EXPERIMENTAL STUDY ON BONE DEFECT REPAIR WITH COMPOSITE OF ATTAPULGITE/COLLAGEN TYPE I/POLY (CAPROLACTONE) IN RABBITS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANGXiaomin ^1,2 , SONGXuewen ¹ , WANGWei ¹ , LIZhenjun ¹ ,  ZHAOHongbin ^1,2

1. Institute of Orthopedics, Lanzhou General Hospital of Lanzhou Military Command of Chinese PLA, Lanzhou Gansu, 730050, P. R. China;
2. Collage of Life Sciences and Technology, Gansu Agriculture University;

Corresponding author：

ZHAOHongbin, Email: zhao761032@163.com

Keywords：

Attapulgite; Collagen type I; Poly (caprolactone); Composite scaffold material; Bone defect; Rabbit

DOI：

10.7507/1002-1892.20160126

Video：

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Objective To investigate the effect of repairing radial bone defect with scaffold material of attapulgite/collagen type I/poly (caprolactone) (ATP/Col I/PCL) in rabbits and the possibility as bone graft substitutes. Methods ATP/Col I/PCL materials were prepared via adding ATP to hexafluoroisopropanol after dissolved Col I/PCL (3∶2), and Col I/PCL materials via dissolving Col I/PCL (3∶2) in hexafluoroisopropanol served as control. The structure of scaffolds was observed under scanning electron microscope (SEM). Twenty-four Japanese white rabbits (male, 2 months old) were used to establish the bilateral radius defect model of 15 mm in length, and randomly divided into group A (6 rabbits, 12 defects), group B (9 rabbits, 18 defects), and group C (9 rabbits, 18 defects); then the Col I/PCL scaffold was implanted in the bone defect area in group B, the ATP/Col I/PCL scaffold in group C, no treatment was done in group A as control. The general condition of rabbits was observed after operation, and bone defect repair was evaluated by X-ray at 4, 8, and 12 weeks. At 12 weeks, the tissue of defect area was harvested for the general, SEM, Micro-CT, histological, and immunohistochemical staining to observe defect repair and material degradation. Results SEM observation showed that two kinds of materials were porous structure, ATP/Col I/PCL structure was more dense than Col I/PCL. All animals survived to the end of experiment, and no incision infection occurred during repair process.X-ray films showed that the bone marrow cavity was re-opened in defect area of group C with time, the repair effect was superior to that of groups A and B. At 12 weeks after operation, general observation showed that scaffold material had good fusion with the surrounding tissue in groups B and C, defect was filled with connective tissue in group A. SEM indicated that the surface and pore of the scaffold were covered with a large number of cells and tissues in groups B and C. Micro-CT demonstrated that the new bone volume, bone mineral content, tissue mineral content, and connectivity density of group C were significantly higher than those of groups A and B (P<0.05). The observation of histology and immunohistochemical staining indicated that there were lots of connective tissues in defect area of group A, and ALP, Col I, and OPN were weakly expressed; there were many collagen fibers in scaffold degradation area in group B, and the expression levels of ALP, Col I, and OPN were higher than those of group A; there was few new bone in group C, the degradation rate of the scaffold was slower than that of group B, and the expression of Col I and OPN were enhanced, while ALP was weakened when compared with groups A and B. Conclusion ATP/Col I/PCL composite scaffold material can degrade in vivo, and has dense three-dimensional porous structure, good biocompatibility, and high potentiality of bone repair, so it can be used as bone substitute material.

Citation： ZHANGXiaomin, SONGXuewen, WANGWei, LIZhenjun, ZHAOHongbin. EXPERIMENTAL STUDY ON BONE DEFECT REPAIR WITH COMPOSITE OF ATTAPULGITE/COLLAGEN TYPE I/POLY (CAPROLACTONE) IN RABBITS. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(5): 626-633. doi: 10.7507/1002-1892.20160126 Copy

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2.	Harada N, Watanabe Y, Sato K, et al. Bone regeneration in a massive rat femur defect through endochondral ossification achieved with chondrogenically differentiated MSCs in a degradable scaffold. Biomaterials, 2014, 35(27):7800-7810.
3.	卫爱林, 刘世清, 彭昊, 等. 磷酸三钙-透明质酸-Ⅰ型胶原-骨髓基质细胞复合修复骨缺损的实验研究. 中国修复重建外科杂志, 2005, 19(6):468-472.
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5.	Long T, Yang J, Shi SS, et al. Fabrication of three-dimensional porous scaffold based on collagen fiber and bioglass for bone tissue engineering. J Biomed Mater Res B Appl Biomater, 2014, 103(7):1455-1464.
6.	Kim JH, Linh NT, Min YK, et al. Surface modification of porous polycaprolactone/biphasic calcium phosphate scaffolds for bone regeneration in rat calvaria defect. J Biomater Appl, 2014, 29(4):624-635.
7.	Ruckh TT, Carroll DA, Weaver GR, et al. Mineralization content alters osteogenic responses of bone marrow stromal cells on hydroxyapatite/polycaprolactone composite nanofiber scaffolds. J Funct Biomater, 2012, 3(4):776-798.
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12.	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.
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14.	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.
15.	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.
16.	杜庆华, 曹均凯, 董溪溪, 等. 镁黄长石浸提液诱导多潜能干细胞的成骨分化. 中国组织工程研究, 2015, 19(14):2236-2242.
17.	Han P, Wu C, Xiao Y. The effect of silicate ions on proliferation, osteogenic differentiation and cell signalling pathways (WNT and SHH) of bone marrow stromal cells. Biomater Sci, 2013, 1(4):379-392.
18.	Wiens M, Wang X, Schröder HC, et al. The role of biosilica in the osteoprotegerin/RANKL ratio in human osteoblast-like cells. Biomaterials, 2010, 31(30):7716-7725.
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20.	Pietak AM, Reid JW, Stott MJ, et al. Silicon substitution in the calcium phosphate bioceramics. Biomaterials, 2007, 28(28):4023-4032.
21.	Jin X, Hu X, Wang Q, et al. Multifunctional cationic polymer decorated and drug intercalated layered silicate (NLS) for early gastric cancer prevention. Biomaterials, 2014, 35(10):3298-3308.
22.	Rezwan K, Chen QZ, Boccaccini JJ. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials, 2006, 27(18):3413-3431.
23.	金光辉, 张馨雯, 孙晓飞, 等. 组织工程化纳米羟基磷灰石/聚己内酯人工骨支架修复兔桡骨大段骨缺损的实验研究. 中华损伤与修复杂志:电子版, 2014, 10(1):43-49.
24.	Han SH, Kim KH, Han JS, et al. Response of osteoblast-like cells cultured on zirconia to bone morphogenetic protein-2. J Periodontal Implant Sci, 2011, 41(5):227-233.
25.	Jensen T, Baas J, Dolathshahi-Pirouz A, et al. Osteopontin functionalization of hydroxyapatite nanoparticles in a PDLLA matrix promotes bone formation. J Biomed Mater Res A, 2011, 99(1):94-101.

1. Pati F, Song TH, Rijal G, et al. Ornamenting 3D printed scaffolds with cell-laid extracellular matri for bone tissue regeneration. Biomaterials, 2014, 37:230-241.
2. Harada N, Watanabe Y, Sato K, et al. Bone regeneration in a massive rat femur defect through endochondral ossification achieved with chondrogenically differentiated MSCs in a degradable scaffold. Biomaterials, 2014, 35(27):7800-7810.
3. 卫爱林, 刘世清, 彭昊, 等. 磷酸三钙-透明质酸-Ⅰ型胶原-骨髓基质细胞复合修复骨缺损的实验研究. 中国修复重建外科杂志, 2005, 19(6):468-472.
4. Prosecká E, Rampichová M, Litvinec A, et al. Collagen/hydroxyapatite scaffold enriched with polycaprolactone nanofibers, thrombocyte-rich solution and mesenchymal stem cells promotes regeneration in large bone defect in vivo. J Biomed Mater Res A, 2015, 103(2):671-682.
5. Long T, Yang J, Shi SS, et al. Fabrication of three-dimensional porous scaffold based on collagen fiber and bioglass for bone tissue engineering. J Biomed Mater Res B Appl Biomater, 2014, 103(7):1455-1464.
6. Kim JH, Linh NT, Min YK, et al. Surface modification of porous polycaprolactone/biphasic calcium phosphate scaffolds for bone regeneration in rat calvaria defect. J Biomater Appl, 2014, 29(4):624-635.
7. Ruckh TT, Carroll DA, Weaver GR, et al. Mineralization content alters osteogenic responses of bone marrow stromal cells on hydroxyapatite/polycaprolactone composite nanofiber scaffolds. J Funct Biomater, 2012, 3(4):776-798.
8. 苏兰, 沈培友, 曹建常, 等. 凹凸棒粘土性能及应用研究. 矿产保护与利用, 2014, (5):43-45.
9. 李增新, 王国明, 王彤, 等. 凹凸棒石负载壳聚糖吸附废水中Cr (Ⅵ)的研究. 安全与环境学报, 2008, 8(3):52-55.
10. 刘元法, 王兴国. 凹凸棒石吸附剂特征及其在油脂脱色过程中的应用研究. 中国油脂, 2006, 31(9):27-30.
11. 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.
12. 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.
13. 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.
14. 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.
15. 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.
16. 杜庆华, 曹均凯, 董溪溪, 等. 镁黄长石浸提液诱导多潜能干细胞的成骨分化. 中国组织工程研究, 2015, 19(14):2236-2242.
17. Han P, Wu C, Xiao Y. The effect of silicate ions on proliferation, osteogenic differentiation and cell signalling pathways (WNT and SHH) of bone marrow stromal cells. Biomater Sci, 2013, 1(4):379-392.
18. Wiens M, Wang X, Schröder HC, et al. The role of biosilica in the osteoprotegerin/RANKL ratio in human osteoblast-like cells. Biomaterials, 2010, 31(30):7716-7725.
19. Liu X, Rahaman MN, Fu Q. Bone regeneration in strong porous bioactive glass (13-93) scaffolds with an oriented microstructure implanted in rat calvarial defects. Acta Biomater, 2013, 9(1):4889-4898.
20. Pietak AM, Reid JW, Stott MJ, et al. Silicon substitution in the calcium phosphate bioceramics. Biomaterials, 2007, 28(28):4023-4032.
21. Jin X, Hu X, Wang Q, et al. Multifunctional cationic polymer decorated and drug intercalated layered silicate (NLS) for early gastric cancer prevention. Biomaterials, 2014, 35(10):3298-3308.
22. Rezwan K, Chen QZ, Boccaccini JJ. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials, 2006, 27(18):3413-3431.
23. 金光辉, 张馨雯, 孙晓飞, 等. 组织工程化纳米羟基磷灰石/聚己内酯人工骨支架修复兔桡骨大段骨缺损的实验研究. 中华损伤与修复杂志:电子版, 2014, 10(1):43-49.
24. Han SH, Kim KH, Han JS, et al. Response of osteoblast-like cells cultured on zirconia to bone morphogenetic protein-2. J Periodontal Implant Sci, 2011, 41(5):227-233.
25. Jensen T, Baas J, Dolathshahi-Pirouz A, et al. Osteopontin functionalization of hydroxyapatite nanoparticles in a PDLLA matrix promotes bone formation. J Biomed Mater Res A, 2011, 99(1):94-101.

Chinese Journal of Reparative and Reconstructive Surgery

EXPERIMENTAL STUDY ON BONE DEFECT REPAIR WITH COMPOSITE OF ATTAPULGITE/COLLAGEN TYPE I/POLY (CAPROLACTONE) IN RABBITS

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