采用乳化剂交联法,以海藻酸钠(SA)为原料,Ca Cl2为物理交联剂,制备了海藻酸钠多孔支架;然后分别以0.2 mol/L Zn(Ac)2、Zn Cl2和Zn(NO3)2为交联剂,在海藻酸钠溶液质量浓度为15 g/L,温度为8℃的条件下,制备了竖直贯通的多孔支架;冷冻干燥后,将支架浸泡在KOH/甲醇/无水乙醇混合溶液中(Zn O量子点原位合成法),制备了荧光竖直定向多孔支架。研究了海藻酸钠的质量浓度(5 g/L、10 g/L、15 g/L)及不同干燥方法(冷冻干燥和乙醇逐级脱水法)对制备海藻酸钠多孔支架的影响;将成纤维细胞与海藻酸钠多孔支架共培养,考察海藻酸钠多孔支架的生物相容性,采用Cy3(Cy–N–羟基琥珀酰亚胺酯)与DAPI(4,6–二脒基–2–苯基吲哚)双荧光染色法观察细胞生长情况及采用MTT(3–(4,5–二甲基噻唑–2)–2,5–二苯基四氢唑溴盐)法定量检测了海藻酸钠多孔支架的细胞毒性;对荧光多孔支架的形貌和荧光性能进行了表征。
The sodium alginate porous scaffolds were prepared by the emulsifier cross-linking method using sodium alginate (SA) as raw material and CaC12 as physical cross-linking agent. The vertical through porous scaffolds were prepared by using 0.2 mol/L Zn(Ac)2, ZnCl2 and Zn(NO3)2 separately as the cross-linking agents when the temperature is 8 ℃, the mass concentration of the sodium alginate solution is 15 g/L. After freeze-drying, the scaffolds were immersed in the KOH/methanol/absolute ethanol mixed solution (ZnO quantum dot in-situ synthesis method), and the fluorescence vertical orientation porous scaffolds were prepared. The influence of sodium alginate mass concentration (5 g/L, 10 g/L, 15 g/L) and different drying methods (freeze-drying method and step-dehydrating method with ethanol) on preparation of the sodium alginate porous scaffolds was investigated. The fibroblast cells and sodium alginate porous scaffolds were co-cultured, the bio-compatibility of the sodium alginate porous scaffolds was investigated. The cell growth was observed by using Cy3 (Cy-N-hydroxysuccinimide ester) and DAPI (4,6-diamidino- 2-phenylindole) double fluorescence staining method, and the cell cytotoxicity of the sodium alginate porous scaffolds was quantitatively detected by using MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrahydro azole bromide salt) method. The morphology and fluorescence performance of the fluorescence porous scaffolds were characterized.