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人骨髓间充质干细胞与脂肪组织间充质干细胞在用于软骨修复的临床相关细胞支架中的整合、生长及软骨形成潜能的体外比较

An In Vitro Comparison of the Incorporation, Growth, and Chondrogenic Potential of Human Bone Marrow versus Adipose Tissue Mesenchymal Stem Cells in Clinically Relevant Cell Scaffolds Used for Cartilage Repair.

作者信息

Kohli Nupur, Wright Karina T, Sammons Rachel L, Jeys Lee, Snow Martyn, Johnson William E B

机构信息

School of Life and Health Sciences, Aston University, Birmingham, UK.

Robert Jones & Agnes Hunt Orthopaedic Hospital, Oswestry, UK.

出版信息

Cartilage. 2015 Oct;6(4):252-63. doi: 10.1177/1947603515589650.

Abstract

AIM

To compare the incorporation, growth, and chondrogenic potential of bone marrow (BM) and adipose tissue (AT) mesenchymal stem cells (MSCs) in scaffolds used for cartilage repair.

METHODS

Human BM and AT MSCs were isolated, culture expanded, and characterised using standard protocols, then seeded into 2 different scaffolds, Chondro-Gide or Alpha Chondro Shield. Cell adhesion, incorporation, and viable cell growth were assessed microscopically and following calcein AM/ethidium homodimer (Live/Dead) staining. Cell-seeded scaffolds were treated with chondrogenic inducers for 28 days. Extracellular matrix deposition and soluble glycosaminoglycan (GAG) release into the culture medium was measured at day 28 by histology/immunohistochemistry and dimethylmethylene blue assay, respectively.

RESULTS

A greater number of viable MSCs from either source adhered and incorporated into Chondro-Gide than into Alpha Chondro Shield. In both cell scaffolds, this incorporation represented less than 2% of the cells that were seeded. There was a marked proliferation of BM MSCs, but not AT MSCs, in Chondro-Gide. MSCs from both sources underwent chondrogenic differentiation following induction. However, cartilaginous extracellular matrix deposition was most marked in Chondro-Gide seeded with BM MSCs. Soluble GAG secretion increased in chondrogenic versus control conditions. There was no marked difference in GAG secretion by MSCs from either cell source.

CONCLUSION

Chondro-Gide and Alpha Chondro Shield were permissive to the incorporation and chondrogenic differentiation of human BM and AT MSCs. Chondro-Gide seeded with BM MSCs demonstrated the greatest increase in MSC number and deposition of a cartilaginous tissue.

摘要

目的

比较骨髓(BM)和脂肪组织(AT)间充质干细胞(MSCs)在用于软骨修复的支架中的黏附、生长及软骨形成潜能。

方法

采用标准方案分离、培养扩增并鉴定人BM和AT MSCs,然后将其接种到两种不同的支架Chondro - Gide或Alpha Chondro Shield中。通过显微镜观察及钙黄绿素AM/碘化丙啶(活/死)染色评估细胞黏附、植入及活细胞生长情况。将接种细胞的支架用软骨形成诱导剂处理28天。分别在第28天通过组织学/免疫组织化学和二甲基亚甲基蓝法检测细胞外基质沉积及可溶性糖胺聚糖(GAG)释放到培养基中的情况。

结果

与Alpha Chondro Shield相比,来自两种来源的更多活MSCs黏附并植入Chondro - Gide。在两种细胞支架中,这种植入的细胞占接种细胞的比例均不到2%。在Chondro - Gide中,BM MSCs有明显增殖,而AT MSCs没有。诱导后,两种来源的MSCs均发生软骨形成分化。然而,在接种BM MSCs的Chondro - Gide中,软骨样细胞外基质沉积最为明显。与对照条件相比,软骨形成条件下可溶性GAG分泌增加。两种细胞来源的MSCs分泌GAG没有明显差异。

结论

Chondro - Gide和Alpha Chondro Shield允许人BM和AT MSCs的植入及软骨形成分化。接种BM MSCs的Chondro - Gide中MSCs数量增加最多,且软骨组织沉积最多。

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