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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

In vitro multilineage differentiation of PBMSCs.(A,B) Calcium deposits, orange-red color stained by alizarin red or black color stained by von kossa staining, were scattered in the osteogenic induced culture. (C,D) ALP and osteocalcin expression in the osteogenic cells were illustrated by ALP staining and immunocytochemistry. (E) aggrecan expressed in chondrogenesis induced cells was demonstrated by Toluidine blue staining. (F) lipid vacuoles in the adipogenesis induced cells were showed by Oil Red O staining. (G,H) induced neurons and Schwann cells were identified by immunocytochemistry with antibodies of β-tubulin III and S100.
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f3: In vitro multilineage differentiation of PBMSCs.(A,B) Calcium deposits, orange-red color stained by alizarin red or black color stained by von kossa staining, were scattered in the osteogenic induced culture. (C,D) ALP and osteocalcin expression in the osteogenic cells were illustrated by ALP staining and immunocytochemistry. (E) aggrecan expressed in chondrogenesis induced cells was demonstrated by Toluidine blue staining. (F) lipid vacuoles in the adipogenesis induced cells were showed by Oil Red O staining. (G,H) induced neurons and Schwann cells were identified by immunocytochemistry with antibodies of β-tubulin III and S100.

Mentions: After being incubated with designed specific chemical inductive media, the PBMSCs differentiated into osteoblast, chondroblast, adipocyte, neuron, and Schwann cell-like cells, respectively. In order to identify the differentiated cells, different staining protocols were performed as described in the Method section. The identifications of each goal-oriented cells are as follows. Calcium deposits, orange-red color stained by alizarin, red or black color stained by von kossa staining, were scattered in the osteogenic induced culture (Fig. 3A,B). ALP, which is an early marker for osseous differentiation and is widely used as osteoblast specific marker, was expressed in the induced cells in varying degrees (Fig. 3C). Additionally, immunocytochemistry and confocal imaging showed osteocalcin positive particles distributing in the osteogenic cells (Fig. 3D). In short, the osteo-differentiation was identified by four methods. Aggrecan expressed in chondrogenesis induced cells was demonstrated by Toluidine blue staining (Fig. 3E) while lipid vacuoles in the adipogenesis induced cells were showed by Oil Red O staining (Fig. 3F). When induced to differentiate into neurons or Schwann cells, these cells were immuno-positive for neuron’s specific maker of β-tubulin III or Schwann cell’s specific maker of S-100 (Fig. 3G,H).


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)

In vitro multilineage differentiation of PBMSCs.(A,B) Calcium deposits, orange-red color stained by alizarin red or black color stained by von kossa staining, were scattered in the osteogenic induced culture. (C,D) ALP and osteocalcin expression in the osteogenic cells were illustrated by ALP staining and immunocytochemistry. (E) aggrecan expressed in chondrogenesis induced cells was demonstrated by Toluidine blue staining. (F) lipid vacuoles in the adipogenesis induced cells were showed by Oil Red O staining. (G,H) induced neurons and Schwann cells were identified by immunocytochemistry with antibodies of β-tubulin III and S100.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: In vitro multilineage differentiation of PBMSCs.(A,B) Calcium deposits, orange-red color stained by alizarin red or black color stained by von kossa staining, were scattered in the osteogenic induced culture. (C,D) ALP and osteocalcin expression in the osteogenic cells were illustrated by ALP staining and immunocytochemistry. (E) aggrecan expressed in chondrogenesis induced cells was demonstrated by Toluidine blue staining. (F) lipid vacuoles in the adipogenesis induced cells were showed by Oil Red O staining. (G,H) induced neurons and Schwann cells were identified by immunocytochemistry with antibodies of β-tubulin III and S100.
Mentions: After being incubated with designed specific chemical inductive media, the PBMSCs differentiated into osteoblast, chondroblast, adipocyte, neuron, and Schwann cell-like cells, respectively. In order to identify the differentiated cells, different staining protocols were performed as described in the Method section. The identifications of each goal-oriented cells are as follows. Calcium deposits, orange-red color stained by alizarin, red or black color stained by von kossa staining, were scattered in the osteogenic induced culture (Fig. 3A,B). ALP, which is an early marker for osseous differentiation and is widely used as osteoblast specific marker, was expressed in the induced cells in varying degrees (Fig. 3C). Additionally, immunocytochemistry and confocal imaging showed osteocalcin positive particles distributing in the osteogenic cells (Fig. 3D). In short, the osteo-differentiation was identified by four methods. Aggrecan expressed in chondrogenesis induced cells was demonstrated by Toluidine blue staining (Fig. 3E) while lipid vacuoles in the adipogenesis induced cells were showed by Oil Red O staining (Fig. 3F). When induced to differentiate into neurons or Schwann cells, these cells were immuno-positive for neuron’s specific maker of β-tubulin III or Schwann cell’s specific maker of S-100 (Fig. 3G,H).

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