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Đánh giá ảnh hưởng của nồng độ Trypsin - Edta đến hiệu quả tách tế bào và bảo toàn cấu trúc da trong chế tạo tấm trung bì da đồng loại không tế bào
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Abstract
Human acellular dermal matrix (hADM) is a biological scaffold derived from human skin in which the epidermis and cellular components of the dermis are completely removed, while preserving the collagen structure of the dermis.
Objective: This study aimed to evaluate the effect of different Trypsin-EDTA concentrations on cell removal efficiency and structural preservation of the dermis, in order to determine the optimal concentration for the preparation of hADM.
Subjects and methods: Thirty donated human skin samples were randomly assigned into three groups (n = 10 each) and treated with different Trypsin-EDTA concentrations: 0.5X (0,125%), 1X (0.25%) and 1.5X (0.375%). All samples underwent repeated freeze-thaw cycles, followed by a two-step enzymatic decellularization process: immersion at 40C for 24 hours, then incubation at 370C for 3 hours. Epidermal separation efficiency and cellular removal were evaluated using epidermal detachment rates and histological analysis.
Results: Increasing Trypsin - EDTA concentrations showed a strong correlation with higher epidermal separation rates after enzymatic treatment at 40C for 24 hours (p < 0.05), with mean 15.24 ± 8.94%, 36.84 ± 7.44%, and 45.99 ± 7.10% for the 0.5X, 1X and 1.5X groups, respectively. After the second enzymatic treatment at 370C for 3 hours, the separation rates continued to rise, reaching 28.25 ± 10.33%, 93.98 ± 13.08%, and 95.47 ± 11.99%, respectively. However, the difference between the 1X and 1.5X groups was not statistically significant (p > 0.05). Histological evaluation confirmed complete cell removal in all groups. However, the 1.5X group exhibited more pronounced disruption of collagen structure, with lower fiber density and looser organization.
Conclusion: Trypsin - EDTA proved to be an effective agent for both epidermal separation and dermal decellularization in hADM preparation. A concentration of 0.25% (1X) was identified as optimal, providing high cell removal efficiency while better preserving collagen structure.
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Keywords
Acellular dermal matrix, Decellularization, Trypsin - EDTA, Allogenic skin, Tissue engineering
References
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