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Background:
Survival and function of the transplanted bone grafts are crucially dependent on rapidly forming blood supply. In order to engineer functional bone tissues successfully, the scaffolds have to be designed to facilitate cellular distribution and guide tissue regeneration in three dimension manner. Pore size and interconnectivity of the pores is a critical determinant of blood vessel ingrowth. In addition to providing tissues with the appropriate architecture and acting as a template, scaffolds can also act as carriers incorporating biological molecules that are known to promote signaling pathways that influence key cell functions such as migration, proliferation and differentiation. The major aim of this study is to create an inductive bone graft with the potential to act as template for seeded cells and as a delivery system for incorporated biological molecules; the aim being to facilitate vascularisation and the synthesis of extracellular matrix and subsequent formation of functional bone tissue.
Aim/Hypothesis:
The aim of this study was to create a 3D porous β calcium metaphosphate (CMP) scaffold with physico-chemical properties similar to natural bone and further attempt to pre-vascularize in vitro by using optimized co-culture condition.
Material and Methods:
Calcium phosphate materials have been shown to interact strongly with bone, due to their similarity to bone mineral. In this study, a block porous scaffolds of β-metacalcium phosphate were produced using monocalcium phosphate mono hydrate (MCP) powder and poly vinyl alcohol (PVA) as the porogen. The MCP and PVA mixes were pressed and sintered at 900 °C to yield the porous scaffolds which were sterilized with γ radiation prior to biological evaluation. Physico-chemical properties were investigated using a number of methods such as FTIR, x-ray diffraction, microcomputed tomography. In vitro biocompatibility tests were performed to confirm that the scaffold was biosafe and able to allow cell survival, cellular ingrowth, and also incorporation of appropriate bioactive growth factors. An in vitro co-culture model with osteoprogenitor cells and endothelial cells was used to exploit the potential of forming, an in vitro, 3D prevascular network in order to facilitate and enhance bone regeneration. The graft was translated in vivo to determine biofunctionality of the bioengineered scaffold. A rabbit model was used to study the healing of the bone graft materials; scaffold with or without BMP-7 was implanted into experimental rabbit maxillary bone defects. Animals were killed and tissue examined with µCT and processed for histomorphometric analysis
Results:
The microstructure of scaffold determined by porosimetry and µCT confirmed a wide range of pore size from macro (>400 µm) to micro (<80 µCT) porosity. Direct and indirect biocompatibility assay showed good cell proliferation and differentiation up to 42 days of studied. Bone morphogenetic protein 7 (BMP-7) and vascular endothelial growth factor (VEGF) were employed, due to their specific effect on bone formation and vascularisation. Various culture conditions were tested in order to optimised the co-culture system. After 14 days, the outgrowth of coculture cells adhered to surface and inside of the porous scaffold. The immunofluorescent and immunohistochemistry staining (CD31+,vWF) showed that cells organised into capillary-like networks inside the pore of scaffold construct. Finally, an in vivo study in critical size maxillary defect model, the results showed large area of newly formed bone were observed at 4 weeks. Osteoclasts were observed around the spaces caused by resorption of the graft material. At the end of 8 weeks, the graft material was no longer present and completely filled with woven bone
Conclusions and clinical implications:
In this study, we have developed a porous β-metacalcium phosphate scaffold and investigated the hypothesis that an in vitro coculture system with osteopregenitor cells and endothelial cells can result in a prevascularised networked for application in bone tissue engineering. Resulted have demonstrated that the developed scaffold has the appropriate properties as an inductive bone graft and in vitro prevascularisation approach could well revolutionize the future therapies of bone regeneration.
Background:
Survival and function of the transplanted bone grafts are crucially dependent on rapidly forming blood supply. In order to engineer functional bone tissues successfully, the scaffolds have to be designed to facilitate cellular distribution and guide tissue regeneration in three dimension manner. Pore size and interconnectivity of the pores is a critical determinant of blood vessel ingrowth. In addition to providing tissues with the appropriate architecture and acting as a template, scaffolds can also act as carriers incorporating biological molecules that are known to promote signaling pathways that influence key cell functions such as migration, proliferation and differentiation. The major aim of this study is to create an inductive bone graft with the potential to act as template for seeded cells and as a delivery system for incorporated biological molecules; the aim being to facilitate vascularisation and the synthesis of extracellular matrix and subsequent formation of functional bone tissue.
Aim/Hypothesis:
The aim of this study was to create a 3D porous β calcium metaphosphate (CMP) scaffold with physico-chemical properties similar to natural bone and further attempt to pre-vascularize in vitro by using optimized co-culture condition.
Material and Methods:
Calcium phosphate materials have been shown to interact strongly with bone, due to their similarity to bone mineral. In this study, a block porous scaffolds of β-metacalcium phosphate were produced using monocalcium phosphate mono hydrate (MCP) powder and poly vinyl alcohol (PVA) as the porogen. The MCP and PVA mixes were pressed and sintered at 900 °C to yield the porous scaffolds which were sterilized with γ radiation prior to biological evaluation. Physico-chemical properties were investigated using a number of methods such as FTIR, x-ray diffraction, microcomputed tomography. In vitro biocompatibility tests were performed to confirm that the scaffold was biosafe and able to allow cell survival, cellular ingrowth, and also incorporation of appropriate bioactive growth factors. An in vitro co-culture model with osteoprogenitor cells and endothelial cells was used to exploit the potential of forming, an in vitro, 3D prevascular network in order to facilitate and enhance bone regeneration. The graft was translated in vivo to determine biofunctionality of the bioengineered scaffold. A rabbit model was used to study the healing of the bone graft materials; scaffold with or without BMP-7 was implanted into experimental rabbit maxillary bone defects. Animals were killed and tissue examined with µCT and processed for histomorphometric analysis
Results:
The microstructure of scaffold determined by porosimetry and µCT confirmed a wide range of pore size from macro (>400 µm) to micro (<80 µCT) porosity. Direct and indirect biocompatibility assay showed good cell proliferation and differentiation up to 42 days of studied. Bone morphogenetic protein 7 (BMP-7) and vascular endothelial growth factor (VEGF) were employed, due to their specific effect on bone formation and vascularisation. Various culture conditions were tested in order to optimised the co-culture system. After 14 days, the outgrowth of coculture cells adhered to surface and inside of the porous scaffold. The immunofluorescent and immunohistochemistry staining (CD31+,vWF) showed that cells organised into capillary-like networks inside the pore of scaffold construct. Finally, an in vivo study in critical size maxillary defect model, the results showed large area of newly formed bone were observed at 4 weeks. Osteoclasts were observed around the spaces caused by resorption of the graft material. At the end of 8 weeks, the graft material was no longer present and completely filled with woven bone
Conclusions and clinical implications:
In this study, we have developed a porous β-metacalcium phosphate scaffold and investigated the hypothesis that an in vitro coculture system with osteopregenitor cells and endothelial cells can result in a prevascularised networked for application in bone tissue engineering. Resulted have demonstrated that the developed scaffold has the appropriate properties as an inductive bone graft and in vitro prevascularisation approach could well revolutionize the future therapies of bone regeneration.
Background:
Survival and function of the transplanted bone grafts are crucially dependent on rapidly forming blood supply. In order to engineer functional bone tissues successfully, the scaffolds have to be designed to facilitate cellular distribution and guide tissue regeneration in three dimension manner. Pore size and interconnectivity of the pores is a critical determinant of blood vessel ingrowth. In addition to providing tissues with the appropriate architecture and acting as a template, scaffolds can also act as carriers incorporating biological molecules that are known to promote signaling pathways that influence key cell functions such as migration, proliferation and differentiation. The major aim of this study is to create an inductive bone graft with the potential to act as template for seeded cells and as a delivery system for incorporated biological molecules; the aim being to facilitate vascularisation and the synthesis of extracellular matrix and subsequent formation of functional bone tissue.
Aim/Hypothesis:
The aim of this study was to create a 3D porous β calcium metaphosphate (CMP) scaffold with physico-chemical properties similar to natural bone and further attempt to pre-vascularize in vitro by using optimized co-culture condition.
Material and Methods:
Calcium phosphate materials have been shown to interact strongly with bone, due to their similarity to bone mineral. In this study, a block porous scaffolds of β-metacalcium phosphate were produced using monocalcium phosphate mono hydrate (MCP) powder and poly vinyl alcohol (PVA) as the porogen. The MCP and PVA mixes were pressed and sintered at 900 °C to yield the porous scaffolds which were sterilized with γ radiation prior to biological evaluation. Physico-chemical properties were investigated using a number of methods such as FTIR, x-ray diffraction, microcomputed tomography. In vitro biocompatibility tests were performed to confirm that the scaffold was biosafe and able to allow cell survival, cellular ingrowth, and also incorporation of appropriate bioactive growth factors. An in vitro co-culture model with osteoprogenitor cells and endothelial cells was used to exploit the potential of forming, an in vitro, 3D prevascular network in order to facilitate and enhance bone regeneration. The graft was translated in vivo to determine biofunctionality of the bioengineered scaffold. A rabbit model was used to study the healing of the bone graft materials; scaffold with or without BMP-7 was implanted into experimental rabbit maxillary bone defects. Animals were killed and tissue examined with µCT and processed for histomorphometric analysis
Results:
The microstructure of scaffold determined by porosimetry and µCT confirmed a wide range of pore size from macro (>400 µm) to micro (<80 µCT) porosity. Direct and indirect biocompatibility assay showed good cell proliferation and differentiation up to 42 days of studied. Bone morphogenetic protein 7 (BMP-7) and vascular endothelial growth factor (VEGF) were employed, due to their specific effect on bone formation and vascularisation. Various culture conditions were tested in order to optimised the co-culture system. After 14 days, the outgrowth of coculture cells adhered to surface and inside of the porous scaffold. The immunofluorescent and immunohistochemistry staining (CD31+,vWF) showed that cells organised into capillary-like networks inside the pore of scaffold construct. Finally, an in vivo study in critical size maxillary defect model, the results showed large area of newly formed bone were observed at 4 weeks. Osteoclasts were observed around the spaces caused by resorption of the graft material. At the end of 8 weeks, the graft material was no longer present and completely filled with woven bone
Conclusions and clinical implications:
In this study, we have developed a porous β-metacalcium phosphate scaffold and investigated the hypothesis that an in vitro coculture system with osteopregenitor cells and endothelial cells can result in a prevascularised networked for application in bone tissue engineering. Resulted have demonstrated that the developed scaffold has the appropriate properties as an inductive bone graft and in vitro prevascularisation approach could well revolutionize the future therapies of bone regeneration.
Background:
Survival and function of the transplanted bone grafts are crucially dependent on rapidly forming blood supply. In order to engineer functional bone tissues successfully, the scaffolds have to be designed to facilitate cellular distribution and guide tissue regeneration in three dimension manner. Pore size and interconnectivity of the pores is a critical determinant of blood vessel ingrowth. In addition to providing tissues with the appropriate architecture and acting as a template, scaffolds can also act as carriers incorporating biological molecules that are known to promote signaling pathways that influence key cell functions such as migration, proliferation and differentiation. The major aim of this study is to create an inductive bone graft with the potential to act as template for seeded cells and as a delivery system for incorporated biological molecules; the aim being to facilitate vascularisation and the synthesis of extracellular matrix and subsequent formation of functional bone tissue.
Aim/Hypothesis:
The aim of this study was to create a 3D porous β calcium metaphosphate (CMP) scaffold with physico-chemical properties similar to natural bone and further attempt to pre-vascularize in vitro by using optimized co-culture condition.
Material and Methods:
Calcium phosphate materials have been shown to interact strongly with bone, due to their similarity to bone mineral. In this study, a block porous scaffolds of β-metacalcium phosphate were produced using monocalcium phosphate mono hydrate (MCP) powder and poly vinyl alcohol (PVA) as the porogen. The MCP and PVA mixes were pressed and sintered at 900 °C to yield the porous scaffolds which were sterilized with γ radiation prior to biological evaluation. Physico-chemical properties were investigated using a number of methods such as FTIR, x-ray diffraction, microcomputed tomography. In vitro biocompatibility tests were performed to confirm that the scaffold was biosafe and able to allow cell survival, cellular ingrowth, and also incorporation of appropriate bioactive growth factors. An in vitro co-culture model with osteoprogenitor cells and endothelial cells was used to exploit the potential of forming, an in vitro, 3D prevascular network in order to facilitate and enhance bone regeneration. The graft was translated in vivo to determine biofunctionality of the bioengineered scaffold. A rabbit model was used to study the healing of the bone graft materials; scaffold with or without BMP-7 was implanted into experimental rabbit maxillary bone defects. Animals were killed and tissue examined with µCT and processed for histomorphometric analysis
Results:
The microstructure of scaffold determined by porosimetry and µCT confirmed a wide range of pore size from macro (>400 µm) to micro (<80 µCT) porosity. Direct and indirect biocompatibility assay showed good cell proliferation and differentiation up to 42 days of studied. Bone morphogenetic protein 7 (BMP-7) and vascular endothelial growth factor (VEGF) were employed, due to their specific effect on bone formation and vascularisation. Various culture conditions were tested in order to optimised the co-culture system. After 14 days, the outgrowth of coculture cells adhered to surface and inside of the porous scaffold. The immunofluorescent and immunohistochemistry staining (CD31+,vWF) showed that cells organised into capillary-like networks inside the pore of scaffold construct. Finally, an in vivo study in critical size maxillary defect model, the results showed large area of newly formed bone were observed at 4 weeks. Osteoclasts were observed around the spaces caused by resorption of the graft material. At the end of 8 weeks, the graft material was no longer present and completely filled with woven bone
Conclusions and clinical implications:
In this study, we have developed a porous β-metacalcium phosphate scaffold and investigated the hypothesis that an in vitro coculture system with osteopregenitor cells and endothelial cells can result in a prevascularised networked for application in bone tissue engineering. Resulted have demonstrated that the developed scaffold has the appropriate properties as an inductive bone graft and in vitro prevascularisation approach could well revolutionize the future therapies of bone regeneration.