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目的:探究新型自组装多肽水凝胶RATEA16支架对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)增殖及其载血管内皮生长因子(vascular endothelial growth factor,VEGF)促进血管形成的作用。方法:制备RATEA16水凝胶,检测其可注射性、微观结构、降解性能及生物相容性等特性;与HUVEC体外三维培养,检测细胞形态及增殖情况;将HUVEC细胞与装载VEGF的RATEA16支架体外三维共培养,倒置显微镜下观察小管形成并通过实时荧光定量PCR检测VEGF-A、血管性假血友病因子(von Willebrand factor,vWF)、基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)和血小板内皮细胞黏附分子1(platelet endothelial cell adhesion molecule-1,PECAM-1)基因的表达,评估载VEGF支架体系对HUVEC分化的影响,并与HUVEC单独培养的阴性对照组进行比较。结果:RATEA16溶液在调节pH值达中性后5 min完成溶胶-凝胶转变;该凝胶可从注射器中轻松推出,具备可注射性;扫描电子显微镜下呈多孔及相互连接的内部结构,支架孔隙率为(67.3±9.4)%;在体外降解4周时剩余质量百分比仍为(82.354±0.006)%;HUVEC在RATEA16水凝胶浸提液中培养24 h后生长良好,与对照组相比HUVEC细胞活力差异无统计学意义(n P>0.05);HUVEC在该水凝胶中三维培养时呈球形,且在14 d内呈持续增殖活力;装载VEGF的RATEA16支架与HUVEC体外三维培养可观察到血管样结构形成,VEGF-A和MMP-9表达是阴性对照组的1.5~2.0倍,vWF表达是阴性对照组的10倍左右,PECAM-1表达是阴性对照组的55倍左右,与阴性对照组相比差异均有统计学意义(n P0.05). HUVEC encapsulated in RATEA16 hydrogel appeared round in shape and exhibited effectively continuous proliferation. When HUVEC were cultured on RATEA16 hydrogel with VEGF for 24 h, the formation of vascular-like structures was observed. The expression of VEGF-A and MMP-9 was 1.5-2.0 times that of control group, and vWF was 10 times and PECAM-1 was 55 times compared with that of the control group (n P<0.05).n Conclusions:The RATEA16 hydrogel used in this study could be prepared by simply adjusting pH to neutral. This hydrogel exhibited good biodegradability, slow degradation and injectability. HUVEC might attach and spread in RATEA16 scaffold. The RATEA16 scaffold with VEGF could promote angiogenic differentiation of HUVEC. The novel scaffold is expected to achieve the critical vascularization process in bone tissue regeneration.