| Objectives To study the effect of MG63 osteoblast-porous tantalum compound on cell proliferation, secreion and expression of COL-I, OC, OPN m RNA and protein, to explore porous tantalum scaffold as bone tissue engineering scaffold and to provide theoretical and experimental basic for bone defects repair and bone tissue physiological structure and function recover.Methods 1 The morphology of porous tantalum was detected by SEM. 2 OB was cultured and identified. α-MEM was prepared as tantalum extracted medium. OB was divided into tantalum extracted liquid group(OB/tantalum extracted liquid), tantalum scaffolds group(OB/porous tantalum), and control group(OB) respectively. 3 The proliferation of OBtantalum compond was detected by CCK-8 assay. 4 The adhesion, proliferation and ultrastructure changes of OB on porous tantalum were detected by inverted phase contrast microscope, SEM and TEM. 5 The secretion of OC and COL-I in OB-porous tantalum compond was examined by ELISA. 6 The expression of COL-I, OC, OPN protein in OBporous tantalum compond was detected by immunocytochemical staining. 7 The expression of COL- I, OC and OPN protein in OB-porous tantalum compond was examined by Western-blot assay. 8 The expression of COL-I, OC, and OPN m RNA in OBporous tantalum compond was examined by RT-q PCR assay.Results 1 SEM showed that porous tantalum had many pores communicating each other like human cancellous bone. 2 The CCK-8 test showed that OB grew well. There was no significant difference among three groups(P>0.05). 3 SEM showed that OB grew into porous tantalum and its surface, and ECM extensively spread on its surface. TEM showed that OB of OB/tantalum extracted liquid group and OB/porous tantalum group had rich cell ultrastructure. 4 The secretion of OC in OB/porous tantalum group was significantly higher than that of OB/tantalum extracted liquid group and control group at 5, 7 day(P<0.05) and there was no significant difference among three groups at 3 day(P>0.05). There was no significant difference in the secretion of COL-I among three groups at 3, 5, 7 day(P>0.05). 5 The expression of OC, OPN protein in OB/ porous tantalum group was significantly higher than that of OB/tantalum extracted liquid group and control groups at 5 day by immunocytochemical staining(P<0.05) and there was no significant difference in the expression of COL-I protein among three groups(P>0.05). 6 The expression of OC, OPNprotein in OB/ porous tantalum group was significantly higher than that of OB/tantalum extracted liquid group and control groups at 5 day by Western-blot(P<0.05) and there was no significant difference in expression of COL-I protein among three groups(P>0.05). 7 The expression of OC, OPN m RNA in OB/ porous tantalum group was significantly higher than that of OB/tantalum extracted liquid group and control groups at 5 day by RT-q PCR(P<0.05) and there was no significant difference in expression of COL-I m RNA among three groups(P>0.05).Conclusions 1 Porous tantalum and its extracted liquid have benefit to the early growth, adhesion and proliferation of human OB, to reveal that porous tantalum has good cell affinity and biocompatibility. 2 Porous tantalum 3D structure can promote adhesion and secretion of OC, to maintain the balance of bone formation and resorption for benefiting to bone mineralization process. The secretion of COL-I is not affected by the spatial structure of porous tantalum, ran through the whole process of bone fomation, Especially that of early. 3 The result of Western-blot and RT-q PCR showed that OB in porous tantalum 3D culture, the expression of OC, OPN m RNA and protein were higher than that of 2D culture.These data suggested that OB-porous tantalum compond under certain conditions could be used as a bone graft material in bone defects repair. |
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