Objective: After using hyaluronic acid (HA) to modify curcumin (CUR), the effects of calcium phosphate cement (CPC) combined with HA/CUR on the proliferation and osteogenesis of osteoblasts were investigated.
Methods: First, HA and CUR were esterified and covalently combined to prepare HA/CUR, and the characteristics were observed and the infrared spectrum was tested. Then, HA, CUR, and HA/CUR were mixed with CPC according to 5% ( W/ W) to prepare HA-CPC, CUR-CPC, and HA/CUR-CPC, respectively. Setting time detection, scanning electron microscope observation, injectable performance test, and compression strength test were conducted; and the CPC was used as a control. Osteoblasts were isolated and cultured from the skull of newborn Sprague Dawley rats, and the 2nd generation cells were cultured with the 4 types of bone cement, respectively. The effects of HA/CUR-CPC on the proliferation and osteogenesis of osteoblasts were estimated by the scanning electron microscopy observation, live/dead cell fluorescence staining, cell counting, osteopontin (OPN) immunofluorescence staining, alkaline phosphatase (ALP) staining,and alizarin red staining.
Results: Infrared spectroscopy test showed that HA and CUR successfully covalently combined. The HA/CUR-CPC group had no significant difference in initial setting time, final setting time, injectable rate, and compressive strength when compared with the other 3 groups ( P>0.05); scanning electron microscope observation showed that HA/CUR was scattered on CPC surface. After co-culture of bone cement and osteoblasts, scanning electron microscopy observation showed that the osteoblasts, which had normal morphology and the growth characteristics of osteoblasts, clustered and adhered to HA/CUR-CPC. There was no significant difference in cell survival rate between HA/CUR-CPC group and other groups ( P>0.05), and the number of cells significantly increased ( P<0.05); the degrees of OPN immunofluorescence staining, ALP staining, and alizarin red staining were stronger than other groups.
Conclusion: HA/CUR-CPC has good biocompatibility and mechanical properties, which can promote the proliferation and osteogenesis of osteoblasts.
目的: 采用透明质酸（hyaluronic acid，HA）对姜黄素（curcumin，CUR）进行改性，探讨磷酸钙骨水泥（calcium phosphate cement，CPC）复合 HA/CUR 后对成骨细胞增殖及成骨能力的影响。.
方法: 首先将 HA 与 CUR 成酯化共价结合制备 HA/CUR，观察其性状并行红外光谱测试。然后，将 HA、CUR 以及 HA/CUR 按照 5%（ W/ W）分别与 CPC 混合，制备 HA-CPC、CUR-CPC、HA/CUR-CPC，行凝结时间检测、扫描电镜观察以及可注射性能、压缩强度测试；以单纯 CPC 作为对照。取新生 SD 大鼠颅骨分离培养成骨细胞，取第 2 代细胞分别与上述骨水泥复合培养，经扫描电镜观察、活/死细胞荧光染色、细胞计数、骨桥蛋白（osteopontin，OPN）免疫荧光染色、ALP 染色以及茜素红染色，评估 HA/CUR-CPC 对成骨细胞增殖及成骨能力的影响。.
结果: 红外光谱测试显示 HA 及 CUR 成功共价结合。骨水泥观测显示，HA/CUR-CPC 组初凝时间、终凝时间、压缩强度以及可注射率与其他组比较，差异均无统计学意义（ P>0.05）；扫描电镜观察示 HA/CUR 散在分布于 CPC 表面。骨水泥与成骨细胞复合培养后，扫描电镜显示成骨细胞黏附于 HA/CUR-CPC 且形态良好，具有成骨细胞的生长特征；HA/CUR-CPC 组细胞存活率与其他组比较差异无统计学意义（ P>0.05），细胞较其他组明显增多（ P<0.05）；OPN 免疫荧光染色、ALP 染色及茜素红染色程度均强于其他组。.
结论: HA/CUR-CPC 具有良好的生物相容性及力学特性，能促进成骨细胞增殖并增强其成骨能力。.
Keywords: Curcumin; calcium phosphate cement; hyaluronic acid; osteoblasts; osteogenesis; proliferation.