[Construction of leptin gene modified tissue engineered composites in vitro].

PURPOSE: To evaluate the feasibility of constructing tissue engineered composites in vitro by combining human leptin (hLEP) gene modified rat bone marrow stromal cells (BMSCs) and guided tissue regeneration collagen membrane (Bio-Gide).

METHODS

BMSCs of SD rats were isolated and cultured by whole bone marrow adherent method. BMSCS were transfected with adenovirus carrying hLEP gene (Ad-hLEP-EGFP) and observed under inverted fluorescence microscope. Enzyme linked immunosorbent assay (ELISA) was used to detect the expression of hLEP. The proliferation activity of transfected cells was assessed by MTT assay. Ad-hLEP-EGFP transfected BMSCs were cultured for 24 h in combination with Bio-Gide collagen membrane, hLEP modified tissue engineered composite was observed under laser scanning confocal microscope (LSCM) and scanning electron microscope (SEM).

RESULTS

Through Ad-hLEP-EGFP transfection, hLEP was overexpressed in BMSCs, which didn't affect the proliferation of cells. SEM showed hLEP modified BMSCs grew well on Bio-Gide collagen membrane and secreted extracellular matrix. LSCM suggested BMSCs could migrate to different scales of Bio-Gide collagen membrane.

CONCLUSIONS

hLEP modified BMSCs can be combined with Bio-Gide collagen membrane and grow well, suggesting that hLEP modified tissue engineered composite can be successfully constructed. The composite might be suitable for periodontal tissue engineering.