Biological tissue matrix material, preparation method therefor and use thereof in otological repair material
AssigneeBeijing Biosis Healing Biological Technology Co., Ltd.
InventorsBo Zhao, Hongquan Wang, Leilei Xia, Yanrui Zhao, Xuejun Li, Jinhui Zhang
Life of this patent
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Abstract
Disclosed are a biological tissue matrix material, a preparation method therefor and the use thereof in an otological repair material. The biological tissue matrix material comprises an extracellular matrix. The extracellular matrix comprises a collagen fiber, a growth factor and fibronectin. The biological tissue matrix material has a low amount of DNA residue, a low immunogenicity, a high anti-infection ability, and a strong repair capability.
Claims11
A preparation method of a biological tissue matrix material, wherein the preparation method comprises the following steps:
- (1) initial treatment of a raw material:
- taking a small intestinal submucosa tissue material, washing, and filtering to dryness;
- (2) virus inactivation:
- soaking the small intestinal submucosa tissue material with a peroxyacetic acid-ethanol solution to carry out virus inactivation;
- (3) washing the small intestinal submucosa tissue material obtained in step (2) under an ultrasonic environment, and then filtering to dryness;
- (4) decellularization, wherein a decellularizing solution comprises trypsin and a PBS solution, the decellularizing solution also comprises EDTA, EDTA-2Na, or EDTA-4Na, and the small intestinal submucosa tissue material obtained in step (3) is treated with the decellularizing solution in a multi-frequency ultrasonic environment to carry out decellularization;
- and
- (5) washing a decellularized small intestinal submucosa tissue material obtained in step (4) under the ultrasonic environment to obtain a decellularized small intestinal submucosa matrix material;
- wherein
- in the peroxyacetic acid-ethanol solution in step (2), peroxyacetic acid has a volume percentage concentration of 0.1% to 5%, ethanol has a volume percentage concentration of 5% to 40%, a volume ratio of the peroxyacetic acid-ethanol solution to the small intestinal submucosa tissue material is (5 to 20):1, an inactivation time is 2 to 4 hours and a temperature range is 10 to 40° C.;
- the trypsin in the decellularizing solution of step (4) has a mass percentace concentration of 0.01% to 0.2%, EDTA, EDTA-2Na or EDTA-4NA a has a concentration of 0.1 to 1 mmol/L, and the decellularizing solution has a pH value of 7.0 to 8.0;
- a volume ratio of the decellularizing solution to the small in intestinal submucosa tissue material is (20 to 40):1, a decellularization process is carried out in a dual-frequency ultrasonic device, in which a low frequency has a frequency range of 20 to 40 KHz, a low-frequency treatment lasts for 5 to 40 min, a high frequency has a frequency of 60 to 90 KHz, a high-frequency treatment lasts for 5 to 40 min the decellularizing solution has a temperature range of 20 to 35° C., and an ultrasonic power is 5000 W or more.
Dependencies are inferred from the claim text.