RESUMEN
OBJECTIVE: To estimate the potential effects of hBMP-7 gene transfected bone marrow stromal cells (BMSC) and to provide a new way for repairing the periodontal defects. METHODS: A total of 30 experimental class II furcation defects in five Beagle dogs were created surgically. The transfected and non-transfected BMSC were seeded in collagen membrane at an initial concentration of 1 x 10(7)/ml and transplanted into experimental class II furcation defects. The animals were sacrificed at 12 weeks post-operation and periodontal regeneration was evaluated by the histological and morphometric analysis. RESULTS: In both experimental and control groups, there were different extents of regeneration. The percentage of new alveolar area in transfected and non-transfected BMSC were significantly higher than the control (P < 0.05). And there was also significant difference between two experimental groups (P < 0.05). The percentage of new cementum length in two experimental groups were significantly higher than the control (P < 0.05). But there was no significant difference between two BMSCs groups (P > 0.05). CONCLUSION: hBMP-7 gene enhanced tissue engineering could be a better way to repair the periodontal defects.
Asunto(s)
Proteína Morfogenética Ósea 7/genética , Regeneración Tisular Guiada Periodontal/métodos , Transfección , Animales , Células de la Médula Ósea , Perros , Terapia Genética/métodos , Células Madre Mesenquimatosas , Enfermedades Periodontales/terapia , Ingeniería de Tejidos/métodosRESUMEN
OBJECTIVE: To explore transient expression of the eukaryotic expression plasmid carrying human platelet-derived growth factor-B (hPDGF-B) in gingival fibroblasts of Beagle dog. METHODS: Plasmid carrying hPDGF-B (EX-A0380-M03) was amplified and identified, and then transfected into gingival fibroblasts of Beagle dog. Reverse transcription polymerase chain reaction (RT-PCR), immunocytochemistry, enzyme-linked immunosorbent assay (ELISA) and Western bolt were choose to detect the expression of hPDGF-B. RESULTS: Target gene carried by EX-A0380-M03 was hPDGF-B. Green fluorescence protein (GFP) expressed by transfected gingival fibroblasts was observed under inverted phase contrast fluorescence microscope (IPCFM) (after 24 hours) and the transfection efficiency was 18%-38% (after 48 hours). Serials other methods (RT-PCR, immunocytochemistry, and ELISA) mentioned above also convinced that cells expressed hPDGF-B, and the protein that was a kind of fusion protein composed of PDGF-BB and GFP was identified by Western blot. CONCLUSION: Eukaryotic expression plasmid carrying hPDGF-B was transfected into gingival fibroblasts successfully, and a kind of fusion protein was expressed.