Culture of human keratinocytes on polypyrrole-based conducting polymers

Variously loaded polypyrrole films, including those containing proteins and polysaccharides, were prepared on gold-coated polycarbonate coverslips. The characteristics of human keratinocytes were studied on these films by microscopy, biochemical assays, and immunocytochemistry. We found keratinocyte viability to be load dependent. For chloride, polyvinyl sulphate, dermatan sulphate, and collagen-loaded polypyrrole films, keratinocyte viability as assessed by the AlamarBlue assay was respectively 47.22, 60.43, 87.71, and 22.65% of tissue culture polystyrene controls after 5 days. This was found to require a previously unreported polymer washing step prior to cell seeding due to the observed toxicity of untreated films. In the case of bare polycarbonate and gold substrates, viability was respectively 75.44 and 61.04% of tissue culture polystyrene controls after 5 days. Keratinocytes stained positive for PCNA (proliferation), K10 (suprabasal differentiation), and K16 (hyperproliferation) markers although cell morphology was poor for organotypical cultures on dermatan- loaded polypyrrole compared with de-epidermalized dermis. From our studies, we concluded that optimized polypyrrole films adequately support keratinocyte growth in submerged cultures with some improvements needed for organotypical cultures. Polypyrrole composites are attractive candidates for tissue-engineering applications since they may incorporate biomolecules and are electrically addressable with the potential to both direct and report on cell activity.