Preparation and In Vitro Characterization of Magnetic CS/PVA/HA/pSPIONs Scaffolds for Magnetic Hyperthermia and Bone Regeneration
Abstract
:1. Introduction
2. Results and Discussion
2.1. Paste Characterization
Paste Rheological Behavior
2.2. Paste Formulations Containing pSPIONs
2.3. Scaffold Characterization
2.3.1. Scanning Electron Microscopy (SEM)
2.3.2. Attenuated Total Reflectance—Fourier Transform Infrared Spectroscopy (ATR-FTIR)
2.3.3. Mechanical Characterization
2.3.4. Magnetic Hyperthermia (MHT)
2.3.5. Swelling Ratio and Scaffold Erosion
2.4. In Vitro Studies
2.4.1. Cell Viability Assays
2.4.2. Cell Adhesion and Proliferation
2.4.3. Relative ALP Expression of Saos-2 Cells on CS/PVA/HA/pSPIONs Scaffolds
2.4.4. Cell Morphology
3. Materials and Methods
3.1. Materials
3.2. Methods
3.2.1. Pristine Superparamagnetic Iron Oxide Nanoparticles (pSPIONs) Synthesis
3.2.2. Paste Preparation
3.2.3. Three-Dimensional (3D) Printing and Post-Printing Treatments
3.2.4. Paste Characterization
Rheological Behavior
3.2.5. Physicochemical, Morphological, and Magnetic Characterization of the Scaffolds
Scanning Electron Microscopy (SEM)
Attenuated Total Reflectance—Fourier Transform Infrared Spectroscopy (ATR-FTIR)
Magnetic Hyperthermia (MHT)
Mechanical Characterization
Porosity
Swelling Ratio and Scaffold Erosion
3.2.6. In Vitro Studies
Cell Viability Assays
Cell Adhesion and Proliferation
Alkaline Phosphatase Assay
Immunofluorescence Study
3.2.7. Statistical Analysis
4. Conclusions
5. Future Perspectives
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Paste Designation | Paste Formulation (wt.%) | Paste Solids Content (wt.%) | |||||
---|---|---|---|---|---|---|---|
CS [a] | PVA/HA | CS:PVA/HA Ratio | CS | PVA | HA | ||
PVA [a] | HA [b] | ||||||
P1 | 6 | 12.5 | 15 | 3:1 | 49.83 | 37.13 | 13.04 |
P2 | 6 | 15.0 | 15 | 3:1 | 45.28 | 41.68 | 13.04 |
P3 | 6 | 17.5 | 15 | 3:1 | 41.28 | 45.68 | 13.04 |
P4 | 6 | 12.5 | 15 | 3:2 | 34.89 | 52.07 | 13.04 |
P5 | 6 | 15.0 | 15 | 3:2 | 30.58 | 56.38 | 13.04 |
P6 | 6 | 17.5 | 15 | 3:2 | 27.04 | 59.92 | 13.04 |
Samples Designation | Samples | CS (wt.%) | PVA (wt.%) | HA (wt.%) | pSPIONs (wt.%) |
---|---|---|---|---|---|
P1_B | CS/PVA/HA | 49.83 | 37.13 | 13.04 | - |
P1_pS1 | CS/PVA/HA/15%pSPIONs | 48.87 | 36.42 | 12.79 | 1.92 |
P1_pS3 | CS/PVA/HA/30%pSPIONs | 47.95 | 35.73 | 12.55 | 3.77 |
P1_pS5 | CS/PVA/HA/45%pSPIONs | 47.07 | 35.07 | 12.32 | 5.54 |
Samples | 18G Needle | P1_B | P1_pS1 | P1_pS3 | P1_pS5 |
---|---|---|---|---|---|
Filament Diameter (µm) | 840 ± 2 | 918 ± 37 | 839 ± 17 | 832 ± 32 | 876 ± 8 |
Samples | Estimated Porosity [a] (%) | Toe Modulus (kPa) | pSPIONs Content (wt. %) |
---|---|---|---|
P1_B | 70 ± 3 | 27 ± 8 | 0.00 |
P1_pS5 | 73 ± 2 | 92 ± 4 | 5.54 |
Samples | Adhesion Ratio (%) | P1_B | P1_pS1 | P1_pS3 | P1_pS5 |
---|---|---|---|---|---|
P1_B | 63 ± 12 | - | * | ** | ** |
P1_pS1 | 77 ± 11 | * | - | ns | ns |
P1_pS3 | 85 ± 8 | ** | ns | - | ns |
P1_pS5 | 86 ± 11 | ** | ns | ns | - |
Samples | Day 1 | Day 4 | Day 7 | PR 1–4 | PR 1–7 |
---|---|---|---|---|---|
P1_B | 0.39 ± 0.09 | 0.46 ± 0.13 | 0.43 ± 0.10 | 1.21 ± 0.10 | 1.11 ± 0.06 |
P1_pS1 | 0.40 ± 0.11 | 0.56 ± 0.13 | 0.58 ± 0.09 | 1.51 ± 0.13 | 1.23 ± 0.08 |
P1_pS3 | 0.38 ± 0.08 | 0.67 ± 0.15 | 0.74 ± 0.12 | 1.96 ± 0.08 | 2.10 ± 0.14 |
P1_pS5 | 0.37 ± 0.10 | 0.65 ± 0.14 | 0.72 ± 0.14 | 1.97 ± 0.20 | 1.87 ± 0.20 |
Cell Control | 1.000 ± 0.004 | 1.87 ± 0.19 | 2.42 ± 0.28 | 1.87 ± 0.19 | 2.42 ± 0.28 |
Samples | P1_B | P1_pS1 | P1_pS3 | P1_pS5 |
---|---|---|---|---|
P1_B | - | **** | **** | **** |
P1_pS1 | **** | - | ns | ns |
P1_pS3 | **** | ns | - | ns |
P1_pS5 | **** | ns | ns | - |
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Tavares, F.J.T.M.; Soares, P.I.P.; Silva, J.C.; Borges, J.P. Preparation and In Vitro Characterization of Magnetic CS/PVA/HA/pSPIONs Scaffolds for Magnetic Hyperthermia and Bone Regeneration. Int. J. Mol. Sci. 2023, 24, 1128. https://doi.org/10.3390/ijms24021128
Tavares FJTM, Soares PIP, Silva JC, Borges JP. Preparation and In Vitro Characterization of Magnetic CS/PVA/HA/pSPIONs Scaffolds for Magnetic Hyperthermia and Bone Regeneration. International Journal of Molecular Sciences. 2023; 24(2):1128. https://doi.org/10.3390/ijms24021128
Chicago/Turabian StyleTavares, Francisco J. T. M., Paula I. P. Soares, Jorge Carvalho Silva, and João Paulo Borges. 2023. "Preparation and In Vitro Characterization of Magnetic CS/PVA/HA/pSPIONs Scaffolds for Magnetic Hyperthermia and Bone Regeneration" International Journal of Molecular Sciences 24, no. 2: 1128. https://doi.org/10.3390/ijms24021128