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CN-121987765-A - Mesenchymal stem cell medicine for promoting knee joint cartilage regeneration and preparation method and application thereof

CN121987765ACN 121987765 ACN121987765 ACN 121987765ACN-121987765-A

Abstract

The invention provides a mesenchymal stem cell medicine for promoting knee joint cartilage regeneration, and a preparation method and application thereof, and belongs to the technical field of medicines. The preparation method comprises the steps of preparing curcumin liposome by reacting polyethylene imine with linoleoyl chloride to prepare linoleoyl chloride modified polyethylene imine, preparing multifunctional liposome by reacting the linoleoyl chloride modified polyethylene imine with multifunctional peptide, mixing the multifunctional liposome with self-assembled RADA16 polypeptide embedded with TGF-beta 1 and BMP-6 and passaged mesenchymal stem cells, adding the mixture into autologous enriched platelet plasma, and preparing the mesenchymal stem cell medicine for promoting knee joint cartilage regeneration. According to the invention, curcumin is embedded in the liposome, and the mesenchymal stem cells can promote the differentiation of the bone formation by using the provided nutrients, and effectively prevent the interference of inflammatory microenvironment. Meanwhile, the self-assembled RADA16 polypeptide wraps TGF-beta 1 and BMP-6, and the synergistic effect is beneficial to regeneration and repair of cartilage tissues, promotes proliferation of bone cells and is beneficial to cartilage repair and regeneration.

Inventors

  • WANG ZHUANG

Assignees

  • 北京青藤谷禧干细胞科技研究院有限公司

Dates

Publication Date
20260508
Application Date
20260225

Claims (10)

  1. 1. A preparation method of a mesenchymal stem cell medicine for promoting knee joint cartilage regeneration is characterized by comprising the steps of preparing curcumin liposome by reacting polyethyleneimine with linoleoyl chloride to prepare linoleoyl chloride modified polyethyleneimine, reacting with multifunctional peptide to prepare multifunctional liposome, mixing the multifunctional liposome with self-assembled RADA16 polypeptide embedded with TGF-beta 1 and BMP-6 and passaged mesenchymal stem cells, adding the self-enriched platelet plasma, and preparing the mesenchymal stem cell medicine for promoting knee joint cartilage regeneration.
  2. 2. The method of manufacturing according to claim 1, comprising the steps of: S1, reacting polyethylene imine with linoleoyl chloride to obtain linoleoyl chloride modified polyethylene imine; S2, preparing liposome from curcumin and linoleoyl chloride modified polyethyleneimine by adopting an ethanol injection method; s3, stirring the multifunctional peptide composition and the liposome for reaction, and separating and purifying to obtain multifunctional liposome; S4, dissolving sucrose and RADA16 polypeptide in water, adding TGF-beta 1 and BMP-6, stirring and mixing uniformly, adding normal saline for crosslinking, washing, and freeze-drying to obtain self-assembled polypeptide; S5, separating and extracting mesenchymal stem cells from human tissues or obtaining the mesenchymal stem cells by commercial purchase, and culturing and passaging the mesenchymal stem cells to 3-5 generations; S6, uniformly mixing the self-assembled polypeptide, the multifunctional liposome, the autologous platelet-enriched plasma and the passaged mesenchymal stem cells to prepare the mesenchymal stem cell medicine for promoting knee joint cartilage regeneration.
  3. 3. The method according to claim 2, wherein the mass ratio of the polyethyleneimine to the oleoyl chloride in the step S1 is 3-4:1, and the number average molecular weight of the polyethyleneimine is 20-30kDa.
  4. 4. The preparation method according to claim 2, wherein the mass ratio of the curcumin to the oleoyl chloride modified polyethyleneimine in the step S2 is 0.3-0.5:1-2.
  5. 5. The method according to claim 2, wherein the mass ratio of the multifunctional peptide composition to the liposome in the step S3 is 1:10-12, the stirring reaction time is 3-5min, and the multifunctional peptide composition comprises Fmoc-SKPPGTSS-Cys and Fmoc-LIANAK-Cys, and the mass ratio is 1:1-3.
  6. 6. The method according to claim 2, wherein the mass ratio of sucrose, RADA16 polypeptide, TGF- β1 and BMP-6 in step S4 is 1:180-200:0.003-0.005:0.001-0.002.
  7. 7. The method according to claim 2, wherein the mesenchymal stem cells in step S5 are derived from at least one of umbilical cord tissue, umbilical cord blood, bone marrow, synovial membrane, placenta tissue or autologous adipose tissue, preferably bone marrow mesenchymal stem cells.
  8. 8. The method according to claim 2, wherein the self-assembled polypeptide, the multifunctional liposome and the passaged mesenchymal stem cells in step S6 are contained in the mesenchymal stem cell drug for promoting cartilage regeneration of knee joint in an amount of 0.1-0.15mg/mL, 0.005-0.01mg/mL and 10 6 -10 7 cells/mL, respectively.
  9. 9. A mesenchymal stem cell medicament for promoting regeneration of knee joint cartilage prepared by the preparation method of any one of claims 1 to 8.
  10. 10. Use of the mesenchymal stem cell medicament for promoting knee joint cartilage regeneration according to claim 9 for preparing a medicament for promoting knee joint cartilage regeneration and treating osteoarthritis.

Description

Mesenchymal stem cell medicine for promoting knee joint cartilage regeneration and preparation method and application thereof Technical Field The invention relates to the technical field of medicines, in particular to a mesenchymal stem cell medicine for promoting knee joint cartilage regeneration, and a preparation method and application thereof. Background The articular cartilage is an important component part of the synovial joint, has the characteristics of low friction, rich elasticity and high permeability, and has the main functional characteristics of buffering and bearing the pressure of a human body, providing compression resistance for cartilage tissues and reducing the friction resistance during joint movement, and has important significance for maintaining joint movement. The weak regeneration potential of articular cartilage is recognized as the biggest obstacle of surgical repair treatment, and mainly because the articular cartilage has the anatomical features of no direct blood supply, no innervation and lymphatic tissue, the articular cavity is an environment of ischemia and hypoxia, and can only rely on the nutrition of joint fluid, so once damaged, the articular cartilage is difficult to repair by itself, even serious dysfunction occurs, osteoarthritis occurs, and the quality of life is seriously affected. At present, the articular cartilage injury caused by trauma or inflammation is very common in clinical orthopedics, and a learner reports that cartilage injury occurs in common people from the age of 40 years. The mature cartilage repair technology at home and abroad at present mainly comprises arthroscope joint cavity flushing, periosteum transplanting, cartilage cleaning forming, autologous or allogenic bone cartilage transplanting, drilling micro fracture, chondrocyte transplanting, joint forming, joint fusion and the like. The method has the advantages and disadvantages of various treatment methods, the primary problem of autologous transplantation is limited source, the method is suitable for small-area defects, local cartilage defects of a supply area can be caused, the tolerance of a tissue structure is very limited, the problem of immune rejection reaction exists in allogeneic transplantation, and the chondrocyte transplantation cannot appear to be capable of large-area cartilage defects. Therefore, these traditional treatments have certain defects, short effective duration and unsatisfactory long-term efficacy, and no method has been available so far to produce hyaline cartilage tissue similar in structure and function to normal cartilage. Thus, there is an urgent need to employ effective new methods to promote self-repair of damaged articular cartilage to rescue joint function. Disclosure of Invention The invention aims to provide a mesenchymal stem cell medicine for promoting knee joint cartilage regeneration, a preparation method and application thereof, wherein curcumin is embedded in a liposome, and the mesenchymal stem cells can promote osteogenic differentiation by using provided nutrients, and effectively protect the interference of inflammatory microenvironment. Meanwhile, the self-assembled RADA16 polypeptide wraps TGF-beta 1 and BMP-6, and the synergistic effect is beneficial to regeneration and repair of cartilage tissues, promotes proliferation of bone cells and is beneficial to cartilage repair and regeneration. The technical scheme of the invention is realized as follows: The invention provides a preparation method of a mesenchymal stem cell medicine for promoting knee joint cartilage regeneration, which comprises the steps of preparing a curcumin liposome by reacting polyethyleneimine with linoleoyl chloride to prepare linoleoyl chloride modified polyethyleneimine, reacting with multifunctional peptide to prepare the multifunctional liposome, mixing the multifunctional liposome with self-assembled RADA16 polypeptide embedded with TGF-beta 1 and BMP-6 and passaged mesenchymal stem cells, adding the mixture into autologous enriched platelet plasma, and preparing the mesenchymal stem cell medicine for promoting knee joint cartilage regeneration. As a further improvement of the invention, the method comprises the following steps: S1, reacting polyethylene imine with oleoyl chloride to obtain the oleoyl chloride modified polyethylene imine, wherein the polyethylene imine is a common cationic drug carrier, but has certain application limit. The invention has the advantages of stable drug release, long drug action time, low drug administration frequency and the like, and obviously reduces the amino content of polyethyleneimine by modifying the hydrophobicity of the polyethyleneimine. S2, preparing liposome from curcumin and oleoyl chloride modified polyethyleneimine by adopting an ethanol injection method, wherein the curcumin is polyphenol extracted from rhizome of turmeric root, has good anti-inflammatory and antibacterial effects, but has extremely low solubility, s