仿生组装纳米羟基磷灰石/壳聚糖骨修复材料的制备及其生物相容性研究

目的 探讨以一种简单、廉价的方法制备纳米羟基磷灰石/壳聚糖(n-HA/CS)复合材料,并评价其理化特征和生物相容性. 方法采用原位沉析和冷冻干燥法制备n-HA/CS支架,通过扫描电镜、组织切片染色、X线衍射和傅立叶红外光谱分析其微观形貌和组成;采用万能材料试验机分析材料的力学性能.采用材料浸提液和表面接种考察n-HA/CS复合材料对第3代人骨髓基质干细胞(hBMSCs)黏附、增殖的影响,评估其细胞相容性.将n-HA/CS复合材料植入新西兰大白兔背部肌袋,经组织学染色后评价其组织相容性. 结果 n-HA/CS复合材料具有多孔结构,孔隙率为(88.65±2.34)%,孔径为(112.63±20.47) μm,HA晶体颗粒长度为200～700 nm,且分散均匀;X线衍射和红外光谱分析表明合成的HA是含CO32-弱结晶纳米晶体.材料的断裂强度为(1.47±0.15)MPa,弹性模量为(37.52±3.43)kPa,可满足非负重部位骨修复要求.n-HA/CS材料浸提液未明显抑制hBMSCs的增殖,直接接种在n-HA/CS复合材料表面的细胞黏附、增殖功能正常;组织相容性实验也表明,植入4周后组织炎性反应明显减轻,12周后材料基本降解并由新生组织爬行替代. 结论采用原位沉析和冷冻干燥法制备的n-HA/CS复合材料具有良好的理化性质和生物相容性,有望应用于组织工程骨的构建.

Biomimetic synthesis and biocompatibility of nano-hydroxyapatite/chitosan scaffold for bone tissue engineering

Objective To evaluate the physicochemical properties and biocompatibility of nano-hydroxyapatite/chitosan (n-HA/CS) scaffolds. Methods n-HA/CS scaffolds were generated by in situ hybridization and freeze-drying technology. The microscopic morphology and components of the composite were analyzed by scanning electron microscopy (SEM), morphology, X-ray diffraction (XRD) examination and Fourier transformed infrared spectroscopy (FTIR) . The effects of u-HA/CS scaffolds on adherence and proliferation of human bone marrow stromal cells (hBMSCs) were evaluated by leaching liquor and scaffolds surface seeding methods. After being implanted into back muscle pockets of New Zealand rabbits, the histo-compatibility of n-HA/CS was observed by histological staining. Results n-HA/CS composite showed abundant homogeneous pores with (112.63 ± 20.47) μm diameter and (88.65 ± 2.34)% porosity. HA particles were distributed on the pore walls homogeneously with (200 ～ 700 nm) nanoscale. XRD and FTIR results showed that the HA crystals were carbonate-substituded and not well-crystallized. Cytocompatibility teat showed that the seeded hBMSCs could adhere to the scaffolds, and their proliferation ability was not affected by n-HA/CS composite and its leaching liquor. In addition, histocompatibility test found that tissue inflammatory reactions of n-HA/CS composite implanted decreased significantly at 4 w, and the composite was degraded mostly and was substituted by new tissue at 12 w. Conclusion The n-HA/CS scaffolds could be successfully used in the bone tissue engineering, as it has good physicochemical properties and fine biocompatibility.