Background and Aim: Ideal scaffold for tissue engineering must be a mimic of extracellular matrices that facilitates proliferation and differentiation of the cells. Biological scaffolds are prepared from decellularization of human or animal tissues. Extracellular matrix of natural tissues can be used as a scaffold for regenerative applications, even xenogenically. In this study, for the first time complete human skin tissue (dermis plus epidermis) was decellularized. This matrix can be used in the clinics for the treatment of severe burns. Unseparated epidermis from the dermis may cause the skin to maintain higher levele of moisture until complete healing.
Material and Methods: Human skin tissues were decellularized by combination of three methods: physical (slow freezing and snap freeze–thaw), enzymatic (trypsin 0.25% for 18 hours) and chemical (sodium dodecyl sulfate- SDS 1% for 12 hours). Then, the prepared decellularized 3D matricies with 5×105 of cells were cultured. Finally, histological studies were performed after 1, 2, 3 and 4 weeks of culture by different stains.
Results: Histological studies confirmed cell elimination and also preservation of collagen fibers in the human skin matrix. In vitro analysis of cellular behaviors showed adhesion and proliferation of L929 fibroblast cell line in the culture in different weeks. Statistical analysis indicated significant induction of cell densities at the end of the 2nd week (p<0.001) compared to the other weeks of our study. The number of cells reduced after the 3rd week.
Conclusion: The results indicated that remnants of ECM of the human skin can be a suitable matrix for adherence, proliferation and differentiation of L929 cells. On the other hand, because of high density of collagen fibers and low porosity of prepared scaffolds, we found no penetration and migration into the dermis.
Keywords: Extracellular matrix, Human skin, Decellularization, L929 fibroblast cells, Scaffold.
Received: Apr 28, 2015 Accepted: Feb 13, 2016 |
