| 摘要: |
| 目的 研究以硅在天然骨中的含量为掺杂量制备的硅-羟基磷灰石(Si-HA)对成骨细胞活性的影响。方法 以水热法制备微量硅掺杂改性的Si-HA;通过X射线衍射(X-ray diffraction,XRD)及扫描电镜观察材料表面构相;MG63细胞在材料表面分别接种培养1、6、12 h及1、4、7、14 d,通过CCK8方法分别检测MG63细胞在材料表面的黏附、增殖情况;通过DAPI染色法验证CCK8检测细胞黏附数量;并对1、4、7、14 d成骨细胞碱性磷酸酶(ALP)活性进行检测;采用RT-PCR检测成骨细胞特异性基因ALP、Ⅰ型胶原(Col-Ⅰ)、骨钙素(OC)在1、4、7、14 d的表达。结果 Si的掺杂未改变HA表面结构及组成;MG63细胞在材料表面培养6、12 h,Si-HA表面细胞附着数量明显高于HA表面(P<0.05);DAPI染色法与CCK8实验结果相同;细胞培养4、7 d时,Si-HA较HA表面的增殖细胞数量增多(P<0.05)。细胞培养4、7 d时,Si-HA材料表面ALP活性及基因表达高于HA表面(P<0.05);4、7、14 d时Col-Ⅰ基因在Si-HA表面的表达量明显高于HA表面(P<0.05)。7、14 d时,OC基因在Si-HA表面的表达量明显高于HA表面(P<0.05)。结论 以硅在天然骨中的含量为掺杂量的Si-HA能够促进MG63成骨细胞黏附、增殖,上调成骨特异性基因的表达,提高了HA材料的生物学活性。 |
| 关键词: 羟基磷灰石类 硅 成骨细胞 生物学活性 |
| DOI：10.16781/j.0258-879x.2016.04.0405 |
| 投稿时间：2015-12-01修订日期：2016-03-29 |
| 基金项目:国家自然科学基金(81271177);军事口腔医学国家重点实验室开放课题(2014KB07);长海医院"1255"学科建设计划科研创新探索项目(CH125541800). |
|
| Preparation of microdosage silicon-doped hydroxyapatite and its effect on functional activity of osteoblasts |
| GAO Jian-yong1,WANG Ming2,TIAN Gang1,WANG Shao-hai1,WANG Da-lin1* |
(1. Department of Stomatology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Key Lab of Inorganic Coating Materials, Shanghai Institute of Ceramics, Chinese Academy of Sciences, Shanghai 200050, China
*Corresponding author) |
| Abstract: |
| Objective To study the effect of silicon-doped hydroxyapatite (Si-HA), which is prepared in accordance with the silicon content in natural bone, on the activity of osteoblasts. Methods Hydrothermal synthesis method was used to prepare hydroxyapatite doped by microdosage silicon. The surface of the material was observed by X-ray diffraction (XRD) and scanning electron microscope. MG63 osteoblast-like cells were cultured on the surface of materials for 1, 6, 12 h and 1, 4, 7, 14 d, respectively; then the adhesion and proliferation of MG63 cells were detected by CCK8 method; and the number of cell adhesion was verified by DAPI staining. Alkaline phosphatase (ALP) activity of the MG63 cells was assessed with ALP assay kit; RT-PCR was used to detect the expression of osteoblast specific genes (ALP, Collagen Ⅰ and osteocalcin) at 1, 4, 7, and 14 d. Results The surface structure and composition of hydroxyapatite were not changed by Si. After cultured for 6 and 12 h, the MG63 cells adhered to Si-HA surface was significantly more than that to HA surface (P<0.05), which was consistent with the findings of DAPI staining. After cultured for 4 and 7 d, the number of cells on Si-HA surface increased significantly compared with that on HA surface(P<0.05). ALP activity in cells cultured on Si-HA surface were significantly higher than those on HA surface (P<0.05); the expression of CollagenⅠgene in Si-HA group was significantly higher than that in HA group on day 4, 7, and 14(P<0.05), and the expression of osteocalcin gene was significantly higher on day 7 and 14(P<0.05). Conclusion Silicon-doped hydroxyapatite in accordance with silicon content in the natural bone can promote the adhesion, proliferation and the expression of osteogenic specific genes of osteoblast MG63 cells, indicating that the prepared material has an improved biological activity compared with pure HA. |
| Key words: hydroxyapatites silicon osteoblasts bioactivity |
