【摘要】：關(guān)節(jié)骨軟骨承載了體重、滑動(dòng)、減震等生理功能,在動(dòng)物體中起著非常重要的作用,同時(shí)由于軟骨組織缺乏血管、淋巴細(xì)胞,導(dǎo)致了關(guān)節(jié)軟骨損傷很難自愈。經(jīng)多年臨床研究,建立了包括拋削術(shù)、清創(chuàng)術(shù)、異體移植術(shù)和自體移植術(shù)在內(nèi)的傳統(tǒng)治療方法。然而,這些方法的治療效果尚不能令人滿意。拋削術(shù)、清創(chuàng)術(shù)會(huì)造成二次傷害,且新生軟骨易發(fā)生退化；異體軟骨移植會(huì)引起免疫反應(yīng),導(dǎo)致?lián)p傷修復(fù)失��；自體軟骨移植受材料來源限制,而不能用于大面積軟骨損傷的修復(fù)。因此,關(guān)節(jié)骨軟骨損傷的治療成為世界性的醫(yī)療難題,組織工程研究為關(guān)節(jié)骨軟骨修復(fù)帶來了希望。 目前,關(guān)節(jié)組織工程研究集中在種子細(xì)胞、支架制備、細(xì)胞生長因子三個(gè)方面,其中制備支架是開展此項(xiàng)研究最基本的條件。在以往骨組織工程支架的研究中研制出了多種骨或軟骨單層支架,單層軟骨支架忽視了軟骨下骨層作為軟骨支撐點(diǎn)并為軟骨提供營養(yǎng)的重要功能。為此,必需設(shè)計(jì)一種仿生骨軟骨一體化支架以滿足骨軟骨損傷組織工程修復(fù)的需要。 本文進(jìn)行了羥基丁酸與羥基辛酸共聚物(PHBHOx)一體化骨軟骨支架的研制,從乳兔提取了骨髓間充質(zhì)干細(xì)胞,并初步進(jìn)行了支架與細(xì)胞的復(fù)合培養(yǎng)研究。 采用溶劑澆注/顆粒瀝濾法,以三氯甲烷為溶劑、PHBHOx為主材料和連續(xù)相,通過分別添加不同粒徑與比例致孔劑,一次性制備由骨層-中間過渡層-軟骨層組成的一體支架。仿照骨軟骨解剖學(xué)特征與物理學(xué)特性,一體支架的各層應(yīng)該結(jié)構(gòu)不同,性能有別,且三層之間彼此連通。經(jīng)過制備條件優(yōu)化,成功制備出骨軟骨一體化支架,其三層孔徑分別約為280μ m,200μ m,≤60μ m,孔隙率分別約為75%,85%,60%,其力學(xué)強(qiáng)度分別為4.23MPa,2.75MPa,6.30MPa,基本上達(dá)到了組織工程修復(fù)骨軟骨損傷的要求。 本文采用全骨髓法提取兔骨髓間充質(zhì)干細(xì)胞(BMSCs),用貼壁法將BMSCs提純。經(jīng)過72小時(shí)的培養(yǎng),貼壁率達(dá)到了60%,六天后BMSCs布滿瓶底,細(xì)胞生長狀態(tài)旺盛,細(xì)胞形態(tài)穩(wěn)定。 本文還進(jìn)行了一體支架與BMSCs細(xì)胞的復(fù)合培養(yǎng)試驗(yàn),在24h后,細(xì)胞粘附率達(dá)到了76%。細(xì)胞形態(tài)良好,表明了一體化支架具有良好的生物相容性。 本文應(yīng)用專利材料PHBHOx首次成功制備了仿生骨層-中間過渡層-軟骨層相連續(xù)的一體化支架,建立了分步操作一次成型支架制作新工藝,獲得了三層結(jié)構(gòu)不同、功能有別的骨軟骨組織工程支架新產(chǎn)品。并進(jìn)行了一體支架與BMSCs細(xì)胞的復(fù)合培養(yǎng)試驗(yàn),證明了該支架有著良好的生物相容性。本文的研究進(jìn)展為關(guān)節(jié)損傷修復(fù)的深入研究提供了一定的理論和實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Articular bone cartilage bears physiological functions such as weight sliding shock absorption and plays a very important role in animals. At the same time because of the lack of blood vessels and lymphocytes in cartilage tissue it is difficult to heal the injury of articular cartilage. After many years of clinical research, traditional treatments including excision, debridement, allograft and autologous transplantation have been established. However, the therapeutic effect of these methods is not satisfactory. Dissection, debridement will cause secondary injury, and new cartilage will degenerate easily; allogenic cartilage transplantation will cause immune response, resulting in damage repair failure; autologous cartilage transplantation is limited by the source of materials. It can not be used to repair large area cartilage injury. Therefore, the treatment of articular cartilage injury has become a worldwide medical problem, tissue engineering research has brought hope for articular cartilage repair. At present, the research of joint tissue engineering focuses on seed cells, scaffold preparation and cell growth factor, among which the preparation of scaffolds is the most basic condition for this study. In previous studies of bone tissue engineering scaffolds, a variety of bone or cartilage monolayer scaffolds have been developed. Monolayer cartilage scaffolds have neglected the important function of subchondral bone layer as the supporting point of cartilage and providing nutrition for cartilage. Therefore, a biomimetic osteochondral scaffold must be designed to meet the need of tissue engineering repair of bone cartilage injury. The bone marrow mesenchymal stem cells (BMSCs) were extracted from neonatal rabbits using hydroxybutyric acid and hydroxyoctanoic acid copolymers (PHBHOx) as bone cartilage scaffolds. The co-culture of bone marrow mesenchymal stem cells (BMSCs) was studied in this paper. A monolithic scaffold composed of bone layer, intermediate transition layer and cartilage layer was prepared by solvent pouring / particle leaching method, using trichloromethane as solvent PHBHOx as main material and continuous phase, respectively, by adding different particle size and proportion of pore-forming agent. According to the anatomical and physical characteristics of osteochondral, each layer of the monolithic scaffold should have different structure and performance, and the three layers should be connected with each other. After optimization of the preparation conditions, the osteochondral scaffolds were successfully prepared. The three-layer pore sizes of the scaffolds were about 280 渭 m ~ 200 渭 m, 鈮，

本文編號(hào)：2154436