Angiogenesis-promoting effect of LIPUS on hADSCs and HUVECs cultured on collagen/hyaluronan scaffolds.

Angiogenesis refers to blood vessel formation through endothelial cell migration and proliferation. Angiogenesis is crucial and beneficial for wound healing and tissue regeneration. In the current study, we prepared porous collagen and collagen/hyaluronan (Col/HA) scaffolds composed of collagen (7 mg/mL) and hyaluronan (HA) (0.5 w%, 1 w%, and 1.5 w%) as culture vehicles for coculture of human adipose-derived stem cells (hADSCs) and human umbilical vein endothelial cells (HUVECs). These scaffolds were combined with low-intensity pulsed ultrasound (LIPUS) to investigate and evaluate angiogenesis in the coculture cell/scaffold constructs in vitro and in vivo. Scaffold porosity decreased (from 74.4% to 60.7%) and readily degraded after addition of various ratios of HA. The porous scaffolds all had high water content (~98%) and similar mechanical properties. The hADSCs alone and hADSCs cocultured with HUVECs exhibited stable proliferative profiles on the Col/HA scaffolds; furthermore, LIPUS significantly enhanced cell growth on the collagen and Col/0.5HA scaffolds by approximately 1.85- and 1.5-fold, respectively, compared with the cells that did not receive LIPUS treatment. In vivo immunohistochemistry results indicated stronger immunofluorescent CD31 presence and vascular endothelial cadherin messenger RNA expression in the hADSCs/HUVECs coculture/scaffold implantation in rats that received LIPUS treatment compared with those that received no such treatment. Our results demonstrated that the hADSCs/HUVECs cocultured on fabricated collagen and Col/HA scaffolds combined with LIPUS treatment had angiogenesis-promoting capability and therapeutic potential when angiogenesis is demanded.