A new approach to the production of a biovital skin graft based on human acellular dermal matrix produced in‐house, in vitro revitalized internally by human fibroblasts and keratinocytes on the surface

Abstract

Patients with extensive and deep burns who do not have enough donor sites for autologous skin grafts require alternative treatment methods. Tissue engineering is a useful tool to solve this problem. The aim of this study was to find the optimal method for the production of a biovital skin substitute based on acellular dermal matrix (ADM) and in vitro cultured fibroblasts and keratinocytes. In this work, nine methods of ADM production were assessed. The proposed methods are based on the use of the following enzymes: Dispase II, collagenase I/ethylenediaminetetraacetic acid (EDTA), collagenase II/EDTA, and mechanical perforation using DermaRoller and mesh dermatome. The obtained ADMs were examined (both on the side of the basement membrane and on the “cut‐off” side) by means of scanning electron microscopy, immunohistochemistry tests and strength tests. ADM was revitalized with human fibroblasts and keratinocytes. The ability of in‐depth revitalization of cultured fibroblasts and their ability to secrete collagen IV was examined. The obtained results indicate that the optimal method of production of live skin substitutes is the colonization of autologous fibroblasts and keratinocytes on the scaffold obtained using two‐step incubation method: Trypsin/EDTA and dispase II.

Influence of electron beam irradiation on extracellular matrix of the human allogeneic skin grafts

“…In the treatment of deep and extensive skin burns, allogeneic human skin grafts are an important element of the therapy (Table 1). [1][2][3][4][5][6]11,20 Such transplants might provide an optimal alternative to autologous skin grafts 1,[3][4][5][6]11,20 and among their many advantages, they can even be considered as life-saving transplants. 1 However, allogeneic human skin grafts are often initially infected with an undefined quantity of hospital strains of microorganisms (possibly multidrug-resistant).…”

“…25 Additionally, a Fourier transform infrared spectroscopy (FTIR) may be used to assess the changes in molecular structure of ECM collagen elements of human skin (e.g., quantity of hydrogen bounds in collagen affected by irradiation). 6,11,[26][27][28] Therefore, changes in the molecular structure of ECM elements might affect the mechanical properties of the tested biomaterial. Thus, performing mechanical tests may be useful in assessing the effect of radiation sterilization on allogeneic human skin.…”

“…T A B L E 3 The range of gamma radiation doses used to sterilize tissue allografts and its impact on extracellular matrix constitution 6,23,25,28,29,52 Gamma The skin was prepared according to SOP and sterilized by radiation (electron beams of 18, 25, or 35 kGy).…”

“…allogeneic nonviable skin grafts, electron beam irradiation, extracellular matrix degradation, sterilization, tissue engineering 1 | BACKGROUND It has been described that the optimal method for the extensive and fullthickness skin burn treatment is a transplantation of autologous split thickness skin grafts (STSG) obtained from an undamaged area of the body (Table 1). [1][2][3][4][5][6] Among many advantages of the autologous STSGs, the compliance with the patient's major histocompatibility complex (MHC) could be listed. This characteristic could be considered as their major advantage.…”

“…19 It has been proposed that the range of irradiation dose used by tissue banks to sterilize tissue allografts is contained between 15 and 35 kGy (Table 3). 20,[23][24][25] However, T A B L E 1 Possible advantages and disadvantages of autologous free split-thickness skin graft (STSG) 1,[3][4][5][6] Autologous free split-thickness skin graft (STSG)…”