Search

CN-122005940-A - Tissue regeneration repair stent and preparation method thereof

CN122005940ACN 122005940 ACN122005940 ACN 122005940ACN-122005940-A

Abstract

The invention provides a tissue regeneration repair stent and a preparation method thereof, and relates to the technical field of medical treatment. The preparation method of the tissue regeneration repair stent comprises the steps of filling a gelatinous acellular matrix in a mold, then contacting the bottom of the mold with a cold source to form a directional temperature gradient field from bottom to top, forming columnar ice crystals by the gelatinous acellular matrix along the direction of the directional temperature gradient field under the directional temperature gradient field, obtaining a porous matrix with directional pore channels after freeze drying, and preparing the tissue regeneration repair stent after crosslinking and solidification by a crosslinking agent. According to the preparation method, a porous structure of the directional pore canal is constructed through a directional freezing strategy, so that a tubular pore channel perpendicular to the joint surface is formed, the columnar structure of natural cartilage can be simulated, and the vertical migration of bone marrow mesenchymal stem cells is facilitated.

Inventors

  • SUN LEI
  • TAO JIANFENG
  • ZHU CHENKAI
  • SHAN YINGHUI

Assignees

  • 首都医科大学附属北京积水潭医院
  • 科林唯傲(北京)生物科技有限公司

Dates

Publication Date
20260512
Application Date
20260305

Claims (10)

  1. 1. The preparation method of the tissue regeneration repair stent is characterized by comprising the following steps: Filling the gel-like acellular matrix into a mold, then contacting the bottom of the mold with a cold source to form a directional temperature gradient field from bottom to top, forming columnar ice crystals by the gel-like acellular matrix along the direction of the directional temperature gradient field under the directional temperature gradient field, obtaining a porous matrix with directional pore channels after freeze drying, and preparing the tissue regeneration repair bracket after crosslinking and curing by a crosslinking agent.
  2. 2. The method of claim 1, wherein the cold source comprises liquid nitrogen; preferably, the temperature of the cold source is-80 ℃ to-50 ℃.
  3. 3. The method of claim 1, wherein the cross-linking agent comprises one or more of formaldehyde, glutaraldehyde, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, and N-hydroxysuccinimide.
  4. 4. The method of claim 1, further comprising the step of treating the porous substrate with a bio-enzyme solution prior to the crosslinking curing; Preferably, the concentration of the biological enzyme solution is 0.01-0.2 wt%, the treatment time of the biological enzyme solution is 1-10 min, and the biological enzyme is one or more of pepsin and pancreatin.
  5. 5. The preparation method of the gel-like acellular matrix according to claim 1, wherein the preparation method comprises the steps of sequentially performing acellular and antigen removal and exogenous DNA removal on animal tissues to obtain the gel-like acellular matrix; the animal tissue includes one or more of pericardial or dermal tissue.
  6. 6. The method according to claim 5, wherein the decellularizing step comprises subjecting the animal tissue matrix to freeze thawing treatment, and then circulating the hypertonic solution treatment and the hypotonic solution treatment; Preferably, the cycle number of the freeze thawing treatment is 3-6; preferably, the freeze thawing treatment is further followed by ultrasonic treatment; Preferably, the hypertonic solution is NaCl solution with the concentration of 1-5 mol/L, and the treatment time of the hypertonic solution is 10-60 minutes; preferably, the hypotonic solution is water, and the treatment time of the hypotonic solution is 10-120 minutes; Preferably, the times of circularly carrying out the hypertonic solution treatment and the hypotonic solution treatment are 1-5 times.
  7. 7. The method according to claim 5, wherein the step of removing the antigen and the foreign DNA comprises immersing the decellularized substrate in a biological enzyme solution; preferably, the biological enzyme comprises one or more of pepsin or trypsin; preferably, the mass concentration of the biological enzyme solution is 0.1% -2.0%; Preferably, the soaking treatment is for 1-4 hours.
  8. 8. The method according to claim 5, wherein the dermis tissue is skin tissue prepared by pretreatment; the pretreatment includes dehairing treatment; The dehairing treatment is to dehairing the skin tissue by adopting a thiosulfate solution; preferably, the concentration of the thiosulfate solution is 2g/100 ml-5 g/100ml, and the pH is 8-10; Preferably, the temperature of the thiosulfate solution treatment is 20-30 ℃; preferably, the time for the treatment of the thiosulfate solution is 2-6 hours.
  9. 9. The method of claim 1, further comprising the step of loading platelet rich plasma or bioactive factors; The bioactive factor includes one or more of TGF-beta, IGF-1 or BMP-2; Preferably, the platelet rich plasma or bioactive factor is loaded by physical adsorption or chemical bonding.
  10. 10. A tissue regeneration repair scaffold characterized by being prepared by the preparation method of any one of claims 1 to 9.

Description

Tissue regeneration repair stent and preparation method thereof Technical Field The invention relates to the technical field of medical treatment, in particular to a tissue regeneration repair stent and a preparation method thereof. Background Tissue defects are a very common disease in the surgical field. Some diseases such as cartilage defect, uterine scar rupture, cerebral spinal cord membrane deficiency caused by surgery and the like are difficult to repair maximally due to the fact that the damaged tissues exceed the regeneration capacity of the brain spinal cord membrane, and the dysfunction of related organs is caused. The regeneration of tissue by regenerating scaffold technology and compounding active substances derived from human body is a method for repairing tissue defect which has been gradually known and used in the last decade. The method is mainly characterized in that the tissue regeneration stent prepared by a specific technology is used as a base for bearing active ingredients collected by normal tissues of a patient, such as autologous Platelet Rich Plasma (PRP), active substances from fat, cells and the like, the active substances are loaded on a stent material according to the final required amount, and then a stent carrier is implanted into a defect part through operation. Under the coordination and control action of the scaffold guidance and the active substance induction, surrounding cells are recruited to gather, proliferate and differentiate at the defect, and the cells regenerate to form natural tissues similar to normal tissues, so that the repair of the defect tissues is completed. Therefore, the formulation design and preparation method of the scaffold material in the strategy are key to determine the tissue repair effect. The ideal tissue regeneration and repair bracket has (1) good biological activity, no toxicity and no teratogenic risk, (2) proper biodegradability, degradation rate and tissue growth adaptation, (3) tissue regeneration induction capability, which can be used as an active substance to bear, provide a good microenvironment for cell growth and proliferation, (4) workability, which is convenient for processing and forming a required shape, (5) structural supportability, and mechanical strength which is enough to maintain the implant in vivo for a certain time and keep the shape stable, (6) a porous structure, the porosity of which is more than 80 percent, which is beneficial to cell attachment and growth, and nutrient component infiltration and metabolite discharge. The tissue matrix of biological origin is mostly the main component of organism connective tissue, and has good biocompatibility. Because of strict requirements of ethical regulations, human tissue sources obtained through human donation and limited human tissue sources cannot meet a great number of clinical requirements, the preparation of biological materials after the removal of immunogenicity by utilizing heterogeneous decellularized dermis to thoroughly remove heterogeneous cells is an important direction of development in the field of medical instruments. Livesey et al, 1995, first reported successful preparation of decellularized dermal matrix, demonstrating the potential of in vivo dermal remodeling by appropriate treatment of allogeneic dermal matrix Transplantation (transformation, 1995,60 (1), 1-9). The tissue repair scaffold material based on the acellular dermal matrix has good effect in clinical application in the fields of repair of burn wound surfaces, oral maxillofacial surfaces, skin, cartilage and the like. However, due to the limitation of the acellular dermal matrix material, the scaffold still faces the problems of poor biological activity, unsatisfactory porosity and the like, and the application of the scaffold in the field of tissue repair is severely limited. The prior art CN102178981A discloses a cartilage repair stent which is successfully prepared by adopting a novel small Niu Zhenpi stent and adopting SDS decellularization, trypsin modification and formaldehyde crosslinking strategies. However, the SDS decellularization and formaldehyde crosslinking strategy may cause the problems that antigen and DNA are difficult to effectively remove, residual formaldehyde inhibits cell growth and the like, and the problem of gap regulation still exists. Therefore, the searching of a preparation method of the animal-derived acellular dermal matrix scaffold for improving the bioactivity and the regeneration function is an important problem to be solved in the field at present. In view of this, the present invention has been made. Disclosure of Invention The first object of the present invention is to provide a method for preparing a tissue regeneration repair stent, so as to solve the above technical problems. A second object of the present invention is to provide a tissue regeneration repair scaffold which has a suitable biocompatibility and degradability, a unique microstructure, ca