β-磷酸三钙复合骨髓间充质干细胞修复山羊骨软骨缺损的实验研究

目的将β-磷酸三钙（β-TCP）与山羊骨髓间充质干细胞（BMSCs）复合后,在生物反应器中分别向成软骨和成骨诱导,并植入骨软骨缺损处,观察软骨修复效果。方法分离、培养山羊BMSCs,在生物反应器中分别向成软骨及成骨诱导2周,植入骨软骨缺损部位。实验组分为A组：旋转力刺激＋成软骨、成骨诱导组（力学刺激组）,B组：单纯成软骨、成骨诱导组（无力学刺激组）,并设空白对照组。术后12周和24周进行大体观察、组织学染色等,并行O＇Driscoll Keeley and Salter评分。结果 A、B组均有新生软骨形成;A组软骨在12周与24周均优于B组（P〈0.05）;术后12、24周A组评分优于B组,差异有统计学意义（P〈0.05）。对照组无新生软骨形成。结论将BMSCs复合于β-TCP,可用于组织工程修复骨软骨缺损;体外培养阶段的旋转力刺激有利于改善组织工程软骨的质量。     

"双相"组织工程软骨修复兔关节骨软骨缺损

目的探讨“双相”异体骨基质明胶（bonematrixgelatin,BMG）作为组织工程软骨载体,与同体骨髓间充质干细胞（marrowmesenchymalstemcells,MSCs）结合,构建组织工程软骨修复兔关节骨软骨缺损的效果。方法4月龄新西兰兔32只,雌雄不限,体重2～3kg。①体外实验：取5只新西兰兔,处死后取髂骨和四肢骨,制备一侧松质骨,一侧皮质骨的“双相”异体BMG载体,扫描电镜观察。另取新西兰兔18只,抽取骨髓,分离MSCs并诱导成软骨分化;将诱导而来的软骨前体细胞与“双相”BMG载体复合构建组织工程软骨,分别于1、3和5周取材行Masson、PAS染色和扫描电镜观察。②体内实验：将抽取骨髓的18只及余下的9只新西兰兔制成双侧股骨内髁骨软骨缺损模型,将前期制备的组织工程软骨同体植入18只兔的右股骨内髁骨软骨缺损（A组）,左侧缺损移植异体BMG（B组）,其余9只双侧软骨缺损未予处理作为空白对照（C组）,分别于术后1、3和6个月取材,行大体、组织学和Ⅱ型胶原mRNA原位杂交观察,改良Wakitani法评分,比较各组修复效果差异。结果①体外实验：“双相”BMG松质骨面孔隙大小100-800μm,细胞于其中增生,形成富含细胞的软骨层;皮质骨面孔隙大小10～40pm,细胞层状覆盖于其表面,可作为起支撑作用的软骨下骨。②体内实验：A组术后1个月即可重建关节骨软骨缺损;修复软骨在观察期内逐渐变薄,但在6个月内始终保持关节面及软骨下骨结构完整。B、C组未能修复缺损,缺损周边软骨磨损加剧。改良Wakitani评分显示A组在3个时间点的各项评分结果,除6个月软骨厚度外,其它指标均优于B、C组,且差异有统计学意义（P〈0．01）。Ⅱ型胶原mRNA原位杂交显示,A组缺损区修复组织中细胞阳性染色率明显高于B、C组,且差异有统计学意义（P〈0．01）。结论“双相”异体BMG可作为组织工程软骨载体材料,其结合自体MSCs诱导的软骨前体细胞制备的组织工程软骨,可修复兔关节软骨和软骨下骨。     

组织工程化软骨修复兔骺板损伤的实验研究

目的　探讨组织工程化软骨细胞 生物载体复合物修复兔骺板损伤的可行性。方法于 8周龄兔胫骨上端骺板缺损模型中 ,A、B、C三组分别植入组织工程化软骨、单纯外消旋聚乳酸(PDLLA)、空白对照 ,术后 4、8、16周时对双下肢行X线摄片、组织学检查。结果　 4、8、16周时双侧胫骨长度、胫骨角之差A组与B、C组相比 ,短缩与成角畸形轻 ,差异有统计学意义 (P <0 .0 1) ,组织学显示缺损区呈薄层骺软骨细胞样结构 ;B、C两组间差异无统计学意义 (P >0 .0 5 )。结论　骨髓间充质干细胞 (MSCs)可定向诱导分化为软骨细胞 ;组织工程化软骨细胞植入可减轻骺板损伤后肢体短缩与成角畸形。     

转壁生物反应器中动态分化组织工程软骨的实验研究

目的 :在转壁生物反应器动态环境中分化骨髓间充质干细胞 (bMSC) ,制备立体组织工程软骨。方法 :穿刺吸取成年兔骨髓 ,分离扩增bMSC ,复合于纤维蛋白凝胶制成圆柱状三维组织 (细胞密度2 0× 10 7/ml) ,置于转壁生物反应器中进行分化培养 ,同时设单纯静止培养组。 5周后观测大体形态和组织学形态、Ⅰ和Ⅱ型胶原免疫组织化学表达 ,测量分化组织的细胞活力。结果 :动态培养可以有效保持材料大体形态 ,甲苯胺兰染色阳性 ,无明显Ⅰ型胶原表达 ,大量表达Ⅱ型胶原 ,人工组织的细胞活力显著高于单纯静止培养方法。结论 :转壁生物反应器培养明显改善组织工程软骨的活力 ,有利于进行三维材料体外软骨分化。     

同种异体脱钙骨复合骨髓间充质干细胞在兔膝关节腔内培养组织工程软骨的研究

目的 比较同种异体脱钙骨和细胞因子诱导骨髓间充质干细胞(BMSC)制成细胞-支架复合物在兔膝关节腔内环境较体外培养出组织工程软骨的优点.方法 BMSC向软骨细胞诱导后与同种异体脱钙骨支架复合分别在体外培养(A组)和成年雄性新西兰白兔(15只)左侧膝关节腔内(B组)培养,右侧膝关节腔内(C组)培养单纯脱钙骨支架做空白对照.每4、8、12周各组标本分别取材,制石蜡切片行苏木素-伊红(HE)染色、甲苯胺蓝染色、Ⅱ型胶原免疫组织化学等组织学观察,并通过形态学分析软件计算免疫组织化学平均光度(A)值.结果 标本石蜡切片HE染色:4周时A组标本见软骨细胞散在分布于支架表面,内部基本未观察到细胞.B组标本支架内软骨细胞数量明显较多,软骨陷窝形成,陷窝周围基质深染,可见由单个细胞分裂形成的同源细胞群.8周时B组标本以成熟软骨细胞为主,细胞数量多,出现柱状排列的同源细胞群,细胞周围分泌着色均一的玻璃样基质,且渗入支架结构内部,脱钙骨支架有部分被吸收.12周时各组支架结构明显被吸收,A组支架内填满软骨细胞,部分已纤维化,但结构排列紊乱.B组支架呈透明软骨样,软骨细胞充分渗透进入支架内,呈一定应力方向排列.Ⅱ型胶原免疫组织化学A值统计学分析比较发现:第4,8,12周B组Ⅱ型胶原免疫组织化学的A值均高于A组,差异有统计学意义(P＜0.01).两组间A值随时间的变化趋势差异有统计学意义(P＜0.01).结论 同种异体脱钙骨支架复合经细胞因子诱导的BMSC,在膝关节腔内进行培养,利用关节腔内低氧、多种生长因子的微环境以及关节活动时的应力刺激等优势,较体外培养可以培养出组织学特点更好的工程软骨.

Abstract：

Objective To compare the superiority of cultivating tissue engineered cartilage by homologous decalcified bone matrix combined with bone-marrow mesenchymal stem cells (BMSCs) in rabbits' knee cavity with culture in vitro. Methods Archiogeneration BMSCs were isolated and purified by adhering to the culture glassware wall from neonatal New Zealand white rabbits. The third generation BMSCs were induced to chondrocytes by transforming growth factor TGF-β1, insulin-like growth factor IGF-1 and vitamin C. Seven days later, the cells were bond to homologous decalcified bone matrix and cultured in vitro (group A) , cultured in rabbits' left knee cavity (group B) , or bond to homologous decalcified bone matrix and cultured in rabbits' right knee cavity ( group C ). Every 4 weeks after cellular transplant, all rabbits in groups B and C were sacrificed and paraffin-embedded sections were made from the specimens.All the section were subjected to H-E stain, toluidine blue stain and type Ⅱ collagen immunohistochemical staining with chromogen diaminobenzidine ( DAB) . Immunohistochemical absorbance ( A) values were calculated by morphology analysis software. Measurement data were expressed as mean ± standard, and satistical analysis was performed by t test, one-way ANOVA using SPSS 15. 0 software. Results After cultivation for 4 weeks, H-E stain showed diffuse distribution of chondrocytes on scaffold' s surface in group A.In group B, there were a great quantity of chondrocytes in the scaffold, and cartilage lacuna, matrix anachromasis and isogenous group were group. At the 8th week, in group B maturated chondrocytes and isogenous groups arranged as column, surrounded by a pond of cellular matrix infiltrating into the scaffolds. The decalcified bone matrix was absorbed partly. At the 12th week, all the scaffolds were absorbed obviously.Chondrocytes filled into the scaffolds in group A, but arranged irregularly. In group B chondrocytes arranged in the stress orientation. At 4th, 8th, and 12th week, A values of type Ⅱ collagen in group B was significantly higher than in group A with the different being significant. Conclusion Compound of Homologous decalcified bone matrix-induced BMSCs cultured in rabbits' knee cavity could yield better histological feature tissue engineered cartilage.     

骨髓间充质干细胞及在软骨组织工程中的研究进展

间充质干细胞 (ｍｅｓｅｎｃｈｙｍａｌｓｔｅｍｃｅｌｌｓ,ＭＳＣｓ)是指能在适宜的条件下分化再生骨、软骨、肌肉、韧带、肌腱、脂肪、骨髓基质等间充质组织的一类细胞 ,具有典型的干细胞特点 ,终身存在于骨髓和其他组织器官中 ,负责组织的修复和更新〔1,2〕。它们具有多向分化潜能和强大的增殖能力 ,并且易于从骨髓中分离及体外扩增纯化 ;因此 ,近年来在组织工程领域倍受重视 ,得到广泛研究。笔者仅就骨髓间充质干细胞的生物学特性及其在软骨组织工程中的研究进展作一综述。1　骨髓间充质干细胞的主要生物学特性　　骨髓中只含有少量间充…     

三种不同方式构建组织工程软骨修复犬膝关节骨软骨缺损对比的实验研究

[目的]通过对比三种不同方式构建的骨软骨复合体修复比格犬膝关节骨软骨缺损的修复情况,探讨构建组织工程化软骨的最佳可行性方案.[方法]本研究分别采用体外单体培养、分体培养、生物反应器内分体培养体内构建三种方式构建骨软骨复合体,模仿马赛克移植术植入比格犬膝关节骨软骨缺损处,通过大体观察、组织学分析观察其修复情况.[结果]术后3个月,3种方式构建的骨软骨复合体均不同程度修复了犬关节软骨缺损,采用生物反应器内分体培养体内构建方式修复骨软骨缺损效果优于前两种方式.[结论]灌注型生物反应器使软骨和成骨细胞在三维载体内存活并增殖,提高细胞在载体内的复合效率,软骨修复情况优于前两种方法.     

经骨向诱导的骨髓间充质干细胞在脱钙骨上生长行为的实验研究

目的:建立成骨细胞-脱钙骨支架复合物,观察其诱导成骨能力,从而探寻组织工程化骨的体外构建方法.方法:运用细胞沉淀法将大鼠BMSCs诱导形成的成骨细胞接种于脱钙骨支架,通过扫描电镜(SEM)、免疫荧光观察脱钙骨表面细胞生长情况,通过上清液碱性磷酸酶(ALP)活性及Ⅰ型前胶原羧端肽(PICP)、骨钙素检测观察成骨细胞活性.结果:扫描电镜示成骨细胞在脱钙骨表面生长良好,上清液ALP、PICP及骨钙素表达均明显高于对照组(P＜0.05),并随时间的增长而显著增加(P＜0.05).结论:采用细胞沉淀法成功将成骨细胞接种于脱钙骨支架,成骨细胞在支架材料中增殖旺盛,细胞活性状态良好,具有良好的骨形成能力,脱钙骨支架可作为骨组织工程中载体支架的较优选择.     

兔骨髓间质干细胞用于构建组织工程软骨组织的初步报告

目的 采用组织工程方法,以培养后的兔骨髓间质干细胞（MSC）制成人工软骨培养物,经体内外培养后发育出成活的软骨组织。方法 抽取兔人经密度梯度离心得到单个核细胞,再经体外分离、培养获得兔骨髓MSC。向MSC培养液内加入地塞米松、转化生长因子－β1（TGF－β1）和维生素C进行软骨起源诱导培养3周,部分细胞开始转变为圆形并分泌基质。将诱导后的细胞与牛Ⅰ型胶原及人纤维蛋白按一定的比例混合,制成软骨样的培养物并分别做体内外培养。结果 体外培养2周后,培养物内大部分细胞已萎缩消失。但剩余的少量细胞成活,形成类似的软骨陷窝并分泌甲苯胺蓝异染的软骨基质。体内移植培养3周后,培养物已发育成颗粒状成熟的软骨组织。结论 骨髓间质干细胞可用于组织工程软骨组织的构建,是一种非常有前途的人工软骨组织构建中的功能细胞。     

以软骨细胞外基质与脱细胞骨基质构建组织工程骨软骨双层支架及其性能的研究

目的 探讨采用软骨细胞外基质(CECM)与脱细胞骨基质(ACBM)为材料制作新型组织工程骨软骨双层支架的可行性,并检测其性能.方法 双层支架的骨部分以犬松质骨制备的ACBM为原料,软骨部分以人CECM为材料,采用冷冻冻干法制备CECM/ACBM双层支架并交联.测定支架孔隙率,采用四甲基偶氮唑盐(MTT)法分析支架浸提液毒性.分离培养犬骨髓基质干细胞(BMSCs),成软骨诱导后种植到支架上,倒置显微镜、电镜、Dead/Live荧光染色观察细胞在支架的生长、分化情况.结果 扫描电镜及Micro-CT观察显示支架内孔洞相互贯通呈海绵状,CECM部分孔径(155±34)μm,孔隙率为91.3%±2.0%;ACBM部分具有大然骨的孔径和空隙率,骨软骨部分结合良好.培养1～6 d不同浓度支架浸提液与对照培养液吸光度值比较羌异均无统计学意义(P＞0.05).倒置显微镜、电镜检查结果表明BMSCs在支架上黏附良好,细胞基质分泌增加,Dead/Live荧光染色表明双层支架内细胞均呈绿色.结论 CECM/ACBM骨软骨双层支架具备良好的孔径和孔隙率,骨、软骨两层间结合良好,无毒,生物相容性良好,可作为支架载体用于组织工程骨软骨复合体的构建.

Abstract：

Objective To fabricate a novel bilayered scaffold constructed with cartilage extracellular matrix (CECM) and acellular bone matrix (ACBM) for osteochondral tissue engineering.Methods The bone layer of the osteochondral scaffold was prepared using canine bone cancellous bone columns, and the cartilage layer was fabricated using CECM.After CECM microfilaments were decellularized, the biphasic scaffolds were fabricated by soaking the ACBM columns into cylindrical silicon moulds with a 30 g/L CECM suspension using simple freeze-drying method.After the scaffolds were cross-linked, the porosity was measureed.MTT test was also done to assess cytotoxicity of the scaffolds.Canine bone marrow-derived mesenchymal stem cells (BMSCs) were induced by chondrogenic medium and seeded into novel scaffold.Cell proliferation and differentiation were analyzed using inverted microscopy, scanning electron microscopy (SEM)and Dead/Live staining method.Results SEM and Micro-CT revealed a 3-D interconnected porous structure, with the CECM pore diameter of 155 ± 34 μm and the porosity of 91.3% ± 2.0%.Cytotoxicity testing with MTT revealed no significant difference in absorbance among different extracts, showing no cytotoxic effect of the scaffold on BMSCs.Inverted microscopy and SEM showed that the novel scaffold could provide a suitable 3-D environment to support the adhesion, proliferation and differentiation of BMSCs to chondroeytes in culture with chondrogenic medium.Confocal microscopy of cell-scaffold constructs revealed cells with green fluorescence.Conclusion Since the novel CECM/ACBM bilayered integrated osteochondral scaffold has good mircostructure, non-toxicity and good biocompatibility, it may be a suitable candidate as an alternative cell-carrier for osteochondral tissue engineering.     

胶原/羟基磷灰石骨软骨一体化支架的生物学特性研究

[目的]对体外条件下胶原/羟基磷灰石一体化复合材料的生物学特性进行评价,探讨其作为骨软骨组织工程支架的可行性。[方法]以胶原和羟基磷灰石为原料,研制出由胶原逐层过渡到羟基磷灰石为主的一体化支架材料,其软骨层主要成分为胶原,骨层主要成分为羟基磷灰石。通过扫描电镜观察材料内部结构及孔径大小,液体位移法测定材料的孔隙率。将材料与兔骨髓基质细胞复合培养,MTT法测定细胞的生长曲线,扫描电镜观察细胞在材料上的生长粘附情况。[结果]胶原/羟基磷灰石一体化复合支架具有疏松多孔结构,其中软骨层孔径大小(90±15)μm,骨层孔径大小(120±20)μm,孔隙率(75±5.0)%。细胞在材料上生长良好。[结论]胶原/羟基磷灰石一体化复合材料具有适宜的孔隙结构和良好的生物相容性,可用作骨软骨组织工程支架材料。     

Biomimetic bilayered gelatin-chondroitin 6 sulfate-hyaluronic acid biopolymer as a scaffold for skin equivalent tissue engineering

In order to develop an adequate scaffold for skin tissue engineering, a bilayered gelatin-chondroitin 6 sulfate-hyaluronic acid membrane with a different pore size on either side was prepared. A rete ridges-like topographic microporous structure, which provided the paracrine crosstalk in the epithelial-mesenchymal interactions, was formed. Chondroitin-6-sulfate and hyaluronic acid were incorporated within the gelatin membrane to mimic skin composition and create an appropriate microenvironment for cell proliferation, differentiation, and migration. In the study, the lower layer of the membrane (pore size: 150 microm) was seeded with dermal fibroblasts and acted as the feeder layer for keratinocyte inoculation. Meanwhile, the upper layer (pore size: 20-50 microm) was seeded with keratinocytes for epidermalization. The dermal fibroblasts were dynamically seeded in a self-designed spinner flask for more even cell distribution. The keratinocytes were cultured in submerged conditions for 5 days and then in an air-liquid interface condition for further differentiation. After being cultured for 21 days, the upper layer, seeded with keratinocytes, developed into an epidermis-like structure while the lower part, which was seeded with dermal fibroblasts developed into a dermis-like structure. A histological examination and immunostain were used to prove that keratinocytes maintain their phenotype and stratified epidermis layers were formed within 21 days. In brief, the bilayered skin substitute with biological dermal analog and epidermal structure was successfully fabricated. From this study, we can suggest that the culture model is suitable for autologous skin equivalent preparation.     

壳聚糖/羟基磷灰石支架修复骨软骨缺损的实验研究

[目的] 探讨双层壳聚糖(chitosan CS)/羟基磷灰石复合支架(hydroxyapatite HA)修复兔骨软骨缺损的可行性.[方法] 采用冻干法和烧结法制作双层壳聚糖(CS)/羟基磷灰石(HA)复合支架,以骨髓间充质干细胞为种子细胞,运用纤维蛋白胶种植技术,以双层壳聚糖(CS)/羟基磷灰石(HA)复合支架为载体,修复骨软骨缺损,实验分3组,A组:BMSc 支架,B组:单纯支架,C组:未处理.将修复材料植入骨软骨缺损模型,分别于6、12周取材,进行大体观察,组织学检测,改良Wakitani法评分,经统计学处理,比较各组修复效果差异(P＜0.05).[结果] (1)CS/HA支架CS层孔隙率为76%± 5.01%,孔径为200～400 μm,平均为300 μm左右,孔相通性好,HA层孔隙率为72%± 4.23%,孔径为200～500 μm,平均为350 μm左右,孔相通性好,结合部结合好;(2)P2骨髓间充质干细胞较纯,扫描电镜观察MSCs附着在复合支架上.大体观察和组织学检测显示, A组基本修复软骨缺损,骨缺损有骨小梁长入.B、C组骨软骨缺损修复不良,组织学检测以纤维性组织或无新生组织形成,软骨及骨缺损均明显存在,改良Wakitani评分显示A组在6周、12周2个时间点的各项评分结果,均优于B、C组,且差异有统计学意义(P＜0.05).[结论] 双层壳聚糖(CS)/羟基磷灰石(HA)复合支架可作为骨软骨组织工程支架,结合BMSc可修复软骨与骨的缺损,重建关节的解剖结构和功能.     

管状膨体聚四氟乙烯用于组织工程支架材料的动物实验研究

目的：研究膨体聚四氟乙烯(ePTFE)植入动物体内后与机体的关系。方法：管状膨体聚四氟乙烯两端闭合，内充DMEM培养液，埋入家兔皮下，观察宿主生命情况，于术后1、2、4、6周取出标本，行大体及光镜下观察。结果：家兔在植入膨体聚四氟乙烯材料及DMEM培养液后存活情况良好，DMEM培养液在1周时即已完全无色透明；4周时膨体聚四氟乙烯与周围组织有粘连，6周时粘连紧密；光镜下观察，各时间点材料间隙均可见着色，1周时可见少量淋巴细胞浸润，无血管及组织细胞。材料内壁未见细胞附着。结论：植入膨体聚四氟乙烯材料及少量DMEM培养液对宿主动物存活无影响；管内外液体可相互交通，管状膨体聚四氟乙烯可作为组织工程化尿道的支架材料。     

不同比例骨软骨镶嵌成形术联合基因增强组织工程方法治疗骨软骨缺损

目的：研究骨软骨镶嵌成形术与联合基因增强组织工程方法治疗急性骨软骨缺损,并观察在不同比例下的组织修复情况,以期发现两者间的最佳组合。方法：对携带hTGF-β1的重组腺病毒转染BMSCs（hTGF-β1转染组）,与采用Adv-βgal转染BMSCs（Adv-βgal转染组）及未转染BMSCs（未转染空白对照组）进行WesternBlot检测hTGF-β1、Ⅱ型胶原及Aggrecan表达。雄性6月龄崇明山羊18只,体重22～25kg,取自体骨髓进行BMSCs分离及培养,传至第3代。每只动物双膝股骨内髁进行实验,分为AR、AL、BR、BL、CR、CL6组。AR为骨软骨移植覆盖面积为44.44%单纯组,AL为44.44%联合组,BR为33.33%单纯组,BL为33.33%联合组,CR为22.22%单纯组,CL为22.22%联合组。于双膝股骨内髁负重区采用骨钻制备直径为9mm,深为3mm的骨软骨缺损后,单纯组采用骨软骨镶嵌成形术的自体骨软骨柱修复,联合组同时将转染hTGF-β1的BMSCs藻酸钠混合液注入空隙,加入氯化钙产生凝胶。术后24周取材,行大体及组织学观察,并参照O＇Driscoll,KeeleyandSalter组织形态学评分标准进行评分,行免疫组织化学及透射电镜观察。结果：hTGF-β1转染组细胞hTGF-β1、Ⅱ型胶原及Aggrecan表达均强于Adv-βgal转染组及未转染空白对照组。大体观察,组织学染色以及透射电镜检查显示各组的缺损有不同程度的修复。33.33%联合组、44.44%单纯组、44.44%联合组的整体修复效果差别无统计学意义,33.33%单纯组、22.22%单纯组、22.22%联合组的整体修复效果差于前3组。结论：骨软骨镶嵌成形术联合基因增强组织工程的方法可以有效修复骨软骨缺损。随着自体骨软骨移植覆盖缺损面积的减少,骨软骨镶嵌成形术联合基因增强组织工程方法的修复优越性可以得到更好的体现。     

以聚羟基丁酸已酯为支架材料的组织工程隆鼻法的实验研究

目的:研究以聚羟基丁酸已酯(PHB)为支架材料形成的组织工程软骨作为隆鼻填充材料的可行性。方法:实验组:取兔的耳软骨细胞,行体外培养扩增,达到一定数量后,再接种于PHB支架上,置于新西兰白兔的鼻背部骨膜下,从大体和组织学方面观察软骨形成情况;对照组:单纯以未接种软骨细胞的支架移植于兔的鼻背部骨膜下,于相同时间点进行检测。结果:实验组:植入4周后该复合物在兔鼻背下支架表层有软骨层形成,并可见软骨细胞突入至支架材料中间。8周后支架材料继续降解,软骨细胞及细胞基质生成明显,伴有少量血管及胶原长入,对照组无相应变化。结论:以聚羟基丁酸已酯(PHB)为支架材料的组织工程软骨可用于隆鼻的填充材料。     

组织工程骨一软骨复合组织体内异位移植的研究

目的：本实验构建组织工程骨软骨复合组织并采用异体异位移植方式,探讨外来因素对移植物的影响,为进一步研究奠定基础。方法：制备明胶一硫酸软骨素一透明质酸及明胶一陶瓷化骨多孔复合支架,复合软骨细胞和骨髓间充质干细胞,将上述组织按照体外培养时间不同分为4纽,植入裸鼠皮下,观察复合组织形成情况。结果：各实验纽在观察期内裸鼠背部均无感染,复合组织表面完整薄层纤维层下可见类似透明样软骨组织,体积均出现不同程度的收缩。HE染色各实验组均可发现,在软骨层和骨层,分别可见成软骨细胞、软骨细胞、骨细胞及相应的细胞外基质,且可以观察到类似正常结构的潮线出现,交界区软骨组织出现骨化,在组织外围,出现了不同程度向纤维样组织转变的现象。结论：经过体外短时间的诱导使得细胞具有分化趋势后,早期植入体内有助于组织工程骨一软骨复合组织的形成。     

Mechanical and structural properties of a novel hybrid heart valve scaffold for tissue engineering

Hybrid heart valve scaffolds were fabricated from decellularized porcine aortic heart valve matrices and enhanced with bioresorbable polymers using different protocols: (i) dip coating of lyophilized decellularized matrices, and (ii) impregnation of wet decellularized matrices. The following polymers were evaluated: poly(4-hydroxybutyrate) and poly(3-hydroxybutyrate-co4-hydroxybutyrate). Tensile tests were conducted to assess the biomechanical behavior of valve leaflet strips. Suture retention strength was evaluated for the adjacent conduit. A pulse duplicator system was used for functional testing of the valves under physiological systemic load conditions. The properties of the hybrid structures were compared with native, decellularized, and glutaraldehyde-fixed specimens. Mechanisms of the polymer impregnation process were studied with IR spectroscopy, fluorescent microscopic imaging, and SEM. Altogether this study demonstrates the feasibility and improved biomechanical function of a novel hybrid heart valve scaffold for an application in tissue engineering.