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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

by Javad Hatami 1,2,†, Sandra G. Silva 1,2,‡, Mariana B. Oliveira 1,2,†, Rui R. Costa 1,2, Rui L. Reis 1,2 and João F. Mano 1,2,*,†
1
3B’s Research Group, Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence of Tissue Engineering and Regenerative Medicine, Avepark—Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco GMR, Portugal
2
ICVS/3B’s, PT Government Associated Laboratory, Braga/Guimarães, Portugal
*
Author to whom correspondence should be addressed.
Current address: Department of Chemistry, CICECO—Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal
Current address: LAQV/REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Science, University of Porto, Rua do campo Alegre s/n, 4169-007 Porto, Portugal
Polymers 2017, 9(9), 440; https://doi.org/10.3390/polym9090440
Received: 10 August 2017 / Revised: 5 September 2017 / Accepted: 6 September 2017 / Published: 13 September 2017
(This article belongs to the Collection Polysaccharides)
The construction of multilayered films with tunable properties could offer new routes to produce biomaterials as a platform for 3D cell cultivation. In this study, multilayered films produced with five bilayers of chitosan and alginate (CHT/ALG) were built using water-soluble modified mesyl and tosyl–CHT via layer-by-layer (LbL) self-assembly. NMR results demonstrated the presences of mesyl (2.83 ppm) and tosyl groups (2.39, 7.37 and 7.70 ppm) in the chemical structure of modified chitosans. The buildup of multilayered films was monitored by quartz-crystal-microbalance (QCM-D) and film thickness was estimated using the Voigt-based viscoelastic model. QCM-D results demonstrated that CHT/ALG films constructed using mesyl or tosyl modifications (mCHT/ALG) were significantly thinner in comparison to the CHT/ALG films constructed with unmodified chitosan (p < 0.05). Adhesion analysis demonstrated that human adipose stem cells (hASCs) did not adhere to the mCHT/ALG multilayered films and formed aggregates with sizes between ca. 100–200 µm. In vitro studies on cell metabolic activity and live/dead staining suggested that mCHT/ALG multilayered films are nontoxic toward hACSs. Multilayered films produced via LbL assembly of ALG and off-the-shelf, water-soluble modified chitosans could be used as a scaffold for the 3D aggregates formation of hASCs in vitro. View Full-Text
Keywords: layer by layer assembly; chitosan; alginate; cytotoxicity; multilayered film; adhesion; 3D culture; spheroid layer by layer assembly; chitosan; alginate; cytotoxicity; multilayered film; adhesion; 3D culture; spheroid
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MDPI and ACS Style

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.

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