Effect of icariin on early steroid-induced osteonecrosis of the femoral head in rabbits_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

SHI Lijun ¹ , MAO Tianli ² , LUO Pan ¹ , LI Tengqi ³ , GAO Fuqiang ⁴ ,  SUN Wei ^1,4 , LI Zirong ⁴

1. Department of Orthopedics, Graduate School of Peking Union Medical College, China-Japan Friendship Hospital, Beijing, 100029, P.R.China;
2. Department of Orthopedics, Peking University Third Hospital, Beijing, 100191, P.R.China;
3. Department of Orthopedics, Peking University China-Japan Friendship School of Clinical Medicine, Beijing, 100029, P.R.China;
4. Center for Osteonecrosis and Joint Preserving & Reconstruction, China-Japan Friendship Hospital, Beijing, 100029, P.R.China;

Corresponding author：

SUN Wei, Email: sun887@163.com

Keywords：

Steroid-induced osteonecrosis of the femoral head; icariin; Micro-CT; trabecula; empty lacuna; rabbit

DOI：

10.7507/1002-1892.201905112

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To explore the effect of icariin on early steroid-induced osteonecrosis of the femoral head in rabbits.Methods Fifty mature New Zealand rabbits (weighing, 2.5-3.0 kg) were randomly divided into control group (n=10), model group (n=20), and experimental group (n=20). The rabbits of model and experimental groups were injected with lipopolysaccharide and methylprednisolone to establish the animal model of early steroid-induced osteonecrosis of the femoral head. The rabbits of experimental group were feeded with icariin solution once a day for 6 weeks since the first injection of methylprednisolone, whereas the rabbits of control and model groups were given normal saline at the same time points. The left femoral heads were removed after 6 weeks and gross morphological features were evaluated. Micro-CT scan was performed to analyze the trabecular microstructure with the following parameters: trabecular bone volume to total volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Tn), and trabecular separation (Tb.Sp). The Micro-CT scan was also converted to three-dimensional reconstruction images for observation. HE staining was applied to observe the trabecular structure and morphological changes of osteocytes and marrow adipocytes. It was also used to determine whether the samples of femoral heads occurred osteonecrosis based on the criteria for pathological diagnosis, and calculate the rate of empty lacunae.Results Seven rabbits died during the study, and 9, 16, and 18 rabbits in the control, model, and experimental groups, respectively, enrolled the final analysis. Compared with control group, the femoral head collapse and trabecular breaks were more obvious, and the trabeculae were sparse with irregular arrangement in the model group according to the results of gross observation, Micro-CT scan, and three-dimensional reconstruction images. But in the experimental group, the surface of femoral head was slight shrinking without obvious collapse, and the degeneration of trabecular structure was mild. According to bone microstructures analysis, the Tb.N, Tb.Tn, and BV/TV of femoral head in model and experimental groups were lower than those in control group, while the Tb.Sp in the model and experimental groups were significantly higher. The Tb.N, Tb.Tn, and BV/TV of femoral head in experimental group were higher than those in model group, while the Tb.Sp in the experimental group was significantly lower. The differences between groups were all significant (P<0.05). In the model group, HE staining showed that the number of osteocytes reduced, the number of empty lacunae increased, and the marrow adipocytes piled up in the space between femoral trabeculae, some even mashed together like a cyst. In the experimental group, the trabecular structure was still relatively complete compared with model group, no obvious apoptosis of osteocytes was observed, the size and number of adipocytes were basically normal. None of the animals in control group occurred osteonecrosis of the femoral head based on the criteria for pathological diagnosis, and the incidence of osteonecrosis were 81.3% (13/16) in the model group and 66.7% (12/18) in the experimental group, and the difference was not significant (P=0.448). The rate of empty lacunae of osteonecrotic femoral heads in the model group was 33.1%±1.4%, which was higher than that in experimental group (18.9%±0.8%) and in control group (12.7%±1.5%), and the differences between groups were significant (P<0.05).Conclusion The icariin has a protective effect on the early steroid-induced osteonecrosis of the femoral head in rabbits, which can decrease osteocytes apoptosis, improve the bone microstructure, and delay such disease processes.

Citation： SHI Lijun, MAO Tianli, LUO Pan, LI Tengqi, GAO Fuqiang, SUN Wei, LI Zirong. Effect of icariin on early steroid-induced osteonecrosis of the femoral head in rabbits. Chinese Journal of Reparative and Reconstructive Surgery, 2020, 34(2): 206-212. doi: 10.7507/1002-1892.201905112 Copy

1.	Yoon BH, Jones LC, Chen CH, et al. Etiologic classification criteria of ARCO on femoral head osteonecrosis part 1: glucocorticoid-associated osteonecrosis. J Arthroplasty, 2019, 34(1): 163-167.
2.	张庆宇, 高福强, 程立明, 等. 淫羊藿苷对骨微血管内皮细胞自噬及外泌体产生的影响. 中国修复重建外科杂志, 2019, 33(5): 568-577.
3.	Li ZR, Cheng LM, Wang KZ, et al. Herbal Fufang Xian Ling Gu Bao prevents corticosteroid-induced osteonecrosis of the femoral head—A first multicentre, randomised, double-blind, placebo-controlled clinical trial. J Orthop Translat, 2018, 12: 36-44.
4.	Xie XH, Wang XL, Yang HL, et al. Steroid-associated osteonecrosis: Epidemiology, pathophysiology, animal model, prevention, and potential treatments (an overview). J Orthop Translat, 2015, 3(2): 58-70.
5.	李子荣, 孙伟, 屈辉, 等. 皮质类固醇与骨坏死关系的临床研究. 中华外科杂志, 2005, 43(16): 1048-1053.
6.	Ikeuchi K, Hasegawa Y, Seki T, et al. Epidemiology of nontraumatic osteonecrosis of the femoral head in Japan. Mod Rheumatol, 2015, 25(2): 278-281.
7.	Mont MA, Pivec R, Banerjee S, et al. High-dose corticosteroid use and risk of hip osteonecrosis: meta-analysis and systematic literature review. J Arthroplasty, 2015, 30(9): 1506-1512.
8.	Issa K, Pivec R, Kapadia BH, et al. Osteonecrosis of the femoral head: the total hip replacement solution. Bone Joint J, 2013, 95-B(11 Suppl A): 46-50.
9.	魏秋实, 杨帆, 陈哓俊, 等. 激素性与酒精性股骨头坏死患者骨标本坏死区域病理与显微结构特点分析. 中国修复重建外科杂志, 2018, 32(7): 866-872.
10.	Cooper MS. Sensitivity of bone to glucocorticoids. Clin Sci (Lond), 2004, 107(2): 111-123.
11.	El Zaoui I, Behar-Cohen F, Torriglia A. Glucocorticoids exert direct toxicity on microvasculature: analysis of cell death mechanisms. Toxicol Sci, 2015, 143(2): 441-453.
12.	Wang Z, Wang D, Yang D, et al. The effect of icariin on bone metabolism and its potential clinical application. Osteoporos Int, 2018, 29(3): 535-544.
13.	宁钰, 秦文, 任亚辉, 等. 载淫羊藿苷/凹凸棒石/Ⅰ 型胶原/聚己内酯复合支架修复兔胫骨缺损的实验研究. 中国修复重建外科杂志, 2019, 33(9): 1181-1189.
14.	Zhang Y, Shen L, Mao Z, et al. Icariin enhances bone repair in rabbits with bone infection during post-infection treatment and prevents inhibition of osteoblasts by vancomycin. Front Pharmacol, 2017, 8: 784.
15.	Tang D, Ju C, Liu Y, et al. Therapeutic effect of icariin combined with stem cells on postmenopausal osteoporosis in rats. J Bone Miner Metab, 2018, 36(2): 180-188.
16.	Liu H, Xiong Y, Zhu X, et al. Icariin improves osteoporosis, inhibits the expression of PPARγ, C/EBPα, FABP4 mRNA, N1ICD and jagged1 proteins, and increases Notch2 mRNA in ovariectomized rats. Exp Ther Med, 2017, 13(4): 1360-1368.
17.	Fan JJ, Cao LG, Wu T, et al. The dose-effect of icariin on the proliferation and osteogenic differentiation of human bone mesenchymal stem cells. Molecules, 2011, 16(12): 10123-10133.
18.	Zhang D, Fong C, Jia Z, et al. Icariin stimulates differentiation and suppresses adipocytic transdifferentiation of primary osteoblasts through estrogen receptor-mediated pathway. Calcif Tissue Int, 2016, 99(2): 187-198.
19.	Wang Y, Wang R, Zhang F. Icariin promotes the proliferation and differentiation of osteoblasts from the rat mandible by the Wnt/β-catenin signalling pathway. Mol Med Rep, 2018, 18(3): 3445-3450.
20.	Huang Z, Cheng C, Wang J, et al. Icariin regulates the osteoblast differentiation and cell proliferation of MC3T3-E1 cells through microRNA-153 by targeting Runt-related transcription factor 2. Exp Ther Med, 2018, 15(6): 5159-5166.
21.	Chen M, Cui Y, Li H, et al. Icariin promotes the osteogenic action of BMP2 by activating the cAMP signaling pathway. Molecules, 2019, 24(21): E3875.
22.	Liu HJ, Liu XY, Jing DB. Icariin induces the growth, migration and osteoblastic differentiation of human periodontal ligament fibroblasts by inhibiting Toll-like receptor 4 and NF-κB p65 phosphorylation. Mol Med Rep, 2018, 18(3): 3325-3331.
23.	Li X, Peng B, Pan Y, et al. Icariin stimulates osteogenic differentiation and suppresses adipogenic differentiation of rBMSCs via estrogen receptor signaling. Mol Med Rep, 2018, 18(3): 3483-3489.
24.	高堪达, 王漱阳, 王秋根. 淫羊藿苷治疗小鼠骨关节炎后血清学及组织学改变的实验研究. 中国修复重建外科杂志, 2017, 31(8): 963-969.
25.	Wu Y, Cao L, Xia L, et al. Evaluation of osteogenesis and angiogenesis of icariin in local controlled release and systemic delivery for calvarial defect in ovariectomized rats. Sci Rep, 2017, 7(1): 5077.
26.	Yu H, Yue J, Wang W, et al. Icariin promotes angiogenesis in glucocorticoid-induced osteonecrosis of femoral heads: In vitro and in vivo studies. J Cell Mol Med, 2019, 23(11): 7320-7330.
27.	余开富, 谭洪波, 徐永清. 股骨头缺血性坏死动物模型制备的研究进展. 中国修复重建外科杂志, 2015, 29(12): 1564-1569.

1. Yoon BH, Jones LC, Chen CH, et al. Etiologic classification criteria of ARCO on femoral head osteonecrosis part 1: glucocorticoid-associated osteonecrosis. J Arthroplasty, 2019, 34(1): 163-167.
2. 张庆宇, 高福强, 程立明, 等. 淫羊藿苷对骨微血管内皮细胞自噬及外泌体产生的影响. 中国修复重建外科杂志, 2019, 33(5): 568-577.
3. Li ZR, Cheng LM, Wang KZ, et al. Herbal Fufang Xian Ling Gu Bao prevents corticosteroid-induced osteonecrosis of the femoral head—A first multicentre, randomised, double-blind, placebo-controlled clinical trial. J Orthop Translat, 2018, 12: 36-44.
4. Xie XH, Wang XL, Yang HL, et al. Steroid-associated osteonecrosis: Epidemiology, pathophysiology, animal model, prevention, and potential treatments (an overview). J Orthop Translat, 2015, 3(2): 58-70.
5. 李子荣, 孙伟, 屈辉, 等. 皮质类固醇与骨坏死关系的临床研究. 中华外科杂志, 2005, 43(16): 1048-1053.
6. Ikeuchi K, Hasegawa Y, Seki T, et al. Epidemiology of nontraumatic osteonecrosis of the femoral head in Japan. Mod Rheumatol, 2015, 25(2): 278-281.
7. Mont MA, Pivec R, Banerjee S, et al. High-dose corticosteroid use and risk of hip osteonecrosis: meta-analysis and systematic literature review. J Arthroplasty, 2015, 30(9): 1506-1512.
8. Issa K, Pivec R, Kapadia BH, et al. Osteonecrosis of the femoral head: the total hip replacement solution. Bone Joint J, 2013, 95-B(11 Suppl A): 46-50.
9. 魏秋实, 杨帆, 陈哓俊, 等. 激素性与酒精性股骨头坏死患者骨标本坏死区域病理与显微结构特点分析. 中国修复重建外科杂志, 2018, 32(7): 866-872.
10. Cooper MS. Sensitivity of bone to glucocorticoids. Clin Sci (Lond), 2004, 107(2): 111-123.
11. El Zaoui I, Behar-Cohen F, Torriglia A. Glucocorticoids exert direct toxicity on microvasculature: analysis of cell death mechanisms. Toxicol Sci, 2015, 143(2): 441-453.
12. Wang Z, Wang D, Yang D, et al. The effect of icariin on bone metabolism and its potential clinical application. Osteoporos Int, 2018, 29(3): 535-544.
13. 宁钰, 秦文, 任亚辉, 等. 载淫羊藿苷/凹凸棒石/Ⅰ 型胶原/聚己内酯复合支架修复兔胫骨缺损的实验研究. 中国修复重建外科杂志, 2019, 33(9): 1181-1189.
14. Zhang Y, Shen L, Mao Z, et al. Icariin enhances bone repair in rabbits with bone infection during post-infection treatment and prevents inhibition of osteoblasts by vancomycin. Front Pharmacol, 2017, 8: 784.
15. Tang D, Ju C, Liu Y, et al. Therapeutic effect of icariin combined with stem cells on postmenopausal osteoporosis in rats. J Bone Miner Metab, 2018, 36(2): 180-188.
16. Liu H, Xiong Y, Zhu X, et al. Icariin improves osteoporosis, inhibits the expression of PPARγ, C/EBPα, FABP4 mRNA, N1ICD and jagged1 proteins, and increases Notch2 mRNA in ovariectomized rats. Exp Ther Med, 2017, 13(4): 1360-1368.
17. Fan JJ, Cao LG, Wu T, et al. The dose-effect of icariin on the proliferation and osteogenic differentiation of human bone mesenchymal stem cells. Molecules, 2011, 16(12): 10123-10133.
18. Zhang D, Fong C, Jia Z, et al. Icariin stimulates differentiation and suppresses adipocytic transdifferentiation of primary osteoblasts through estrogen receptor-mediated pathway. Calcif Tissue Int, 2016, 99(2): 187-198.
19. Wang Y, Wang R, Zhang F. Icariin promotes the proliferation and differentiation of osteoblasts from the rat mandible by the Wnt/β-catenin signalling pathway. Mol Med Rep, 2018, 18(3): 3445-3450.
20. Huang Z, Cheng C, Wang J, et al. Icariin regulates the osteoblast differentiation and cell proliferation of MC3T3-E1 cells through microRNA-153 by targeting Runt-related transcription factor 2. Exp Ther Med, 2018, 15(6): 5159-5166.
21. Chen M, Cui Y, Li H, et al. Icariin promotes the osteogenic action of BMP2 by activating the cAMP signaling pathway. Molecules, 2019, 24(21): E3875.
22. Liu HJ, Liu XY, Jing DB. Icariin induces the growth, migration and osteoblastic differentiation of human periodontal ligament fibroblasts by inhibiting Toll-like receptor 4 and NF-κB p65 phosphorylation. Mol Med Rep, 2018, 18(3): 3325-3331.
23. Li X, Peng B, Pan Y, et al. Icariin stimulates osteogenic differentiation and suppresses adipogenic differentiation of rBMSCs via estrogen receptor signaling. Mol Med Rep, 2018, 18(3): 3483-3489.
24. 高堪达, 王漱阳, 王秋根. 淫羊藿苷治疗小鼠骨关节炎后血清学及组织学改变的实验研究. 中国修复重建外科杂志, 2017, 31(8): 963-969.
25. Wu Y, Cao L, Xia L, et al. Evaluation of osteogenesis and angiogenesis of icariin in local controlled release and systemic delivery for calvarial defect in ovariectomized rats. Sci Rep, 2017, 7(1): 5077.
26. Yu H, Yue J, Wang W, et al. Icariin promotes angiogenesis in glucocorticoid-induced osteonecrosis of femoral heads: In vitro and in vivo studies. J Cell Mol Med, 2019, 23(11): 7320-7330.
27. 余开富, 谭洪波, 徐永清. 股骨头缺血性坏死动物模型制备的研究进展. 中国修复重建外科杂志, 2015, 29(12): 1564-1569.

Chinese Journal of Reparative and Reconstructive Surgery

Effect of icariin on early steroid-induced osteonecrosis of the femoral head in rabbits

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content