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.	王魯云, 郭林, 田豐德. 兔自體松質骨顆粒修復關節軟骨損傷. 臨床骨科雜志, 2016, 19(4): 500-503, 507.
3.	趙家寧, 徐寶山, 楊強. 組織工程骨軟骨復合組織構建研究進展. 中國矯形外科雜志, 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.	李運濤, 程科, 胡德志, 等. 柚皮苷對經 TNF-α 誘導體外培養兔關節軟骨細胞增殖及 SOD、NOS 影響. 遼寧中醫藥大學學報, 2008, 10(5): 175-177.
8.	蘇友新, 陳寶軍, 王藝茹, 等. 壯骨健膝方含藥血清中藥物成分骨碎補柚皮苷對 IL-1β 誘導兔退變軟骨細胞" caveolin-p38MAPK”信號通路的影響. 中國中西醫結合雜志, 2014, 34(12): 1492-1498.
9.	童華, 劉婷婷, 楊范莉, 等. 柚皮苷口腔黏附片的制備及體外釋放度研究. 西北藥學雜志, 2014, 29(5): 500-502.
10.	王慎東. 自體及異體骨軟骨移植與骨缺損修復. 中國組織工程研究與臨床康復, 2008, 12(40): 7905-7908.
11.	張佼佼, 王冬青, 王志強, 等. 組織工程軟骨支架材料的應用選擇. 中國組織工程研究與臨床康復, 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.	林建華, 賴建明, 林文平, 等. 膠原-殼聚糖/膠原-納米羥基磷灰石仿生支架材料的制備及其生物相容性檢測. 中國修復重建外科雜志, 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, 歐陽麗娜, 丁勁松. 凹土棒石黏土及其應用進展. 中南藥學, 2009, 7(6): 441-445.
18.	張曉敏, 王世勇, 李根, 等. Ⅰ 型膠原/聚己內酯/凹凸棒石復合支架材料體外誘導成骨的研究. 中國生物工程雜志, 2016, 36(5): 27-33.
19.	張曉敏, 宋學文, 王維, 等. 凹凸棒石/Ⅰ 型膠原/聚己內酯復合修復兔骨缺損的實驗研究. 中國修復重建外科雜志, 2016, 30(5): 626-633.
20.	王長昇, 林建華, 吳朝陽. TGF-β₁ 和 IGF-1 對人骨髓基質干細胞體外增殖和分化的影響. 臨床和實驗醫學雜志, 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. 王魯云, 郭林, 田豐德. 兔自體松質骨顆粒修復關節軟骨損傷. 臨床骨科雜志, 2016, 19(4): 500-503, 507.
3. 趙家寧, 徐寶山, 楊強. 組織工程骨軟骨復合組織構建研究進展. 中國矯形外科雜志, 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. 李運濤, 程科, 胡德志, 等. 柚皮苷對經 TNF-α 誘導體外培養兔關節軟骨細胞增殖及 SOD、NOS 影響. 遼寧中醫藥大學學報, 2008, 10(5): 175-177.
8. 蘇友新, 陳寶軍, 王藝茹, 等. 壯骨健膝方含藥血清中藥物成分骨碎補柚皮苷對 IL-1β 誘導兔退變軟骨細胞" caveolin-p38MAPK”信號通路的影響. 中國中西醫結合雜志, 2014, 34(12): 1492-1498.
9. 童華, 劉婷婷, 楊范莉, 等. 柚皮苷口腔黏附片的制備及體外釋放度研究. 西北藥學雜志, 2014, 29(5): 500-502.
10. 王慎東. 自體及異體骨軟骨移植與骨缺損修復. 中國組織工程研究與臨床康復, 2008, 12(40): 7905-7908.
11. 張佼佼, 王冬青, 王志強, 等. 組織工程軟骨支架材料的應用選擇. 中國組織工程研究與臨床康復, 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. 林建華, 賴建明, 林文平, 等. 膠原-殼聚糖/膠原-納米羥基磷灰石仿生支架材料的制備及其生物相容性檢測. 中國修復重建外科雜志, 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, 歐陽麗娜, 丁勁松. 凹土棒石黏土及其應用進展. 中南藥學, 2009, 7(6): 441-445.
18. 張曉敏, 王世勇, 李根, 等. Ⅰ 型膠原/聚己內酯/凹凸棒石復合支架材料體外誘導成骨的研究. 中國生物工程雜志, 2016, 36(5): 27-33.
19. 張曉敏, 宋學文, 王維, 等. 凹凸棒石/Ⅰ 型膠原/聚己內酯復合修復兔骨缺損的實驗研究. 中國修復重建外科雜志, 2016, 30(5): 626-633.
20. 王長昇, 林建華, 吳朝陽. TGF-β₁ 和 IGF-1 對人骨髓基質干細胞體外增殖和分化的影響. 臨床和實驗醫學雜志, 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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