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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 healing of bony defect and new bone formation.(A,B) gross images of the samples harvested at 12 weeks after the cranial defect was transplanted with SAP/PLGA or PBMSCs/SAP/PLGA scaffold. The images of cross-section in the midline of defect area (A1,B1), views of intracranial side (A2,B2) and extracranial side (A3,B3) indicated both of SAP/PLGA and PBMSCs/SAP/PLGA grafts were integrated smoothly with the host. Micro-CT images showing the minerazation in PBMSCs/SAP/PLGA (C) and SAP/PLGA (D) grafted subjects at 12 weeks post surgery. (E,F) Statistical quantifications of the bone mineral density (BMD) and bone volume(BV) in the defect area assessed by micro-CT. (G,g,H,h) the whole and local images of H&E staining of cranial defect at 12 weeks post surgery (G,g), SAP/PLGA grafted; (H,h) PBMSCs/SAP/PLGA grafted), (I) Statistical quantifications of the percentage of new formed bone in the defect area assessed by H&E staining.
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f5: The healing of bony defect and new bone formation.(A,B) gross images of the samples harvested at 12 weeks after the cranial defect was transplanted with SAP/PLGA or PBMSCs/SAP/PLGA scaffold. The images of cross-section in the midline of defect area (A1,B1), views of intracranial side (A2,B2) and extracranial side (A3,B3) indicated both of SAP/PLGA and PBMSCs/SAP/PLGA grafts were integrated smoothly with the host. Micro-CT images showing the minerazation in PBMSCs/SAP/PLGA (C) and SAP/PLGA (D) grafted subjects at 12 weeks post surgery. (E,F) Statistical quantifications of the bone mineral density (BMD) and bone volume(BV) in the defect area assessed by micro-CT. (G,g,H,h) the whole and local images of H&E staining of cranial defect at 12 weeks post surgery (G,g), SAP/PLGA grafted; (H,h) PBMSCs/SAP/PLGA grafted), (I) Statistical quantifications of the percentage of new formed bone in the defect area assessed by H&E staining.

Mentions: As designed, the PLGA membranes provided physical support for the inner soft and gel-like scaffold of SAP seeded with or without PBMSCs. Moreover, the membranes are apt to be trimmed to cater to the size of the calvarial defect. Twelve weeks after transplantation the graft was integrated smoothly with the host, regardless if the graft was PBMSCs/SAP/PLGA(Fig. 5A) or SAP/PLGA(Fig. 5B). Micro-CT and 3D reconstruction revealed minerazation was found in the grafted scaffold 12 weeks post surgery. The images showed that the bone formation occurred only around the edge of the calvarical defect in the SAP/PLGA group (Fig. 5C), while the new formed bone filled the majority of the defect area in the PBMSCs/SAP/PLGA group (Fig. 5D). Notably, there were some bony islands scattered in the defect area, which indicated these bony islands resulted from osteogenesis of grafted PBMSCs but not from the osteoblasts of host tissue. In order to quantify the new bone formation in the defects, BV and BMD in the defect area were measured with the built-in software of the Micro-CT system. As showed in Fig. 5E,F, both BV and BMD were significantly higher in PBMSCs/SAP/PLGA group than that of SAP/PLGA group. Micro-CT findings were further confirmed by histological evidence. H&E staining demonstrated the new bone formation mainly localized by the bony borders in the SAP/PLGA group (Fig. 5G,g), while PBMSCs/SAP/PLGA group displayed both peripheral and central distribution (Fig. 5H,h). The difference of the percentage of new bone area quantified from the H&E stained samples was significant between the two groups (Fig. 5I).


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 healing of bony defect and new bone formation.(A,B) gross images of the samples harvested at 12 weeks after the cranial defect was transplanted with SAP/PLGA or PBMSCs/SAP/PLGA scaffold. The images of cross-section in the midline of defect area (A1,B1), views of intracranial side (A2,B2) and extracranial side (A3,B3) indicated both of SAP/PLGA and PBMSCs/SAP/PLGA grafts were integrated smoothly with the host. Micro-CT images showing the minerazation in PBMSCs/SAP/PLGA (C) and SAP/PLGA (D) grafted subjects at 12 weeks post surgery. (E,F) Statistical quantifications of the bone mineral density (BMD) and bone volume(BV) in the defect area assessed by micro-CT. (G,g,H,h) the whole and local images of H&E staining of cranial defect at 12 weeks post surgery (G,g), SAP/PLGA grafted; (H,h) PBMSCs/SAP/PLGA grafted), (I) Statistical quantifications of the percentage of new formed bone in the defect area assessed by H&E staining.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: The healing of bony defect and new bone formation.(A,B) gross images of the samples harvested at 12 weeks after the cranial defect was transplanted with SAP/PLGA or PBMSCs/SAP/PLGA scaffold. The images of cross-section in the midline of defect area (A1,B1), views of intracranial side (A2,B2) and extracranial side (A3,B3) indicated both of SAP/PLGA and PBMSCs/SAP/PLGA grafts were integrated smoothly with the host. Micro-CT images showing the minerazation in PBMSCs/SAP/PLGA (C) and SAP/PLGA (D) grafted subjects at 12 weeks post surgery. (E,F) Statistical quantifications of the bone mineral density (BMD) and bone volume(BV) in the defect area assessed by micro-CT. (G,g,H,h) the whole and local images of H&E staining of cranial defect at 12 weeks post surgery (G,g), SAP/PLGA grafted; (H,h) PBMSCs/SAP/PLGA grafted), (I) Statistical quantifications of the percentage of new formed bone in the defect area assessed by H&E staining.
Mentions: As designed, the PLGA membranes provided physical support for the inner soft and gel-like scaffold of SAP seeded with or without PBMSCs. Moreover, the membranes are apt to be trimmed to cater to the size of the calvarial defect. Twelve weeks after transplantation the graft was integrated smoothly with the host, regardless if the graft was PBMSCs/SAP/PLGA(Fig. 5A) or SAP/PLGA(Fig. 5B). Micro-CT and 3D reconstruction revealed minerazation was found in the grafted scaffold 12 weeks post surgery. The images showed that the bone formation occurred only around the edge of the calvarical defect in the SAP/PLGA group (Fig. 5C), while the new formed bone filled the majority of the defect area in the PBMSCs/SAP/PLGA group (Fig. 5D). Notably, there were some bony islands scattered in the defect area, which indicated these bony islands resulted from osteogenesis of grafted PBMSCs but not from the osteoblasts of host tissue. In order to quantify the new bone formation in the defects, BV and BMD in the defect area were measured with the built-in software of the Micro-CT system. As showed in Fig. 5E,F, both BV and BMD were significantly higher in PBMSCs/SAP/PLGA group than that of SAP/PLGA group. Micro-CT findings were further confirmed by histological evidence. H&E staining demonstrated the new bone formation mainly localized by the bony borders in the SAP/PLGA group (Fig. 5G,g), while PBMSCs/SAP/PLGA group displayed both peripheral and central distribution (Fig. 5H,h). The difference of the percentage of new bone area quantified from the H&E stained samples was significant between the two groups (Fig. 5I).

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