title: Development of a new method for the preparation of an acellular allodermis, quality control and cytotoxicity testing.
abstract: Demand for use of acellular allodermis is high but commercially appropriate products are not used routinely because of very high price and limited availability. These facts did motivate us to prepare acellular allodermis using a new, simple and less expensive method. We have developed a original method for preparation of acellular allogeneic dermis based on action of a proteolytic enzyme in combination with distilled water. Hypotonic environment in comparison with SDS or Triton ansure no toxicity of the final product. Trials for determination of optimal trypsin concentrations, temperature and time of action were performed. According to our results, the use of 2.5% trypsin/EDTA solution overnight at +4 °C was proving to be optimal. The histology confirmed absence of cells in the prepared dermis. No toxicity of final acellular dermis was confirmed by three independent tests (agar diffusion test contact cytotoxicity test and grow curve). The prepared acellular dermis seems to be suitable not only for direct clinical use, but it can be used as a scaffold for cell cultivation as well.
authors.affiliation: Department of Burns and Reconstructive Surgery, Faculty of Medicine, Comenius University Bratislava, Bratislava, Slovak Republic.
authors.forename: J
authors.initials: J
authors.lastname: Dragúňová
authors.affiliation: Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University, Bratislava, Slovak Republic. virupepo@savba.sk.|Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Bratislava, Slovak Republic. virupepo@savba.sk.
authors.forename: P
authors.initials: P
authors.lastname: Kabát
authors.affiliation: Institute of Pathological Anatomy, Faculty of Medicine, Comenius University, Bratislava, Slovak Republic.
authors.forename: P
authors.initials: P
authors.lastname: Babál
authors.affiliation: Institute of Pathological Anatomy, Faculty of Medicine, Comenius University, Bratislava, Slovak Republic.
authors.forename: H
authors.initials: H
authors.lastname: Mrázová
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Faculty of Medicine, Comenius University, Bratislava, Slovak Republic.
authors.forename: M
authors.initials: M
authors.lastname: Boháč
authors.affiliation: Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University, Bratislava, Slovak Republic.
authors.forename: Ľ
authors.initials: Ľ
authors.lastname: Krajčíová
authors.affiliation: Department of Burns and Reconstructive Surgery, Faculty of Medicine, Comenius University Bratislava, Bratislava, Slovak Republic.
authors.forename: J
authors.initials: J
authors.lastname: Koller
mesh_terms: D016475:3T3 Cells; D062088:Acellular Dermis / Q000009:adverse effects* / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods; D004492:Edetic Acid / Q000378:metabolism; D006801:Humans; D051379:Mice; D009997:Osmotic Pressure; D059748:Proteolysis; D011786:Quality Control; D016038:Skin Transplantation; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000009:adverse effects* / Q000737:chemistry; D020858:Tissue and Organ Harvesting / Q000379:methods; D014357:Trypsin / Q000378:metabolism; D014867:Water / Q000737:chemistry
chemical_list: D014867:Water; D004492:Edetic Acid; D014357:Trypsin
keywords: Cadaverous allodermis; Decellularization; New preparation procedure