Chondrocyte proliferation and tissue formation enhanced by stromal cell derived factor-1 modified poly-L-lactic acid porous microspheres

doi:10.12307/2025.443

Abstract

Abstract: BACKGROUND: The proliferation and phenotypic maintenance of chondrocytes are limited under two-dimensional culture conditions. Porous microspheres serve as scaffolds, providing a three-dimensional culture environment that better mimics in vivo growth conditions. Stromal cell derived factor-1, a homeostatic cytokine with potent chemotactic effects, facilitates cell adhesion and proliferation.
OBJECTIVE: To investigate the impact of stromal cell derived factor-1 grafted poly-L-lactic acid porous microspheres on the biological characteristics of chondrocytes and the formation of cartilage tissue.
METHODS: (1) The effects of different concentrations of stromal cell derived factor-1 on rabbit chondrocyte proliferation, migration, and phenotypic maintenance were investigated in an in vitro setting. (2) Poly-L-lactic acid porous microspheres were prepared by double emulsion method. Stromal cell derived factor-1 was grafted onto poly-L-lactic acid porous microspheres through carbodiimide reaction. The grafting was verified by enzyme-linked immunosorbent assay and incubation with stromal cell derived factor-1-specific fluorescent antibodies. (3) Rabbit chondrocytes were inoculated on poly-L-lactic acid porous microspheres and grafted on stromal cell derived factor-1 poly-L-lactic acid porous microspheres to detect cell proliferation and adhesion. (4) The methylacrylamide-gelatin-chondrocyte complex (control group), poly-L-lactic acid porous microsphere-methylacrylamide-gelatin-chondrocyte complex (porous microsphere group), and grafted stromal cell derived factor-1 poly-L-lactic acid porous microsphere-methylacrylamide-gelatin-chondrocyte complex (porous microsphere modified group) were implanted under the skin of the back of nude mice, respectively. Samples were collected 8 weeks later and detected using histological staining and qRT-PCR for chondroblast related genes.
RESULTS AND CONCLUSION: (1) Compared with 0 and 1 000 ng/mL stromal cell derived factor-1, 1 and 500 ng/mL stromal cell derived factor 1 could promote the proliferation and migration of chondrocytes, and enhance the mRNA expression levels of type II collagen, elastin, proliferating cell nuclear antigen, and Bcl-2 in chondrocytes. (2) Stromal cell derived factor-1 was successfully grafted onto poly-L-lactic acid porous microspheres with a grafting rate of 93.75%. (3) Compared with poly-L-lactic acid porous microspheres, grafted stromal cell derived factor-1 poly-L-lactic acid porous microspheres promoted the proliferation and adhesion of chondrocytes. (4) After 8 weeks of subcutaneous implantation in nude mice, compared with the control group and the porous microsphere group, the porous microsphere modified group had clearer cartilage lacunae structure, more chondro-specific matrix and type II collagen deposition, and increased expression of elastin, type II collagen, proliferating cell nuclear antigen, and Bcl-2 mRNA. These findings indicate that stromal cell derived factor-1 grafted poly-L-lactic acid porous microspheres are beneficial to chondrocyte adhesion, proliferation, phenotypic maintenance, and the formation of cartilage tissue in vivo.

Key words: poly-L-lactic acid porous microsphere, stromal cell derived factor-1, chondrocyte, three-dimensional cell culture, tissue engineered cartilage, composite scaffold

CLC Number:

Ma Yue, Tan Shiyu, Chu Feiyang, Chen Zhuoqi, Liu Siyu, Liu Wenshuai, Liu Xia. Chondrocyte proliferation and tissue formation enhanced by stromal cell derived factor-1 modified poly-L-lactic acid porous microspheres[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(22): 4653-4662.

Figures/Tables 8

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