小肠黏膜下层促血管内皮细胞体外增殖及成血管化作用.

焦自钊; 薛武军; 安茂竹; 付海霞; 李 杨; 李凤楼; 盛练芬

Objective To investigate the role of small intestinal submucosa (SIS) in promoting the proliferation and vascularization of umbilical vein endothelial cells (UVECs) in vitro. Methods UVECs were isolated by collagenase digestion method, and then cultured and identified by morphology and immunofluorescence. SIS was prepared by tissue engineering technique; then its components were analyzed by immunohistochemistry and enzyme linked immunosorbent assay (ELISA) methods. The co-culture system of UVECs and SIS was established; UVECs cultured alone were set as the control group. The proliferation and vascularization of UVECs were compared between the two groups. Results Immunohistochemical test showed that the prepared SIS was mainly composed of the extracellular matrix components type Ⅰ and Ⅲ collagens. The concentrations of vascular endothelial growth factor (VEGF) and basic fibroblast growth factors (b-FGF) in SIS incubation solution detected by ELISA were (48.83±5.67)ng/mL and (72.36±8.71)ng/mL, respectively. When cultured in vitro, there were angiogenesis and high cell density in UVECs and SIS co-cultured group than in UVECs cultured alone group. Growth curve of UVECs obtained by UVECs count showed that the proliferation ability of UVECs in UVECs and SIS co-cultured group was higher than that in the cultured alone group. Growth curve of UVECs showed at days 4 and 5 of logarithmic phase, UVECs count in UVECs and SIS co-cultured group was (7.91±0.42)×104 and (9.15±0.48)×104, while that in the UVECs cultured alone group was (4.26±0.27)×104 and (6.71±0.31)×104(P<0.01). Population doubling time of UVECs in UVECs and SIS co-cultured group was shorter than that in the UVECs cultured alone group (P< 0.01). Conclusion SIS can promote the proliferation and vascularization of UVECs in vitro by its extracellular matrix components type I and III collagens, and its components VEGF and b-FGF.

Journal of Xi'an Jiaotong University (Medical Sciences), 2018, Vol 39, Issue 6, p805

1671-8259

Academic Journal

10.7652/jdyxb201806007