EXPERIMENTAL STUDY ON CHITOSAN/ALLOGENEIC BONE POWDER COMPOSITE POROUS SCAFFOLD TO REPAIR BONE DEFECTS IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

KANGXiangang ¹ , ZHAOZhiyuan ¹ , WUXuzhi ¹ , SHENQingxin ¹ ,  WANGZhiqiang ² , KANGYue ³ , XINGZhenguang ¹ , ZHANGTao ¹

1. The First Department of Orthopedics, the Second Affiliated Hospital of Xingtai Medical College, Xingtai Hebei, 054000, P.R.China;
2. Department of Orthopedics, the North China University of Science and Technology Affiliated Hospital;
3. Shanxi Medical Tissue Bank;

Corresponding?author：

WANGZhiqiang, Email: wzhqde@163.com

Keywords：

Bone tissue engineering; Scaffold material; Chitosan; Allogeneic bone; Rat

DOI：

10.7507/1002-1892.20160059

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Objective To explore the feasibility of chitosan/allogeneic bone powder composite porous scaffold as scaffold material of bone tissue engineering in repairing bone defect. Methods The composite porous scaffolds were prepared with chitosan and decalcified allogeneic bone powder at a ratio of 1∶5 by vacuum freeze-drying technique. Chitosan scaffold served as control. Ethanol alternative method was used to measure its porosity, and scanning electron microscopy (SEM) to measure pore size. The hole of 3.5 mm in diameter was made on the bilateral femoral condyles of 40 adult Sprague Dawley rats. The composite porous scaffolds and chitosan scaffolds were implanted into the hole of the left femoral condyle (experimental group) and the hole of the right femoral condyle (control group), respectively. At 2, 4, 8, and 12 weeks after implantation, the tissues were harvested for gross observation, histological observation, and immunohistochemical staining. Results The composite porous scaffold prepared by vacuum freeze-drying technique had yellowish color, and brittle and easily broken texture; pore size was mostly 200-300μm; and the porosity was 76.8%±1.1%, showing no significant difference when compared with the porosity of pure chitosan scaffold (78.4%±1.4%) (t=-2.10, P=0.09). The gross observation and histological observation showed that the defect area was filled with new bone with time, and new bone of the experimental group was significantly more than that of the control group. At 4, 8, and 12 weeks after implantation, the bone forming area of the experimental group was significantly larger than that of the control group (P < 0.05). The immunohistochemical staining results showed that osteoprotegerin (OPG) positive expression was found in the experimental group at different time points, and the positive expression level was significantly higher than that in the control group (P < 0.05). Conclusion Chitosan/allogeneic bone powder composite porous scaffold has suitable porosity and good osteogenic activity, so it is a good material for repairing bone defect, and its bone forming volume and bone formation rate are better than those of pure chitosan scaffold.

Citation： KANGXiangang, ZHAOZhiyuan, WUXuzhi, SHENQingxin, WANGZhiqiang, KANGYue, XINGZhenguang, ZHANGTao. EXPERIMENTAL STUDY ON CHITOSAN/ALLOGENEIC BONE POWDER COMPOSITE POROUS SCAFFOLD TO REPAIR BONE DEFECTS IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(3): 298-302. doi: 10.7507/1002-1892.20160059 Copy

1.	郭睿, 農曉琳.復合支架材料在口腔骨組織工程領域的研究與進展.口腔生物醫學, 2012, 3(3): 155-158.
2.	魏冀榮, 章瑩.骨移植支架材料的理論研究與臨床應用.中國組織工程, 2012, 16(47): 8880-8884.
3.	Holt GE, Halpern JL, Dovan TT, et al. Evolution of an in vivo bioreactor. J Orthop Res, 2005, 23(4): 916-923.
4.	Xu HH, Simon CG Jr. Fast setting calcium phosphate-chitosan scaffold: mechanical properties and biocompatibility. Biomaterials, 2005, 26(12): 1337-1348.
5.	Hsu FY, Tsai SW, Lan CW, et al. An in vivo study of a bone grafting material consisting of hydroxyapatite and reconstituted collagen. J Mater Sci Mater Med, 2005, 16(4): 341-345.
6.	楊志明.組織工程基礎與臨床.成都:四川科學技術出版社, 2000: 1-11.
7.	宋雪蓮, 于靜濤, 劉揚, 等.殼聚糖在骨組織工程中應用研究進展.中國實用口腔科雜志, 2013, 6(5): 306-310.
8.	張屹, 陳建常, 史振滿.異體骨支架及其衍生材料治療骨缺損的研究現狀及進展.中國矯形外科雜志, 2014, 22(14): 1277-1279.
9.	楊樹青, 蘇立新, 王志強, 等.兩種不同比例殼聚糖復合同種異體顆粒骨修復兔橈骨節段性缺損的對比研究.中國修復重建外科雜志, 2011, 25(7): 877-883.
10.	陳海霞, 謝志剛.骨組織工程支架材料:脫礦骨基質.中國組織工程研究, 2014, 18(3): 426-431.
11.	Wang Y, Huang YC, Gertzman AA, et al. Endogenous regeneration of critical-size chondral defects in immunocompromised rat xiphoid cartilage using decellularzed human bone matrix scaffolds. Tissue Eng Part A, 2012, 18(21-22): 2332-2342.
12.	Kirk JF, Ritter G, Waters C, et al. Osteoconductivity and osteoinductivity of NanoFUSE(?) DBM. Cell Tissue Bank, 2012, 14(1): 33-44.
13.	Kumar JP, Lakshmi L, Jyothsna V, et al. Synthesis and characterization of diopside particles and their suitability along with chitosan matrix for bone tissue engineering in vitro and in vivo. J Biomed Nanotechnol, 2014, 10(6): 970-981.
14.	Yasuda H, Shima N, Nakagawa N, et al. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci U S A, 1998, 95(7): 3597-3602.
15.	Simonet WS, Lacey DL, Dumstan CR, et al. Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell, 1997, 89(2): 309-319.
16.	劉健, 宋暉.骨保護素對破骨細胞的影響程度分析.中國生化藥物雜志, 2015, 35(3): 65-68.
17.	鄭揚, 李危石, 劉忠軍.骨組織3D打印:骨再生的未來.北京大學報(醫學版), 2015, 47(2): 203-206.
18.	Yang J, Cai H, Lv J, et al. In vivo study of a self-stabilizing artificial vertebral body fabricated by electron beam melting. Spin (Phila Pa 1976), 2014, 39(8): E486-E492.

1. 郭睿, 農曉琳.復合支架材料在口腔骨組織工程領域的研究與進展.口腔生物醫學, 2012, 3(3): 155-158.
2. 魏冀榮, 章瑩.骨移植支架材料的理論研究與臨床應用.中國組織工程, 2012, 16(47): 8880-8884.
3. Holt GE, Halpern JL, Dovan TT, et al. Evolution of an in vivo bioreactor. J Orthop Res, 2005, 23(4): 916-923.
4. Xu HH, Simon CG Jr. Fast setting calcium phosphate-chitosan scaffold: mechanical properties and biocompatibility. Biomaterials, 2005, 26(12): 1337-1348.
5. Hsu FY, Tsai SW, Lan CW, et al. An in vivo study of a bone grafting material consisting of hydroxyapatite and reconstituted collagen. J Mater Sci Mater Med, 2005, 16(4): 341-345.
6. 楊志明.組織工程基礎與臨床.成都:四川科學技術出版社, 2000: 1-11.
7. 宋雪蓮, 于靜濤, 劉揚, 等.殼聚糖在骨組織工程中應用研究進展.中國實用口腔科雜志, 2013, 6(5): 306-310.
8. 張屹, 陳建常, 史振滿.異體骨支架及其衍生材料治療骨缺損的研究現狀及進展.中國矯形外科雜志, 2014, 22(14): 1277-1279.
9. 楊樹青, 蘇立新, 王志強, 等.兩種不同比例殼聚糖復合同種異體顆粒骨修復兔橈骨節段性缺損的對比研究.中國修復重建外科雜志, 2011, 25(7): 877-883.
10. 陳海霞, 謝志剛.骨組織工程支架材料:脫礦骨基質.中國組織工程研究, 2014, 18(3): 426-431.
11. Wang Y, Huang YC, Gertzman AA, et al. Endogenous regeneration of critical-size chondral defects in immunocompromised rat xiphoid cartilage using decellularzed human bone matrix scaffolds. Tissue Eng Part A, 2012, 18(21-22): 2332-2342.
12. Kirk JF, Ritter G, Waters C, et al. Osteoconductivity and osteoinductivity of NanoFUSE(?) DBM. Cell Tissue Bank, 2012, 14(1): 33-44.
13. Kumar JP, Lakshmi L, Jyothsna V, et al. Synthesis and characterization of diopside particles and their suitability along with chitosan matrix for bone tissue engineering in vitro and in vivo. J Biomed Nanotechnol, 2014, 10(6): 970-981.
14. Yasuda H, Shima N, Nakagawa N, et al. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci U S A, 1998, 95(7): 3597-3602.
15. Simonet WS, Lacey DL, Dumstan CR, et al. Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell, 1997, 89(2): 309-319.
16. 劉健, 宋暉.骨保護素對破骨細胞的影響程度分析.中國生化藥物雜志, 2015, 35(3): 65-68.
17. 鄭揚, 李危石, 劉忠軍.骨組織3D打印:骨再生的未來.北京大學報(醫學版), 2015, 47(2): 203-206.
18. Yang J, Cai H, Lv J, et al. In vivo study of a self-stabilizing artificial vertebral body fabricated by electron beam melting. Spin (Phila Pa 1976), 2014, 39(8): E486-E492.

Chinese Journal of Reparative and Reconstructive Surgery

EXPERIMENTAL STUDY ON CHITOSAN/ALLOGENEIC BONE POWDER COMPOSITE POROUS SCAFFOLD TO REPAIR BONE DEFECTS IN RATS

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