Techniques for obtaining dermal extracellular matrix scaffold

Despite advancements in modern surgery in the treatment of cutaneous injuries, the search for new methods of ensuring faster and more effective wound healing appears especially urgent today. Tissue engineering is undoubtedly of interest when it comes to developing such technologies. Objective: to determine the optimal protocol for obtaining a decellularized dermal matrix scaffold for subsequent development of tissue-engineered skin. Materials and methods. One Landrace piglet was used as the experimental animal. After preliminary skin treatment with dermatome, 0.3 cm thick samples were taken. Two decellularization protocols were considered: protocol No. 1 was based on the use of Triton X-100 and deoxycholate, protocol No. 2 was only based on deoxycholate. There were 5 processing cycles in total for the 2 protocols. After treatment, acellular matrix scaffolds were examined through histological analysis and quantitative determination of DNA concentration. Next, static recellarization of the matrix scaffolds was carried out with porcine dermal fibroblasts. After that, the matrix scaffolds were tested for cytotoxicity using XTT test and differential staining test to differentiate between live and dead cells. Results. Comparative analysis of the two protocols for porcine dermis decellularization showed that both protocols effectively remove cells and nuclear material, while maintaining the architectonics of the intercellular substance intact, since fibrous structures are not destroyed. But when assessing the biocompatibility of matrix scaffolds based on analysis of cell viability according to data obtained from XTT test and cell–matrix adhesion, the matrix scaffold processed under protocol No. 1, shows advantages. Conclusion. In this study, a decellularization protocol based on Triton X-100 and deoxycholate was noted. The results obtained mark the first stage towards developing a tissue-engineered skin.

regenerative medicine, skin, decellularization, tissue‑engineered graft

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