Abstract
Scaffolds constitute an important element in vascularized tissues and are therefore investigated for providing the desired mechanical stability and enabling vasculogenesis and angiogenesis. In this study, supplementation of hydrogels containing either MatrigelTM and rat tail collagen I (MatrigelTM/rCOL) or human collagen (hCOL) with SeaPlaqueTM agarose were analyzed with regard to construct thickness and formation and characteristics of endothelial cell (EC) networks compared to constructs without agarose. Additionally, the effect of increased rCOL content in MatrigelTM/rCOL constructs was studied. An increase of rCOL content from 1 mg/mL to 3 mg/mL resulted in an increase of construct thickness by approximately 160%. The high rCOL content, however, impaired the formation of an EC network. The supplementation of MatrigelTM/rCOL with agarose increased the thickness of the hydrogel construct by approximately 100% while supporting the formation of a stable EC network. The use of hCOL/agarose composite hydrogels led to a slight increase in the thickness of the 3D hydrogel construct and supported the formation of a multi-layered EC network compared to control constructs. Our findings suggest that agarose/collagen-based composite hydrogels are promising candidates for tissue engineering of vascularized constructs as cell viability is maintained and the formation of a stable and multi-layered EC network is supported.
Highlights
Tissue engineering aims to develop in vitro tissues to improve or replace impaired biological tissues in vivo
GFP-human umbilical vein endothelial cells (HUVECs) exhibited a round morphology in 3 mg/mL rCOL (Figure 1b)
After 9 days of culture, a formation of a stable HUVEC network was observed in control construct (Figure 1c), while a low GFP-HUVECs cell number was observed in 3 mg/mL rCOL hydrogels developing a more elongated morphology compared to day 2 and forming only short vascular-like structures (Figure 1d)
Summary
Tissue engineering aims to develop in vitro tissues to improve or replace impaired biological tissues in vivo. Clinical applications of tissue engineering are limited to tissues which do not require vascularization nor anastomosis [1,2,3,4]. The 3D engineered tissues exceeding 200 μm (the effective diffusion limit of oxygen), which are densely populated with cells, quickly develop a necrotic core, if they are not vascularized [5]. The vascularization of the engineered tissue is a major challenge in tissue engineering. Research advanced in pre-vascularization strategies with the aim to create vascular networks resembling the physiological capillary bed.
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