骨形态发生蛋白2载体材料在脊柱融合方面的研究与应用

doi:10.3969/j.issn.2095-4344.2015.52.022

摘要/Abstract

摘要：

背景：骨形态发生蛋白2在临床中单独应用存在一些不良反应，因此迫切需要研制理想的载体材料，来减少骨形态发生蛋白2引起的不良反应。

目的：全面了解不同骨形态发生蛋白2载体材料的性能，综述其在脊柱融合方面的研究进展。

方法：以“tissue engineering，Bone morphogenetic protein-2，spinal fusion，scaffold”或“组织工程，骨形态发生蛋白2、脊柱融合、支架”等作为检索词，由第一作者检索PubMed及万方数据库1995-07-01/2015-07-01有关脊柱融合中骨形态发生蛋白2支架方面的文献，排除重复性研究。

结果与结论：骨形态发生蛋白2的载体材料主要包括4大类，天然生物材料、人工合成有机材料、人工合成无机材料及复合材料，每一类都有几种代表性的支架材料，用于动物脊柱融合中各有缺点，比如天然生物材料普遍力学性能较差，人工合成有机材料有可能引起炎症反应，人工合成无机材料机械强度较差，复合支架材料制作工艺较复杂等，因此骨形态发生蛋白2载体材料的选择仍需进一步的实验研究。

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

关键词: 生物材料, 骨生物材料, 骨形态发生蛋白2, 脊柱融合, 支架, 综述, 国家自然科学基金

Abstract:

BACKGROUND: There are some adverse reactions when bone morphogenetic protein-2 is applied independently in clinic; therefore, it is urgent to develop an ideal carrier material to reduce the incidence of adverse reactions induced by bone morphogenetic protein-2.

OBJECTIVE: To comprehensively understand the performance of different kinds of bone morphogenetic protein-2 carrier materials, and to summarize their research progress in the field of spinal fusion.

METHODS: The literatures about bone morphogenetic protein-2 scaffolds in spinal fusion were retrieved by the first author from PubMed and Wanfang datebase durings July 1, 1995 to July 1, 2015 with the key words of“tissue engineering, bone morphogenetic protein-2, spinal fusion, scaffold” in English and Chinese, respectively. Repetitive studies were excluded.

RESULTS AND CONCLUSION: Bone morphogenetic protein-2 carrier materials mainly include four categories: natural biological materials, synthetic organic materials, synthetic inorganic materials and composite materials, each of which has several representative scaffold materials. Each category has their own shortcomings when applied in animal spinal fusion, for instance, the mechanical properties of natural biological materials are generally poor; synthetic organic materials may cause inflammatory reactions; the mechanical strength of synthetic inorganic materials is poor and the production process of composite materials is relatively complicated. Therefore, the selection of bone morphogenetic protein-2 carrier materials still needs further experimental study.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

中图分类号:

R318

蒋杰，陈亮，顾勇，彭展. 骨形态发生蛋白2载体材料在脊柱融合方面的研究与应用[J]. 中国组织工程研究, 2015, 19(52): 8487-8491.

Jiang Jie, Chen Liang, Gu Yong, Peng Zhan. Bone morphogenetic protein-2 carrier materials in the spinal fusion[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(52): 8487-8491.

参考文献

[1] Alsaleh KA,Tougas CA,Roffey DM,et al.Osteoconductive bone graft extenders in posterolateral thoracolumbar spinal fusion: a systematic review.Spine (Phila Pa 1976). 2012; 37(16): E993-1000.

[2] Bessa PC,Casal M,Reis RL.Bone morphogenetic proteins in tissue engineering: the road from laboratory to clinic, part II (BMP delivery).J Tissue Eng Regen Med. 2008; 2(2-3): 81-96.

[3] Lee SS, Huang BJ, Kaltz SR, et al.Bone regeneration with low dose BMP-2 amplified by biomimetic supramolecular nanofibers within collagen scaffolds.Biomaterials.2013; 34(2): 452-459.

[4] 欧阳毅,于博.骨形态发生蛋白-2载体材料在骨缺损修复方面的研究进展[J].广东医学, 2013,34(9):1454-1456.

[5] 吴军成,霍然,吕仁荣.软骨组织工程中支架材料的文献回顾[J].中国组织工程研究,2012, 16(3): 522-6.

[6] Venkatesan J, Bhatnagar I, Manivasagan P, et al. Alginate composites for bone tissue engineering: a review. Int J Biol Macromol.2015;72:269-281.

[7] Abbah SA, Liu J, Goh JC,et al.Enhanced control of in vivo bone formation with surface functionalized alginate microbeads incorporating heparin and human bone morphogenetic protein-2.Tissue Eng Part A. 2013;19(3-4): 350-359.

[8] 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[J].中国组织工程研究, 2011,15(29):5467-5470.

[9] 陈书军,裴国献,郭刚,等.纤维蛋白在软骨组织工程中的应用[J].国外医学:生物医学工程分册, 2005,28(4):249-253.

[10] Koo KH,Yeo do H, Ahn JM,et al.Lumbar posterolateral fusion using heparin-conjugated fibrin for sustained delivery of bone morphogenic protein-2 in a rabbit model.Artif Organs. 2012; 36(7):629-634.

[11] Ben-David D, Srouji S, Shapira-Schweitzer Ket al.Low dose BMP-2 treatment for bone repair using a PEGylated fibrinogen hydrogel matrix. Biomaterials. 2013; 34(12): 2902-2910.

[12] 梁卫东,王宏伟,王志强.不同骨组织工程支架材料的生物安全性及性能[J].中国组织工程研究,2010,14(34):6385-6388.

[13] 韩佳珺,周赞,顾燕柄,等.胶原支架负载BMP2促进大鼠脊柱横突融合的临床观察[J].中国现代医药杂志,2012,14(11):1-4.

[14] 张鸣,李翔.脱钙骨基质的临床应用与研究进展[J].医学综述, 2013,19(14):2540-2542.

[15] Biswas D, Bible JE, Whang PH,et al.Augmented demineralized bone matrix: x a potential alternative for posterolateral lumbar spinal fusion.Am J Orthop (Belle Mead NJ). 2010;39(11):531-538.

[16] Velasco MA, Narváez-Tovar CA, Garzón-Alvarado DA.Design, materials, and mechanobiology of biodegradable scaffolds for bone tissue engineering. Biomed Res Int.2015; 2015(729076.

[17] 张娜,郭圣荣.聚己酸内酯及其在医药领域的应用[J].生物医学工程学杂志,2003,20(4): 746-749.

[18] Yong MR,Saifzadeh S,Woodruff M,et al.Biological performance of a polycaprolactone-based scaffold plus recombinant human morphogenetic protein-2 (rhBMP-2) in an ovine thoracic interbody fusion model.Eur Spine J.2013;23(3): 650-657.

[19] 李梦雪.聚乳酸羟基乙酸共聚物的制备及应用[J].黑龙江医药, 2012,25(3):410-412.

[20] Fischgrund JS, James SB, Chabot MC,et al.Augmentation of autograft using rhBMP-2 and different carrier media in the canine spinal fusion model.J Spinal Disord.1997;10(6): 467-472.

[21] 秦超师,冯高科,蒋学俊,等.无定形磷酸钙应用在生物医学中的特点[J].中国组织工程研究, 2014,18(39):6353-6358.

[22] Liu B,Lun DX.Current application of beta-tricalcium phosphate composites in orthopaedics.Orthop Surg. 2012; 4(3):139-144.

[23] Sterling JA, Guelcher SA.Biomaterial scaffolds for treating osteoporotic bone. Curr Osteoporos Rep.2014;12(1):48-54.

[24] Pelletier MH, Oliver RA, Christou C,et al.Lumbar spinal fusion with beta-TCP granules and variable Escherichia coli-derived rhBMP-2 dose.Spine J.2014;14(8):1758-1768.

[25] 冯晓娜,杨宪园,卢婷利,等.羟基磷灰石微载体的应用及制备[J].材料导报,2013,27(3): 87-90.

[26] Kong CB, Lee JH, Baek HR,et al.Posterolateral lumbar fusion using Escherichia coli-derived rhBMP-2/hydroxyapatite in the mini pig.Spine J.2014;14(12): 2959-2967.

[27] Abbah SA, Lam CX, Ramruttun AK,et al.Fusion performance of low-dose recombinant human bone morphogenetic protein 2 and bone marrow-derivedmultipotent stromal cells in biodegradable scaffolds: a comparative study in a large animal model of anterior lumbar interbody fusion.Spine (Phila Pa 1976).2011;36(21):1752-1759.

[28] Khan SN, Toth JM, Gupta K,et al. Early-term and mid-term histologic events during single-level posterolateral intertransverse process fusion with rhBMP-2/collagen carrier and a ceramic bulking agent in a nonhuman primate model: implications for bone graft preparation.J Spinal Disord Tech. 2014;27(4):212-219.

[29] Gu Y, Chen L, Yang HLet al Evaluation of an injectable silk fibroin enhanced calcium phosphate cement loaded with human recombinant bone morphogenetic protein-2 in ovine lumbar interbody fusion.J Biomed Mater Res A. 2011;97(2): 177-185.

[30] Matsumoto T, Toyoda H, Dohzono S,et al.Efficacy of interspinous process lumbar fusion with recombinant human bone morphogenetic protein-2 delivered with a synthetic polymer and beta-tricalcium phosphate in a rabbit model.Eur Spine J .2012; 21(7):1338-1345.

[31] Pilipchuk SP, Plonka AB, Monje A, et al.Tissue engineering for bone regeneration and osseointegration in the oral cavity. Dent Mater.2015;31(4):317-338.

[32] Claros S, Rico-Llanos GA, Becerra J,et al.A novel human TGF-beta1 fusion protein in combination with rhBMP-2 increases chondro-osteogenic differentiation of bone marrow mesenchymal stem cells.Int J Mol Sci. 2014; 15(7): 11255- 11274.

[33] Haidar ZS, Hamdy RC, Tabrizian M.Delivery of recombinant bone morphogenetic proteins for bone regeneration and repair. Part B: Delivery systems for BMPs in orthopaedic and craniofacial tissue engineering.Biotechnol Lett. 2009;31(12): 1825-1835.

[34] Francis CS, Mobin SS,Lypka MA,et al.rhBMP-2 with a demineralized bone matrix scaffold versus autologous iliac crest bone graft for alveolar cleft reconstruction. Plast Reconstr Surg 2013;131(5):1107-1115.

[35] Koo KH, Lee JM, Ahn JM,et al.Controlled delivery of low-dose bone morphogenetic protein-2 using heparin-conjugated fibrin in the posterolateral lumbar fusion of rabbits.Artif Organs. 2013;37(5):487-494.

[36] Romagnoli C,D'Asta F,Brandi ML.Drug delivery using composite scaffolds in the context of bone tissue engineering. Clin Cases Miner Bone Metab. 2013;10(3):155-161.

[37] Walmsley GG,McArdle A,Tevlin R,et al.Nanotechnology in bone tissue engineering. Nanomedicine. 2015; 11(5): 1253-1263 .

[38] Chung YI,Ahn KM,Jeon SH,et al.Enhanced bone regeneration with BMP-2 loaded functional nanoparticle- hydrogel complex.J Control Release. 2007; 121(1-2):91-99.

[39] Sowjanya JA,Singh J,Mohita T,et al.Biocomposite scaffolds containing chitosan/alginate/nano-silica for bone tissue engineering.Colloids Surf B Biointerfaces. 2013; 109: 294-300.

[40] He X,Dziak R,Mao K,et al.Integration of a novel injectable nano calcium sulfate/alginate scaffold and BMP2 gene-modified mesenchymal stem cells for bone regeneration.Tissue Eng Part A.2013;19(3-4):508-518.

引言

材料方法

讨论

骨形态发生蛋白首先被Urist发现并命名，它是转化生长因子β超家族一员^[31]，具有诱导成骨和成软骨分化、促进骨组织形成和异位成骨的作用，在已知的43种骨形态发生蛋白中，以骨形态发生蛋白2的成骨作用最强^[32]。尽管美国FDA早在2002年就批准重组骨形态发生蛋白2用于脊柱融合^[33-34]，但由于骨形态发生蛋白2的半衰期较短、结构不稳定、需要大剂量运用，这会导致包括异位骨形成、骨溶解、炎症反应等不良反应。因此，有必要研制出理想的载体系统，以此降低骨形态发生蛋白2的用量，减少不良反应^[35]。

理想的支架载体系统需要具有以下的物理、生物性质：可提供临时的机械支撑作用，能够在不失活前提下消毒，含有多孔结构，促进骨细胞迁移作用，骨诱导性，骨结合性，降解可控性，产生无毒性降解产物，不发生慢性炎症反应，具备承载生物活性因子性质^[36]。以上介绍了多种支架材料，每一种都具有一些优点，可作为骨形态发生蛋白2的载体用于动物脊柱融合。但它们又都存在一些缺陷，比如天然生物材料普遍力学性能较差，人工合成有机材料有可能会引起炎症反应，人工合成无机材料机械强度较差，复合支架材料制作工艺较复杂，它们目前还不是最理想的骨形态发生蛋白2支架。

近年来，纳米生物材料逐渐成为骨组织工程的一个研究热点，已被广泛运用到各种动物模型中^[37]。Chung等^[38]用肝素功能化的聚乳酸-羟基乙酸共聚物纳米微粒和纤维蛋白水凝胶，负载骨形态发生蛋白2用在大鼠颅骨缺损上，成骨效果显著。Sowjanya等^[39]通过实验证明壳聚糖、海藻酸钠和纳米二氧化硅的复合支架材料具有多孔结构，无毒性，生物相容性好，可用于骨组织工程研究。He等^[40]构建了一种具有生物相容性和生物可降解性的纳米硫酸钙/藻酸盐可注射支架材料，并且在小鼠颅骨缺损实验中，发挥了良好的载体作用。但目前这些纳米支架材料在脊柱融合方面的研究还较少。

想要研制出人体脊柱融合的理想骨形态发生蛋白2载体，还需要更广泛、更深入、更细致的研究，尤其要注重纳米科学、智能生物材料学、组织工程学、分子生物学等交叉学科领域的研究，逐步由实验向临床过渡。相信随着研究的进一步深入，在不远的将来能够开发出一种最理想的骨形态发生蛋白2载体运用到人类脊柱融合手术中。

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程