Conducting cryogel scaffold as a potential biomaterial for cell stimulation and proliferation

The aim of the study was to demonstrate the potential of the cryogelation technique for the synthesis of the conducting cryogel scaffolds which would encompass the advantages of the cryogel matrix, like the mechanical strength and interconnected porous network as well as the conductive properties of the incorporated conducting polymeric material, polypyrrole. The cryogels were synthesized using different combinations of oxidizing agents and surfactants like, sodium dodecyl sulfate (SDS)/ammonium persulfate (APS), SDS/iron chloride (FeCl3), cetyl trimethyl ammonium bromide (CTAB)/APS, and CTAB/FeCl3. The synthesized gels were characterized by scanning electron microscopic analysis for morphology, Fourier transform infrared spectroscopy for analyzing the presence of the polypyrrole (0.5–4 %) as nano-fillers in the gel. It was observed that the presence of these nano-fillers increased the swelling ratio by approximately 50 %. The synthesized conducting cryogels displayed high stress bearing capacity without being deformed as analysed by rheological measurements. The degradation studies showed 12–15 % degradation in 4 weeks time. In vitro studies with conducting and non-conducting cryogel scaffold were carried out to optimize the stimulation conditions for the two cell lines, neuro2a and cardiac muscle C2C12. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed approximately 25 and 15 % increase in the cell proliferation rate for neuro2a and C2C12 cell line, respectively. This was observed at a specific voltage of 100 mV and 2 V, for a specified duration of 2 h and 1 min, respectively for the conducting scaffold as compared to the control. This can play an important role in tissue engineering applications for cell lines where acquiring a high cell number and functionality is desired.

[1]  Yan Huang,et al.  In vitro characterization of chitosan-gelatin scaffolds for tissue engineering. , 2005, Biomaterials.

[2]  Daniel H. Chen,et al.  Polypyrrole-Titania Nanocomposites Derived from Different Oxidants , 2011 .

[3]  G. Xue,et al.  Preparation of a porous conducting polymer film by electrochemical synthesis–solvent extraction method , 2004 .

[4]  M. Cicerone,et al.  Alternating current electric field effects on neural stem cell viability and differentiation , 2010, Biotechnology progress.

[5]  T. Arinzeh,et al.  Neural differentiation of human neural stem/progenitor cells on piezoelectric scaffolds , 2010, Proceedings of the 2010 IEEE 36th Annual Northeast Bioengineering Conference (NEBEC).

[6]  Kurt E. Geckeler,et al.  ENHANCED ELECTRICAL CONDUCTIVITY OF POLYPYRROLE PREPARED BY CHEMICAL OXIDATIVE POLYMERIZATION: EFFECT OF THE PREPARATION TECHNIQUE AND POLYMER ADDITIVE , 2000 .

[7]  N. Gharavi,et al.  The effect of nanofiller on electrical and mechanical properties of silicone rubber , 2010 .

[8]  Ashok Kumar,et al.  Thermoresponsive poly(N-vinylcaprolactam) cryogels: synthesis and its biophysical evaluation for tissue engineering applications , 2010, Journal of materials science. Materials in medicine.

[9]  I. Cianga,et al.  Review paper: Progress in the Field of Conducting Polymers for Tissue Engineering Applications , 2011, Journal of biomaterials applications.

[10]  Deepti Singh,et al.  Proliferation of Myoblast Skeletal Cells on Three-Dimensional Supermacroporous Cryogels , 2010, International journal of biological sciences.

[11]  Pen-Hsiu Grace Chao,et al.  Effects of Applied DC Electric Field on Ligament Fibroblast Migration and Wound Healing , 2007, Connective tissue research.

[12]  Nigel H Lovell,et al.  Impact of co-incorporating laminin peptide dopants and neurotrophic growth factors on conducting polymer properties. , 2010, Acta biomaterialia.

[13]  Ioannis S. Chronakis,et al.  Conductive polypyrrole nanofibers via electrospinning: Electrical and morphological properties , 2006 .

[14]  Jaroslav Stejskal,et al.  Synthesis and structural study of polypyrroles prepared in the presence of surfactants , 2003 .

[15]  Seeram Ramakrishna,et al.  Applications of conducting polymers and their issues in biomedical engineering , 2010, Journal of The Royal Society Interface.

[16]  Sumrita Bhat,et al.  Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering , 2011, Journal of The Royal Society Interface.

[17]  L. A. Geddes,et al.  The discovery of bioelectricity and current electricity The Galvani-Volta controversy , 1971, IEEE Spectrum.

[18]  Y. Wan,et al.  Porous-conductive chitosan scaffolds for tissue engineering II. in vitro and in vivo degradation , 2005, Journal of materials science. Materials in medicine.

[19]  S. Hsu,et al.  Chitosan as Scaffold Materials: Effects of Molecular Weight and Degree of Deacetylation , 2004 .

[20]  James D. Weiland,et al.  Electrodeposition and Characterization of Thin-Film Platinum-Iridium Alloys for Biological Interfaces , 2011 .

[21]  S. J. Kim,et al.  Biphasic electrical targeting plays a significant role in schwann cell activation. , 2011, Tissue engineering. Part A.

[22]  Kang Wang Enzyme Immobilization on Chitosan-Based Supports , 2011 .

[23]  Alison P McGuigan,et al.  Modular tissue engineering: fabrication of a gelatin‐based construct , 2007, Journal of Tissue Engineering and Regenerative Medicine.

[24]  Eytan Modiano,et al.  Reliability in Layered Networks with Random Link Failures , 2010, INFOCOM 2010.

[25]  Christine E. Schmidt,et al.  Conducting polymers in biomedical engineering , 2007 .

[26]  Milica Radisic,et al.  Electrical stimulation systems for cardiac tissue engineering , 2009, Nature Protocols.

[27]  Y. Wan,et al.  Preparation and characterization of porous conducting poly(dl-lactide) composite membranes , 2005 .

[28]  J. Waldman,et al.  Dielectric properties of polypyrrole doped with tosylate anion in the far infrared and microwave , 1991 .

[29]  R Langer,et al.  Stimulation of neurite outgrowth using an electrically conducting polymer. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[30]  K. Leong,et al.  Chitosan nanoparticles for oral drug and gene delivery , 2006, International journal of nanomedicine.

[31]  M. Mariatti,et al.  Thermal properties and moisture absorption of nanofillers-filled epoxy composite thin film for electronic application , 2012 .

[32]  W. Prissanaroon-Ouajai,et al.  Synthesis of Highly Conductive Polypyrrole Nanoparticles via Microemulsion Polymerization , 2008 .

[33]  D. Mcclements,et al.  Influence of pH, ionic strength, and temperature on self-association and interactions of sodium dodecyl sulfate in the absence and presence of chitosan. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[34]  S. Mikhalovsky,et al.  Gelatin-fibrinogen cryogel dermal matrices for wound repair: preparation, optimisation and in vitro study. , 2010, Biomaterials.

[35]  Y. Wan,et al.  Porous-conductive chitosan scaffolds for tissue engineering, 1. Preparation and characterization. , 2004, Macromolecular bioscience.

[36]  K. Kar,et al.  Synthesis and characterization of elastic and macroporous chitosan-gelatin cryogels for tissue engineering. , 2009, Acta biomaterialia.

[37]  Jiurong Liu,et al.  Synthesis and characterization of polypyrrole/Au nanocomposites by microemulsion polymerization , 2012 .

[38]  Zhongfan Liu,et al.  Controllable synthesis of conducting polypyrrole nanostructures. , 2006, The journal of physical chemistry. B.

[39]  Zaiping Guo,et al.  Low-temperature synthesis of polypyrrole-coated LiV3O8 composite with enhanced electrochemical properties , 2007 .

[40]  Hossein Baharvand,et al.  Application of conductive polymers, scaffolds and electrical stimulation for nerve tissue engineering , 2011, Journal of tissue engineering and regenerative medicine.

[41]  C. Schmidt,et al.  Design of a Novel Electrically Conducting Biocompatible Polymer with Degradable Linkages for Biomedical Applications , 2006 .

[42]  Hyoungshin Park,et al.  Effects of electrical stimulation in C2C12 muscle constructs , 2008, Journal of tissue engineering and regenerative medicine.

[43]  M. Levin Bioelectric mechanisms in regeneration: Unique aspects and future perspectives. , 2009, Seminars in cell & developmental biology.