Tuning Nanopore Diameter of Titanium Surfaces to Improve Human Gingival Fibroblast Response
Abstract
:1. Introduction
2. Results and Discussion
2.1. Surface Characterization
2.2. Cell Response to Nanoporous Surfaces
3. Materials and Methods
3.1. Preparation of Nanoporous Layers on Ti Foil
3.2. Characterization of Ti Nanopore Arrays
3.2.1. SEM
3.2.2. AFM
3.2.3. Contact Angle or Surface Wettability
3.2.4. Nanoparticle Release
3.3. Cell Culture
3.4. Cell Cytotoxicity
3.5. Cell Adhesion and Metabolic Activity
3.6. Gene Expression by Real-Time Polymerase Chain Reaction (RT-PCR)
3.7. Collagen Quantification
3.8. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
hGF | Human gingival fibroblasts |
MSCs | Mesenchymal stem cells |
Ti | Titanium |
TiO2 | Titanium dioxide |
NP | Nanopore |
SEM | Scanning electronic microscope |
Ra | Average surface roughness |
Rq | Root Mean Square (RMS) roughness |
Rmax | Maximum surface roughness |
Rskw | Skewness |
Rkur | Kurtosis |
Rpc | Peak counts |
Rsa | Surface Area difference |
AFM | Atomic force microscope |
LDH | Lactic dehydrogenase |
NADH | Nicotinamide adenine dinucleotide |
DLS | Dynamic light scattering |
S.E.M. | Standard Error of the Mean |
ANOVA | Analysis of variance |
LSD | Least significant difference |
ECM | Extracellular matrix |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
COL1A1 | α-1 type I collagen |
COL3A1 | α-1 type III collagen |
DCN | Decorin |
TCP | Tissue culture plastic |
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Parameters | Ti Control | NP-S | NP-B |
---|---|---|---|
Pore diameter (nm) 1 | (-) | 48.2 ± 1.2 | 74.0 ± 3.3 † |
Ra (nm) | 51.7 ± 5.71 | 54.7 ± 1.4 | 41.6 ± 5.5 |
Rskw (-) | 0.123 ± 0.337 | 0.069 ± 0.162 | −0.310 ± 0.196 |
Rkur (-) | 3.62 ± 0.39 | 2.62 ± 0.15 * | 2.99 ± 0.15 |
Rpc (-) | 29.8 ± 5.0 | 22.0 ± 4.0 | 156 ± 34 * † |
Srf. Area (µm2) | 26.4 ± 0.2 | 26.4 ± 0.5 | 30.4 ± 0.4 * † |
Rsa (%) | 5.68 ± 0.86 | 5.41 ± 0.21 | 21.6 ± 1.6 * † |
Contact angle (°) | 53.2 ± 2.5 | 78.6 ± 2.2 * | 65.5 ± 5.8 † |
TiO2 Structure | First Anodizing Step 30 min | Second Anodizing Step 10 min | Interspace |
---|---|---|---|
NP-S | 35 V 1 | 1 V | 2.5 cm |
NP-B | 60 V | 60 V | 5 cm |
Gene | Primer Sequence (5′–3′) | Product Size (bp) | GenBank ID |
---|---|---|---|
Collagen I α1 (COL1A1) | S: AGAGCATGACCGATGGATTC A: TTCTTGAGGTTGCCAGTC | 122 | NM_000088.3 |
Collagen III α1 (COL3A1) | S: GGCCTACTGGGCCTGGTGGT A: CCACGTTCACCAGGGGCACC | 190 | NM_000090.3 |
Decorin (DCN) | S: ATCTCAGCTTTGAGGGCTCC A: GCCTCTCTGTTGAAACGGTC | 146 | NM_001920.3 |
Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) | S: TGCACCACCAACTGCTTAGC A: AAGGGACTTCCTGTAACAA | 87 | NM_002046.3 |
β-Actin (ACTBL2) | S: CTGGAACGGTGAAGGTGACA A: AAGGGACTTCCTGTAACAA | 140 | NM_001101.3 |
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Ferrà-Cañellas, M.D.M.; Llopis-Grimalt, M.A.; Monjo, M.; Ramis, J.M. Tuning Nanopore Diameter of Titanium Surfaces to Improve Human Gingival Fibroblast Response. Int. J. Mol. Sci. 2018, 19, 2881. https://doi.org/10.3390/ijms19102881
Ferrà-Cañellas MDM, Llopis-Grimalt MA, Monjo M, Ramis JM. Tuning Nanopore Diameter of Titanium Surfaces to Improve Human Gingival Fibroblast Response. International Journal of Molecular Sciences. 2018; 19(10):2881. https://doi.org/10.3390/ijms19102881
Chicago/Turabian StyleFerrà-Cañellas, Maria Del Mar, Maria Antonia Llopis-Grimalt, Marta Monjo, and Joana Maria Ramis. 2018. "Tuning Nanopore Diameter of Titanium Surfaces to Improve Human Gingival Fibroblast Response" International Journal of Molecular Sciences 19, no. 10: 2881. https://doi.org/10.3390/ijms19102881