Effect Of Yunnan Baiyao On The MRNA Expression Of Osteogenic Related Markers By HPDLFs-Bio-Oss Collagen In Vivo

[Objective] The HPDLFs-Bio-Oss Collagen complex was constructed in vitro. The complexes were added to a solution of Yunnan Baiyao. The composite was implanted subcutaneously in nude mice. The expression of mRNAs for bone sialoprotein, osteocalcin, osteopontin and type-I collagen were detected. This study was to evaluate the feasibility of using Yunnan Baiyao.[Materials and Methods] Human Periodontal ligament fibroblasts (HPDLFs) were obtained using an explant culture method. By using the4Passages for the experiment, cell morphology was observed.Taking in vitro build successful generation of cell-material complex. The cell-material complex were divided into four groups:one group was exposed to an concentration of100μg/ml Yunnan Baiyao solution; one group was exposed to an concentration of50μg/ml Yunnan Baiyao solution; one group was exposed to1OOng/ml of recombinant human bone morphogenetic protein-2(rhBMP-2) as positive control; The final group was exposed to a-MEM containing1%fetal bovine serum as negative control. Comeplexes are implanted subcutaneously in nude mice. Take out the compounds after eight weeks. The expression levels of OCN、OPN、BSP、type-I collagen was observed by Using real-time quantitative RT-PCR. Observe the impact of HPDLFs differentiation which cultured in the Bio-Oss Collagen in vivo with Yunnan Baiyao. Finally the experimental data were statistically analyzed.[Result] OCN mRNA statistical results:The mean expression level of Yunnan Baiyao group at concentration of100μg/ml was higher than the positive control group, the control group and Yunnan Baiyao group at concentration of50μg/ml(P<0.05); The average expression levels of Yunnan Baiyao group at concentration of50μg/ml was lower than the positive control group, Yunnan Baiyao group at concentration of100μg/ml (P<0.05); The average expression level of the positive control group was higher than the control group (P<0.05). BSP mRNA statistical results:The mean expression level of Yunnan Baiyao group at concentration of100μg/ml was higher than the positive control group, the control group and Yunnan Baiyao group at concentration of50μg/ml(P<0.05); The average expression levels of Yunnan Baiyao group at concentration of50μg/ml was lower than the positive control group, Yunnan Baiyao group at concentration of100μg/ml (P<0.05); The average expression level of the positive control group was higher than the control group (P<0.05). OPN mRNA statistical results:The mean expression level of Yunnan Baiyao group at concentration of100μg/ml was higher than the positive control group and Yunnan Baiyao group at concentration of50μg/ml(P<0.05); The average expression levels of Yunnan Baiyao group at concentration of50μg/ml was lower than the control group and Yunnan Baiyao group at concentration of100μg/ml (P<0.05); The average expression level of the control group was higher than the positive control group (P<0.05). Type-I collagen mRNA statistical results:The mean expression level of Yunnan Baiyao group at concentration of100μg/ml was higher than the positive control group, the control group and Yunnan Baiyao group at concentration of50μg/ml(P<0.05); The average expression levels of Yunnan Baiyao group at concentration of50μg/ml was lower than the positive control group, Yunnan Baiyao group at concentration of100μg/ml (P<0.05); The average expression level of the positive control group was higher than the control group (P<0.05).[Conclusion] Yunnan Baiyao at concentration of100μg/ml was able to significantly increase HPDLFs differentiation in vivo.