Search

CN-122005939-A - Tissue engineering blood vessel and preparation method and application thereof

CN122005939ACN 122005939 ACN122005939 ACN 122005939ACN-122005939-A

Abstract

The application relates to the technical fields of biological medicine and tissue engineering, in particular to a tissue engineering blood vessel, a preparation method and application thereof. The tissue engineering blood vessel is obtained by modifying interleukin 24 (IL-24) on the surface of a decellularized blood vessel, the tissue engineering blood vessel is transplanted into a body, the cell number of CD31+ and CD34+ can be obviously improved, the infiltration of M2 type macrophages (CD 163+) on the surface of the blood vessel is obviously increased, the expression of HIF-1 alpha (hypoxia marker) and MMP9 is obviously reduced, the expression of nerve specific protein S-100 is obviously increased, the thrombosis and intimal hyperplasia are effectively inhibited, and the patency rate of the transplanted blood vessel is obviously improved.

Inventors

  • XIAO LI
  • KONG XIANGRUI
  • XIE LIXIN
  • CHEN WEN
  • ZHU MENGYANG
  • CHEN Jin

Assignees

  • 中国人民解放军总医院第八医学中心

Dates

Publication Date
20260512
Application Date
20260302

Claims (10)

  1. 1. A tissue engineering blood vessel, which is characterized in that the tissue engineering blood vessel is obtained by modifying interleukin 24 (IL-24) on the surface of decellularized blood vessel.
  2. 2. The tissue engineering blood vessel according to claim 1, wherein the tissue engineering blood vessel is obtained by further crosslinking IL-24 after crosslinking collagen by decellularized blood vessel.
  3. 3. The tissue engineering blood vessel according to claim 2, wherein the cross-linking comprises using a cross-linking agent, preferably comprising one or more of glutaraldehyde, succinaldehyde, dialdehyde starch, genipin, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), butanediol glycidyl ether (BDDE), N' -methylenebisacrylamide (BIS), EDC/NHS cross-linking agent, ethylene Glycol Dimethacrylate (EGDMA) or disuccinimidyl suberate (DSS).
  4. 4. A method of preparing a tissue engineering blood vessel according to any one of claims 1 to 3, wherein the method comprises modifying IL-24 on the surface of a decellularized blood vessel; Preferably, the preparation method comprises the step of further crosslinking IL-24 after the decellularized vascular crosslinked collagen.
  5. 5. The method according to claim 4, wherein the method for preparing a decellularized blood vessel comprises rupture of a membrane after taking a blood vessel, and then shaking in a detergent, preferably for 20-100 hours; Preferably, the detergent comprises Sodium Dodecyl Sulfate (SDS) and/or Sodium Deoxycholate (SDC).
  6. 6. The method according to claim 4 or 5, wherein crosslinking the collagen comprises incubating the decellularized blood vessel in a collagen solution, and adding a crosslinking agent to crosslink by shaking; preferably, the incubation time in the collagen solution is 10-40 hours; Preferably, the temperature of incubation in the collagen solution is 25-50 ℃; preferably, the concentration of the collagen solution is 1-10mg/mL; preferably, the time of oscillation crosslinking in the collagen solution is 10-40 hours; Preferably, the temperature of the oscillating crosslinking in the collagen solution is 25-50 ℃; preferably, the rate of oscillation crosslinking in the collagen solution is 100-1000 revolutions per minute.
  7. 7. The method according to any one of claims 4 to 6, wherein crosslinking the IL-24 comprises placing the decellularized blood vessel after crosslinking the collagen in an IL-24 solution, and then adding a crosslinking agent to crosslink by oscillation; Preferably, the time of shaking crosslinking in the IL-24 solution is 10-40 hours; preferably, the temperature of the oscillating crosslinking in the IL-24 solution is 25-50 ℃; preferably, the concentration of the IL-24 solution is 1-10 mug/mL; preferably, the rate of oscillation crosslinking in the IL-24 solution is from 100 to 1000 revolutions per minute.
  8. 8. The method of any one of claims 4-7, wherein the cross-linking agent used to cross-link collagen or cross-link IL-24 is the same cross-linking agent or a different cross-linking agent.
  9. Use of il-24 in the preparation of tissue engineering blood vessels.
  10. 10. Use of a tissue engineering blood vessel according to any one of claims 1 to 3 or a tissue engineering blood vessel obtained by a method of preparation according to any one of claims 4 to 8, characterized in that said use comprises: A) The use in the manufacture of medical devices, preferably medical devices including those for vascular grafts, or, B) The application of the composition in preparing products for treating vascular injury, vascular stenosis or vascular occlusion.

Description

Tissue engineering blood vessel and preparation method and application thereof Technical Field The invention relates to the technical fields of biological medicine and tissue engineering, in particular to a tissue engineering blood vessel, a preparation method and application thereof. Background Vascular grafts are an important means of treating vascular injury, stenosis or occlusion. Although large-caliber artificial blood vessels are widely applied, small-caliber artificial blood vessels with diameters smaller than 6mm are transplanted and then face risks of acute thrombosis, intimal hyperplasia, infection and the like, so that the long-term patency rate is low. The absence of functional endothelial cells is a key cause of graft failure. Local tissue damage caused by vascular grafting creates an immune microenvironment for ischemic, hypoxic, and inflammatory responses. IL-24 is known to play a role in tumor suppression and inflammatory diseases as a multifunctional cytokine, but modification of tissue engineering blood vessels with IL-24 has not been reported yet, and the present application has been specifically proposed. Disclosure of Invention In order to solve the problems of slow endothelialization and low patency rate of the existing tissue engineering after vascular grafting. The application promotes macrophage to M2 phenotype polarization by modifying IL-24 on the surface of decellularized blood vessel to form tissue engineering blood vessel, enhances the expression of IL-8 and IL-10, thereby obviously improving the proliferation and migration capacity of endothelial cells. The tissue engineering blood vessel obtained by the application can obviously improve the vascular patency rate after transplantation, promote the generation of micro blood vessels and the reconstruction of nerves, and improve the anoxic microenvironment. Specific: in a first aspect of the present invention, there is provided a tissue engineering blood vessel obtained by modifying interleukin 24 (IL-24) on the surface of a decellularized blood vessel. Preferably, the IL-24 is modified on the inner wall of decellularized blood vessels. Preferably, the tissue engineering blood vessel is obtained by further crosslinking IL-24 after crosslinking collagen through decellularized blood vessel. Preferably, the crosslinking includes the use of a crosslinking agent, preferably including one or more of glutaraldehyde, succinaldehyde, dialdehyde starch, genipin, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), butanediol glycidyl ether (BDDE), N' -methylenebisacrylamide (BIS), EDC/NHS crosslinking agent, ethylene Glycol Dimethacrylate (EGDMA), or disuccinimide suberate (DSS). In one embodiment of the invention, the crosslinking agent is EDC. Preferably, the tissue engineering blood vessel is a small-caliber tissue engineering blood vessel. Preferably, the minor caliber comprises a diameter of less than 10mm, more preferably less than 6mm, still more preferably less than 1mm, for example less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1mm, preferably 0.5-1.0mm. In a second aspect of the present invention, there is provided a method for preparing a tissue engineering blood vessel according to the first aspect, wherein the method comprises modifying IL-24 on the surface of a decellularized blood vessel. Preferably, the preparation method comprises the step of further crosslinking IL-24 after the decellularized vascular crosslinked collagen. Preferably, the decellularized blood vessel is prepared by rupture of the membrane after removal of the blood vessel, followed by shaking treatment in a detergent, preferably for 20-100 hours, such as 20, 24, 30, 40, 48, 50, 60, 70, 72, 75, 80, 90, 96 or 100 hours. Preferably, a step of changing the liquid (detergent) is also included, for example, once every 12-24 hours. Preferably, the blood vessel may be a venous blood vessel or an arterial blood vessel, such as the common carotid artery. Preferably, the rupture of the membrane comprises rupture of the membrane under hypotonic conditions, and more preferably, the hypotonic conditions comprise placing the vessel in distilled water. Preferably, the detergent comprises Sodium Dodecyl Sulfate (SDS) and/or Sodium Deoxycholate (SDC). Wherein the concentration of Sodium Dodecyl Sulfate (SDS) is 0.1% -5%, e.g. 0.1%, 0.5%, 1%, 2%, 3%, 4% or 5%. The concentration of Sodium Deoxycholate (SDC) is 0.1% -5%, e.g. 0.1%, 0.5%, 1%, 2%, 3%, 4% or 5%. In one embodiment of the invention, the detergent is 0.1% -5% SDS and 0.1% -5% SDC. Preferably, the method of preparing a decellularized blood vessel further comprises a step of washing, further preferably with PBS for 12-30 hours, e.g., 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 hours. Preferably, the cross-linking of the collagen comprises incubating the decellularized blood vessel in a collagen solution, and then adding a cross-linking agent for o