Effect of the Polymer Modification on the Cytocompatibility of Human and Rat Cells
Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable, semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H2O and CO2. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1131806Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 406
 M. I. Santos, K. Tuzlakoglu, S. Fuchs, M. E. Gomes, K. Peters, R. E. Unger, E. Piskin, R. L. Reis, J. Kirkpatrick, “Endothelial cell colonization and angiogenic of potential on combined nano- and micro-fibrous scaffolds for bone tissue engineering”, Biomaterials, vol. 29, pp. 4306-4313, 2008.
 A. Wang, Q. Ao, Y. Wei, K. Gong, X. Liu, N. Zhao, Y. Gong, X. Zhang, “Physical properties and biocompatibility of a porous chitosan-baset fiberreinforced conduit for nerve regeneration”, Biotechnol. Lett., vol. 29, pp. 1697-1702, 2007.
 G. S. Sailaja, P. Ramesh, T. V. Kumary, H. K. Varma, “Synthetic structure of industrial plastics”, Acta Biomaterialia, vol. 2, pp. 651-657, 2006.
 M. O. Montjovent, S. Mark, L. Mathieu, C. Scaletta, A. Scherberich, C. Delabarde, P. Y. Zambelli, P. E. Bourban, L. H. Applegate, D. P. Pioletti, “Human fetal bone cells associated with ceramic reinforced PLA scaffolds for tissue engineering”, Bone, vol. 42, pp. 554-464, 2008.
 C. Y. Hsieh, S. P. Tsai, D. M. Wang, Y. N. Chang, H. J. Hsieh, “Preparation of γ-PGA/chitosan composite tissue engineering matrices”, Biomaterials, vol. 26, pp. 5617-5623, 2005.
 L. Bačákova, E. Filová, F. Rypáček, V. Švorčík, V. Starý, “Cell adhesion on artificial materials for tissue engineering”, Physiol. Res., vol. 53, pp. 35-45, 2004.
 L. Bačáková, V. Švorčík, “Cell colonization control by physical and chemical modification of materials”, In Cell Growth Processes: New Research; D. Kimura, Ed., Nova Science Publishers, Inc.: Hauppauge, New York, 2008, pp. 5-56.
 M. T. Khorasani, H. Mirzadeh, S. Irani, “Plasma surface modification of poly (l-lactic acid) and poly (lactic-co-glycolic acid) films for improvement of nerve cells”, Radiat. Phys. Chem., vol. 77, pp. 280-287, 2008.
 Y. Wan, J. Yang, J. Bei, S. Wang, “Cell adhesion on gaseous plasma modified poly-(l-lactide) surface under shear stress field”, Biomaterials, vol. 24, pp. 3757-3764, 2003.
 R. Mikulíková, S. Moritz, T. Gumpenberger, M. Olbrich, C. Romanin. L. Bačáková, V. Švorčík, J. Heitz, “Cell microarrays on photochemically modified PTFE”, Biomaterials, vol. 26, pp. 5572-5580, 2005.
 J. Y. Lim, H. J. Donahue, “Cell Sensing and Response to Micro- and Nanostructured Surfaces Produced by Chemical and Topographic Patterning”, Tissue Eng., vol. 13. pp. 1879-1891, 2007.
 L. Bačáková, V. Starý, O. Kofroňová, V. LISÁ, “Polishing and coating carbon fibre-reinforced carbon composites with a carbon-titanium layer enhances adhesion and growth of osteoblast-like MG63 cells and vascular smooth muscle cells in vitro”, J. Biomed. Mater. Res., vol. 54, pp. 567-578, 2001.
 L. Bačáková, V. Starý, J. Horník, P. Glogar, V. Lisá, O. Kofroňová, “Osteoblast-like MG63 cells in cultures on carbon fibre-reinforced carbon composites”, Eng. Biomater., vol. 4, pp. 17-19, 2001.
 L. Bačáková, V. Starý, P Glogar, V. Lisá, “Adhesion, differentiation and immune activation of human osteogenic cells in cultures on carbon-fibre reinforced carbon composites.”, Eng. Biomater., vol. 6, pp. 30-33, 2003.
 B. Dvořánková, Z. Holíková, J. Vacík, R. Königová, Z. Kapounková, J. Michálek, M. Přádný, K. Smetana jr., “Reconstruction of epidermis by grafting of keratinocytes cultured on polymer support - clinical study”, Int. J. Dermatol., vol. 42, pp. 219-223, 2003.
 Y. Zhang, K. Dilaware, J. Delling, E. Tobiasch, “Mechanisms Underlying the Osteo- and Adipo-Differentiation of Human Mesenchymal Stem Cells”, The Scientific World Journal, Vol. 2012, pp. 1-14, 2012.
 M. Tobita, H. Orbay, H. Mizuno, “Adipose-derived stem cells: current findings and future perspectives”, Discov. Med., vol. 11, pp. 160-170, 2011.
 M. Witkowska-Zimny, K. Walenko, “Stem cells from adipose tissue”, Cell. Mol. Biol. Lett., vol. 16, pp. 236-257, 2011.
 M. Barba, C. Cicione, C. Bernardini, F. Michetti, W. Lattanzi, “Adipose-Derived Mesenchymal Cells for Bone Regereneration: State of the Art”, Biomed. Res. Int., vol. 2013, pp. 1-10, 2013.
 L. E. Kokai, K. Marra, J. P. Rubin, “Adipose stem cells: Biology and clinical application for tissue repair and regeneration”, Transl. Res., vol. 134, pp. 399-408, 2014.
 W. K. Ong, S. Sugii, “Adipose-derived stem cells: Fatty potentials for therapy”, Int. J. Biochem. Cell B., vol. 45, pp. 1083-1086, 2013.
 V. Svorcik, V. Rybka, V. Hnatowicz, K. Smetana, “Structure and biocompatibility of ion beam modified polyethylene”, J. Appl. Mater. Sci. Mater. Med., vol. 8, pp. 435-440, 1997.
 M. Parizek, N. Kasalkova, L. Bacakova, P. Slepicka, V. Lisa, M. Blazkova, V. Svorcik, “Improved Adhesion, Growth and Maturation of Vascular Smooth Muscle Cells on Polyethylene Grafted with Bioactive Molecules and Carbon Particles”, Int. J. Mol. Sci., vol 10, pp. 4352-4374, 2009.
 M. Vandrovcova, J. Vacik, V. Svorcik, P. Slepicka, N. Kasalkova, V. Vorlicek, V. Lavrentiev, V. Vosecek, L. Grausova, V. Lisa, L. Bacakova, “Fullerene C60 and hybrid C60/Ti films as substrates for adhesion and growth of bone cells”, Phys. stat. sol. (a), vol. 205, pp. 2252–2261, 2008.