Zhu Chenhui, Fan Daidi, Duan Zhiguang, Xue Wenjiao, Shang Longan, Chen Fulin, Luo Yane
Northwest University, Xi'an, Shaanxi, China.
J Biomed Mater Res A. 2009 Jun;89(3):829-40. doi: 10.1002/jbm.a.32256.
With the increasing occurrence of vascular diseases and poor long-term patency rates of current small diameter vascular grafts, it becomes urgent to pursuit biomaterial as scaffold to mimic blood vessel morphologically and mechanically. In this study, novel human-like collagen (HLC, produced by recombinant E. coli)/chitosan tubular scaffolds were fabricated by cross-linking and freeze-drying process. The scaffolds were characterized by scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), and tensile test, respectively. Human venous fibroblasts were expanded and seeded onto the scaffolds in the density of 1 x 10(5) cells/cm(2). After a 15-day culture under static conditions, the cell-polymer constructs were observed using SEM, confocal laser scanning microscopy (CLSM), histological examination, and biochemical assays for cell proliferation and extracellular matrix production (collagen and glycosaminoglycans). Furthermore, the scaffolds were implanted into rabbits' livers to evaluate their biocompatibility. The results indicated that HLC/chitosan tubular scaffolds (1) exhibited interconnected porous structure; (2) achieved the desirable levels of pliability (elastic up to 30% strain) and stress of 300 +/- 16 kPa; (3) were capable of enhancing cell adhesion and proliferation and ECM secretion; (4) showed superior biocompatibility. This study suggested the feasibility of HLC/chitosan composite as a promising candidate scaffold for blood vessel tissue engineering.
随着血管疾病的发病率不断上升以及当前小口径血管移植物的长期通畅率较低,迫切需要寻找一种生物材料作为支架,在形态和力学上模拟血管。在本研究中,通过交联和冷冻干燥工艺制备了新型类人胶原蛋白(HLC,由重组大肠杆菌产生)/壳聚糖管状支架。分别通过扫描电子显微镜(SEM)、X射线光电子能谱(XPS)和拉伸试验对支架进行了表征。将人静脉成纤维细胞进行扩增,并以1×10(5)个细胞/cm(2)的密度接种到支架上。在静态条件下培养15天后,使用SEM、共聚焦激光扫描显微镜(CLSM)、组织学检查以及细胞增殖和细胞外基质产生(胶原蛋白和糖胺聚糖)的生化分析对细胞-聚合物构建体进行观察。此外,将支架植入兔肝脏以评估其生物相容性。结果表明,HLC/壳聚糖管状支架(1)呈现相互连接的多孔结构;(2)具有理想的柔韧性(应变高达30%时具有弹性)和300±16 kPa的应力;(3)能够增强细胞黏附、增殖和细胞外基质分泌;(4)表现出优异的生物相容性。本研究表明HLC/壳聚糖复合材料作为血管组织工程有前景的候选支架具有可行性。