Abstract
IntroductionThe main aim of this study is to evaluate potential human stem cells, such as dental pulp stem cells and amniotic fluid stem cells, combined with collagen scaffold to reconstruct critical-size cranial bone defects in an animal model.MethodsWe performed two symmetric full-thickness cranial defects on each parietal region of rats and we replenished them with collagen scaffolds with or without stem cells already seeded into and addressed towards osteogenic lineage in vitro. After 4 and 8 weeks, cranial tissue samples were taken for histological and immunofluorescence analysis.ResultsWe observed a new bone formation in all of the samples but the most relevant differences in defect correction were shown by stem cell–collagen samples 4 weeks after implant, suggesting a faster regeneration ability of the combined constructs. The presence of human cells in the newly formed bone was confirmed by confocal analysis with an antibody directed to a human mitochondrial protein. Furthermore, human cells were found to be an essential part of new vessel formation in the scaffold.ConclusionThese data confirmed the strong potential of bioengineered constructs of stem cell–collagen scaffold for correcting large cranial defects in an animal model and highlighting the role of stem cells in neovascularization during skeletal defect reconstruction.
Highlights
The main aim of this study is to evaluate potential human stem cells, such as dental pulp stem cells and amniotic fluid stem cells, combined with collagen scaffold to reconstruct critical-size cranial bone defects in an animal model
To study the bone-forming ability of amniotic fluid stem cells (AFSC) and dental pulp stem cells (DPSC) on collagen scaffolds, a critical-size bone defect was obtained by a full-thickness dissection removing both the internal and external tables of compact bone and the trabecular diploë constituting the parietal skeletal segment
The stem cell–scaffold constructs were induced in osteogenic medium for 7 days, in order to have a cell population committed to osteogenic differentiation yet able to proliferate
Summary
The main aim of this study is to evaluate potential human stem cells, such as dental pulp stem cells and amniotic fluid stem cells, combined with collagen scaffold to reconstruct critical-size cranial bone defects in an animal model. Over 2 million bone replacement procedures are performed every year worldwide requiring the use of bone graft materials. This makes bone second only to blood on the list of transplanted materials. A critical-size bone defect will not heal spontaneously and tissue engineering strategies are a very promising option [5]. For this strategy, therapeutic scaffolds have been developed from various classes of natural materials including collagen, fibroin [6] and hyaluronic acid-based hydrogel [7]. Cell-free scaffolds as tissue graft substitutes have been used in the human clinical setting since Yannas and colleagues developed the collagen glycosaminoglycan scaffold for skin grafting [8,9]
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