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A scaffold material capable of recruiting endogenous mesenchymal stem cells and its preparation method and application

A technology of scaffold material and mesenchymal stem cells, which is applied in prosthetics, tissue regeneration, medical science, etc., can solve the problems of low immunogenicity, fast degradation of hyaluronic acid, lack of cell adhesion sites, etc., and achieve mechanical properties and degradation Appropriate performance, conducive to the delivery of nutrients, and beneficial to the effect of maintaining the phenotype of cells

Active Publication Date: 2022-04-01
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hyaluronic acid hydrogel has good biocompatibility, biodegradability, high water retention and the characteristics of promoting cartilage formation, but the degradation rate of hyaluronic acid is fast, the mechanical strength is poor, and it lacks cell adhesion sites. Acid-forming biomaterials inhibit cell attachment and fail to meet application requirements
Silk fibroin is a natural polymer material, non-toxic, non-irritating, low immunogenicity, good biodegradability and biocompatibility, but silk fibroin lacks cell growth adhesion sites, and it is harmful to cells Unfavorable growth, which makes it unable to meet the needs of practical applications
At present, the scaffold materials based on hyaluronic acid or silk fibroin have the shortcomings of insufficient ability to recruit and retain endogenous cells. Insufficient exogenous risk of

Method used

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  • A scaffold material capable of recruiting endogenous mesenchymal stem cells and its preparation method and application
  • A scaffold material capable of recruiting endogenous mesenchymal stem cells and its preparation method and application
  • A scaffold material capable of recruiting endogenous mesenchymal stem cells and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] This example prepares thiolated hyaluronic acid (HA-SH), the steps are as follows:

[0055] (1) Dissolve sodium hyaluronate with a molecular weight of 0.3MDa in deionized water, add N-hydroxysuccinimide (NHS), fully dissolve, and then add 1-ethyl-(3-dimethylaminopropyl ) carbodiimide hydrochloride (EDC HCl), fully dissolved, adjust the pH value of the mixture to 4.75 with 1mol / L NaOH solution and 1mol / L HCl solution, react at room temperature for 2h, and then add cysteine The aqueous solution of ammonia hydrochloride (CSH·HCl) was reacted at room temperature for 24 hours, and then the pH value of the reaction solution was adjusted to 8.5 with 1 mol / L NaOH solution, and dithiothreitol (DTT) solution was added to react at room temperature for 12 hours.

[0056] In this step, the molar ratio of sodium hyaluronate, NHS, EDC·HCl, CSH·HCl and DTT is 1:2:4:4:12.

[0057] (2) Adjust the pH value of the reaction solution obtained in step (1) to 3.0 to 3.5 with 1mol / L HCl soluti...

Embodiment 2

[0060] This embodiment prepares dopamine-modified silk fibroin (SF-DOPA), and the steps are as follows:

[0061] (1) Cut the silkworm cocoon into pieces, and use 0.02mol / L Na in a bath ratio of 1:100 2 CO 3 Boil in the solution for 2h to remove sericin, wash and dry, then immerse in 9.3mol / L LiBr solution and dissolve for 4h, transfer the resulting reaction solution into a dialysis bag, and dialyze for 72h with deionized water to obtain silk fibroin (SF ) solution.

[0062] (2) Add to the SF solution obtained in step (1), add EDC·HCl and NHS, adjust the pH value of the mixture to 4.75 with 1mol / L NaOH solution and 1mol / L HCl solution, activate for 2h, then add hydrochloric acid Dopamine was reacted at room temperature for 10 h, and the obtained reaction liquid was transferred into a dialysis bag, and dialyzed with deionized water for 72 h to obtain an aqueous solution of SF-DOPA.

[0063] In step (2), the molar ratio of SF, NHS, EDC·HCl and dopamine hydrochloride is control...

Embodiment 3

[0067] In this embodiment, the steps of preparing freeze-dried sponge support are as follows:

[0068] (1) Dissolve the HA-SH prepared in Example 1 in deionized water to form a HA-SH solution with a concentration of 1wt.% to 10wt.%; adjust the concentration of the SF-DOPA aqueous solution prepared in Example 2 to SF-DOPA The concentration is 1wt.%~10wt.%, to obtain SF-DOPA solution.

[0069](2) Mix the HA-SH solution and the SF-DOPA solution according to the volume ratio of (1~10):(1~10) under ice bath conditions, and use 0.5mol / L NaOH solution and 0.1mol / L NaOH solution The HCl solution adjusted the pH value to 7.4, injected into the silica gel mold, and stood at 37° C. for 5 minutes to form a composite hydrogel. The composite hydrogel was freeze-dried to obtain a freeze-dried sponge scaffold.

[0070] In this example, by adjusting the concentration and volume ratio in steps (1) and (2), a total of five groups of experiments were carried out, and five kinds of freeze-dried s...

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Abstract

The invention provides a scaffold material capable of recruiting endogenous mesenchymal stem cells. The scaffold material is a hydrogel with a composite three-dimensional network structure formed by cross-linking thiolated hyaluronic acid and dopamine-modified silk fibroin The freeze-dried sponge scaffold formed after freeze-drying is sequentially soaked in a solution of maleimide polyethylene glycol active ester and a solution of E7 peptide connected to cysteine ​​at one end, and the E7 peptide is treated with maleimide polyethylene glycol The diol active ester is grafted on the freeze-dried sponge support and formed by drying, and has an interpenetrating three-dimensional porous structure. The invention also provides a preparation method of the scaffold material and its application as a three-dimensional scaffold for cartilage tissue engineering or a three-dimensional cell scaffold for cartilage tissue engineering. The scaffold material provided by the invention can provide a suitable microenvironment and suitable mechanical properties for cell growth, and has the property of recruiting endogenous mesenchymal stem cells.

Description

technical field [0001] The invention belongs to the technical field of biomaterials, and relates to a scaffold material capable of recruiting endogenous mesenchymal stem cells and a preparation method and application thereof. The scaffold material has good application prospects in the field of tissue repair. Background technique [0002] Articular cartilage defect is a common orthopedic disease. Since articular cartilage has no blood vessels, lymph nodes, and nerves, it is difficult to repair itself once it is damaged. Current treatment methods include microfractures, autologous chondrocyte transplantation, etc., but limited by the shortage of donor cells and limited long-term repair effects, the clinical use of these methods is limited. As a cutting-edge technology, tissue engineering has attracted the attention of many researchers in the field of cartilage repair. The three elements of tissue engineering include scaffold materials, cells, and growth factors. The most basi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/20A61L27/22A61L27/18A61L27/38A61L27/50A61L27/54A61L27/56A61L27/58C08J9/42C08L5/08C08L89/00C08L71/02C08H1/00
CPCA61L27/20A61L27/227A61L27/18A61L27/3834A61L27/3852A61L27/50A61L27/54A61L27/56A61L27/58C08J9/42C08H1/00A61L2430/06A61L2300/412C08J2305/08C08J2389/00C08J2471/02C08L5/08C08L89/00C08L71/02
Inventor 孙勇祝嘉懿樊渝江
Owner SICHUAN UNIV
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