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pH responsive nanohydrogels based on poly(HEMA-co-DMAEMA) are able to keep the drug molecules (DOX) in physiological pH, circulate in blood vessels for a longer time due to their size (<200 nm), enter and accumulate the tumor microenvironment by enhanced permeability and retention (EPR) effect and release their cargo after swelling is triggered by acidic pH (pH: 5.5) in tumor microenvironment.Successful RAFT synthesis of a pH responsive nanohydrogel.In vitro pH responsive drug release by the nanohydrogel.Cellular uptake and tumor cell cytotoxicity of the drug adsorbed nanohydrogel.Safety and hemocompatibility assessment of the developed nanohydrogel.The advent of smart nanohydrogel has revealed new opportunities for scientists to develop the most efficient anti-cancer vehicles with safe and biocompatible profile. In this experiment, using reversible addition-fragmentation chain transfer polymerization method as a novel, safe and smart pH responsive formulation of poly (hydroxyethyl methacrylate-co-N,N-dimethylaminoethyl methacrylate) and poly (ethylene glycol)-diacrylate as cross-linker were synthesized. The synthesized structure was confirmed by Fourier-transform infrared spectroscopy and proton nuclear magnetic resonance methods. The pH responsive behavior of the synthesized particles was checked by size measurement in two different pH values (5.5 and 7.4) by dynamic light scattering and transmission electron microscopy. The prepared structure had nanometer sizes of 180 in medium with pH of 7.4, when it encountered acidic medium (e.g. pH 5.5), the particles swelled to about 400nm. The efficiency of the prepared pH responsive nanohydrogels was tested as a drug delivery system. An anti-cancer drug, doxorubicin successfully interacted with this material. The release profiles of nanoparticles carrying drug molecules were checked in two different simulated pH of healthy organs (7.4) and tumor site (5.5). Despite lower release in pH of 7.4 (˜20%), an increased drug release of 80% was obtained in pH of 5.5. The in vitro toxicity assay, apoptosis evaluation and uptake experiments were performed on breast cancer cell line (MCF-7), which showed a time dependency cellular entrance, an enhanced cytotoxicity and apoptosis induction by the doxorubicin loaded nanoparticles. Hemolysis assays confirmed the safety and hemocompatibility of the developed nanohydrogel. The suitable size (<200nm), pH responsive behavior, anti-proliferative activity and apoptosis induction in cancer cells and hemocompatibility were the noticeable features of the developed doxorubicin adsorbed nanoparticle, which introduced this formulation as an ideal vehicle in anti-cancer drug delivery.