Transinfection of rabbit chondrocytes via chitosan microsphere as gene carriers with recombined human IL-1Ra gene and TGF-β1 gene / 中华显微外科杂志

ABSTRACT

Objective To explore transinfection of rabbit chondrocytes via chitosan microsphere with human IL-1Ra and TGF-β1 gene. Methods Chitosan-DNA microspheres carrying plasmids with IL-1 Ra and TGF-β1 genes were prepared.The encapsulation efficiency,DNA-released kinetics and lysozyme degradation in vitro were performed.Articular rabbit chondrocytes were co-transinfected with the plasmids with IL-1Ra and TGF-β1 genes via chitosan-DNA mierosphere,evaluated by fluorescence microscope,TaqMan real-time PCR assay and MTF test. Results The chitosan microspheres with IL-1Ra and TGF-β1 genes were(2.8±0.2)μm and(2.6±0.1)μm in diameters respectively.The encapsulation efficiency were(88.3±4.1)%and(87.2±2.6)%.During the degradation,significant morphological changes were noticed.The plasmids could be released in a multiphasic fashion.Enhanced green fluorescent protein and Real-Time PCR analysis showed that genes were expressed in chondrocytes.lasting near 30 days.MTT indicated that the cotransinfection promoted the chondrocytes'proliferation. Conclusion IL-1Ra and TGF-β1 genes cotransfected into chondrocytes via chitosan-DNA microsphere could be expressed in a long term and cotransinfection promoted the chondroeytes'proliferation,which is the base of inhibiting the degeneration of cartilage and promote cartilage repair.