Chitosan Covalently Functionalized with Peptides Mapped on Vitronectin and BMP-2 for Bone Tissue Engineering
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Peptide Synthesis and Characterization
2.2.1. S-X-HVP
2.2.2. GBMP1a
2.3. S-X-HVP N-Terminal Conversion in an Alpha-oxo-Aldheyde
2.4. Scaffold Preparation
2.4.1. Chitosan Functionalization with ald-X-HVP and GBMP1a
2.4.2. Chitosan Spongy Scaffold Preparation
2.5. Scaffold Characterization
2.5.1. Fourier Transform Infrared Spectroscopy (FT-IR) Spectroscopy
2.5.2. X-ray Photoelectron Spectroscopy (XPS)
2.5.3. Nuclear Magnetic Resonance analysis (NMR) of Chitosan, Chit-HVP, and Chit-GBMP1a in Solution
2.5.4. Scanning Electron Microscope Analysis (SEM)
2.6. Biological Assays
2.6.1. Cell Culture
2.6.2. Cell Viability Assay
2.6.3. Cell Proliferation Assay
2.6.4. Quantitative Real-Time Polymerase Chain Reaction
2.6.5. Calcium Assay
2.7. Statistical Analysis
3. Results
3.1. Scaffold Characterization
3.1.1. FT-IR
3.1.2. XPS Spectroscopy
3.1.3. NMR Analysis of Chitosan, Chit-HVP, and Chit-GBMP1a in Solution
3.1.4. SEM Analysis
3.2. Biological Assays
3.2.1. Functionalized Matrices Are Nontoxic for Osteoblast Cells and Support Cell Proliferation
3.2.2. Functionalized Matrices Induce Differentiation in Cultured Osteoblast Cells
4. Discussion
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Sequence |
---|---|
GAPDH | Fw: 5′-agtgccagcctcgtcccgta-3′ Rv: 5′-caggcgcccaatacggccaa-3′ |
RUNX2 | Fw: 5′-cagtgacaccatgtcagcaa-3′ Rv: 5′-gctcacgtcgctcattttg-3′ |
VTN | Fw: 5′- ggaggacatcttcgagcttct-3′ Rv: 5′- gctaatgaactggggctgtc-3′ |
SPP1 | Fw: 5′-aagtttcgcagacctgacatc-3′ Rv: 5′-ggctgtcccaatcagaagg-3′ |
Sample | N/C Ratio |
---|---|
Chit | 0.07 |
Chit-HVP | 0.15 |
Chit-GBMP1a | 0.10 |
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Brun, P.; Zamuner, A.; Cassari, L.; D’Auria, G.; Falcigno, L.; Franchi, S.; Contini, G.; Marsotto, M.; Battocchio, C.; Iucci, G.; et al. Chitosan Covalently Functionalized with Peptides Mapped on Vitronectin and BMP-2 for Bone Tissue Engineering. Nanomaterials 2021, 11, 2784. https://doi.org/10.3390/nano11112784
Brun P, Zamuner A, Cassari L, D’Auria G, Falcigno L, Franchi S, Contini G, Marsotto M, Battocchio C, Iucci G, et al. Chitosan Covalently Functionalized with Peptides Mapped on Vitronectin and BMP-2 for Bone Tissue Engineering. Nanomaterials. 2021; 11(11):2784. https://doi.org/10.3390/nano11112784
Chicago/Turabian StyleBrun, Paola, Annj Zamuner, Leonardo Cassari, Gabriella D’Auria, Lucia Falcigno, Stefano Franchi, Giorgio Contini, Martina Marsotto, Chiara Battocchio, Giovanna Iucci, and et al. 2021. "Chitosan Covalently Functionalized with Peptides Mapped on Vitronectin and BMP-2 for Bone Tissue Engineering" Nanomaterials 11, no. 11: 2784. https://doi.org/10.3390/nano11112784