Search

Background: Chitosan is known as a biodegradable and non-toxic natural polymer that enhances wound healing and bone formation. The aims of this study are to evaluate the biocompatibility of chitosan nanofiber membranes and to examine the effect of the chitosan nanofiber membranes on bone regeneration in rabbit calvarial defects. Methods: In vitro cell proliferation tests using human osteosarcoma cell line MG63 and reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of alkaline phosphatase (ALP), collagen, osteocalcin (OCN), and GAPDH were done on chitosan nanofiber membranes. Chitosan nanofiber membranes were implanted in subcutaneous connective tissue and observed for 2, 4, and 6 weeks in rats. Ten-millimeter diameter round cranial defects were made in rabbits and covered by chitosan nanofiber membranes for 4 weeks. Specimens were decalcified and observed by a light microscope. Results: MG63 cells proliferated for 28 days on the chitosan nanofiber membranes and expressed ALP, collagen, OCN, and GAPDH at 2 weeks. Chitosan nanofiber membranes that were grafted into rat subcutaneous tissue maintained their shape and space for bone regeneration for as long as 6 weeks. No inflammation could be seen on the membrane surface or in the surrounding tissues. In the rabbit calvarial defects, new bone filled the defects and fused to the original old bone in 4 weeks. Conclusions: The biocompatibility of the chitosan nanofiber membrane was confirmed, with enhanced bone regeneration and no evidence of an inflammatory reaction. This experiment shows that the novel biodegradable chitosan nanofiber membrane may be useful as a tool for guided bone regeneration. [ABSTRACT FROM AUTHOR]

Background: The present study was undertaken to determine the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded biodegradable membranes on bone augmentation in a rabbit calvarial model. Methods: Five µg of rhBMP-2 was loaded into a stiff hemi-spherical dome membrane made of poly(L-lactide) and tricalcium phosphate (PLLA/TCP). The release kinetics of rhBMP-2 from the membrane were determined in vitro using a human BMP-2 immunoassay. Twelve rhBMP-2-loaded dome membranes (test group) and 12 control dome membranes (control group) were placed on the partial-thickness calvarial defects of 24 rabbits. The animals were sacrificed at 4 and 8 weeks, and undecalcified ground sections were prepared. Newly formed bone area and height were measured histomorphometrically and calculated by percentage ratio to the total submembranous space area and height below the dome. Results: In vitro release results demonstrated that rhBMP-2 was released consistently over a 4-week period following a high initial burst release on the first day. At both 4 and 6 weeks, histomorphometric analysis revealed that the test group showed significantly higher newly formed bone heights and areas than the control group (P <0.01). In the control group, new bone height was 35.3% of the dome height and the new bone area reached 8.2% of the submembranous space area at 8 weeks, while the test group reached 87.3% and 35.4%, respectively. Conclusion: These results suggest that the use of rhBMP-2-loaded PLLA/TCP membranes can result in additional bone augmentation, which is due to the osteoinductive properties of rhBMP-2 released from the membrane during healing. [ABSTRACT FROM AUTHOR]

Background; In order to achieve optimal effects, growth factors including platelet-derived growth factor (PDGF] should be delivered with a biodegradable carrier that will release therapeutic concentrations over a sufficient length of time. The purpose of this study was to evaluate the bone regenerative effect of PDGF-BB delivered with a chitosan/tricalcium phosphate (TCP) sponge carrier in a rat calvarial defect model. Methods: The PDGF-BB-loaded chitosan/TCP sponge carrier was fabricated by freeze-drying a mixture of chitosan solution and TCP powder and soaking in a PDGF-BB solution. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with 125l-labeled PDGF-BB. Chitosan/TCP sponges with and without PDGF-BB were implanted into 8 mm calvarial defects in rats. Rats were sacrificed at 2 and 4 weeks following implantation, and histologic and histomorphometrical examinations were performed. Results: In vitro evaluation demonstrated that an effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. In the histologic examination, the chitosan/TCP sponge carrier promoted osseous healing of the rat calvarial defects as compared to controls. The addition of PDGF-BB to the carrier further enhanced bone regeneration. Evidence of the degraded sponge matrix was observed mingled within the newly formed bone without connective tissue encapsulation. Conclusions: The results of this study support the use of chitosan/TCP sponges as a delivery system for growth factors and demonstrate that PDGF-BB loaded onto chitosan/TCP sponge carriers has an osteogenic effect on bone regeneration in vivo. [ABSTRACT FROM AUTHOR]

Background: Chitosan is a biodegradable natural polymer that has been shown to improve wound healing. This study aimed to develop chitosan/tricalcium phosphate (TCP) sponges as tissue engineering scaffolds for bone formation by three-dimensional osteoblast culture. Methods: The sponges were prepared by freeze-drying and cross-linking a mixture of chitosan solution with TCP. Fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 56 days. Cell proliferation, alkaline phosphatase (ALPase) activity, and calcium deposition in the cell-sponge constructs were measured at 1, 7, 14, 28, and 56 days. Histologic examination was performed with light microscopy and scanning electron microscopy. Results: Chitosan/TCP sponges supported the proliferation of osteoblastic cells as well as their differentiation as indicated by high ALPase activities and deposition of mineralized matrices by the cells. Light and scanning electron microscopic examination indicated that seeded osteoblastic ceils were well attached to sponge matrices and proliferated in a multi-layer fashion. Small bone-like spicules were observed on the sponge matrix at 14 days. Seeded cells appeared to be embedded in the newly formed tissue matrix, which is characteristic of the osteoblast differentiation and their progression into osteocytic cells. The amount of mineralized tissue formed in the sponge at 56 days was significant. Conclusions; These results suggest that the chitosan/TCP sponge is a feasible tool as a scaffolding material to grow osteoblast in a three-dimensional structure for transplantation into a site for bone regeneration. [ABSTRACT FROM AUTHOR]