Preparation method for decellularized matrix biomaterial

A method for preparing a decellularized matrix biomaterial, consisting essentially of the following steps:

• (a) rinsing off a residue from a surface layer of an animal tissue with purified water,
• (b) placing the animal tissue in a solution of 10-40 wt % NaCl and shaking for 1-6 h,
• (c) placing in a solution of 0.05-1 wt % NaCl and shaking for 12-20 h,
• (d) removing a muscle layer and an epidermal layer of the animal tissue by physical scraping to obtain a matrix material, wherein the animal tissue is selected from the group consisting of bovine pericardium, porcine pericardium, sheep stomach mucosa, porcine peritoneum, porcine bladder membrane, and porcine small intestine mucos;
• (e) mixing the matrix material with a degreasing reagent according to a mass ratio of matrix material to solution of 1:5-1:20 and shaking for 30-180 min;
• wherein the degreasing reagent is selected from the group consisting of a solution of 2-10 wt % sodium dodecyl sulfonate, a solution of 0.1-1 wt % sodium hydroxide, a solution of 10-100 wt % isopropanol, a solution of 20-80 wt % n-propanol, a mixed solution of 20-60 wt % trichloromethane and 40-80 wt % ethanol, a mixed solution of 30-50 wt % methylene chloride and 50-70 wt % ethanol, a solution of 10-65 wt % petroleum ether, and a combination thereof;
• (f) mixing the matrix material with a disinfectant according to a mass ratio of matrix material to solution of 1:3-1:10 and shaking for 30-180 min;
• wherein the disinfectant is selected from the group consisting of a mixed solution of 0.02-0.2 wt % peroxyacetic acid and 10-40 wt % ethanol, a solution of 0.02-0.2 wt % peroxyacetic acid, a solution of 5-10 wt % hydrogen peroxide, a solution of 0.25-2.5 wt % sodium hypochlorite and a solution of 75-95 wt % ethanol;
• (g) mixing the matrix material with a solution of 0.01-0.5 wt % alkaline protease at a mass ratio of matrix material to solution of 1:3-1:10 and shaking for 60-240 min;
• (h) performing an operation selected from the group consisting of repeated freeze-thaw rinsing, mechanical crushing rinsing, rinsing with an α-galactosidase solution, and a combination thereof;
• and
• (i) sterilizing the matrix material by irradiation sterilization or ethylene oxide sterilization to obtain the decellularized matrix biomaterial.