Multilayered Films Produced by Layer-by-Layer Assembly of Chitosan and Alginate as a Potential Platform for the Formation of Human Adipose-Derived Stem Cell aggregates
"> Figure 1
<p>NMR characterization of mesyl–CHT (<b>A</b>) and tosyl–CHT (<b>B</b>). Peaks of mesyl (2.83 ppm) and tosyl groups (2.39, 7.37 and 7.70 ppm) are highlighted by arrows.</p> "> Figure 2
<p>Representative quartz microbalance with dissipation monitoring (QCM-D) acquisition graphics depicting 5th overtone variations of frequency (descending curves) and dissipation (ascending curves) during construction of the multilayered films using normal–CHT/alginate (ALG) (normal), mesyl–CHT/ALG (mesyl) and tosyl–CHT/ALG (tosyl) (<b>A</b>); Thickness of films after buildup of 5 bilayers was estimated using Voigt-based viscoelastic model (Average ± SD; * <span class="html-italic">p</span> < 0.05) (<b>B</b>); Representative cumulative thickness evolution of the multilayer films as a function of the number of deposition layers. Lines represent linear trend lines with <span class="html-italic">R</span><sup>2</sup> = 0.99 (normal-CHT), <span class="html-italic">R</span><sup>2</sup> = 0.97 (tosyl–CHT) and <span class="html-italic">R</span><sup>2</sup> = 0.97 (mesyl–CHT) (<b>C</b>).</p> "> Figure 3
<p>Representative fluorescence images of live (green) and dead (red) hASCs cultured on 24-well plates with the extract of mesyl–CHT/ALG, tosyl–CHT/ALG and normal–CHT/ALG multilayered films and latex. (<b>A</b>) (Scale bar = 200 µm); Normalized metabolic activity of hASCs was measured in contact with the extract of CHT/ALG multilayered films as well as negative (24-well plate) and positive (Latex) controls (<b>B</b>). Results are presented as average ± SD.</p> "> Figure 4
<p>Representative fluorescence images of the hASCs cultured on top of normal–CHT/ALG (<b>A</b>); mesyl–CHT/ALG (<b>B</b>); tosyl–CHT/ALG (<b>C</b>); multilayered films and glass substrate (<b>D</b>). Multilayered films included subsequent layers of CHT and ALG with top layer (11th layer) was CHT. Scale bar: 200 µm. Cell nuclei appeared bright blue and F-actin filaments appeared bright red due to DAPI and phalloidin dyes, respectively.</p> "> Scheme 1
<p>Chemical structure of mesyl–chitosan (CHT) (<b>A</b>) and tosyl–CHT (<b>B</b>).</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chitosan Purification, Modification and Analysis by 1H NMR
2.2. Construction and Characterization of mCHT/ALG Multilayered Films by Quartz-Crystal Microbalance
2.3. Estimation of the Film Thickness and Properties
2.4. Human Adipose Stem Cell (hASC) Culture
2.5. Preparation of Multilayered Films for Cell Culture Studies
2.6. Cytotoxicity and Viability Analysis
2.7. Cell Adhesion Studies on mCHT/ALG Multilayered Film
2.8. Statistical Analysis
3. Results and Discussion
3.1. Synthesis and Characterization of the Mesyl–CHT and Tosyl–CHT
3.2. LbL Build-Up of mCHT/ALG Multilayered Films at pH = 5.5
3.3. Cytotoxicity of mCHT/ALG Multilayered Films
3.4. Aggregate Formation of hASCs on the mCHT/ALG Multilayered Films
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Polyelectrolyteolyte | ζ-Potentialtial |
---|---|
Normal–CHT | +20 ± 1.3 |
Mesyl–CHT | +22.3 ± 1.6 |
Tosyl–CHT | +19.8 ± 1.1 |
Alginate | −28.6 ± 1.7 |
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Hatami, J.; Silva, S.G.; Oliveira, M.B.; Costa, R.R.; Reis, R.L.; Mano, J.F. Multilayered Films Produced by Layer-by-Layer Assembly of Chitosan and Alginate as a Potential Platform for the Formation of Human Adipose-Derived Stem Cell aggregates. Polymers 2017, 9, 440. https://doi.org/10.3390/polym9090440
Hatami J, Silva SG, Oliveira MB, Costa RR, Reis RL, Mano JF. Multilayered Films Produced by Layer-by-Layer Assembly of Chitosan and Alginate as a Potential Platform for the Formation of Human Adipose-Derived Stem Cell aggregates. Polymers. 2017; 9(9):440. https://doi.org/10.3390/polym9090440
Chicago/Turabian StyleHatami, Javad, Sandra G. Silva, Mariana B. Oliveira, Rui R. Costa, Rui L. Reis, and João F. Mano. 2017. "Multilayered Films Produced by Layer-by-Layer Assembly of Chitosan and Alginate as a Potential Platform for the Formation of Human Adipose-Derived Stem Cell aggregates" Polymers 9, no. 9: 440. https://doi.org/10.3390/polym9090440