聚乳酸/壳聚糖纳米纤维/纳米羟基磷灰石支架与兔软骨细胞的相容性

背景：传统的支架材料存在疏水性强,材料表面缺乏细胞表面受体特异结合的生物活性分子,材料的酸性降解产物易引发无菌性炎性反应等不足。根据仿生原理及软骨真实结构和构成来选择和制备组织工程软骨支架能够获得理想效果。 目的：制备聚乳酸/壳聚糖纳米纤维/纳米羟基磷灰石支架,评价其与兔膝关节软骨细胞的生物相容性,探讨其应用于关节软骨组织工程的可行性。 方法：采用二次相分离技术制备聚乳酸/壳聚糖纳米纤维/纳米羟基磷灰石复合支架,将第3代新西兰兔软骨细胞接种至复合支架材料上复合培养,倒置相差显微镜下观察细胞生长情况。细胞-支架复合物在24孔板中培养5 d以后,将其植入裸鼠皮下8周。 结果与结论：聚乳酸/壳聚糖纳米纤维/纳米羟基磷灰石支架材料经化学合成后,具有合适的三维多孔结构,孔隙率为90%,孔径300~450 μm;植入裸鼠皮下8周后Ⅱ型胶原免疫组织化学染色和甲苯胺蓝染色显示细胞-支架复合物中的软骨细胞可以像天然软骨一样分泌黏多糖和Ⅱ型胶原。提示生物材料聚乳酸/壳聚糖纳米纤维/纳米羟基磷灰石对于兔软骨细胞有良好的生物相容性,可作为生物组织工程支架。

Biocompatibility between polylactic acid/chitosan nano-fiber/nano-hydroxyapatite scaffolds and rabbit chondrocytes

BACKGROUND: Traditional scaffold materials have the disadvantages of strong hydrophobic property, lack of bioactive molecules for specific binding of cell surface receptors on their surfaces, easy to lead to aseptic inflammation by their acidic degradation products, and so on. The ideal scaffolds for cartilage tissue engineering can be prepared based on bionic principle and real structure and composition of cartilage. OBJECTIVE: To prepare the polylactic acid/chitosan nano-fiber/nano-hydroxyapatite (PLLA/CSNF/nHA) scaffolds, evaluate their biocompatibility rabbit knee chondrocytes, and explore their application in articular cartilage tissue engineering. METHODS: The PLLA/CSNF/nHA scaffolds were prepared by two steps separation technology. Chondrocytes from New Zealand rabbits were seeded on the scaffolds for compound culture. The cell growth was observed under inverted phase contrast microscope. Cell-scaffold compounds were cultured in 24-well plates for 5 days, and then the compounds were implanted under the skin of nude mice for 8 weeks. RESULTS AND CONCLUSION: The prepared PLLA/CSNF/nHA scaffolds had an appropriate three-dimensional porous structure. The porosity was 90% and the pore size was 300-450 µm. Chondrocytes in cell-scaffold compounds could secrete proteoglycan and type Ⅱ collagen like natural cartilage tested by immunohistochemical staining and toluidine blue staining. It is indicated that PLLA/CSNF/nHA has a good biocompatibility with rabbit chondrocytes and can be used as a type of bio-engineered stent.