om
CVDen us g oup (56.2% o he samples om he g oup). In NSK g oup he sco e exhibi ing he
highes incidence was “2” wi h a pe cen age o 43.8%. Con a iwise o ma ginal in eg i y, s a is ically
signi ican di e ences (U = 50.50; Z = −3.093; P = 0.003) we e ound be ween he es ed g oups
ega ding he p esence o deb is (Figu e 3b and Figu e 4).
Figu e 4. Scanning elec on mic oscopy (SEM) images ollowing oo -end p epa a ion: (a) CVDen us
specimen exhibi ing an in acanal mic oc ack (a ow); (b) NSK specimen exhibi ing an in acanal
mic oc ack (a ow) and i egula oo -end ca i y ma gins, as well as showing isible accumula ion o
deb is (as e isk) esul ing om he oo -end p epa a ion.
3.4. Time Requi emen s
Conside ing he ime equi emen s o pe o m apical p epa a ion (Table 4), no s a is ically
signi ican di e ences could be de ec ed be ween he es ed g oups (U = 120.00; Z = −0.302; P = 0.780).
Figu e 3.
Sco e dis ibu ion wi hin he es ed g oups ega ding (
a
) ma ginal in eg i y (Mann—Whi ney
es ; p=0.102) and (b) p esence o deb is. (Mann—Whi ney es ; p=0.003).
3.3. P esence o Deb is (Walls Quali y)
Conce ning he p esence o deb is ollowing oo -end p epa a ion (Table 3b), he maximum alue
o “3” was ound in one sample om NSK g oup (6.2% o he samples om he e e ed expe imen al
g oup). The minimum egis e ed sco e was “0” mos ly e i ied in specimens om CVDen us g oup
(56.2% o he samples om he g oup). In NSK g oup he sco e exhibi ing he highes incidence was
“2” wi h a pe cen age o 43.8%. Con a iwise o ma ginal in eg i y, s a is ically signi ican di e ences
(U =50.50; Z =
−
3.093; p=0.003) we e ound be ween he es ed g oups ega ding he p esence o
deb is (Figu es 3b and 4).
Biomedicines 2020,8, 383 9 o 19
Biomedicines 2020, 8, x FOR PEER REVIEW 8 o 18
Figu e 3. Sco e dis ibu ion wi hin he es ed g oups ega ding (a) ma ginal in eg i y (Mann—
Whi ney es ; P = 0.102) and (b) p esence o deb is. (Mann—Whi ney es ; P = 0.003).
3.3. P esence o Deb is (Walls Quali y)
Conce ning he p esence o deb is ollowing oo -end p epa a ion (Table 3b), he maximum
alue o “3” was ound in one sample om NSK g oup (6.2% o he samples om he e e ed
expe imen al g oup). The minimum egis e ed sco e was “0” mos ly e i ied in specimens om
CVDen us g oup (56.2% o he samples om he g oup). In NSK g oup he sco e exhibi ing he
highes incidence was “2” wi h a pe cen age o 43.8%. Con a iwise o ma ginal in eg i y, s a is ically
signi ican di e ences (U = 50.50; Z = −3.093; P = 0.003) we e ound be ween he es ed g oups
ega ding he p esence o deb is (Figu e 3b and Figu e 4).
Figu e 4. Scanning elec on mic oscopy (SEM) images ollowing oo -end p epa a ion: (a) CVDen us
specimen exhibi ing an in acanal mic oc ack (a ow); (b) NSK specimen exhibi ing an in acanal
mic oc ack (a ow) and i egula oo -end ca i y ma gins, as well as showing isible accumula ion o
deb is (as e isk) esul ing om he oo -end p epa a ion.
3.4. Time Requi emen s
Conside ing he ime equi emen s o pe o m apical p epa a ion (Table 4), no s a is ically
signi ican di e ences could be de ec ed be ween he es ed g oups (U = 120.00; Z = −0.302; P = 0.780).
Figu e 4.
Scanning elec on mic oscopy (SEM) images ollowing oo -end p epa a ion: (
a
) CVDen us
specimen exhibi ing an in acanal mic oc ack (a ow); (
b
) NSK specimen exhibi ing an in acanal
mic oc ack (a ow) and i egula oo -end ca i y ma gins, as well as showing isible accumula ion o
deb is (as e isk) esul ing om he oo -end p epa a ion.
3.4. Time Requi emen s
Conside ing he ime equi emen s o pe o m apical p epa a ion (Table 4), no s a is ically
signi ican di e ences could be de ec ed be ween he es ed g oups (U =120.00; Z =
−
0.302; p=0.780).
Table 4.
Mean, s anda d de ia ion (SD), and minimum and maximum alues o each expe imen al
g oup ega ding he ime o apical p epa a ion.
Time o Apical P epa a ion
Mean SD Minimum Maximum
CVDen us 60.56 33.18 30.0 160.0
NSK 55.56 20.41 34.0 96.0
N=32; Mann—Whi ney es ; p=0.780.
3.5. Roo -End P epa a ion Tip Wea —Quali a i e Analysis
Figu es 5and 6depic images o he ul asonic ips ob ained by scanning elec on mic oscopy.
(SEM) examina ion, showing ip wea a e oo -end p epa a ion. No ul asonic ip ac u es we e
egis e ed in any o he g oups.
Biomedicines 2020,8, 383 16 o 19
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