Abstract:Objective:To construct the TGF-β3 protein with targeted therapy function in which latency associated peptide, MMP enzyme and TGF-β3 play the role of latent, targeting and therapeutic effect, respectively. In addition, the specific targeted therapeutic effect was investigated by transferring TGF-β3 gene to chondrogenic progenitor cells(CPCs) from perichondrium. Methods:The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and TGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP, respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3, which was then transferred to chondrogenic progenitor cells. The genetically modified CPCs were cultured with medium with or without MMP-1. The expression of Collagen II, Aggrecan and TIMP were detected at day 7,14 and 21. Results:pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed and identified by enzyme digestion and sequencing analysis. Only the medium with MMP enzyme can promote the chondrogenesis and the matrix production of genetically modified CPCs. Conclusion:The engineering TGF-β3 could have targeted therapy for cartilage repair in future.