Abstract
Identification of a suitable cell source combined with an appropriate 3D scaffold is an essential prerequisite for successful engineering of skeletal tissues. Both osteogenesis and angiogenesis are key processes for bone regeneration. This study investigated the vascularization potential of a novel combination of human dental pulp stromal cells (HDPSCs) with 45S5 Bioglass® scaffolds for tissue-engineered mineral constructs in vivo and in vitro. 45S5 Bioglass scaffolds were produced by the foam replication technique with the standard composition of 45 wt% SiO2, 24.5 wt% Na2O, 24.5 wt% CaO, and 6 wt% P2O5. HDPSCs were cultured in monolayers and on porous 45S5 Bioglass scaffolds under angiogenic and osteogenic conditions for 2–4 weeks. HDPSCs expressed endothelial gene markers (CD34, CD31/PECAM1, and VEGFR2) under both conditions in the monolayer. A combination of HDPSCs with 45S5 Bioglass enhanced the expression of these gene markers. Positive immunostaining for CD31/PECAM1 and VEGFR2 and negative staining for CD34 supported the gene expression data, while histology revealed evidence of endothelial cell-like morphology within the constructs. More organized tubular structures, resembling microvessels, were seen in the constructs after 8 weeks of implantation in vivo. In conclusion, this study suggests that the combination of HDPSCs with 45S5 Bioglass scaffolds offers a promising strategy for regenerating vascularized bone grafts.
Highlights
Vascularization of tissue-engineered constructs is essential to allow diffusion of oxygen and nutrients to the constructs’ center, avoiding lack of perfusion that might otherwise lead to central necrosis.[1]
We have investigated the expression of endothelial cell markers by Human dental pulp stem/stromal cells (HDPSCs) in monolayer culture, 3D culture, and after in vivo implantation to test the hypothesis that this novel combination might promote and support construct vascularization
A standard set of characterization techniques was used to investigate the properties of these 45S5 Bioglass scaffolds to ascertain whether they were suitable for the cell culture work and to prove that they aligned with previous results.[25,26]
Summary
Vascularization of tissue-engineered constructs is essential to allow diffusion of oxygen and nutrients to the constructs’ center, avoiding lack of perfusion that might otherwise lead to central necrosis.[1]. Human dental pulp stem/stromal cells (HDPSCs) are a population of cells present in the pulp, the vital part of adult and deciduous teeth They are multipotent, highly proliferative, and capable of producing mineralized nodules in vitro or forming a pseudo pulp/dentin complex and/or bone in vivo depending on the site of implantation.[4,5,6] HDPSCs are known to express several stem cell surface antigen markers, among which are vascular-associated and smooth muscle markers, such as vascular cell adhesion molecule (VCAM), alpha smooth muscle actin, and melanoma-associated antigen/mucin-18 (MUC-18)/CD146, which identifies stem cells of the endothelial lineage. They are potentially predisposed to default toward angiogenic differentiation.[8,9,10,11]
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