牵张力调控3D打印组织血管形成的初步研究  

A preliminary study on the regulation of 3D printing tissue angiogenesis by stretch tension

作　　者：曹高标 顾诚 乐颖影 巨积辉 张广亮 侯瑞兴 Cao Gaobiao;Gu Cheng;Le Yingying;Ju Jihui;Zhang Guangliang;Hou Ruixing(Department of Hand Surgery,Suzhou Ruihua Orthopedic Hospital,Suzhou 215104,China)

机构地区：[1]扬州大学教学医院苏州瑞华骨科医院手外科,苏州215104

出　　处：《中华手外科杂志》2022年第3期219-224,共6页Chinese Journal of Hand Surgery

基　　金：国家自然科学基金项目(31900969);苏州市第二批科技发展计划(SYSD2020052);苏州瑞兴医院集团科研项目(RX2020002)。

摘　　要：目的探究牵张力对3D打印组织血管形成的调控作用及其涉及的分子机制。方法以脐静脉内皮细胞、成纤维细胞为种子细胞,明胶、透明质酸、纤维蛋白为支架材料,进行3D打印。以聚己内酯(polycaprolactone,PCL)支架固定打印组织建立牵张力调控血管形成模型。用活/死细胞染色试剂盒检测打印组织内细胞的活性。通过免疫荧光染色CD31观察血管形成,实时定量PCR检测CDC42表达。结果3D打印组织培养14 d,细胞活性为(96.7±0.7)%。与对照组打印组织相比,牵张力组打印组织血管生长方向与牵张力方向基本一致,血管分支数量显著减少。进一步研究发现,牵张力组打印组织CDC42表达显著降低,用含CDC42激动剂的培养基培养牵张力组打印组织能显著增加血管分支数量。结论牵张力对3D打印组织血管分支形成具有抑制作用,可能通过下调CDC42表达减少血管分支形成。该研究可能为组织工程皮肤血管化的调控提供有效方法。Objective To explore the regulatory effect of stretch tension on 3D printing tissue angiogenesis and its molecular mechanism.Methods Umbilical vein endothelial cells and fibroblasts were used as seed cells,and gelatin,hyaluronic acid and fibrin were used as scaffold materials for 3D printing.Polycaprolactone(PCL)scaffold was used to fix the printed tissue to establish the model of angiogenesis regulated by stretch tension.The activity of cells in the printed tissue was tested using a live/dead cell staining kit.Angiogenesis was observed by immunofluorescence staining for CD31,and the expression of CDC42 was detected by real-time quantitative PCR.Results The cell viability of 3D printed tissue cultured for 14 days was(96.7±0.7)%.Compared with the control group,the vascular growth direction of the stretch tension group was basically the same as that of stretch tension,and the number of vascular branches was significantly reduced.Further studies showed that CDC42 expression was significantly decreased in the stretch tension group,and the number of vascular branches was significantly increased in the stretch tension group cultured in media containing CDC42 agonists.Conclusions The stretch tension can inhibit the formation of vascular branches in 3D printing tissues,and may reduce the formation of vascular branches by down-regulating CDC42 expression.This study may provide an effective method for the regulation of tissue engineering skin vascularization.

关 键 词：内皮细胞 透明质酸 3D打印 牵张力 血管形成 CDC42 

分 类 号：R318.08[医药卫生—生物医学工程]