Microfluidic Engineering for the Development of Ozonized Oil Microemulsions Aimed at Canine Leishmania Treatment: Implementation of 3D Skin Models
Lopes, Inês; Tavares, Vitória; Cerqueira, Débora; Weber, Juliana; Martins, Liege; CIMETTA, Elisa; Micheli, Sara; Zanrè, Eleonora; Souza, Vinicius RC; Santos-Gomes, Gabriela; Rodrigues, Armanda; Martins, Verónica C
- Publisher
- Zenodo
- Language
- en
Abstract
The skin is the primary interface between the host and Leishmania parasites, where initial interactions can lead to severe cutaneous lesions [1]. In vitro 3D skin models allow for studying these interactions and exploring new therapies like ozone therapy, recognized for its antimicrobial and regenerative properties [2, 3]. However, ensuring the stability and bioavailability of ozonederived compounds in ozonated vegetable oils remains a challenge, requiring innovative engineering approaches. This study aims to establish and evaluate the viability of canine skin explants in vitro as a platform for testing the efficacy of ozonized oil against Leishmania infection. Additionally, we explore different approaches for developing ozonized oil microemulsions. In this initial phase, a pre-existing microfluidic device was used to evaluate formulation stability. Skin explants from five dogs (n=5) were maintained in culture and assessed at T0, T3, T5, T7, and T10. Metabolic Activity was analyzed using the resazurin assay, while histological integrity was assessed through hematoxylin-eosin staining. In parallel, a microfluidic-based device was used to produce microemulsions of ozonized olive oil at 400 IP and 800 IP in an aqueous solution of surfactant (Tween 80 at 2.5%) and study formulation stability. Preliminary results demonstrated that the culture system maintained viable skin explants for at least five days, supporting its feasibility for Leishmania infection studies. Microfluidic systems showed promise in developing ozonized oil microemulsions, with further optimisation needed. Future applications may extend beyond cutaneous treatments, potentially exploring ocular and other therapeutic uses.
Full text
Corresponding/Presenting Author: Inês Lopes,mailto:a2100201[email protected] Microfluidic Engineering for the Development of Ozonized Oil Microemulsions Aimed at Canine Leishmania Treatment: Implementation of 3D Skin Models Lopes, Inês MC1; Tavares, Vitória1; Cerqueira, Débora1; Weber, Juliana1; Martins, Liege2; Cimetta, Elisa3; Micheli Sara3; Zanrè, Eleonora3; Souza, Vinicius RC 2; SantosGomes, Gabriela1; Rodrigues, Armanda1; Martins, Verónica C4 1Global Health and Tropical Medicine, GHTM, Associate Laboratory in Translation and Innovation Towards Global Health, LA-REAL, Inst. de Higiene e Medicina Tropical, Univ. Nova de Lisboa, UNL, Portugal; 2Faculdade de Medicina Veterinária, Universidade Lusófona, Lisboa, Portugal; 3Department of Industrial Engineering (DII), University of Padua, Via Marzolo 9, 35131 Padova, Italy ; 42M Pharma Lda, Sobreda, Almada, Portugal The skin is the primary interface between the host and Leishmania parasites, where initial interactions can lead to severe cutaneous lesions [1]. In vitro 3D skin models allow for studying these interactions and exploring new therapies like ozone therapy, recognized for its antimicrobial and regenerative properties [2, 3]. However, ensuring the stability and bioavailability of ozonederived compounds in ozonated vegetable oils remains a challenge, requiring innovative engineering approaches. This study aims to establish and evaluate the viability of canine skin explants in vitro as a platform for testing the efficacy of ozonized oil against Leishmania infection. Additionally, we explore different approaches for developing ozonized oil microemulsions. In this initial phase, a pre-existing microfluidic device was used to evaluate formulation stability. Skin explants from five dogs (n=5) were maintained in culture and assessed at T0, T3, T5, T7, and T10. Metabolic Activity was analyzed using the resazurin assay, while histological integrity was assessed through hematoxylin-eosin staining. In parallel, a microfluidic-based device was used to produce microemulsions of ozonized olive oil at 400 IP and 800 IP in an aqueous solution of surfactant (Tween 80 at 2.5%) and study formulation stability. Preliminary results demonstrated that the culture system maintained viable skin explants for at least five days, supporting its feasibility for Leishmania infection studies. Microfluidic systems showed promise in developing ozonized oil microemulsions, with further optimisation needed. Future applications may extend beyond cutaneous treatments, potentially exploring ocular and other therapeutic uses. Funding/ acknowledgements: This work was supported by FCT-Foundation for Science and Technology (FCT) through research grants miT3D-DOI 10.54499/EXPL/CVT-CVT/0175/2021; DogIPMDOI 10.54499/PTDC/CVT-CVT/0228/2020, 2022.00499.CEECIND/CP1725/CT0023, studentships, 2023.02612.BDANA, National funds GHTM—UID/04413/2020 and LA-REAL— LA/P/0117/2020, and European funds Ipamena, H2020-MSCA-RISE-2019 GA ID: 872662, doi: 10.3030/872662. References [1] F. Sasani, J. Javanbakht, R. Samani, and D. Shirani, Canine cutaneous leishmaniasis,Journal of Parasitic Diseases, vol. 40, no. 1, pp. 57–60, doi: 10.1007/s12639-014-0444-4. [2] T. F. Moda et al., Effects of ozone therapy on hematological and serum biochemical parameters in dogs affected by visceral leishmaniasis, Research, Society and Development, vol. 11, no. 7, p. e16711729886, doi: 10.33448/rsd-v11i7.29886. [3] I. L. Cabral et al., Aqueous ozone therapy improves the standard treatment of leishmaniasis lesions in animals leading to local and systemic alterations, Parasitol Res, vol. 119, no. 12, pp. 4243–4253, , doi: 10.1007/s00436-020-06925-8.
