Cerium nanoparticles synthesized using aqueous extract of Centella asiatica: characterization, determination of free radical scavenging activity and evaluation of ecacy against cardiomyoblast hypertrophy Vandana Sankar,Palayyan SalinRaj, Raj Athira, Rema Sreenivasan Soumya and Kozhiparambil Gopalan Raghu * Cerium nanoparticles (CeNPs) are used in biomedical applications due to their autocatalytic properties. Even though their use in the management of neurodegenerative diseases, diabetes, etc. has been reported, their role in cardiovascular disease management remains largely unexploited. Moreover, conventional procedures for synthesizing metal nanoparticles are non- environmentally friendly and require high-priced chemicals. Therefore, there is a need for eco friendly and cost eective green technologiesfor CeNP synthesis. This study is the rst of its kind to green synthesize, characterize, and evaluate the ecacy of CeNPs against cardiomyoblast hypertrophy. CeNPs were synthesized using 1 : 5 volume ratios of Centella asiatica aqueous extract and an aqueous solution of cerium ammonium nitrate, and were characterized by various methods such as UV-visible spectroscopy, DLS, SEM and HRTEM, EDX, etc. The particles were demonstrated to be spherical and monodispersed with 8 nm diameter. Zeta potential was found to be negative. Superoxide and hydroxyl radical scavenging activities of CeNPs were found to be higher when compared to bulk cerium and the extract. H9c2 rat cardiomyoblasts were used for cell line studies. Flow cytometry and uorescence imaging were employed for the cell line experiments. High cellular uptake and viability were revealed for CeNPs when compared to macro counterparts. Intracellular superoxide anion generation was less on incubation with higher concentrations of CeNPs. Moreover, preliminary results provided an insight into the potential of CeNPs in attenuating isoproterenol-stimulated cardiomyoblast hypertrophy and has been linked to the inhibition of intracellular calcium overload and reactive oxygen species. 1. Introduction Cerium is the most abundant among the lanthanide series of elements and is widely used for various applications such as in catalytic converters, solid oxide fuel cells, and as excellent oxygen buers. Unlike the other elements of the lanthanide series, which exhibit a trivalent state (+3), the cerium atom can exist in either the +3 or +4 state. This is due to the requisite of only small amounts of energy to change the relative occupancy of the 4f and 5d orbitals. Cerium nanoparticles (CeNPs) demonstrated strikingly dierent autocatalytic properties compared to their macro counterparts, and the ability of nanoceria to switch between oxidation states is comparable to that of biological antioxidants. Cerium atom also exhibits oxygen vacancies or defects in the lattice structure; these arise through loss of oxygen and or its electrons, alternating between CeO 2 and CeO 2x during redox reactions. This property has attracted lots of attention among researchers worldwide for potential application in nanomedicine as a free radical scav- enger. 1 Although cerium nanoparticles are perceived as impressive antioxidants for therapeutic applications, very little consideration has been given to the fact that one needs to synthesize biocompatible cerium nanoparticles with better stability and biocompatibility in physiological solutions. 2 The conventional methods that have been employed in the synthesis of nanoparticles require several processing steps, high-priced chemicals, expensive instruments and moreover, may generate hazardous toxic chemicals to the ecosystem. 3,4 This has prompted the researchers to develop safe and eco- Agroprocessing and Natural Products Division, Council of Scientic and Industrial Research (CSIR) -National Institute for Interdisciplinary Science and Technology (NIIST), Industrial Estate P.O, Pappanamcode, Thiruvananthapuram-695019, Kerala, India. E-mail: raghukgopal2009@gmail.com; Fax: +91-471- 2491712/2490186; Tel: +91-9495902522 Electronic supplementary information (ESI) available. See DOI: 10.1039/c4ra16893c These authors contributed equally to the work and deserve to have the status of rst author. Cite this: RSC Adv. , 2015, 5, 21074 Received 23rd December 2014 Accepted 16th February 2015 DOI: 10.1039/c4ra16893c www.rsc.org/advances 21074 | RSC Adv. , 2015, 5, 2107421083 This journal is © The Royal Society of Chemistry 2015 RSC Advances PAPER