Large-Pore Platelet-Rich Fibrin with a Mg Ring to Allow MC3T3-E1 Preosteoblast Migration and to Improve Osteogenic Ability for Bone Defect Repair
<p>Large-pore platelet-rich fibrin and a magnesium scaffold improved cell migration and osteogenic differentiation for critical-size bone defect repair.</p> "> Figure 2
<p>Platelet-rich fibrin scaffold preparation and analysis. After collection, the platelet-rich fibrin was subjected to repeated freeze drying to remove the water naturally present within it. It was then immersed in water and dried using three cycles of the repeated freeze drying method to expand the pores, thus enabling cell migration and ingrowth. Large-pore platelet-rich fibrin was thereby produced. (<b>a</b>) Micro-CT images of platelet-rich fibrin after several iterations of freeze drying; (<b>b</b>) scanning electron microscopy images of platelet-rich fibrin; (<b>c</b>) images of platelet-rich fibrin; (<b>d</b>) volume of platelet-rich fibrin; (<b>e</b>) porosity of platelet-rich fibrin; (<b>f</b>) volume of platelet-rich fibrin pores; (<b>g</b>) pore diameter of top-section platelet-rich fibrin; (<b>h</b>) pore diameter of middle-section platelet-rich fibrin; and (<b>i</b>) pore diameter of bottom-section platelet-rich fibrin. (Individual data points are shown as means ± SD; N = 5 per group; ** <span class="html-italic">p</span> < 0.01 and **** <span class="html-italic">p</span> < 0.001).</p> "> Figure 3
<p>Growth factors released from platelet-rich fibrin before and after repeated freeze drying: (<b>a</b>) vascular endothelial growth factor (VEGF), (<b>b</b>) insulin-like growth factor-1 (IGF-1) and (<b>c</b>) transforming growth factor beta (TGF-β). (Individual data points are shown as means ± SD; N = 5 per group; * <span class="html-italic">p</span> < 0.05 and ** <span class="html-italic">p</span> < 0.01).</p> "> Figure 4
<p>Degradation behavior of the pure magnesium ring: (<b>a</b>) dimensions of the metal ring, (<b>b</b>) weight change of the pure magnesium ring, and (<b>c</b>) pH change of Hank’s solution. (N = 5 per group).</p> "> Figure 5
<p>Surface morphology of magnesium and titanium rings after 3, 7 and 14 days of immersion in Hank’s solution.</p> "> Figure 6
<p>Cell proliferation of MC3T3-E1 preosteoblasts cultured with platelet-rich fibrin, large-pore platelet-rich fibrin, magnesium ring and titanium ring precipitate media and their mixtures for (<b>a</b>) 1, (<b>b</b>) 3 and (<b>c</b>) 7 days. (N = 5 per group; * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, and *** <span class="html-italic">p</span> < 0.005).</p> "> Figure 7
<p>Migration capacity of MC3T3-E1 preosteoblasts cultured with platelet-rich fibrin, large-pore platelet-rich fibrin, magnesium ring and titanium ring precipitate media and their mixtures. (<b>a</b>) Cell migration at first scratch and after 24 h of incubation; reduction of the gap due to migration of MC3T3-E1 preosteoblasts after (<b>b</b>) 4, (<b>c</b>) 8 and (<b>d</b>) 24 h of incubation. (N = 5 per group; * <span class="html-italic">p</span> < 0.05 and ** <span class="html-italic">p</span> < 0.01).</p> "> Figure 8
<p>Migration capacity of MC3T3-E1 preosteoblasts cultured in large-pore platelet-rich fibrin membrane and platelet-rich fibrin membrane for 8 h. (<b>a</b>) Images of migrated cells stained with Hoechst 33,342 in large-pore platelet-rich fibrin and platelet-rich fibrin at 0, 1, 2, 4, 6 and 8 h after seeding, (<b>b</b>) Combination of the images obtained at 0 and 8 h showing cell migration in large-pore platelet-rich fibrin and platelet-rich fibrin and (<b>c</b>) Number of migrated cells against time.</p> "> Figure 9
<p>Scanning electron microscopy images of MC3T3-E1 preosteoblasts cultured on (<b>a</b>,<b>b</b>) large-pore platelet-rich fibrin and (<b>c</b>,<b>d</b>) platelet-rich fibrin captured with different magnification.</p> "> Figure 10
<p>Calcium deposition with platelet-rich fibrin, large-pore platelet-rich fibrin, magnesium ring and titanium ring precipitate media and their mixtures. (N = 5 per group; ** <span class="html-italic">p</span> < 0.01 and *** <span class="html-italic">p</span> < 0.005).</p> "> Figure 11
<p>(<b>a</b>) Calcium deposition of MC3T3-E1 preosteoblasts on platelet-rich fibrin and large-pore platelet-rich fibrin with magnesium and titanium rings, as detected through alizarin red S staining on the scaffold with 5-μm-thick sections. The location of calcium deposits is indicated by a yellow arrow. (<b>b</b>) Quantitative results of stain marks, obtained using Image J software. (N = 4 per group; ** <span class="html-italic">p</span> < 0.01, and *** <span class="html-italic">p</span> < 0.005).</p> "> Figure 12
<p>Large-pore platelet-rich fibrin preparation procedure and the molecular basis of why water expands when frozen.</p> ">
Abstract
:1. Introduction
2. Results
2.1. Fundamental Properties of Modified PRF
2.2. Degradation Behavior of Metallic Rings
2.3. Cell Viability of MC3T3-E1 Preosteoblasts Treated with PRF, LPPRF, Mg Ring and Ti Ring Precipitate Media and Corresponding Mixtures
2.4. Migration Capacity of MC3T3-E1 Preosteoblasts Treated with PRF, LPPRF, Mg Ring and Ti Ring Precipitate Media and Their Mixtures
2.5. Migration Capacity of MC3T3-E1 Preosteoblasts on PRF and LPPRF Sheets
2.6. Cell Morphology of MC3T3-E1 Preosteoblasts on PRF and LPPRF
2.7. Calcium Deposition of MC3T3-E1 Preosteoblasts Treated with PRF, LPPRF, Mg Ring and Ti Ring Precipitate Media and Their Mixtures
2.8. Calcium Deposition of MC3T3-E1 Preosteoblasts on PRF and LPPRF
3. Discussion
4. Materials and Methods
4.1. Study Design
4.2. PRF Collection and Analysis
4.3. Analysis of PRF and LPPRF Structures
4.4. Growth Factor Tests
4.5. Preparation of Mg and Ti scaffolds
4.6. Degradation Behavior of the Metallic Ring
4.7. Surface Morphology Observation of Metallic Ring and MC3T3-E1 Preosteoblasts When Attached to PRF and LPPRF
4.8. Precipitate Medium Preparation
4.9. Cell Viability of MC3T3-E1 Preosteoblasts
4.10. Analysis of MC3T3-E1 Cell Migration Capacity through a Scratch Assay
4.11. Migration Capacity of MC3T3-E1 Cells in PRF and LPPRF Sheets
4.12. Extracellular Matrix Calcium Deposition of MC3T3-E1 Cells with Precipitate Medium Culture
4.13. Extracellular Matrix Calcium Deposition of MC3T3-E1 Cells on PRF and LPPRF Scaffolds with Mg and Ti Rings
4.14. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Wong, P.-C.; Wang, C.-Y.; Jang, J.S.-C.; Lee, C.-H.; Wu, J.-L. Large-Pore Platelet-Rich Fibrin with a Mg Ring to Allow MC3T3-E1 Preosteoblast Migration and to Improve Osteogenic Ability for Bone Defect Repair. Int. J. Mol. Sci. 2021, 22, 4022. https://doi.org/10.3390/ijms22084022
Wong P-C, Wang C-Y, Jang JS-C, Lee C-H, Wu J-L. Large-Pore Platelet-Rich Fibrin with a Mg Ring to Allow MC3T3-E1 Preosteoblast Migration and to Improve Osteogenic Ability for Bone Defect Repair. International Journal of Molecular Sciences. 2021; 22(8):4022. https://doi.org/10.3390/ijms22084022
Chicago/Turabian StyleWong, Pei-Chun, Chen-Yun Wang, Jason Shian-Ching Jang, Chian-Her Lee, and Jia-Lin Wu. 2021. "Large-Pore Platelet-Rich Fibrin with a Mg Ring to Allow MC3T3-E1 Preosteoblast Migration and to Improve Osteogenic Ability for Bone Defect Repair" International Journal of Molecular Sciences 22, no. 8: 4022. https://doi.org/10.3390/ijms22084022