Morphological characteristics of acellular dermal matrix manufacturing

Abstract

The aim is to develop a method of the acellular dermal matrix manufacturing from pig’s skin dermis while preserving the native structure. Materials and methods: Combination of physical and chemical effects on the dermis underlies in the process of an acellular dermal matrix manufacturing. Dermal collection of 1.0-1.3 mm thickness in pigs under 1 year of age from the back and partially from the lateral parts of the body was carried out. The 0.3-0.4 mm thickness layer of skin was previously removed from the relevant areas with help of a dermatome, which was physically and chemically treated. The maximum acellularization of the dermal matrix was achieved step-by-step and included four stages of skin processing: 1 – freeze-thaw process; 2 – glycerin dehydration; 3 – osmotic stress; 4 – cell residue removal by detergent. Results: Histological analysis of the of the pig’s skin dermis revealed that after freeze-thaw cycles the collagen scaffold of the dermal matrix maintains its structural organization that was obtained as a result of the first stage of decellularization. On the second stage of decullalarization, the decreased number of fibroblastic cells was indicated. By means of this, the connective tissue elements that are represented by collagen fibers’ multidirectional bundles retained their structural organization. Fibroblasts lysis as basophilic stained elements was revealed in small amount of dermis on the third stage of the decellularization. Washing of lyophilized skin with nonionic detergent sodium dodecyl sulfate the complete absence of fibroblasts, epidermocytes in the hair follicles, endothelial cells in the wall of blood vessels was detected indicating the effectiveness of this reagent in removing residual products. Conclusions: Suggested protocol for decullalarization of the pig’s skin dermis is effective in removing nuclear and cellular structures from dermis. Particular protocols can be modified by increasing the temperature difference or changing the number of freeze-thaw cycles. KEY WORDS: pig’s skin, derma, acellular dermal matrix, morphology.