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Published on: October 28, 2013
Sustainable Primary Cell Banking for Topical Compound Cytotoxicity Assays: Protocol Validation on Novel Biocides and
Zhifeng Liao1, Nicolas Laurent1, Nathalie Hirt-Burri1,2
1Plastic, Reconstructive and Hand Surgery Service, Lausanne University Hospital, University of Lausanne, CH-1011 Lausanne, Switzerland.
Abstract:
Thorough biological safety testing of topical therapeutic compounds and antimicrobials is a critical prerequisite for appropriate cutaneous wound care. Increasing pathogen resistance rates to traditional antibiotics and antifungals are driving the development and registration of novel chemical entities. Although they are notably useful for animal testing reduction, the gold standard in vitro cytotoxicity assays in continuous cell lines (HaCaT keratinocytes, 3T3 fibroblasts) may be discussed from a translational relevance standpoint. The aim of this study was thus to establish and validate a sustainable primary cell banking model with a view to performing optimized in vitro cytotoxicity assay development. Primary dermal fibroblasts and adipose-derived stem cell (ASC) types were established from four infant polydactyly sources. A multi-tiered primary cell banking model was then applied to prepare highly sustainable and standardized dermal fibroblast and ASC working cell banks (WCBs), potentially allowing for millions of biological assays to be performed. The obtained cellular materials were then validated for use in cytotoxicity assays through in vitro biosafety testing of topical antiseptics (chlorhexidine, hypochlorous acid) and an antifungal compound (AR-12) of interest for optimized burn wound care. The experimental results confirmed that IC50 values were comparable between cytotoxicity assays, which were performed with cell lines and with primary cells. The results also showed that hypochlorous acid (HOCl) displayed an enhanced toxicological profile as compared to the gold standard chlorhexidine (CLX). Generally, this study demonstrated that highly sustainable primary cell sources may be established and applied for consistent topical compound biological safety assessments with enhanced translational relevance. Overall, the study underscored the safety-oriented interest of functionally benchmarking the products that are applied on burn patient wounds for the global enhancement of burn care quality.
Insights
Developing a sustainable primary cell banking model enhances in vitro cytotoxicity testing for topical therapeutics. This approach offers improved translational relevance for biological safety assessments in wound care.
Area of Science:
- Biomedical Science
- Cell Biology
- Dermatology
Background:
- Rising antimicrobial resistance necessitates novel topical therapeutics.
- In vitro cytotoxicity assays are crucial for biological safety testing of wound care products.
- Current reliance on continuous cell lines may limit translational relevance in preclinical safety assessments.
Purpose of the Study:
- To establish and validate a sustainable primary cell banking model for in vitro cytotoxicity assay development.
- To enhance the translational relevance of biological safety testing for topical therapeutic compounds.
- To optimize preclinical safety assessments for burn wound care products.
Main Methods:
- Established primary dermal fibroblasts and adipose-derived stem cells (ASCs) from infant polydactyly tissue.
- Developed a multi-tiered primary cell banking model to create standardized working cell banks (WCBs).
- Validated the model using in vitro biosafety testing of topical antiseptics (chlorhexidine, hypochlorous acid) and an antifungal (AR-12).
Main Results:
- Primary cell banks provided sustainable and standardized cellular material for millions of assays.
- Cytotoxicity assay results (IC50 values) using primary cells were comparable to those using established cell lines.
- Hypochlorous acid demonstrated a more favorable toxicological profile than chlorhexidine in primary cell assays.
Conclusions:
- Sustainable primary cell sources can be reliably established for consistent biological safety assessments of topical compounds.
- This primary cell banking model offers enhanced translational relevance for in vitro cytotoxicity testing.
- Benchmarking product safety using functionally relevant models is vital for improving burn care quality.
