Effects of calcium phosphate cement combined with hyaluronic acid/curcumin on the proliferation and osteogenesis of osteoblasts_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Ying ¹ , JIA Shuaijun ² , TIAN Fang ² , GAO Xinlin ² , WANG Zhiyuan ² , ZHU Lei ² ,  HAO Dingjun ^1,2

1. Shaanxi University of Traditional Chinese Medicine, Xianyang Shaanxi, 712046, P.R.China;
2. Department of Spine Surgery, Honghui Hospital Affiliated to Medical School of Xi’an Jiaotong University, Xi’an Shaanxi, 710054, P.R.China;

Corresponding author：

HAO Dingjun, Email: haodingjun@163.com

Keywords：

Curcumin; hyaluronic acid; calcium phosphate cement; osteoblasts; proliferation; osteogenesis

DOI：

10.7507/1002-1892.202007088

Video：

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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.

Citation： ZHANG Ying, JIA Shuaijun, TIAN Fang, GAO Xinlin, WANG Zhiyuan, ZHU Lei, HAO Dingjun. Effects of calcium phosphate cement combined with hyaluronic acid/curcumin on the proliferation and osteogenesis of osteoblasts. Chinese Journal of Reparative and Reconstructive Surgery, 2021, 35(1): 104-110. doi: 10.7507/1002-1892.202007088 Copy

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11.	He J, Yang XF, Liu F, et al. The impact of curcumin on bone osteogenic promotion of MC3T3 cells under high glucose conditions and enhanced bone formation in diabetic mice. Coating, 2020, 10: 258.
12.	徐新, 施丹萍, 叶玟希, 等. 交联透明质酸衍生物的制备进展. 广东化工, 2012, 39(5): 99-100.
13.	韩愈, 杜池. 淫羊藿苷与磷酸钙骨水泥形成蛋白载体修复骨缺损的相关性分析. 临床口腔医学杂志, 2020, 36(10): 605-608.
14.	董航, 黄嘉华, 麦喆钘, 等. 载骨碎补总黄酮磷酸钙骨水泥对骨缺损模型大鼠诱导膜中成骨细胞分化的影响及其机制研究. 中国药房, 2019, 30(10): 1321-1327.
15.	Landeros JM, Belmont-Bernal F, Pérez-González AT, et al. A two-step synthetic strategy to obtain a water-soluble derivative of curcumin with improved antioxidant capacity and in vitro cytotoxicity in C6 glioma cells. Mater Sci Eng C Mater Biol Appl, 2017, 71: 351-362.
16.	Ogurkowska MB, Błaszczyk A. Variation in human vertebral body strength for vertebral body samples from different locations in segments L1-L5. Clin Biomech (Bristol, Avon), 2018, 60: 66-75.
17.	Xiong Y, Zhao B, Zhang W, et al. Curcumin promotes osteogenic differentiation of periodontal ligament stem cells through the PI3K/AKT/Nrf2 signaling pathway. Iran J Basic Med Sci, 2020, 23(7): 954-960.

1. Yang D, Zhang Y, Ma X, et al. Resources utilisation and economic burden of percutaneous vertebroplasty or percutaneous kyphoplasty for treatment of osteoporotic vertebral compression fractures in China: a retrospective claim database study. BMC Musculoskelet Disord, 2020, 21(1): 255-268.
2. 陆俭, 唐天驷, 杨惠林. 椎体成形术治疗骨质疏松性椎体压缩骨折的进展. 中国脊柱脊髓杂志, 2002, 12(4): 302-304.
3. Mehbod A, Aunoble S, Le Huec JC. Vertebroplasty for osteoporotic spine fracture: prevention and treatment. Eur Spine J, 2003, 12(Suppl 2): S155-162.
4. Brown WE, Chow LC. A new calcium phosphate water setting cement//Brown PW. Cements research progress. Westerville: American Ceramic Society, 1986: 352-379.
5. Costantino PD, Friedman CD, Jones K, et al. Experimental hydroxyapatite cement cranioplasty. Plast Reconstr Surg, 1992, 90(2): 174-185.
6. Dolci LS, Panzavolta S, Torricelli P, et al. Modulation of Alendronate release from a calcium phosphate bone cement: An in vitro osteoblast-osteoclast co-culture study. Int J Pharm, 2019, 554: 245-255.
7. Wu T, Yang S, Lu T, et al. Strontium ranelate simultaneously improves the radiopacity and osteogenesis of calcium phosphate cement. Biomed Mater, 2019, 14(3): 035005.
8. 宋莉平, 王宇. 姜黄素药理作用及机制研究进展. 中国医药导报, 2020, 17(20): 29-33.
9. Chen YC, Shie MY, Wu YA, et al. Anti-inflammation performance of curcumin-loaded mesoporous calcium silicate cement. J Formos Med Assoc, 2017, 116(9): 679-688.
10. 熊艺璇, 赵斌, 贾凌璐, 等. 姜黄素通过 Nrf2 信号通路促进炎症状态下牙周膜干细胞的成骨分化. 山东大学学报 (医学版), 2020, 58(5): 19-26.
11. He J, Yang XF, Liu F, et al. The impact of curcumin on bone osteogenic promotion of MC3T3 cells under high glucose conditions and enhanced bone formation in diabetic mice. Coating, 2020, 10: 258.
12. 徐新, 施丹萍, 叶玟希, 等. 交联透明质酸衍生物的制备进展. 广东化工, 2012, 39(5): 99-100.
13. 韩愈, 杜池. 淫羊藿苷与磷酸钙骨水泥形成蛋白载体修复骨缺损的相关性分析. 临床口腔医学杂志, 2020, 36(10): 605-608.
14. 董航, 黄嘉华, 麦喆钘, 等. 载骨碎补总黄酮磷酸钙骨水泥对骨缺损模型大鼠诱导膜中成骨细胞分化的影响及其机制研究. 中国药房, 2019, 30(10): 1321-1327.
15. Landeros JM, Belmont-Bernal F, Pérez-González AT, et al. A two-step synthetic strategy to obtain a water-soluble derivative of curcumin with improved antioxidant capacity and in vitro cytotoxicity in C6 glioma cells. Mater Sci Eng C Mater Biol Appl, 2017, 71: 351-362.
16. Ogurkowska MB, Błaszczyk A. Variation in human vertebral body strength for vertebral body samples from different locations in segments L1-L5. Clin Biomech (Bristol, Avon), 2018, 60: 66-75.
17. Xiong Y, Zhao B, Zhang W, et al. Curcumin promotes osteogenic differentiation of periodontal ligament stem cells through the PI3K/AKT/Nrf2 signaling pathway. Iran J Basic Med Sci, 2020, 23(7): 954-960.

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