METHOD OF PRODUCING ACELLULAR DERMAL MATRIX Russian patent published in 2020 - IPC A61L27/36 

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to regenerative medicine, and can be used in reconstructive surgery to create a technology for obtaining and using bioengineering matrix as a graft for practical purposes. That is ensured by sampling pig's dermis 3 mm thick after preliminary removal of the epidermis by dermatome, then samples are frozen to temperature of -80 °C, after defrosting samples are poured with Trypsin-Versene solution and placed in shaker-incubator in mode of 100 rpm at 37 °C for 18 hours; at the second stage, the samples are placed on a rotating platform in mode of 170 rpm and subjected to sequential cyclic action of detergent solutions: 1 % Triton X100 solution for 2 hours and 4 % sodium deoxycholate in combination with 0.002 M Na2-EDTA for 2 hours, total number of treatment cycles is equal to 5. After each detergent samples are washed in deionised water for 10 minutes. At the third stage, the samples are treated with a solution of porcine pancreatic DNase I (2,000 units are dissolved in 200 ml of a phosphate buffer with calcium and magnesium) in a shaker incubator in mode of 100 rpm at 37 °C for 4 hours. Decellularization is terminated by exposure to 10 % chlorhexidine bigluconate in phosphate buffer for 24 hours with solution change every 6 hours. That is followed by confirming the quality of decellulyarisation by histological examination, molecular-biological analysis and culture methods.

EFFECT: method enables reducing exposure time of decellularising solutions, reducing the level of residual DNA in tissue up to 60 ng/mg of tissue in a wet sample.

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