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Osteogenesis of peripheral blood mesenchymal stem cells in self assembling peptide nanofiber for healing critical size calvarial bony defect.

Wu G, Pan M, Wang X, Wen J, Cao S, Li Z, Li Y, Qian C, Liu Z, Wu W, Zhu L, Guo J - Sci Rep (2015)

Bottom Line: Herein, PBMSCs were seeded into a nanofiber scaffold of self-assembling peptide (SAP) and cultured in osteogenic medium.Furthermore, the SAP seeded with the induced PBMSCs was splinted by two membranes of poly(lactic)-glycolic acid (PLGA) to fabricate a composited scaffold which was then used to repair a critical-size calvarial bone defect model in rat.To our knowledge this is the first report with solid evidence demonstrating PBMSCs can survive in the bone defect area and directly contribute to new bone formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou 510515, China.

ABSTRACT
Peripheral blood mesenchymal stem cells (PBMSCs) may be easily harvested from patients, permitting autologous grafts for bone tissue engineering in the future. However, the PBMSC's capabilities of survival, osteogenesis and production of new bone matrix in the defect area are still unclear. Herein, PBMSCs were seeded into a nanofiber scaffold of self-assembling peptide (SAP) and cultured in osteogenic medium. The results indicated SAP can serve as a promising scaffold for PBMSCs survival and osteogenic differentiation in 3D conditions. Furthermore, the SAP seeded with the induced PBMSCs was splinted by two membranes of poly(lactic)-glycolic acid (PLGA) to fabricate a composited scaffold which was then used to repair a critical-size calvarial bone defect model in rat. Twelve weeks later the defect healing and mineralization were assessed by H&E staining and microcomputerized tomography (micro-CT). The osteogenesis and new bone formation of grafted cells in the scaffold were evaluated by immunohistochemistry. To our knowledge this is the first report with solid evidence demonstrating PBMSCs can survive in the bone defect area and directly contribute to new bone formation. Moreover, the present data also indicated the tissue engineering with PBMSCs/SAP/PLGA scaffold can serve as a novel prospective strategy for healing large size cranial defects.

No MeSH data available.


Related in: MedlinePlus

The survival and osteogenesis of grafted PBMSCs in the bony defect area.(A) Lots of GFP positive graft cells were detected in the bony defect area 2 weeks post-implantation (A1), and these cells were positive for osteocalcin (A2), A3 is the merged image of A1 and A2. (B) BrdU immunohistochemistry showing lots of BrdU labeled graft cells were embedded by new formed bone matrix. In order to show the details in different areas, local images were step by step amplified and showed in (B’–B”’) and (b’–b”’). In the central area, almost all of the osteocytes in the new formed bone were BrdU positive (arrows),while in the edge of defect area, both of BrdU positive (arrows) and negative (arrow heads) osteocytes were located in the new formed bone.
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f6: The survival and osteogenesis of grafted PBMSCs in the bony defect area.(A) Lots of GFP positive graft cells were detected in the bony defect area 2 weeks post-implantation (A1), and these cells were positive for osteocalcin (A2), A3 is the merged image of A1 and A2. (B) BrdU immunohistochemistry showing lots of BrdU labeled graft cells were embedded by new formed bone matrix. In order to show the details in different areas, local images were step by step amplified and showed in (B’–B”’) and (b’–b”’). In the central area, almost all of the osteocytes in the new formed bone were BrdU positive (arrows),while in the edge of defect area, both of BrdU positive (arrows) and negative (arrow heads) osteocytes were located in the new formed bone.

Mentions: Even though both micro-CT and H&E staining showed the PBMSCs/SAP/PLGA scaffold promoting the new bone formation in the cranial defect, it still cannot declare that PBMSCs survived in the grafted scaffold and contributed to the new bone formation. At 2 weeks post-implantation, immunohistochemistry and confocal microscopy revealed plenty of GFP positive cells survived within the grafts of PBMSCs/SAP/PLGA scaffolds, while the osteocalcin immunoreactive signal was overlapped with majority of the GFP positive cells (Fig. 6A). The above data strongly support the transplanted PBMSCs could survive and differentiate into osteoblasts in the bony defect area. Moreover, BrdU assay performed at 12 weeks post-implantation showed that a majority of osteoctyes embedded in the new formed bone were positive for BrdU (Fig. 6B). This meant the cells in this area were derived from the PBMSCs and these cells directly contribute to form new bone matrix in the defect area.


Osteogenesis of peripheral blood mesenchymal stem cells in self assembling peptide nanofiber for healing critical size calvarial bony defect.

Wu G, Pan M, Wang X, Wen J, Cao S, Li Z, Li Y, Qian C, Liu Z, Wu W, Zhu L, Guo J - Sci Rep (2015)

The survival and osteogenesis of grafted PBMSCs in the bony defect area.(A) Lots of GFP positive graft cells were detected in the bony defect area 2 weeks post-implantation (A1), and these cells were positive for osteocalcin (A2), A3 is the merged image of A1 and A2. (B) BrdU immunohistochemistry showing lots of BrdU labeled graft cells were embedded by new formed bone matrix. In order to show the details in different areas, local images were step by step amplified and showed in (B’–B”’) and (b’–b”’). In the central area, almost all of the osteocytes in the new formed bone were BrdU positive (arrows),while in the edge of defect area, both of BrdU positive (arrows) and negative (arrow heads) osteocytes were located in the new formed bone.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645224&req=5

f6: The survival and osteogenesis of grafted PBMSCs in the bony defect area.(A) Lots of GFP positive graft cells were detected in the bony defect area 2 weeks post-implantation (A1), and these cells were positive for osteocalcin (A2), A3 is the merged image of A1 and A2. (B) BrdU immunohistochemistry showing lots of BrdU labeled graft cells were embedded by new formed bone matrix. In order to show the details in different areas, local images were step by step amplified and showed in (B’–B”’) and (b’–b”’). In the central area, almost all of the osteocytes in the new formed bone were BrdU positive (arrows),while in the edge of defect area, both of BrdU positive (arrows) and negative (arrow heads) osteocytes were located in the new formed bone.
Mentions: Even though both micro-CT and H&E staining showed the PBMSCs/SAP/PLGA scaffold promoting the new bone formation in the cranial defect, it still cannot declare that PBMSCs survived in the grafted scaffold and contributed to the new bone formation. At 2 weeks post-implantation, immunohistochemistry and confocal microscopy revealed plenty of GFP positive cells survived within the grafts of PBMSCs/SAP/PLGA scaffolds, while the osteocalcin immunoreactive signal was overlapped with majority of the GFP positive cells (Fig. 6A). The above data strongly support the transplanted PBMSCs could survive and differentiate into osteoblasts in the bony defect area. Moreover, BrdU assay performed at 12 weeks post-implantation showed that a majority of osteoctyes embedded in the new formed bone were positive for BrdU (Fig. 6B). This meant the cells in this area were derived from the PBMSCs and these cells directly contribute to form new bone matrix in the defect area.

Bottom Line: Herein, PBMSCs were seeded into a nanofiber scaffold of self-assembling peptide (SAP) and cultured in osteogenic medium.Furthermore, the SAP seeded with the induced PBMSCs was splinted by two membranes of poly(lactic)-glycolic acid (PLGA) to fabricate a composited scaffold which was then used to repair a critical-size calvarial bone defect model in rat.To our knowledge this is the first report with solid evidence demonstrating PBMSCs can survive in the bone defect area and directly contribute to new bone formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou 510515, China.

ABSTRACT
Peripheral blood mesenchymal stem cells (PBMSCs) may be easily harvested from patients, permitting autologous grafts for bone tissue engineering in the future. However, the PBMSC's capabilities of survival, osteogenesis and production of new bone matrix in the defect area are still unclear. Herein, PBMSCs were seeded into a nanofiber scaffold of self-assembling peptide (SAP) and cultured in osteogenic medium. The results indicated SAP can serve as a promising scaffold for PBMSCs survival and osteogenic differentiation in 3D conditions. Furthermore, the SAP seeded with the induced PBMSCs was splinted by two membranes of poly(lactic)-glycolic acid (PLGA) to fabricate a composited scaffold which was then used to repair a critical-size calvarial bone defect model in rat. Twelve weeks later the defect healing and mineralization were assessed by H&E staining and microcomputerized tomography (micro-CT). The osteogenesis and new bone formation of grafted cells in the scaffold were evaluated by immunohistochemistry. To our knowledge this is the first report with solid evidence demonstrating PBMSCs can survive in the bone defect area and directly contribute to new bone formation. Moreover, the present data also indicated the tissue engineering with PBMSCs/SAP/PLGA scaffold can serve as a novel prospective strategy for healing large size cranial defects.

No MeSH data available.


Related in: MedlinePlus