负载BMP的新型组织工程骨的构建及骨缺损修复实验

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (11): 1369-1374.

负载BMP的新型组织工程骨的构建及骨缺损修复实验

胡稷杰¹, 金丹¹, 全大萍², 钟世镇³, 陈炅昊¹, 魏宽海¹, 赵洁², 裴国献¹

1. 南方医科大学南方医院创伤骨科, 广东, 广州, 510515;
2. 中山大学高分子研究所, 广东, 广州, 510275;
3. 南方医科大学临床解剖研究所, 广东, 广州, 510515

出版日期:2005-11-20 发布日期:2005-11-20
基金资助:
收稿日期:2005-5-20。
基金项目:国家高新技术发展项目(2003AA205010)
作者简介:胡稷杰(1979-),硕士,研究方向:骨组织工程,电话:020-61641745;61360586,E-mail:hjj0610@163.com
通讯作者:裴国献,教授,博士生导师,电话:020-61641741

Bone defect repair with a new tissue-engineered bone carrying bone morphogenetic protein in rabbits

HU Ji-jie¹, JIN dan¹, QUAN Da-ping², ZHONG Shi-zhen³, CHEN Jiong-hao¹, WEI Kuan-hai¹, ZHAO Jie², PEI Guo-xian¹

1. 南方医科大学南方医院创伤骨科, 广东, 广州, 510515;
2. 中山大学高分子研究所, 广东, 广州, 510275;
3. 南方医科大学临床解剖研究所, 广东, 广州, 510515

Online:2005-11-20 Published:2005-11-20

摘要/Abstract

摘要： 目的探讨以聚乳乙醇酸(poly-(D,L-lactide-co-glycolide),PLGA)、骨形态发生蛋白(BMP)、骨髓基质干细胞(BM-SCs)构建成新型组织工程骨并观察其在动物体内的成骨能力。方法制作新西兰大白兔右桡骨中段15mm骨缺损实验模型,随机分为实验组、对照组和空白组,实验组植入同时负载5mgBMP及1×10⁶个已向成骨细胞诱导的BMSCs的PLGA、对照组植入负载1×10⁶个已向成骨细胞诱导的BMSCs的PLGA、空白组仅植入PLGA。术后对动物进行大体观察、摄X线片观察各组不同时相骨缺损修复情况、比较不同时相的骨缺损区X线阻射密度、并于术后第4、8、12周取出骨缺损区标本进行大体观察和组织学切片观察,图像分析骨小梁的生成数量。结果实验组在12周内骨缺损完全修复,且同时期内新生骨的数量和质量显著优于对照组,空白材料组骨缺损主要由纤维结缔组织填充。结论利用含BMP的PLGA支架与BMSCs复合构建的新型组织工程骨具有良好的骨缺损修复能力。

Abstract: Objective To construct a new tissue-engineered bone with poly (D, L-lactide-co-glycolide) (PLGA), bone morphogenetic protein (BMP) and bone marrow-derived stem cells (BMSCs) and observe its effect in repairing segmental bone defects. Methods A 15-mm bone defect in the right radius was induced in New Zealand white rabbits, and the models were randomized into three groups to receive implantation of the tissue-engineered bone grafts constructed with PLGA carrying 5 mg BMP and about 1×10⁶ BMSCs (experimental group), grafts of PLGA with about 1×10⁶ BMSCs (control group), or grafts of exclusive PLGA (blank control group), respectively. The osteogenesis in the bone defect after the implantation on was evaluated X-ray films, and the histological changes of the tissues sampled from the bone defect 4, 8, and 12 weeks after operation were observed and new bone formation was measured by image analysis. Results The bone defect was completely repaired in the experimental group 12 weeks after the implantation, showing the best results among the 3 groups. The bone defects in the blank control group was filled with only fibrous and connective tissues at 12 weeks. Conclusion This tissue-engineered bone constructed with PLGA, BMP and BMSCs possesses good ability in repairing segmental bone defect.

中图分类号:

R322.71

胡稷杰¹, 金丹¹, 全大萍², 钟世镇³, 陈炅昊¹, 魏宽海¹, 赵洁², 裴国献¹. 负载BMP的新型组织工程骨的构建及骨缺损修复实验[J]. 南方医科大学学报, 2005, 25(11): 1369-1374.

HU Ji-jie¹, JIN dan¹, QUAN Da-ping², ZHONG Shi-zhen³, CHEN Jiong-hao¹, WEI Kuan-hai¹, ZHAO Jie², PEI Guo-xian¹. Bone defect repair with a new tissue-engineered bone carrying bone morphogenetic protein in rabbits[J]. Journal of Southern Medical University, 2005, 25(11): 1369-1374.

参考文献

[1] Bruder SP, Fox BS. Tissue engineering of bone. Cell based strategies[J]. Clin Orthop, 1999, (367 Suppl): 68-83.,
[2] Carl A. Kirker H. Potential application sand delivery strategies for bone morphogenetic proteins[J]. Advanced Drug Delivery Rev,2000, 43(1): 65-92.
[3] 胡稷杰,裴国献,全大萍,等.新型聚乳酸-羟基乙酸支架的细胞相容性研究[J].中华创伤骨科杂志,2005,7(4):358-62.Hu JJ, Pei GX, Quan DP, et al. Cellular compatibility of a new type ofpolylactic glycolic acid (PLGA) scaffold[J]. Chin J Orthop Trauma, 2005, 7(4): 358-62.
[4] 曹谊林,商庆新.软骨、骨组织工程的现状与趋势[J].中华创伤杂志,2001,17(1):7-9.Cao YL, Shang QX. The status quo and tendency of cartilage and bone tissue engineering[J]. Chin J Traumatol, 2001, 17(1): 7-9.
[5] Quirk RA, Chan WC, Davies MC, et al. poly(l-lysine)-GRGDS as abiomimetic surface modifier for poly (lacticacid)[J]. Biomaterials,2001, 22(8): 865-972.
[6] Chang SL, Yang JW, Rifas L, et al. Differentiation of human bone marrow osteogenic stromal cells in vitro: induction of the osteoblast phenotypebydexamethasone[J]. Endocrinology, 1994,134(1): 277-86.
[7] 金丹,裴国献,王前,等.骨髓基质细胞体外培养骨发生潜能及条件研究[J].中国矫形外科杂志,2000,8(6):565-6.
[8] Wakitani S. Repair of large full-thickness articular cartilage de fects with allograft articular chondricytes embededinacollagengel[J].Tissue Engineering, 1998, 4(4): 429-44.
[9] Garcia AJ, Ducheyne P, Boettiger D, et al. Effect of surface reaction stage on fi-broncection mediated adhesion of osteoblast-like cell to bioactive glass[J]. J Biomed Mater Res, 1998, 40(1): 48-56.
[10] Robinson BP, Hollinger JO, Szachowicz EH, et al. Calvarial bone repair with porous D.L Polylactide[J]. Otalaryngol Head Neck Surg, 1995, (1126): 707-713.
[11] 罗丙红,全大萍,廖凯荣,等.低热-高压法制备PLGA多孔支架及其体外降解研究[J].功能高分子学报,2003,(16)2:149-55.Luo BH, Quan DP, Liao KR, et al. Scaffolds with high porosity for tissue engineering fabricated by low heat-h igh pressure and there degradation in vitro[J]. J Funct Polymers, 2003, (16)2: 149-55.
[12] Agrawal CM, Best J, Heckman JD, et al. Protein release kinetics of a biodegradable implant for fracture nonunions[J]. Biomaterials,1995, 16(16): 1255-60.
[13] Zellin G, Linde A. Importance of delivery system for growth-stimulatory factors in combination with osteopromotive membranes. An experimental study using rhBMP-2 in rat mandibular defects[J]. J Biomed Mater Res, 1997, 35(2): 181-90.
[14] Piattelli A, Scarano A, Coraggio F, et al. Early tissue reactions to polylactic acid resorbable membranes: a histological and histochemical study in rabbit[J]. Biomaterials, 1998, 19(10): 889-96.
[15] 谭祖键,李起鸿,许建中,等.聚乳酸作为骨形态发生蛋白载体修复骨缺损的实验研究[J].中华骨科杂志,2000,20(12):742-7.Tan ZJ, Li QH, Xu JZ, et al. An experimental study of bone defects repaired with bone morphogenetic protein using polyactic acid as carrier[J]. Chin J Orthop, 2000, 20(12): 742-7.
[16] Takashi K. Potentiation of the activity of bone morphogenetic protein-2 in bone.regeneration by a PLA-PEG/hydroxyapatite composite[J]. Biomaterials 26 (2005): 73-9.