组织工程与重建外科杂志 ›› 2025, Vol. 21 ›› Issue (6): 555-.

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脱钙骨-明胶复合支架结合BMSC-软骨细胞共培养技术体内再生软骨的研究

  

  • 出版日期:2025-12-25 发布日期:2025-12-30

Study on in vivo cartilage regeneration using decalcified bone matrix-gelatin composite scaffold combined with bmsc-chondrocyte co-culture technology

  • Online:2025-12-25 Published:2025-12-30

摘要:

目的 探索使用脱钙骨-明胶(DBM-GT)复合支架结合BMSC-软骨细胞共培养技术实现体内稳定软骨再生的可行性。

方法 通过明胶冻干交联技术制备DBM-GT复合支架,并接种BMSC明确其细胞负载效率;通过对支架负载细胞进行活死细胞染色,明确软骨细胞调控BMSC增殖的作用;随后将BMSC与软骨细胞按1∶4的比例混合接种于复合支架中作为实验组,将软骨细胞接种于复合支架作为对照组,将实验组与对照组体外培养5 d,然后植入裸鼠皮下,4周后取材,组织学检测比较实验组与对照组间的软骨再生效果。结果 DBM-GT支架相较于 DBM支架具有更好的孔隙结构;活死细胞染色与DNA定量结果显示软骨细胞可有效促进BMSC增殖;复合支架负载BMSC-软骨细胞共培养技术体内再生软骨相较于支架负载软骨细胞再生软骨之间无明显差异。

结论 成功制备DBM-GT明胶支架,并证实其结合BMSC-软骨细胞共培养技术可实现裸鼠皮下稳定软骨再生。

关键词:

Abstract:

Objective To explore the feasibility of achieving stable in vivo cartilage regeneration using demineralized bone matrix-gelatin (DBM-GT) composite scaffolds combined with BMSC-chondrocyte co-culture technology. Methods  DBM-GT composite scaffolds were prepared via gelatin freeze-drying cross-linking technology, and bone marrow mesenchymal stem cells (BMSCs) were seeded to determine cell loading efficiency. Live/dead staining of scaffold-loaded cells was performed to evaluate the role of chondrocytes in regulating BMSC proliferation. Subsequently, a mixture of BMSCs and chondrocytes at a 1∶4 ratio was seeded onto the composite scaffold as the experimental group, while chondrocytes alone were seeded as the control group. After 5 days of in vitro culture, both groups were implanted subcutaneously into nude mice. Post-harvest histological examination was conducted to assess differences in cartilage regeneration between the experimental and control groups. Results The DBM-GT scaffold exhibited superior porous structure compared to the DBM scaffold. Live/ dead staining and DNA quantitative analysis demonstrated that chondrocytes effectively promoted BMSC proliferation. No significant difference in cartilage regeneration was observed between the experimental group (BMSC-chondrocyte co-culture) and the control group (chondrocyte-only) in vivo. Conclusion The DBM-GT gelatin scaffold can be successfully 

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