FIELD: medicine.
SUBSTANCE: invention refers to medicine, namely to medical biotechnologies, and aims at producing a biological-based implantation material having a regenerative potential, as well as a biocarrier when creating tissue engineering structures and organ prototypes. Method for producing dermal matrix involves de-epithelialization of skin flap, decellularisation, sterilization, packaging and preservation, as well as quality control. Initial raw material is swine skin, from which necessary flaps are cut out, washed, subjected to three cycles of freezing and thawing, after that, at the first stage, flap de-epithelialization is carried out in solutions of the declared composition and efficiency of de-epithelialization is evaluated visually by absence of epithelial fragments on the derma surface . At the second stage, for decellularisation, a separate three-component decellularizing system of the declared composition is used to obtain a target lipid content, which is complete absence. After that, the flaps are immersed into the DNA-ase solution of the declared composition until the DNA content in the samples is not more than 200 ng/mg. Further, the samples are washed twice with deionized water and three times by three procedures with phosphate-salt solution pH 7.2–7.4 on a shaker in mode of 250 beats per minute at room temperature, wherein flushing solutions are changed every 12 hours. Control samples are subject to morphological examination; the qualitative composition of the dermal matrix is examined using time-of-flight mass spectrometry with electrospray ionization. Target parameter achieved is removal of the main pool of cell proteins and identification of matrimonial proteins in the matrix composition. When the target parameters are achieved, the dermal matrix is subject to chemical sterilization, packaging and cryopreservation.
EFFECT: invention provides the possibility of using animal skins as raw material with no potential risks of transmission of dangerous infections.
1 cl, 1 tbl
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RU2504334C1 |
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Authors
Dates
2019-07-16—Published
2018-11-09—Filed