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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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.
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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. 姜黃素藥理作用及機制研究進展. 中國醫藥導報, 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.	熊藝璇, 趙斌, 賈凌璐, 等. 姜黃素通過 Nrf2 信號通路促進炎癥狀態下牙周膜干細胞的成骨分化. 山東大學學報 (醫學版), 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, 葉玟希, 等. 交聯透明質酸衍生物的制備進展. 廣東化工, 2012, 39(5): 99-100.
13.	韓愈, 杜池. 淫羊藿苷與磷酸鈣骨水泥形成蛋白載體修復骨缺損的相關性分析. 臨床口腔醫學雜志, 2020, 36(10): 605-608.
14.	董航, 黃嘉華, 麥喆钘, 等. 載骨碎補總黃酮磷酸鈣骨水泥對骨缺損模型大鼠誘導膜中成骨細胞分化的影響及其機制研究. 中國藥房, 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, B?aszczyk 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. 陸儉, 唐天駟, 楊惠林. 椎體成形術治療骨質疏松性椎體壓縮骨折的進展. 中國脊柱脊髓雜志, 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. 姜黃素藥理作用及機制研究進展. 中國醫藥導報, 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. 熊藝璇, 趙斌, 賈凌璐, 等. 姜黃素通過 Nrf2 信號通路促進炎癥狀態下牙周膜干細胞的成骨分化. 山東大學學報 (醫學版), 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, 葉玟希, 等. 交聯透明質酸衍生物的制備進展. 廣東化工, 2012, 39(5): 99-100.
13. 韓愈, 杜池. 淫羊藿苷與磷酸鈣骨水泥形成蛋白載體修復骨缺損的相關性分析. 臨床口腔醫學雜志, 2020, 36(10): 605-608.
14. 董航, 黃嘉華, 麥喆钘, 等. 載骨碎補總黃酮磷酸鈣骨水泥對骨缺損模型大鼠誘導膜中成骨細胞分化的影響及其機制研究. 中國藥房, 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, B?aszczyk 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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