Tissue engineered cartilage from hTGF beta2 transduced human adipose derived stem cells seeded in PLGA/alginate compound in vitro and in vivo

Xiao Bing Jin, Yong Sheng Sun, Ke Zhang, Jing Wang, Tai Ping Shi, Xiao Dong Ju, Si Quan Lou
Journal of Biomedical Materials Research. Part A 2008 September 15, 86 (4): 1077-87
Chondrogenic potential of human adipose derived stem cells (hASCs) makes them a possible source of seeding cells for cartilage tissue engineering. We aim to examine the chondrogenic differentiation of human transforming growth factor beta2 (hTGF beta2) transduced hASCs seeded in three-dimensional scaffold in vitro and in vivo. In this study, hASCs were isolated from human subcutaneous adipose tissue and transduced with a replication deficient adenovirus carrying hTGF beta2 (Ad5-hTGF beta2), and then the transduced cells were seeded and cultured in PLGA/alginate compounds. RT-PCR analysis revealed that Ad5-hTGF beta2 transduced hASCs produced aggrecan and collagen type II after 7-day induction in vitro and continued throughout the culture period; this was also demonstrated by the positive staining of Alcian blue and immunohistochemistry for collagen type II. For in vivo study, Ad5-hTGF beta2 transduced hASCs seeded in PLGA/alginate compounds were implanted in subcutaneous pockets of nude mice; after 12 weeks, the implants were harvested and examined by haematoxylin and eosin staining, AB-PAS staining, and immunohistochemical analysis, and the results demonstrated the formation of cartilage tissue. As a control, all these were not observed in the constructs with Ad5-EGFP transduced hASCs. In conclusion, our study demonstrates that adenovirus-mediated hTGF beta2 gene transfer is able to induce the hASCs into chondrogenic lineage both in vitro and in vivo. Ad5-hTGF beta2 transduced hASCs combined with three-dimensional PLGA/alginate compound may be a viable method in treating injuries of cartilage.

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