髓核组织工程支架的构建及其生物相容性Construction of nucleus pulposus tissue engineering scaffold and its biocompatibility
徐子昂;刘晨;肖良;王效;徐宏光;
摘要(Abstract):
[目的]制备脱细胞髓核基质,并鉴定其物理特性,制备壳聚糖凝胶组织支架,探讨其生物相容性。[方法]分离、培养SD大鼠骨髓间充质干细胞(MSC),并研究其成骨、成软骨、成脂肪多向分化能力。分离家猪髓核组织,逐步经过胰酶、核酶及TritonX-100处理,得到脱细胞髓核基质,观察脱细胞效果及其形态特点,并检测分析脱细胞髓核基质的主要成分。将脱细胞髓核基质溶于醋酸,制备成壳聚糖联合脱细胞髓核基质,将大鼠MSC接种于基质膜上培养。在第1、2、3、4、5 d采用CCK-8检测大鼠MSC在基质膜上的增殖情况,计算细胞相对增殖率(RGR)来评价其生物相容性,并计算其孔隙率。[结果]观察到SD大鼠MSC具有成骨、成软骨和成脂肪分化能力。制备出的家猪脱细胞髓核基质肉眼呈乳白色糜状,质较软,扫描电镜下观察发现,由无规则排列的网状胶原纤维组成,表面无细胞残留。红外光谱仪分析表明脱细胞髓核基质的主要成分是胶原蛋白。通过CCK-8检测,5 d内细胞毒性分级在0级与2级之间,说明大鼠MSC与壳聚糖联合脱细胞髓核基质具有很好的生物相容性。壳聚糖联合脱细胞髓核基质支架的孔隙率为(87.52±0.53)%。[结论]壳聚糖联合脱细胞髓核基质具有良好的生物相容性及孔隙率,符合生物工程支架选材标准。
关键词(KeyWords): 组织工程支架;生物相容性;脱细胞基质;壳聚糖
基金项目(Foundation): 国家自然科学基金面上项目(编号:81572185);; 国家青年科学基金项目(编号:81702158);; 安徽省自然科学基金面上项目(编号:1708085MH185)
作者(Author): 徐子昂;刘晨;肖良;王效;徐宏光;
Email:
DOI:
参考文献(References):
- [1]舒剑臣,唐小穗.P物质、白介素6与腰椎间盘突出症所致腰腿痛的相关性研究[J].中国骨与关节损伤杂志,2013,(9):825-827.
- [2]Upadhyayula PS,Curtis EI,Yue JK,et al.Anterior versus transforaminal lumbar interbody fusion:perioperative risk factors and 30-day outcomes[J].Int J Spine Surg,2018,12(5):533-542.
- [3]Zhang ZQ,Wang CS,Yang P,et al.Mesenchymal stem cells induced by microencapsulated chondrocytes on repairing of intervertebral disc degeneration[J].Orthop Surg,2018,10(4):328-336.
- [4]Gawri R,Shiba T,Pilliar R,et al.Inorganic polyphosphates enhances nucleus pulposus tissue formation in vitro[J].J Orthop Res,2017,35(1):41-50.
- [5]董视师,李景峰,周逸驰,等.腰椎间盘退变:组织工程修复及生物力学评价[J].中国组织工程研究,2015,(37):6024-6030.
- [6]王新宏,沈忆新,范志海,等.组织工程支架材料在脊髓损伤修复中的应用[J].中国组织工程研究与临床康复,2011,15(25):4719-4722.
- [7]Chaudhuri R,Ramachandran M,Moharil P,et al.Biomaterials and cells for cardiac tissue engineering:Current choices[J].Mater Sci Eng C Mater Biol Appl,2017,79:950-957.
- [8]Sadeghi-Ataabadi M,Mostafavi-Pour Z,Vojdani Z,et al.Fabrication and characterization of platelet-rich plasma scaffolds for tissue engineering applications[J].Mater Sci Eng C Mater Biol Appl,2017,71:372-380.
- [9]Kumbhar SG,Pawar SH.Synthesis and characterization of chitosan-alginate scaffolds for seeding human umbilical cord derived mesenchymal stem cells[J].Biomed Mater Eng,2016,27(6):561-575.
- [10]Ikeda T,Ikeda K,Yamamoto K,et al.Fabrication and characteristics of chitosan sponge as a tissue engineering scaffold[J].Biomed Res Int,2014,2014:786892.
- [11]Oh HJ,Kim SH,Cho JH,et al.Mechanically Reinforced Extracellular Matrix Scaffold for Application of Cartilage Tissue Engineering[J].Tissue Eng Regen Med,2018,15(3):287-299.
- [12]Barbeck M,Serra T,Booms P,et al.Analysis of the in vitro degradation and the in vivo tissue response to bi-layered 3D-printed scaffolds combining PLA and biphasic PLA/bioglass componentsGuidance of the inflammatory response as basis for osteochondral regeneration[J].Bioact Mater,2017,2(4):208-223.
- [13]Escudero-Castellanos A,Ocampo-García BE,Domínguez-García MV,et al.Hydrogels based on poly(ethylene glycol)as scaffolds for tissue engineering application:biocompatibility assessment and effect of the sterilization process[J].J Mater Sci Mater Med,2016,27(12):176.
- [14]Jin GZ,Park JH,Wall I,et al.Isolation and culture of primary rat adipose derived stem cells using porous biopolymer microcarriers[J].Tissue Eng Regen Med,2016,13(3):242-250.
- [15]Liu S,Hou KD,Yuan M,et al.Characteristics of mesenchymal stem cells derived from Wharton's jelly of human umbilical cord and for fabrication of non-scaffold tissue-engineered cartilage[J].J Biosci Bioeng,2014,117(2):229-235.
- [16]Salloum RH,Rubin JP,Marra KG.The role of steroids in mesenchymal stem cell differentiation:molecular and clinical perspectives[J].Horm Mol Biol Clin Investig,2013,14(1):3-14.
- [17]Skoronski E,Fernandes M,Magalh?es ML,et al.Substrate specificity and enzyme recycling using chitosan immobilized laccase[J].Molecules,2014,19(10):16794-809.
- [18]Merchant Z,Taylor KM,Stapleton P,et al.Engineering hydrophobically modified chitosan for enhancing the dispersion of respirable microparticles of levofloxacin[J].Eur J Pharm Biopharm,2014,88(3):816-29.
- [19]Han F,Dong Y,Su Z,et al.Preparation,characteristics and assessment of a novel gelatin-chitosan sponge scaffold as skin tissue engineering material[J].Int J Pharm,2014,476(1-2):124-33.