透明质酸改性壳聚糖-胶原-羟基磷灰石复合支架力学特性及成骨细胞的增殖(英文)

Mechanical properties of hyaluronic acid modifying chitosan/collagen/nano-hydroxyapatite composite scaffold and its effect on osteoblast proliferation

背景:组织工程中,种子细胞需依赖于细胞外基质的存在才能发挥功能。因此支架材料的选择具有重大意义。目的:制备一种新型改性壳聚糖-胶原-羟基磷灰石复合支架,优化易于细胞黏附的组织工程支架材料工艺。方法:壳聚糖与透明质酸进行交联,红外和差示扫描量热图谱检测其结构;改性壳聚糖与胶原按1:2,1:1和2:1制备3种改性壳聚糖-胶原-羟基磷灰石复合支架,将复合支架与成骨细胞MC3T3-E1联合培养,CCK-8法检测增殖,绘制生长曲线。结果与结论:透明质酸和壳聚糖以酰胺键形成交联的新化合物,孔径在50～250μm之间,孔隙率随着胶原水平、弹性模量的增加而增加,而密度则减少;增加胶原的含量在细胞联合培养初期有利于细胞对支架的黏附和增殖,但从第10天开始,3种样品中细胞数量相差不大,均出现平台期;苏木精-伊红染色发现成骨细胞在培养初期沿着支架材料内部空隙贴壁生长,随着培养天数的增加,贴壁细胞呈集落样生长,可明显看到细胞间连接。说明透明质酸改性壳聚糖/胶原/纳米羟基磷灰石复合材料可以作为骨支架材料供成骨细胞黏附、增殖,其中胶原与壳聚糖的体积比为1:1为较优配比。

BACKGROUND：Seed cell exerting its function is required to depend on the extracellular matrix in tissue engineering,so that biocompatible material is important to be selected.OBJECTIVE：To prepare a novel composite scaffolds of chitosan/collagen/nano-hydroxyapatite（HA-CS/Col/nHAP） and to optimize the technology of tissue engineered-stents according to the circumstances of cell adhesion.METHODS：Chitosan was modified by hyaluronate acid.The structure was observed by differential scanning calorimetry and the Fourier transformed infrared spectroscopy.Three composites of HA-CS/Col/nHAP according to different ratio of chitosan and collagen solution（1：2;1：1 and 2：1） were prepared.The composite scaffolds were co-cultured with osteoblast MC3T3-E1,and the proliferation and cell growth curve were measured by CCK-8 method.RESULTS AND CONCLUSION：Hyaluronic acid and chitosan were crosslinked with amide linkage.Pore size was on the range from 50 μm to 250 μm.Porosity was increased with increased collagen level and elastic modulus,but density was reduced.Increased collagen content was beneficial for cell adhesion and proliferation on stent in the primary phase of cell co-culture.However,from day 10,no significant difference was determined among three samples.At the beginning of cell culture,cells adhered to the airspace insides the composite scaffolds.In the following days,cells grew in a colony manner,and cell-cell junction could be easily observed.These indicate that HA-CS/Col /nHAP composite scaffolds can improve the adhesion and proliferation of osteoblast.The ratio of chitosan to collagen volume at 1：1 was optimal.