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AIM:To develop an in vitro three-dimensional(3-D)angiogenesis system to analyse the capillary sprouts inducedin response to the concentration ranges of basic fibroblastgrowth factor(bFGF)and vascular endothelial growth factor(VEGF)and to quantify their synergistic activity.METHODS:Microcarriers(MCs)coated with humanmicrovascular endothelial cells(HMVECs)were embeddedin fibrin gel and cultured in 24-well plates with assaymedia.The growth factors bFGF,or VEGF,or both wereadded to the system.The wells(n=8/group)were digitallyphotographed and the average length of capillary-like sprouts(ALS)from each microcarrier was quantitated.RESULTS:In aprotinin-stabilized fibrin matrix,humanmicrovascular endothelial cells on the MCs invaded fibrin,forming sprouts and capillary networks with lumina.Theangiogenic effects of bFGF or VEGF were dose-dependent inthe range from 10 to 40 ng/mL.At d 1,10 ng/mL of bFGF andVEGF induced angiogenesis with an ALS of 32.13±16.6 μmand 43.75:1:27.92 IJm,respectively,which were significantlyhigher than that of the control(5.88±4.45 μm,P<0.01),and the differences became more significant as the timeincreased.In addition,the combination of 20 ng/mL ofbFGF and VEGF each induced a more significant effect thanthe summed effects of bFGF(20 ng/mL)alone and VEGF(10 ng/mL)alone when analyzed using SPSS system forgeneral linear model(GLM)(P= 0.011),and that also exceededthe effects by 20 ng/mL of either bFGF or VEGF.CONCLUSION:A microcarrier-based in vitro three-dimensional angiogenesis model can be developed in fbrin.It offers a unique system for quantitative analysis ofangiogenesis.Both bFGF and VEGF exert their angiogeniceffects on HMVECs synergistically and in a dose-dependentmanner.
AIM: To develop an in vitro three-dimensional (3-D) angiogenesis system to analyze the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) and to quantify their synergistic activity. METHODS: Microcarriers (MCs) coated with human microvascular endothelial cells (HMVECs) were embedded in fibrin gel and cultured in 24-well plates with assay media. The growth factors bFGF, or VEGF, or both wereadded to the system.The wells (n = group) were digitallyphotographed and the average length of capillary-like sprouts (ALS) from each microcarrier was quantitated .RESULTS: In aprotinin-stabilized fibrin matrix, human microvascular endothelial cells on the MCs invaded fibrin, forming sprouts and capillary networks with lumina.Theangiogenic effects of bFGF or VEGF were dose-dependent in ranges from 10 to 40 ng / mL. At d 1, 10 ng / mL of bFGF and VEGF-induced angiogenesis with an ALS of 32.13 ± 16.6 μmand 43.75: 1: 27.92 IJm, respectively , which were significantlyhigher than that of the control (5.88 ± 4.45 μm, P <0.01), and the differences became more significant as the time increments. In addition, the combination of 20 ng / mL of bFGF and VEGF each induced a more significant effect thanthe summed effects of bFGF (20 ng / mL) alone and VEGF (10 ng / mL) alone when used using the SPSS system for general linear model (GLM) (P = 0.011), and that also exceeded the effects by 20 ng / mL of either bFGF or VEGF. CONCLUSION: A microcarrier-based in vitro three-dimensional angiogenesis model can be developed in fbrin. It offers a unique system for quantitative analysis of angiogenesis. BothGFA and VEGF exert their angiogeniceffects on HMVECs synergistically and in a dose-dependentmanner.