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目的:观察大鼠牙髓干细胞(dental pulp stem cells,DPSCs)在微弧氧化(micro-arc oxidation,MAO)钛片表面粘附和分化情况,初步评价MAO钛片对DPSCs的成骨诱导性能。方法:实验组:DPSCs与MAO钛片共培养,阳性对照组:DPSCs成骨诱导培养,空白对照组:DPSCs无诱导普通培养。扫描电镜(SEM)观察DPSCs在MAO钛片表面粘附情况,碱性磷酸酶(ALP)活力检测,实时定量PCR分析Runx2、Osterix、DSPP和DMP-1基因表达和钙结节茜素红染色,鉴定DPSCs的成骨能力。结果:SEM观察MAO钛片表面粗糙多孔,DPSCs在MAO钛片表面粘附紧密,有触角向微孔内伸展。ALP活力检测和钙结节染色,实验组与阳性对照组无显著差异(P>0.05),与空白对照组有显著差异(P<0.05)。实验组Runx2、Osterix表达增强,DSPP和DMP-1表达无明显变化。结论:粗糙多孔的MAO钛片与DPSCs有良好的生物相容性,DPSCs在MAO钛片表面粘附紧密,具有向成骨细胞系分化的能力。
OBJECTIVE: To observe the adhesion and differentiation of rat dental pulp stem cells (DPSCs) on the surface of micro-arc oxidation (MAO) titanium plate and to evaluate the osteoinductivity of MAO titanium plate on DPSCs. Methods: Experimental group: DPSCs co-cultured with MAO titanium plate, positive control group: osteogenic induction of DPSCs, blank control group: DPSCs without induction of normal culture. Scanning electron microscopy (SEM) was used to observe the adhesion of DPSCs on the surface of MAO titanium plate, the activity of alkaline phosphatase (ALP) and the expression of Runx2, Osterix, DSPP and DMP-1 mRNA and alizarin red staining were detected by real- Identification of osteogenic ability of DPSCs. Results: The surface of the MAO titanium sheet was rough and porous. The DPSCs adhered tightly to the surface of the MAO titanium sheet, and the antennae extended into the micropores. ALP activity and calcium staining, there was no significant difference between the experimental group and the positive control group (P> 0.05), which was significantly different from that of the blank control group (P <0.05). The experimental group Runx2, Osterix expression increased, DSPP and DMP-1 expression did not change significantly. CONCLUSION: The coarse and porous MAO titanium plate has good biocompatibility with DPSCs. DPSCs adhere closely to the surface of MAO titanium plate and have the ability to differentiate into osteoblast lines.