Abstract
The initial step of bone regeneration requires the migration of osteogenic cells to defective sites. Our previous studies suggest that a salmon DNA-based scaffold can promote the bone regeneration of calvarial defects in rats. We speculate that the salmon DNA may possess osteoinductive properties, including the homing of migrating osteogenic cells. In the present study, we investigated the influence of the salmon DNA on osteoblastic differentiation and induction of osteoblast migration using MG63 cells (human preosteoblasts) in vitro. Moreover, we analyzed the bone regeneration of a critical-sized in vivo calvarial bone defect (CSD) model in rats. The salmon DNA enhanced both mRNA and protein expression of the osteogenesis-related factors, runt-related transcription factor 2 (Runx2), alkaline phosphatase, and osterix (OSX) in the MG63 cells, compared with the cultivation using osteogenic induction medium alone. From the histochemical and immunohistochemical assays using frozen sections of the bone defects from animals that were implanted with DNA disks, many cells were found to express aldehyde dehydrogenase 1, one of the markers for mesenchymal stem cells. In addition, OSX was observed in the replaced connective tissue of the bone defects. These findings indicate that the DNA induced the migration and accumulation of osteogenic cells to the regenerative tissue. Furthermore, an in vitro transwell migration assay showed that the addition of DNA enhanced an induction of osteoblast migration, compared with the medium alone. The implantation of the DNA disks promoted bone regeneration in the CSD of rats, compared with that of collagen disks. These results indicate that the salmon DNA enhanced osteoblastic differentiation and induction of migration, resulting in the facilitation of bone regeneration.
Highlights
Recent strategies for bone tissue engineering incorporate an interactive triad of viable osteocompetent cells, soluble osteoinductive factors, and osteoconductive scaffolds, with the aim of achieving satisfactory bone regeneration within the defects [1,2]
We have examined the effects of salmon DNA on the migration and differentiation of MG63 cells in vitro, as well as bone regeneration of calvarial defects in an in vivo rat model
A potential scaffold for bone regeneration is required for the migration of osteogenic cells from the surrounding tissues and the formation of a mineralized tissue
Summary
Recent strategies for bone tissue engineering incorporate an interactive triad of viable osteocompetent cells, soluble osteoinductive factors, and osteoconductive scaffolds, with the aim of achieving satisfactory bone regeneration within the defects [1,2]. The use of optimal scaffolds as osteoconductive constructs, are required for the delivery of osteogenic cells from the host tissue to the replacement tissue in the bone defects. Our group reported that the control of cell viability, flowability, soft tissue response, and biodegradation rate was dependent on the molecular weight of the DNA within the DNA and protamine complexes [5,6]. These results suggest that DNA is a potent tissue engineering candidate for use in biomaterials. The role of these complexes in the acceleration of osteogenesis remains undetermined
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