Gradient Chitosan Hydrogels Modified with Graphene Derivatives and Hydroxyapatite: Physiochemical Properties and Initial Cytocompatibility Evaluation
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization of the Graphene Materials
2.2. Rheological Characterization of Hydrogels
2.3. Microstructure
2.4. Cross-Linking Degree
2.5. Chemical Stability
2.6. Structural Analysis
2.7. Mechanical Properties
2.8. In Vitro Bioactivity
2.9. Cytocompatibility
2.9.1. The Influence of Liquid Extracts of Chitosan on hUC-MSCs Morphology
2.9.2. The Impact of Liquid Extracts of Chitosan on hUC-MSCs Cytotoxicity
2.9.3. The Influence of Liquid Extracts of Chitosan on hUC-MSCs Proliferation
3. Materials and Methods
3.1. Materials
3.2. Synthesis of Graphene Oxide (GO), Reduced Graphene Oxide (rGO) and Poly(Ethylene Glycol) Grafted Graphene Oxide (GO-PEG)
3.3. Preparation of Chitosan Hydrogels Modified with GO, rGO and GO-PEG
3.4. Characterization
3.4.1. X-Ray Photoelectron Spectroscopy (XPS)
3.4.2. X-Ray Diffractometry (XRD)
3.4.3. Attenuated Total Reflection Infrared Spectroscopy (ATR-FTIR)
3.4.4. Microstructural Analysis
3.4.5. Rheology
3.4.6. Determination of Cross-Linking Degree
3.4.7. Mechanical Properties
3.4.8. In Vitro Chemical Stability
3.4.9. Bioactivity Test
3.4.10. Cytocompatibility
Isolation and Culture of hUC-MSCs
Preparation of Liquid Extracts of Chitosan
Cytotoxicity Test
Proliferation Test
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
CS | chitosan |
GO | graphene oxide |
rGO | reduced graphene oxide |
GO-PEG | poly(ethylene glycol) grafted graphene oxide |
HAp | hydroxyapatite |
PBS | phosphate buffered saline |
SBF | simulated body fluid |
hUC-MSCs | human umbilical cord Wharton’s jelly mesenchymal stem cells |
TPP | sodium tripolyphosphate |
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Material | Wavenumber (cm−1) | Assignment |
---|---|---|
Chitosan | ≈3400 | N-H and O-H stretching |
≈2800 | C-H symmetric and asymmetric stretching | |
1719 | C=O stretching in amide I (residual N-acetyl groups) | |
1576 | N-H bending in primary amine | |
1450 | CH2 bending | |
1414 | CH3 symmetrical deformations in amide groups | |
1371 | N-H bending in amide II | |
1219 | OH bending | |
1120 | C-O-C bridge-asymmetric stretching | |
1080 | C-O stretching | |
1070 | ||
1027 | ||
855 | CH bending out of the plane of the ring | |
TPP | 1212 | P=O stretching |
1136 | O-P=O symmetric and asymmetric stretching | |
1090 | PO3 symmetric and asymmetric stretching | |
884 | P-O-P bridge asymmetric stretching | |
HAp | 1015 | P-O asymmetric stretching in PO4 |
TAc | 1702 | C=O stretching |
1443 | -C-Caromatic- stretching | |
1177 | C-O stretching | |
753 | C-C distortion in benzene ring | |
GO | ≈3400 | O-H stretching |
1725 | Carboxyl C=O stretching | |
1626 | C=C skeletal vibrations | |
1391 | C-OH deformation | |
1073 | C-O stretching | |
Hydrogels | ≈3400 | N-H stretching overlapped with O-H stretching |
1632 | N-H bending | |
1534 | N-O-P stretching |
Sample | GO | GO-PEG | rGO | HAp |
---|---|---|---|---|
H0 (ref. CS only) | – | – | – | – |
H1 | 0.5% | – | – | – |
H2 | – | 0.5% | – | – |
H3 | – | – | 0.5% | – |
H4 | 0.5% | – | – | 10% |
H5 | – | 0.5% | – | 10% |
H6 | – | – | 0.5% | 10% |
Attempt | 1st Bath | Time | 2nd Bath | Time |
---|---|---|---|---|
1 | 5% TPP | 2 h | 10% NaOH | 10 min |
2 | 5% TPP | 4 h | 10% NaOH | 10 min |
3 | 5% NaCl + 0.5% TPP | 24 h | – | – |
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Kosowska, K.; Domalik-Pyzik, P.; Sekuła-Stryjewska, M.; Noga, S.; Jagiełło, J.; Baran, M.; Lipińska, L.; Zuba-Surma, E.; Chłopek, J. Gradient Chitosan Hydrogels Modified with Graphene Derivatives and Hydroxyapatite: Physiochemical Properties and Initial Cytocompatibility Evaluation. Int. J. Mol. Sci. 2020, 21, 4888. https://doi.org/10.3390/ijms21144888
Kosowska K, Domalik-Pyzik P, Sekuła-Stryjewska M, Noga S, Jagiełło J, Baran M, Lipińska L, Zuba-Surma E, Chłopek J. Gradient Chitosan Hydrogels Modified with Graphene Derivatives and Hydroxyapatite: Physiochemical Properties and Initial Cytocompatibility Evaluation. International Journal of Molecular Sciences. 2020; 21(14):4888. https://doi.org/10.3390/ijms21144888
Chicago/Turabian StyleKosowska, Karolina, Patrycja Domalik-Pyzik, Małgorzata Sekuła-Stryjewska, Sylwia Noga, Joanna Jagiełło, Magdalena Baran, Ludwika Lipińska, Ewa Zuba-Surma, and Jan Chłopek. 2020. "Gradient Chitosan Hydrogels Modified with Graphene Derivatives and Hydroxyapatite: Physiochemical Properties and Initial Cytocompatibility Evaluation" International Journal of Molecular Sciences 21, no. 14: 4888. https://doi.org/10.3390/ijms21144888