OBJECTIVES: Bioactive peptides derived from receptor binding motifs of native proteins are a potent source of bioactive molecules that can induce signalling pathways. These peptides could substitute for osteogenesis promoting supplements. The work presented here compares three kinds of bioactive peptides derived from collagen III, bone morphogenetic protein 7 (BMP-7) and BMP-2 with their potential osteogenic activity on the model of porcine mesenchymal stem cells (pMSCs). MATERIALS AND METHODS: pMSCs were cultured on electrospun polycaprolactone nanofibrous scaffolds with different concentrations of the bioactive peptides without addition of any osteogenic supplement. Analysis of pMSCs cultures included measurement of the metabolic activity and proliferation, immunofluorescence staining and also qPCR. RESULTS: Results showed no detrimental effect of the bioactive peptides to cultured pMSCs. Based on qPCR analysis, the bioactive peptides are specific for osteogenic differentiation with no detectable expression of collagen II. Our results further indicate that peptide derived from BMP-2 protein promoted the expression of mRNA for osteocalcin (OCN) and collagen I significantly compared to control groups and also supported deposition of OCN as observed by immunostaining method. CONCLUSION: The data suggest that bioactive peptide with an amino acid sequence of KIPKASSVPTELSAISTLYL derived from BMP-2 protein was the most potent for triggering osteogenic differentiation of pMSCs.

• Keywords
• bioactive peptides, electrospun scaffold, mesenchymal stem cells, osteogenic differentiation,
• MeSH
• Cell Differentiation drug effects   MeSH
• Cell Culture Techniques MeSH
• Collagen Type II genetics   metabolism   MeSH
• Collagen Type I genetics   metabolism   MeSH
• Microscopy, Confocal MeSH
• Cells, Cultured MeSH
• Mesenchymal Stem Cells cytology   drug effects   metabolism   MeSH
• Microscopy, Electron, Scanning MeSH
• Swine, Miniature MeSH
• Osteogenesis drug effects   MeSH
• Osteocalcin genetics   metabolism   MeSH
• Peptides chemistry   pharmacology   MeSH
• Swine MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Collagen Type II MeSH
• Collagen Type I MeSH
• Osteocalcin MeSH
• Peptides MeSH

OBJECTIVES: We prepared 3D poly (ε-caprolactone) (PCL) nanofibre scaffolds and tested their use for seeding, proliferation, differentiation and migration of mesenchymal stem cell (MSCs). MATERIALS AND METHODS: 3D nanofibres were prepared using a special collector for common electrospinning; simultaneously, a 2D PCL nanofibre layer was prepared using a classic plain collector. Both scaffolds were seeded with MSCs and biologically tested. MSC adhesion, migration, proliferation and osteogenic differentiation were investigated. RESULTS: The 3D PCL scaffold was characterized by having better biomechanical properties, namely greater elasticity and resistance against stress and strain, thus this scaffold will be able to find broad applications in tissue engineering. Clearly, while nanofibre layers of the 2D scaffold prevented MSCs from migrating through the conformation, cells infiltrated freely through the 3D scaffold. MSC adhesion to the 3D nanofibre PCL layer was also statistically more common than to the 2D scaffold (P < 0.05), and proliferation and viability of MSCs 2 or 3 weeks post-seeding, were also greater on the 3D scaffold. In addition, the 3D PCL scaffold was also characterized by displaying enhanced MSC osteogenic differentiation. CONCLUSIONS: We draw the conclusion that all positive effects observed using the 3D PCL nanofibre scaffold are related to the larger fibre surface area available to the cells. Thus, the proposed 3D structure of the nanofibre layer will find a wide array of applications in tissue engineering and regenerative medicine.

• MeSH
• Cell Differentiation * MeSH
• Cell Culture Techniques instrumentation   methods   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Mesenchymal Stem Cells cytology   metabolism   MeSH
• Nanofibers chemistry   ultrastructure   MeSH
• Osteogenesis MeSH
• Osteocalcin metabolism   MeSH
• Cell Movement MeSH
• Polyesters chemistry   MeSH
• Surface Properties MeSH
• Cell Proliferation MeSH
• Elasticity MeSH
• Regenerative Medicine MeSH
• Integrin-Binding Sialoprotein metabolism   MeSH
• Tissue Engineering MeSH
• Tissue Scaffolds * MeSH
• Cell Survival MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Osteocalcin MeSH
• polycaprolactone MeSH Browser
• Polyesters MeSH
• Integrin-Binding Sialoprotein MeSH