Department of Periodontology, School of Dentistry, Wonkwang University, Iksan, Jeonbuk 570-749, Korea.
J Biomed Mater Res A. 2013 Jun;101(6):1550-8. doi: 10.1002/jbm.a.34456. Epub 2012 Nov 7.
Scaffolds can be used for tissue engineering because they can serve as templates for cell adhesion and proliferation for tissue repair. In this study, chitosan/hydroxyapatite (CS/HAp) composites were prepared by coprecipitation synthesis. Then, CS and CS/HAp fabrics were prepared by wet spinning. CS fibers with a diameter of 15 ± 1.3 μm and CS/HAp fibers with a diameter of 22 ± 1.2 μm were successfully produced; incorporation of HAp into the CS/HAp fibers was confirmed by X-ray diffraction analysis. Biological in vitro evaluations showed that human mesenchymal stem cells (hMSCs) cultured on CS/HAp fabric showed increased proliferation compared to those cultured on pure CS fabric, which was observed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, DNA content assay, and [(3) H] thymidine incorporation assay. Neither the CS nor CS/HAp scaffold exhibited any cytotoxicity to hMSCs, as shown by viability staining and cytotoxicity fluorescence image assays. After 10 days of culturing, the attachment of cells onto the scaffold was observed by scanning electron microscopy. Furthermore, under osteogenic differentiation conditions, alkaline phosphatase (ALP) activity and calcium accumulation was higher in cells cultured on the CS/HAp scaffold than in cells cultured on the CS scaffold. The mRNA expression of osteoblast markers, including ALP, osteocalcin, Co1Ia1, and runt-related transcription factor 2, was higher in cells cultured on CS/HAp than in cells cultured on the CS fabric. The results of this study indicate that the CS/HAp composite fabric may serve as a good scaffold for bone tissue engineering applications.
支架可用于组织工程,因为它们可以作为细胞黏附和增殖的模板,用于组织修复。在本研究中,通过共沉淀合成法制备壳聚糖/羟基磷灰石(CS/HAp)复合材料。然后,通过湿法纺丝制备 CS 和 CS/HAp 纤维织物。成功制备了直径为 15±1.3μm 的 CS 纤维和直径为 22±1.2μm 的 CS/HAp 纤维;通过 X 射线衍射分析证实了 HAp 掺入 CS/HAp 纤维中。体外生物学评价表明,与培养在纯 CS 纤维织物上的细胞相比,培养在 CS/HAp 纤维织物上的人骨髓间充质干细胞(hMSCs)增殖增加,这通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐比色法、DNA 含量测定和[(3)H]胸苷掺入测定进行观察。细胞活力染色和细胞毒性荧光图像分析表明,无论是 CS 还是 CS/HAp 支架对 hMSCs 均无细胞毒性。培养 10 天后,通过扫描电子显微镜观察细胞在支架上的附着情况。此外,在成骨分化条件下,在 CS/HAp 支架上培养的细胞的碱性磷酸酶(ALP)活性和钙积累高于在 CS 支架上培养的细胞。在 CS/HAp 上培养的细胞的成骨细胞标志物(包括 ALP、骨钙素、Co1Ia1 和 runt 相关转录因子 2)的 mRNA 表达高于在 CS 纤维织物上培养的细胞。本研究结果表明,CS/HAp 复合纤维织物可用作骨组织工程应用的良好支架。