Cell engineering: nanometric grafting of poly-N-isopropylacrylamide onto polystyrene film by different doses of gamma radiation

Abstract

Esmaeil Biazar1, Reza Zeinali2, Naser Montazeri1, Khalil Pourshamsian1, Mahmoud Jabarvand Behrouz3, Azadeh Asefnejad2, Ahad Khoshzaban3, Gholamreza Shahhosseini4, Mostafa Soleimannejad Najafabadi5, Reza Abyani2, Hamidreza Jamalzadeh6, Mahdi Fouladi1, Sasan Rahbar F Hagh7, Aylar Shams Khamaneh1, Soudabeh Kabiri1, Saeed Heidari Keshel3, Ana Mansourkiaei61Department of Chemistry, 6Department of Biology, 7Young Researchers Club, Islamic Azad University, Tonekabon Branch, Mazandaran, Iran; 2Biomedical Engineering Faculty, Islamic Azad University, Science and Research Branch, Tehran, Iran; 3Stem Cell Preparation Unit, Farabi Hospital, Tehran University Medical Science, Tehran, Iran; 4Agricultural, Medical and Industrial Research School, Nuclear Science and Techniques Research Institute, Karaj, Iran; 5Material Engineering Faculty, Islamic Azad University, Najafabad Branch, Isfahan, IranAbstract: Poly-N-isopropylacrylamide was successfully grafted onto a polystyrene cell culture dish and γ-preirradiated in air. In this study, the effect of a γ-pre-irradiation dose of radiation (radiation absorbed dosages of 10, 20, 30, 40 KGy) under appropriate temperature and grafting conditions was investigated. The Fourier transform infrared spectroscopy analysis showed the existence of the graft poly-N-isopropylacrylamide (PNIPAAm) on the substrate. The optimal value of the dose for grafting was 40 KGy at 50°C. The scanning electron microscopy and atomic force microscopy (AFM) images clearly showed that increasing the absorbed dose of radiation would increase the amount of grafting. Surface topography and graft thickness in AFM images of the radiated samples showed that the PNIPAAm at the absorbed dose of radiation was properly grafted. The thickness of these grafts was about 50–100 nm. The drop water contact angles of the best grafted sample at 37°C and 10°C were 55.3 ± 1.2° and 61.2 ± 0.9° respectively, which showed the hydrophilicity and hydrophobicity of the grafted surfaces. Differential scanning calorimetry analysis also revealed the low critical solution temperature of the grafted sample to be 32°C. Thermoresponsive polymers were grafted to dishes covalently which allowed fibroblast cells to attach and proliferate at 37°C; the cells also detached spontaneously without using enzymes when the temperature dropped below 32°C. This characteristic proves that this type of grafted material has potential as a biomaterial for cell sheet engineering.Keywords: Nanometric grafting, PNIPAAm, polystyrene film, gamma ray, dose, cell engineering