Improved Adhesion, Growth and Maturation of Vascular Smooth Muscle Cells on Polyethylene Grafted with Bioactive Molecules and Carbon Particles
Abstract
:1. Introduction
2. Results and Discussion
2.1. Physicochemical Properties of Polymer Samples
2.2. Initial Adhesion of VSMC on Polymer Samples
2.3. Spreading of VSMC on Polymer Samples
2.4. Proliferation of VSMC on Polymer Samples
2.5. Distribution and Concentration of Molecular Markers of Adhesion and Maturation in VSMCs
3. Experimental Section
3.1. Preparation of the Polymer Samples
3.2. Evaluation of the Physical and Chemical Properties of the Polymer Samples
3.3. Cell Source and Culture Conditions
3.4. Evaluation of the Cell Number and Morphology
3.5. Immunofluorescence Staining
3.6. Enzyme-Linked Immunosorbent Assay (ELISA)
3.7. Statistics
4. Conclusions
Acknowledgments
References and Notes
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Sample | O concentration (1016 at cm−2) |
---|---|
PE/plasma | 2.63 |
PE/plasma/H2O | 1.23 |
PE/plasma/PEG | 2.19 |
Protein/PE modification | Talin | Vinculin | Paxillin | α-Actinin | β-Actin | α-Actin | SM1 SM2 myosins |
---|---|---|---|---|---|---|---|
Pristine | 100 ± 9 | 100 ± 14 | 100 ± 28 | 100 ± 15 | 100 ± 8 | 100 ± 4 | 100 ± 16 |
Plasma | 134 ± 12 | 154 ± 17 | 98 ± 2 | 129 ± 28 | 77 ± 10 | 91 ± 10 | 69 ± 18 |
Gly | 109 ± 12 | 140 ± 18 | 136 ± 23 | 50 ± 8$ | 68 ± 3 | 119 ± 6C, * | 78 ± 15 |
PEG | 202 ± 16#, $ | 123 ± 8 | 152 ± 30 | 52 ± 10$ | 107 ± 8* | 123 ± 6$, C, * | 128 ± 14 |
BSA | 127 ± 10 | 151 ± 13 | 143 ± 27 | 37 ± 9$ | 95 ± 7* | 115 ± 7 | 73 ± 17 |
C | 112 ± 4 | 133 ± 9 | 104 ± 15 | 89 ± 8 | 111 ± 17* | 87 ± 3 | 87 ± 16 |
BSA + C | 201 ± 14#, * | 185 ± 14#, * | 114 ± 18 | 72 ± 12 | 110 ± 12* | 111 ± 7 | 108 ± 13 |
PS dish | 155 ± 29 | 113 ± 12 | 175 ± 37 | 93 ± 7 | 55 ± 9 | 89 ± 10 | 103 ± 16 |
Antibody against | Developed in, type | Company, Cat. No. | Dilution | Incubation |
---|---|---|---|---|
Chicken talin | Mouse, monoclonal | Sigma a T 3287 | Immf.: 1:200 ELISA: 1:500 | Immf.: Overnight, 4 °C ELISA: 60 min, RT c |
Human vinculin | Mouse, monoclonal | Sigma a V9131 | Immf.: 1:200 ELISA: 1:400 | Immf.: Overnight, 4 °C ELISA: 60 min, RT c |
Recombinant human paxilin | Rabbit polyclonal | Chemicon b P1093 | Immf.: 1:200 ELISA: 1:400 | Immf.: Overnight, 4 °C ELISA: 60 min, RT c |
Chicken α-actinin | Rabbit, polyclonal | Sigma a A2543 | Immf.: 1:200 ELISA: 1:500 | Immf.: Overnight, 4 °C ELISA: 60 min, RT c |
Synthetic peptide of α-smooth muscle actin | Mouse, monoclonal | Sigma a A2547 | Immf.: 1:200 ELISA: 1:400 | Immf.: Overnight, 4 °C ELISA: 60 min, RT c |
Synthetic peptide of β-actin | Mouse, monoclonal | Sigma a A 5441 | Immf.: 1:200 ELISA: 1:400 | Immf.: Overnight, 4 °C ELISA: 60 min, RT c |
Human smooth muscle myosin SM1 and SM2 | Mouse, monoclonal | Sigma a M7786 | Immf.: 1:200 ELISA: 1:500 | Immf.: Overnight, 4 °C ELISA: 60 min, RT c |
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Parizek, M.; Kasalkova, N.; Bacakova, L.; Slepicka, P.; Lisa, V.; Blazkova, M.; Svorcik, V. Improved Adhesion, Growth and Maturation of Vascular Smooth Muscle Cells on Polyethylene Grafted with Bioactive Molecules and Carbon Particles. Int. J. Mol. Sci. 2009, 10, 4352-4374. https://doi.org/10.3390/ijms10104352
Parizek M, Kasalkova N, Bacakova L, Slepicka P, Lisa V, Blazkova M, Svorcik V. Improved Adhesion, Growth and Maturation of Vascular Smooth Muscle Cells on Polyethylene Grafted with Bioactive Molecules and Carbon Particles. International Journal of Molecular Sciences. 2009; 10(10):4352-4374. https://doi.org/10.3390/ijms10104352
Chicago/Turabian StyleParizek, Martin, Nikola Kasalkova, Lucie Bacakova, Petr Slepicka, Vera Lisa, Martina Blazkova, and Vaclav Svorcik. 2009. "Improved Adhesion, Growth and Maturation of Vascular Smooth Muscle Cells on Polyethylene Grafted with Bioactive Molecules and Carbon Particles" International Journal of Molecular Sciences 10, no. 10: 4352-4374. https://doi.org/10.3390/ijms10104352