Department of Orthopaedics, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China.
Department of Orthopaedics, Beijing Tongren Hospital, The Affiliated Hospital of Capital University of Medical Sciences, Beijing 100730, China.
Biochem Biophys Res Commun. 2019 Jan 1;508(1):25-30. doi: 10.1016/j.bbrc.2018.11.097. Epub 2018 Nov 20.
HA modified by bisphosphonate (BP) (HA-BP) was synthesized by chemical reaction and possessed promising properties such as self-healing, injection ability, and strong adhesion. The main aim of this study was to confirm its role in promoting osteogenic differentiation in vitro and bone regeneration in vivo.
The cell biocompatibility of this material was determined using the CCK-8 assay. Alkaline phosphatase (ALP), osteocalcin (OT), vascular endothelial growth factor (VEGF), and collagen I were assessed by quantitative real-time polymerase chain reaction (Q-PCR) in the treated group. The number and density of calcium nodules and ALP were evaluated by Alizarin Red staining and ALP staining. We have successfully developed an animal model simulating osteonecrosis of the femoral head (ONFH). Utilizing this animal model, the impact of HA-BP/CaP on bone formation was assessed. The amount of bone regeneration at 1 and 2 months after HA-BP/CaP injection was estimated by micro-computed tomography (micro-CT) analysis and H&E, collagen I, and periostin staining.
The number of cells gradually increased in the experimental group over time and was close to that of the blank control group. ALP, collagen I, and VEGF expression was significantly higher in the experimental group than in the blank group (VEGF, ALP, both **p < 0.01; collagen I, ***p<0.001). In addition, the number and density of calcium nodules and ALP was clearly greater in the material group than in the control group. The quantification analysis showed that the mineral contents of regenerated bone at 1 and 2 months after HA-BP/CaP injection were significantly greater than those in the control group, according to micro-CT evaluation (**p<0.01). The amount of organic components in the HA-BP/CaP group was greater than that in the control group after decalcification and H&E staining. In addition, collagen I and periostin staining further confirmed the results of H&E staining.
This material can boost proliferation and osteogenic differentiation of MC3T3-E1 cells in vitro. It can intensely accelerate bone regeneration in vivo, which is a promising strategy for tissue engineering.
通过化学反应合成了经双膦酸盐（BP）修饰的 HA（HA-BP），其具有自修复、可注射和强附着力等有前景的特性。本研究的主要目的是确认其在体外促进成骨分化和体内骨再生中的作用。
通过 CCK-8 测定法确定该材料的细胞生物相容性。通过定量实时聚合酶链反应（Q-PCR）评估处理组中碱性磷酸酶（ALP）、骨钙素（OT）、血管内皮生长因子（VEGF）和胶原蛋白 I 的表达。通过茜素红染色和 ALP 染色评估钙结节的数量和密度以及 ALP 的活性。我们成功地建立了模拟股骨头坏死（ONFH）的动物模型。利用该动物模型评估了 HA-BP/CaP 对骨形成的影响。通过微计算机断层扫描（micro-CT）分析和 H&E、胶原蛋白 I 和骨膜蛋白染色评估 HA-BP/CaP 注射后 1 个月和 2 个月时的骨再生量。
实验组细胞数量随时间逐渐增加，接近空白对照组。实验组中 ALP、胶原蛋白 I 和 VEGF 的表达明显高于空白组（VEGF、ALP，均 **p<0.01；胶原蛋白 I，***p<0.001）。此外，材料组钙结节和 ALP 的数量和密度明显大于对照组。定量分析显示，根据 micro-CT 评价，HA-BP/CaP 注射后 1 个月和 2 个月时再生骨的矿物质含量明显大于对照组（**p<0.01）。脱钙和 H&E 染色后，HA-BP/CaP 组的有机成分含量大于对照组。此外，胶原蛋白 I 和骨膜蛋白染色进一步证实了 H&E 染色的结果。
该材料可促进 MC3T3-E1 细胞在体外的增殖和成骨分化。它可以强烈促进体内骨再生，这是组织工程的一种有前途的策略。
