Experimental study on loading naringin composite scaffolds for repairing rabbit osteochondral defects_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

HUANG Junbo ^1,2 , WANG Shiyong ² , ZHANG Xiaomin ² , LI Gen ² , JI Puzhong ² ,  ZHAO Hongbin ^1,2

1. Provincial-Level Key Laboratory for Molecular Medicine of Major Diseases and the Prevention and Treatment with Traditional Chinese Medicine Research in Gansu Colleges and Universities, Gansu University of Traditional Chinese Medicine, Lanzhou Gansu, 730000, P.R.China;
2. Institute of Orthopedics, Lanzhou General Hospital of Lanzhou Military Command of Chinese PLA, Lanzhou Gansu, 730050, P.R.China;

Corresponding author：

ZHAO Hongbin, Email: zhao761032@163.com

Keywords：

Naringin; attpulgite; collagen; microsphere; osteochondral composite scaffold; osteochondral defect; rabbit

DOI：

10.7507/1002-1892.201611112

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Objective To investigate the performance of loading naringin composite scaffolds and its effects on repair of osteochondral defects. Methods The loading naringin and unloading naringin sustained release microspheres were prepared by W/O/W method; with the materials of the attpulgite and the collagen type I, the loading naringin, unloading naringin, and loading transforming growth factor β₁ (TGF-β₁) osteochondral composite scaffolds were constructed respectively by " 3 layers sandwich method”. The effect of sustained-release of loading naringin microspheres, the morphology of the composite scaffolds, and the biocompatibility were evaluated respectively by releasingin vitro, scanning electron microscope, and cell counting kit 8. Forty Japanese white rabbits were randomly divided into groups A, B, C, and D, 10 rabbits each group. After a osteochondral defect of 4.5 mm in diameter and 4 mm in depth was made in the intercondylar fossa of two femurs. Defect was not repaired in group A (blank control), and defect was repaired with unloading naringin composite scaffolds (negative control group), loading naringin composite scaffolds (experimental group), and loading TGF-β₁ composite scaffolds (positive control group) in groups B, C, and D respectively. At 3 and 6 months after repair, the intercondylar fossa was harvested for the general, HE staining, and toluidine blue staining to observe the repair effect. Western blot was used to detect the expression of collagen type II in the new cartilage. Results Loading naringin microspheres had good effect of sustained-release; the osteochondral composite scaffolds had good porosity; the cell proliferation rate on loading naringin composite scaffold was increased significantly when compared with unloading naringin scaffold (P<0.05). General observation revealed that defect range of groups C and D was reduced significantly when compared with groups A and B at 3 months after repair; at 6 months after repair, defects of group C were covered by new cartilage, and new cartilage well integrated with the adjacent cartilage in group D. The results of histological staining revealed that defects were filled with a small amount of fibrous tissue in groups A and B, and a small amount of new cartilage in groups C and D at 3 months after repair; new cartilage of groups C and D was similar to normal cartilage, but defects were filled with a large amount of fibrous tissue in groups A and B at 6 months after repair. The expression of collagen type II in groups C and D was significantly higher than that in groups A and B (P<0.05), but no significant difference was found between groups C and D (P>0.05). Conclusion Loading naringin composite scaffolds have good biocompatibility and effect in repair of rabbit articular osteochondral defects.

Citation： HUANG Junbo, WANG Shiyong, ZHANG Xiaomin, LI Gen, JI Puzhong, ZHAO Hongbin. Experimental study on loading naringin composite scaffolds for repairing rabbit osteochondral defects. Chinese Journal of Reparative and Reconstructive Surgery, 2017, 31(4): 489-496. doi: 10.7507/1002-1892.201611112 Copy

1.	Cui W, Wang Q, Chen G, et al. Repair of articular cartilage defects with tissue-engineered osteochondral composites in pigs. J BiosciBioeng, 2011, 111(4): 493-500.
2.	王鲁云, 郭林, 田丰德. 兔自体松质骨颗粒修复关节软骨损伤. 临床骨科杂志, 2016, 19(4): 500-503, 507.
3.	赵家宁, 徐宝山, 杨强. 组织工程骨软骨复合组织构建研究进展. 中国矫形外科杂志, 2014, 22(23): 2156-2160.
4.	Reyes R, Delgado A, Solis R, et al. Cartilage repair by local delivery of transforming growth factor-β₁ or bone morphogenetic protein-2 from a novel, segmented polyurethane/polylactic-co-glycolic bilayered scaffold. J Biomed Mater Res A, 2014, 102(4): 1110-1120.
5.	Li B, Yang J, Ma L, et al. Influence of the molecular weight of poly (lactide-co-glycolide) on thein vivo cartilage repair by a construct of poly (lactide-co-glycolide)/fibrin gel/mesenchymal stem cells/transforming growth factor-β₁. Tissue Eng Part A, 2014, 20(1-2): 1-11.
6.	Yin F, Cai J, Zen W, et al. Cartilage Regeneration of Adipose-Derived Stem Cells in the TGF-β₁-Immobilized PLGA-Gelatin Scaffold. Stem Cell Rev, 2015, 11(3): 453-459.
7.	李运涛, 程科, 胡德志, 等. 柚皮苷对经 TNF-α 诱导体外培养兔关节软骨细胞增殖及 SOD、NOS 影响. 辽宁中医药大学学报, 2008, 10(5): 175-177.
8.	苏友新, 陈宝军, 王艺茹, 等. 壮骨健膝方含药血清中药物成分骨碎补柚皮苷对 IL-1β 诱导兔退变软骨细胞" caveolin-p38MAPK”信号通路的影响. 中国中西医结合杂志, 2014, 34(12): 1492-1498.
9.	童华, 刘婷婷, 杨范莉, 等. 柚皮苷口腔黏附片的制备及体外释放度研究. 西北药学杂志, 2014, 29(5): 500-502.
10.	王慎东. 自体及异体骨软骨移植与骨缺损修复. 中国组织工程研究与临床康复, 2008, 12(40): 7905-7908.
11.	张佼佼, 王冬青, 王志强, 等. 组织工程软骨支架材料的应用选择. 中国组织工程研究与临床康复, 2011, 15(29): 5467-5470.
12.	Müller WE, Neufurth M, Wang S, et al. Morphogenetically active scaffold for osteochondral repair (polyphosphate/alginate/N, O-carboxymethyl chitosan). Eur Cell Mater, 2016, 31: 174-190.
13.	林建华, 赖建明, 林文平, 等. 胶原-壳聚糖/胶原-纳米羟基磷灰石仿生支架材料的制备及其生物相容性检测. 中国修复重建外科杂志, 2012, 26(8): 1001-1006.
14.	Da H, Jia SJ, Meng GL, et al. The impact of compact layer in biphasic scaffold on osteochondral tissue engineering. PLoS One, 2013, 8(1): e54838.
15.	Li W, Wang C, Peng J, et al. Naringin inhibits TNF-α induced oxidative stress and inflammatory response in HUVECs via Nox4/NF-κB and PI3K/Akt pathways. Curr Pharm Biotechnol, 2014, 15(12): 1173-1182.
16.	Zhao Y, Li Z, Wang W, et al. Naringin Protects Against Cartilage Destruction in Osteoarthritis Through Repression of NF-κB Signaling Pathway. Inflammation, 2016, 39(1): 385-392.
17.	徐世希, 欧阳丽娜, 丁劲松. 凹土棒石黏土及其应用进展. 中南药学, 2009, 7(6): 441-445.
18.	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.
19.	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.
20.	王长昇, 林建华, 吴朝阳. TGF-β₁ 和 IGF-1 对人骨髓基质干细胞体外增殖和分化的影响. 临床和实验医学杂志, 2008, 7(1): 3-4.
21.	Fang J, Xu L, Li Y, et al. Roles of TGF-beta 1 signaling in the development of osteoarthritis. Histol Histopathol, 2016, 31(11): 1161-1167.

1. Cui W, Wang Q, Chen G, et al. Repair of articular cartilage defects with tissue-engineered osteochondral composites in pigs. J BiosciBioeng, 2011, 111(4): 493-500.
2. 王鲁云, 郭林, 田丰德. 兔自体松质骨颗粒修复关节软骨损伤. 临床骨科杂志, 2016, 19(4): 500-503, 507.
3. 赵家宁, 徐宝山, 杨强. 组织工程骨软骨复合组织构建研究进展. 中国矫形外科杂志, 2014, 22(23): 2156-2160.
4. Reyes R, Delgado A, Solis R, et al. Cartilage repair by local delivery of transforming growth factor-β₁ or bone morphogenetic protein-2 from a novel, segmented polyurethane/polylactic-co-glycolic bilayered scaffold. J Biomed Mater Res A, 2014, 102(4): 1110-1120.
5. Li B, Yang J, Ma L, et al. Influence of the molecular weight of poly (lactide-co-glycolide) on thein vivo cartilage repair by a construct of poly (lactide-co-glycolide)/fibrin gel/mesenchymal stem cells/transforming growth factor-β₁. Tissue Eng Part A, 2014, 20(1-2): 1-11.
6. Yin F, Cai J, Zen W, et al. Cartilage Regeneration of Adipose-Derived Stem Cells in the TGF-β₁-Immobilized PLGA-Gelatin Scaffold. Stem Cell Rev, 2015, 11(3): 453-459.
7. 李运涛, 程科, 胡德志, 等. 柚皮苷对经 TNF-α 诱导体外培养兔关节软骨细胞增殖及 SOD、NOS 影响. 辽宁中医药大学学报, 2008, 10(5): 175-177.
8. 苏友新, 陈宝军, 王艺茹, 等. 壮骨健膝方含药血清中药物成分骨碎补柚皮苷对 IL-1β 诱导兔退变软骨细胞" caveolin-p38MAPK”信号通路的影响. 中国中西医结合杂志, 2014, 34(12): 1492-1498.
9. 童华, 刘婷婷, 杨范莉, 等. 柚皮苷口腔黏附片的制备及体外释放度研究. 西北药学杂志, 2014, 29(5): 500-502.
10. 王慎东. 自体及异体骨软骨移植与骨缺损修复. 中国组织工程研究与临床康复, 2008, 12(40): 7905-7908.
11. 张佼佼, 王冬青, 王志强, 等. 组织工程软骨支架材料的应用选择. 中国组织工程研究与临床康复, 2011, 15(29): 5467-5470.
12. Müller WE, Neufurth M, Wang S, et al. Morphogenetically active scaffold for osteochondral repair (polyphosphate/alginate/N, O-carboxymethyl chitosan). Eur Cell Mater, 2016, 31: 174-190.
13. 林建华, 赖建明, 林文平, 等. 胶原-壳聚糖/胶原-纳米羟基磷灰石仿生支架材料的制备及其生物相容性检测. 中国修复重建外科杂志, 2012, 26(8): 1001-1006.
14. Da H, Jia SJ, Meng GL, et al. The impact of compact layer in biphasic scaffold on osteochondral tissue engineering. PLoS One, 2013, 8(1): e54838.
15. Li W, Wang C, Peng J, et al. Naringin inhibits TNF-α induced oxidative stress and inflammatory response in HUVECs via Nox4/NF-κB and PI3K/Akt pathways. Curr Pharm Biotechnol, 2014, 15(12): 1173-1182.
16. Zhao Y, Li Z, Wang W, et al. Naringin Protects Against Cartilage Destruction in Osteoarthritis Through Repression of NF-κB Signaling Pathway. Inflammation, 2016, 39(1): 385-392.
17. 徐世希, 欧阳丽娜, 丁劲松. 凹土棒石黏土及其应用进展. 中南药学, 2009, 7(6): 441-445.
18. 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.
19. 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.
20. 王长昇, 林建华, 吴朝阳. TGF-β₁ 和 IGF-1 对人骨髓基质干细胞体外增殖和分化的影响. 临床和实验医学杂志, 2008, 7(1): 3-4.
21. Fang J, Xu L, Li Y, et al. Roles of TGF-beta 1 signaling in the development of osteoarthritis. Histol Histopathol, 2016, 31(11): 1161-1167.

Chinese Journal of Reparative and Reconstructive Surgery

Experimental study on loading naringin composite scaffolds for repairing rabbit osteochondral defects

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