Toxicological Assessment of 2-Hydroxychalcone-Mediated Photodynamic Therapy: Comparative In Vitro and In Vivo Approaches

Author(s)

Bila, Niura Madalena ; Vaso, Carolina Orlando ; Belizario, Jenyffie Araujo ; Assis, Leticia Ribeiro ; Regasini, Luis Octavio ; Fontana, Carla Raquel ; Fusco-Almeida, Ana Marisa ; Costa-Orlandi, Caroline Barcelos ; Mendes-Giannini, Maria Jose Soares

• Document type: Journal article
• Source: PHARMACEUTICS; v. 16, n. 12, p. 14-pg., 2024-12-01.

Abstract

Background: Photodynamic therapy (PDT) is a treatment modality that uses light to activate a photosensitizing agent, destroying target cells. The growing awareness of the necessity to reduce or eliminate the use of mammals in research has prompted the search for safer toxicity testing models aligned with the new global guidelines and compliant with the relevant regulations. Objective: The objective of this study was to assess the impact of PDT on alternative models to mammals, including in vitro three-dimensional (3D) cultures and in vivo, in invertebrate animals, utilizing a potent photosensitizer, 2-hydroxychalcone. Methods: Cytotoxicity was assessed in two cellular models: monolayer (2D) and 3D. For this purpose, spheroids of two cell lines, primary dermal fibroblasts (HDFa) and adult human epidermal cell keratinocytes (HaCat), were developed and characterized following criteria on cell viability, shape, diameter, and number of cells. The survival percentages of Caenorhabditis elegans and Galleria mellonella were evaluated at 1 and 7 days, respectively. Results: The findings indicated that all the assessed platforms are appropriate for investigating PDT toxicity. Furthermore, 2-hydroxychalcone demonstrated low toxicity in the absence of light and when mediated by PDT across a range of in vitro (2D and 3D cultures) and in vivo (invertebrate animal models, including G. mellonella and C. elegans) models. Conclusion: There was a strong correlation between the in vitro and in vivo tests, with similar toxicity results, particularly in the 3D models and C. elegans, where the concentration for 50% viability was approximately 100 mu g/mL. (AU)

• FAPESP's process: 20/15586-4 - Establishment of a three-dimensional model to determine the pharmacological efficacy and safety of nitrofuran and indoles derivatives and evaluation of Histoplasma capsulatum infection
• Grantee: Carolina Orlando Vaso
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 19/22188-8 - Evaluation of the interaction of biofilms mono and dual-species of Candida spp and dermatophytes photodynamic therapy combined with 2-chalcone
• Grantee: Níura Madalena Bila
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 21/03805-6 - Monospecies and mixed biofilms of Trichophyton and Staphylococcus: influence of nutrients and presence of persister cells
• Grantee: Jenyffie Araujo Belizario
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 18/02785-9 - Dermatophytes and Dermatophytosis: biofilm formation and development of control strategies.
• Grantee: Maria José Soares Mendes Giannini
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/18388-6 - Role of biofilm in the pathogenesis of dermatophytosis and development of combat strategies
• Grantee: Caroline Barcelos Costa Orlandi
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 23/03556-1 - Study of the pathogenesis of mono/polymicrobial dermatophytes biofilms in ex vivo models and application of control strategies.
• Grantee: Maria José Soares Mendes Giannini
• Support Opportunities: Regular Research Grants