Published online Dec 26, 2018. doi: 10.4252/wjsc.v10.i12.212
Peer-review started: August 9, 2018
First decision: August 31, 2018
Revised: October 18, 2018
Accepted: November 7, 2018
Article in press: November 7, 2018
Published online: December 26, 2018
To evaluate the angiogenic effect of platelet-rich plasma (PRP)-preconditioned adipose-derived stem cells (ADSCs) both in vitro and in a mouse ischemic hindlimb model.
ADSCs were divided based on culture medium: 2.5% PRP, 5% PRP, 7.5% PRP, and 10% PRP. Cell proliferation rate was analyzed using the MTS assay. The gene expression of CD31, vascular endothelial growth factor, hypoxia-inducible factors, and endothelial cell nitric oxide synthase was analyzed using reverse transcription polymerase chain reaction. Cell markers and structural changes were assessed through immunofluorescence staining and the tube formation assay. Subsequently, we studied the in vivo angiogenic capabilities of ADSCs by a mouse ischemic hindlimb model.
The proliferation rate of ADSCs was higher in the 2.5%, 5%, and 7.5% PRP groups. The expression of hypoxia-inducible factor, CD31, vascular endothelial growth factor, and endothelial cell nitric oxide synthase in the 5% and 7.5% PRP groups increased. The 5%, 7.5%, and 10% PRP groups showed higher abilities to promote both CD31 and vascular endothelial growth factor production and tubular structure formation in ADSCs. According to laser Doppler perfusion scan, the perfusion ratios of ischemic limb to normal limb were significantly higher in 5% PRP, 7.5% PRP, and human umbilical vein endothelial cells groups compared with the negative control and fetal bovine serum (FBS) groups (0.88 ± 0.08, 0.85 ± 0.07 and 0.81 ± 0.06 for 5%, 7.5% PRP and human umbilical vein endothelial cells compared with 0.42 ± 0.17 and 0.54 ± 0.14 for the negative control and FBS, P < 0.01).
PRP-preconditioned ADSCs presented endothelial cell characteristics in vitro and significantly improved neovascularization in ischemic hindlimbs. The optimal angiogenic effect occurred in 5% PRP- and 7.5% PRP-preconditioned ADSCs.
Core tip: We reported the in vitro angiogenic effect of platelet-rich plasma (PRP) treated adipose-derived stem cells (ADSCs) and the neovascularization ability of these cells in animal models. This is significant because we demonstrated that ADSCs presented endothelial cell characteristics after PRP treatment. We were the first to observe that treatment with PRP-preconditioned ADSCs significantly enhanced circulation in mouse ischemic hindlimbs models. Our result further showed that 5% and 7.5% PRP exerted the optimal effect on promoting angiogenesis of ADSCs and improving perfusion. We developed a stem cell-based, safe, and efficient way to promote peripheral circulation in animal model.