In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin Models
<b>Background/Objectives</b>: Drugs for human use require several studies for the assessment of their efficacy and safety. An important property is cytotoxicity, which should be tested in different environments and models in closer proximity to the final use of the drug, with greater rel...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-07-01
|
| Series: | Antibiotics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2079-6382/14/7/691 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849418621624778752 |
|---|---|
| author | Marcel Nani Leite Natália Aparecida de Paula Leandra Náira Zambelli Ramalho Marco Andrey Cipriani Frade |
| author_facet | Marcel Nani Leite Natália Aparecida de Paula Leandra Náira Zambelli Ramalho Marco Andrey Cipriani Frade |
| author_sort | Marcel Nani Leite |
| collection | DOAJ |
| description | <b>Background/Objectives</b>: Drugs for human use require several studies for the assessment of their efficacy and safety. An important property is cytotoxicity, which should be tested in different environments and models in closer proximity to the final use of the drug, with greater reliability. Thus, we proposed to evaluate the toxicity of rifampicin, the only bactericidal drug in the anti-leprosy multidrug therapy, using skin cells and skin explant cultures. <b>Methods</b>: Cell viability was tested by the MTT method using primary keratinocytes and fibroblasts and immortalized skin cells (HaCaT and 3T3) at 24, 48, and 72 h of treatment. For the skin explant, we used the TTC assay to determine viability (24, 48, 72, and 96 h), hematoxylin and eosin staining to analyze the structure and architecture of the tissue, and TUNEL to assess apoptotic cells at 3, 6, 12, 24, 48, 72, and 96 h. <b>Results</b>: Regarding the toxicity of primary and immortalized cells, viability was above 70% up to a concentration of 50 μg/mL at 24, 48, and 72 h, and at the concentration of 200 μg/mL, all cells showed greater sensitivity, especially at 72 h. Tissue viability analysis revealed a high percentage (above 96%) of viable tissue at the concentrations of 100, 150, and 200 μg/mL at the time points studied. Histological analysis showed that tissue architecture was maintained, with no apoptotic cells being observed. <b>Conclusions</b>: Thus, our results showed the importance of evaluating drug toxicity using different cell types, with the ex vivo skin model proving to be an alternative to animal use. |
| format | Article |
| id | doaj-art-7f6739ff7fb04ff3baad16228cb831a4 |
| institution | Kabale University |
| issn | 2079-6382 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj-art-7f6739ff7fb04ff3baad16228cb831a42025-08-20T03:32:24ZengMDPI AGAntibiotics2079-63822025-07-0114769110.3390/antibiotics14070691In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin ModelsMarcel Nani Leite0Natália Aparecida de Paula1Leandra Náira Zambelli Ramalho2Marco Andrey Cipriani Frade3Division of Dermatology, Department of Internal Medicine, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14040-900, São Paulo, BrazilDivision of Dermatology, Department of Internal Medicine, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14040-900, São Paulo, BrazilDepartment of Pathology and Legal Medicine, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14040-900, São Paulo, BrazilDivision of Dermatology, Department of Internal Medicine, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14040-900, São Paulo, Brazil<b>Background/Objectives</b>: Drugs for human use require several studies for the assessment of their efficacy and safety. An important property is cytotoxicity, which should be tested in different environments and models in closer proximity to the final use of the drug, with greater reliability. Thus, we proposed to evaluate the toxicity of rifampicin, the only bactericidal drug in the anti-leprosy multidrug therapy, using skin cells and skin explant cultures. <b>Methods</b>: Cell viability was tested by the MTT method using primary keratinocytes and fibroblasts and immortalized skin cells (HaCaT and 3T3) at 24, 48, and 72 h of treatment. For the skin explant, we used the TTC assay to determine viability (24, 48, 72, and 96 h), hematoxylin and eosin staining to analyze the structure and architecture of the tissue, and TUNEL to assess apoptotic cells at 3, 6, 12, 24, 48, 72, and 96 h. <b>Results</b>: Regarding the toxicity of primary and immortalized cells, viability was above 70% up to a concentration of 50 μg/mL at 24, 48, and 72 h, and at the concentration of 200 μg/mL, all cells showed greater sensitivity, especially at 72 h. Tissue viability analysis revealed a high percentage (above 96%) of viable tissue at the concentrations of 100, 150, and 200 μg/mL at the time points studied. Histological analysis showed that tissue architecture was maintained, with no apoptotic cells being observed. <b>Conclusions</b>: Thus, our results showed the importance of evaluating drug toxicity using different cell types, with the ex vivo skin model proving to be an alternative to animal use.https://www.mdpi.com/2079-6382/14/7/691rifampicincell cultureskin explant modeltoxicityhistological analysis |
| spellingShingle | Marcel Nani Leite Natália Aparecida de Paula Leandra Náira Zambelli Ramalho Marco Andrey Cipriani Frade In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin Models Antibiotics rifampicin cell culture skin explant model toxicity histological analysis |
| title | In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin Models |
| title_full | In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin Models |
| title_fullStr | In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin Models |
| title_full_unstemmed | In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin Models |
| title_short | In Vitro and Ex Vivo Evaluation of Rifampicin Cytotoxicity in Human Skin Models |
| title_sort | in vitro and ex vivo evaluation of rifampicin cytotoxicity in human skin models |
| topic | rifampicin cell culture skin explant model toxicity histological analysis |
| url | https://www.mdpi.com/2079-6382/14/7/691 |
| work_keys_str_mv | AT marcelnanileite invitroandexvivoevaluationofrifampicincytotoxicityinhumanskinmodels AT nataliaaparecidadepaula invitroandexvivoevaluationofrifampicincytotoxicityinhumanskinmodels AT leandranairazambelliramalho invitroandexvivoevaluationofrifampicincytotoxicityinhumanskinmodels AT marcoandreyciprianifrade invitroandexvivoevaluationofrifampicincytotoxicityinhumanskinmodels |