Effect of icariin/attapulgite/collagen type <b>Ⅰ</b>/polycaprolactone composite scaffold in repair of rabbit tibia defect_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

NING Yu ^1,2 , QIN Wen ² , REN Yahui ² , LI Chenkai ² , CHEN Wenyang ² ,  ZHAO Hongbin ^1,2

1. School of Basic Medical Sciences, Gansu University of Traditional Chinese Medicine, Lanzhou Gansu, 730000, P.R.China;
2. Department of Orthopaedic Laboratory, Changzhou Second People’s Hospital, Changzhou Jiangsu, 213000, P.R.China;

Corresponding author：

ZHAO Hongbin, Email: zhao761032@163.com

Keywords：

Icariin; attapulgite; collagen typeⅠ; polycaprolactone; composite scaffold; tissue engineered bone

DOI：

10.7507/1002-1892.201902044

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Objective To investigate the effect of icarin/attapulgite/collagen type Ⅰ/polycaprolactone (ICA/ATP/Col Ⅰ/PCL) composite scaffold in repair of rabbit tibia defect. Methods The ICA/20%ATP/Col Ⅰ/PCL (scaffold 1), ICA/30%ATP/Col Ⅰ/PCL (scaffold 2), 20%ATP/Col Ⅰ/PCL (scaffold 3), and 30%ATP/Col Ⅰ/PCL (scaffold 4) composite scaffolds were constructed by solution casting-particle filtration method. The structure characteristics of the scaffold 2 before and after cross-linking were observed by scanning electron microscopy, and the surface contact angles of the scaffold 2 and the scaffold 4 were used to evaluate the water absorption performance of the material. The in vitro degradation test was used to evaluate the sustained-release effect of the scaffold 2. Thirty male Japanese white rabbits, weighing (2.0±0.1) kg, were randomly divided into groups A, B, C, D, and E, 6 in each group. After making a 1 cm- diameter bilateral tibial defects model, group A was the defect control group without any material implanted. Groups B, C, D, and E were implanted with scaffolds 3, 4, 1, and 2 at the defect sites, respectively. At 4, 8, and 12 weeks after operation, the repairing effects of 4 scaffolds were observed by gross observation, histological observation of HE and Masson staining, and immunohistochemical staining of osteogenic specific transcription factor (runt-related transcription factor 2, RUNX2), osteogenic related transcription factor [Osterix (OSX), Col Ⅰ, osteopontin (OPN)]. Results Scanning electron microscopy observation showed that the scaffolds were all porous. The structure of the material was loose before and after cross-linking. The surface contact angle showed that the scaffold was hydrophobic, and the scaffold 2 was more hydrophobic than scaffold 4. The sustained-release effect in vitro showed that the drug could be released in a micro and long-term manner. In the animal implantation experiment, the gross observation showed that the defects were significantly smaller in groups D and E than in groups A, B, and C at 4 and 12 weeks after operation. HE and Masson staining showed that the defect of group A was full of connective tissue at 4 weeks after operation, a large number of fibers were seen in groups B and C, and the new bone formation was observed in groups D and E. The increase of new bone was observed in each group at 8 weeks after operation. The defect of group A was still dominated by connective tissue at 12 weeks after operation, and a small amount of new bone tissue was observed in groups B and C, and a large number of new bone tissue was observed in groups D and E, especially in group E, and most of the materials degraded. Immunohistochemical staining showed that the expressions of RUNX2 and OSX in the new tissues of groups D and E were significantly higher than those of the other groups at 4 weeks after operation. The expression of RUNX2 decreased at 8 and 12 weeks after operation. After 8 weeks and 12 weeks, the expressions of Col Ⅰand OPN increased than in 4 weeks. And the expressions of Col Ⅰ and OPN in the new tissues of groups D and E were significantly more than those of the other groups. Conclusion ICA/ATP/Col I/PCL composite scaffolds have good porosity and biocompatibility, can promote bone formation, and have good bone regeneration and repair effect.

Citation： NING Yu, QIN Wen, REN Yahui, LI Chenkai, CHEN Wenyang, ZHAO Hongbin. Effect of icariin/attapulgite/collagen type Ⅰ/polycaprolactone composite scaffold in repair of rabbit tibia defect. Chinese Journal of Reparative and Reconstructive Surgery, 2019, 33(9): 1181-1189. doi: 10.7507/1002-1892.201902044 Copy

1.	郭宜姣, 李文华. 骨缺损修复生物工程研究进展. 中国骨质疏松杂志, 2014, 20(8): 988-993.
2.	赵兴绪, 唐俊杰, 李根, 等. 胶原蛋白与黏土混合材料的性能研究. 中国组织工程研究, 2014, 18(47): 7573-7578.
3.	Faundez A, Tournier C, Garcia M, et al. Bone morphogenetic protein use in spine surgery-complications and outcomes: a systematic review. Int Orthop, 2016, 40(6): 1309-1319.
4.	Lykissas M, Gkiatas I. Use of recombinant human bone morphogenetic protein-2 in spine surgery. World J Orthop, 2017, 8(7): 531-535.
5.	刘艳辉, 谢利娜, 张晓娟, 等. 淫羊藿在骨膜附加活性材料 nHAC/PLA 修复颌骨缺损中的影响. 广州中医药大学学报, 2014, 31(5): 791-794, 850.
6.	赵宏霞. 载药壳聚糖/羟基磷灰石复合材料在骨组织工程中的研究进展. 功能材料, 2014, 13(45): 13006-13012.
7.	吴涛, 徐俊昌, 南开辉, 等. 淫羊藿苷促进羊骨髓间充质干细胞的增殖和成骨分化. 中国组织工程研究与临床康复, 2009, 13(19): 3725-3729.
8.	Li M, Gu Q, Chen M, et al. Controlled delivery of icariin on small intestine submucosa for bone tissue engineering. Mater Sci Eng C Mater Biol Appl, 2017, 71: 260-267.
9.	Wu T, Shu T, Kang L, et al. Icaritin, a novel plant-derived osteoinductive agent, enhances the osteogenic differentiation of human bone marrow- and human adipose tissue-derived mesenchymal stem cells. Int J Mol Med, 2017, 39(4): 984-992.
10.	张晓敏, 王世勇, 李根, 等. Ⅰ型胶原/聚己内酯/凹凸棒石复合支架材料体外诱导成骨的研究. 中国生物工程杂志, 2016, 36(5): 27-33.
11.	张晓敏, 宋学文, 王维, 等. 凹凸棒石/Ⅰ型胶原/聚己内酯复合修复兔骨缺损的实验研究. 中国修复重建外科杂志, 2016, 30(5): 626-633.
12.	He F, Chen Y, Li J, et al. Improving bone repair of femoral and radial defects in rabbit by incorporating PRP into PLGA/CPC composite scaffold with unidirectional pore structure. J Biomed Mater Res A, 2015, 103(4): 1312-1324.
13.	朱淑倩, 何家才. 人工骨替代材料修复种植体周围骨缺损的研究进展. 医学综述, 2010, 16(6): 863-865.
14.	Chen SH, Lei M, Xie XH, et al. PLGA/TCP composite scaffold incorporating bioactive phytomolecule icaritin for enhancement of bone defect repair in rabbits. Acta Biomater, 2013, 9(5): 6711-6722.
15.	李斯日古楞, 胡晓文. 纳米羟基磷灰石/明胶仿生复合材料的制备及其细胞相容性. 中南大学学报 (医学版), 2014, 39(9): 949-958.
16.	Li X, Li Y, Zuo Y, et al. Osteogenesis and chondrogenesis of biomimetic integrated porous PVA/gel/V-n-HA/pa6 scaffolds and BMSCs construct in repair of articular osteochondral defect. J Biomed Mater Res A, 2015, 103(10): 3226-3236.
17.	古玉梅, 徐长青, 洪铭范, 等. 淫羊藿苷对实验性自身免疫性脑脊髓炎小鼠的治疗作用. 广东药学院学报, 2015, 31(5): 617-620.
18.	Chen KM, Ma HP, Ge BF, et al. Icariin enhances the osteogenic differentiation of bone marrow stromal cells but has no effects on the differentiation of newborn calvarial osteoblasts of rats. Pharmazie, 2007, 62(10): 785-789.
19.	鲍远, 黄俊明, 靖兴志, 等. 淫羊藿苷促进骨髓间充质干细胞成骨分化. 中国组织工程研究, 2016, 20(24): 6501-3507.
20.	李根, 李志宏, 王世勇, 等. 改性淫羊藿苷共价结合壳聚糖/聚羟基丁酸酯-羟基戊酸酯构建组织诱导型骨修复支架材料. 复合材料学报, 2016, 33(8): 1645-1653.
21.	Komori T. Roles of Runx2 in skeletal development. Adv Exp Med Biol, 2017, 962: 83-93.
22.	Zhang C. Transcriptional regulation of bone formation by the osteoblast-specific transcription factor Osx. J Orthop Surg Res, 2010, 5: 37.

1. 郭宜姣, 李文华. 骨缺损修复生物工程研究进展. 中国骨质疏松杂志, 2014, 20(8): 988-993.
2. 赵兴绪, 唐俊杰, 李根, 等. 胶原蛋白与黏土混合材料的性能研究. 中国组织工程研究, 2014, 18(47): 7573-7578.
3. Faundez A, Tournier C, Garcia M, et al. Bone morphogenetic protein use in spine surgery-complications and outcomes: a systematic review. Int Orthop, 2016, 40(6): 1309-1319.
4. Lykissas M, Gkiatas I. Use of recombinant human bone morphogenetic protein-2 in spine surgery. World J Orthop, 2017, 8(7): 531-535.
5. 刘艳辉, 谢利娜, 张晓娟, 等. 淫羊藿在骨膜附加活性材料 nHAC/PLA 修复颌骨缺损中的影响. 广州中医药大学学报, 2014, 31(5): 791-794, 850.
6. 赵宏霞. 载药壳聚糖/羟基磷灰石复合材料在骨组织工程中的研究进展. 功能材料, 2014, 13(45): 13006-13012.
7. 吴涛, 徐俊昌, 南开辉, 等. 淫羊藿苷促进羊骨髓间充质干细胞的增殖和成骨分化. 中国组织工程研究与临床康复, 2009, 13(19): 3725-3729.
8. Li M, Gu Q, Chen M, et al. Controlled delivery of icariin on small intestine submucosa for bone tissue engineering. Mater Sci Eng C Mater Biol Appl, 2017, 71: 260-267.
9. Wu T, Shu T, Kang L, et al. Icaritin, a novel plant-derived osteoinductive agent, enhances the osteogenic differentiation of human bone marrow- and human adipose tissue-derived mesenchymal stem cells. Int J Mol Med, 2017, 39(4): 984-992.
10. 张晓敏, 王世勇, 李根, 等. Ⅰ型胶原/聚己内酯/凹凸棒石复合支架材料体外诱导成骨的研究. 中国生物工程杂志, 2016, 36(5): 27-33.
11. 张晓敏, 宋学文, 王维, 等. 凹凸棒石/Ⅰ型胶原/聚己内酯复合修复兔骨缺损的实验研究. 中国修复重建外科杂志, 2016, 30(5): 626-633.
12. He F, Chen Y, Li J, et al. Improving bone repair of femoral and radial defects in rabbit by incorporating PRP into PLGA/CPC composite scaffold with unidirectional pore structure. J Biomed Mater Res A, 2015, 103(4): 1312-1324.
13. 朱淑倩, 何家才. 人工骨替代材料修复种植体周围骨缺损的研究进展. 医学综述, 2010, 16(6): 863-865.
14. Chen SH, Lei M, Xie XH, et al. PLGA/TCP composite scaffold incorporating bioactive phytomolecule icaritin for enhancement of bone defect repair in rabbits. Acta Biomater, 2013, 9(5): 6711-6722.
15. 李斯日古楞, 胡晓文. 纳米羟基磷灰石/明胶仿生复合材料的制备及其细胞相容性. 中南大学学报 (医学版), 2014, 39(9): 949-958.
16. Li X, Li Y, Zuo Y, et al. Osteogenesis and chondrogenesis of biomimetic integrated porous PVA/gel/V-n-HA/pa6 scaffolds and BMSCs construct in repair of articular osteochondral defect. J Biomed Mater Res A, 2015, 103(10): 3226-3236.
17. 古玉梅, 徐长青, 洪铭范, 等. 淫羊藿苷对实验性自身免疫性脑脊髓炎小鼠的治疗作用. 广东药学院学报, 2015, 31(5): 617-620.
18. Chen KM, Ma HP, Ge BF, et al. Icariin enhances the osteogenic differentiation of bone marrow stromal cells but has no effects on the differentiation of newborn calvarial osteoblasts of rats. Pharmazie, 2007, 62(10): 785-789.
19. 鲍远, 黄俊明, 靖兴志, 等. 淫羊藿苷促进骨髓间充质干细胞成骨分化. 中国组织工程研究, 2016, 20(24): 6501-3507.
20. 李根, 李志宏, 王世勇, 等. 改性淫羊藿苷共价结合壳聚糖/聚羟基丁酸酯-羟基戊酸酯构建组织诱导型骨修复支架材料. 复合材料学报, 2016, 33(8): 1645-1653.
21. Komori T. Roles of Runx2 in skeletal development. Adv Exp Med Biol, 2017, 962: 83-93.
22. Zhang C. Transcriptional regulation of bone formation by the osteoblast-specific transcription factor Osx. J Orthop Surg Res, 2010, 5: 37.

Chinese Journal of Reparative and Reconstructive Surgery

Effect of icariin/attapulgite/collagen type Ⅰ/polycaprolactone composite scaffold in repair of rabbit tibia defect

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