International Journal of Science and Research (IJSR) ISSN (Online): 2319-7064 Index Copernicus Value (2015): 78.96 | Impact Factor (2015): 6.391 Volume 6 Issue 6, June 2017 www.ijsr.net Licensed Under Creative Commons Attribution CC BY Effect of Electric Fields on Normal Fibroblast (BJ) Cells Omat Rachmat 1 , Ilma Fidayanti 2 , Dewi Ratih 3 , Endang Sutedja 1 , Darmadji Ismono 1 , Nucki N Hidajat 1 1, 4, 5, 6 Faculty of Medicine, University of Padjadjaran, Bandung 40163, West Java, Indonesia 2, 3 Faculty of Medicine, University of Jenderal Achmad Yani, Cimahi, West Java, Indonesia Abstract: Background : Electric fields (EFs)has been reported to affect living tissues.Effects of EFsas anticancer have beenrecently developed. Designing a therapy that specifically recognizeand kills cancer cells without affecting normal cells, has been a recentobjective in theranostics. Thus, this study aimed to evaluate the effect of EFs on normal fibroblast cells (BJ cells). Method : Cells were incubated for 2, 4, and 6 days with frequency of 50 KHz, 100 KHz, 150 KHz, 200 KHz, 300 KHz. Parameters measured were cells number and viability. Cells number and viability assesment was performed with MTS. Results : Number of BJ cells increased each day, compared to 0 day. Number of cells in each day showed no significant difference among treatment of EFs. There was no significant difference of cells viability among treatments each day, and among treatments-0 days, that indicate EFs showed no toxicity toward BJ cells. Conclusion : The result of present study showed there was no significant difference among treatment of EFs in normal fibroblast cells. These results indicate EFs did not affect proliferation in normal cell. Keywords: Electric fields, fibroblast 1. Introduction Electric fields (EFs)has been reported to affect living tissues. Their activity ranged from stimulating excitable tissues such as nerve, muscle or heart in frequency-dependent manner [Pol, 1995; Palti, 1966], through stimulating bone growth and accelerating fracture healing [Besset, 1985], to using it for diathermy and radiofrequency tumour ablation [Chou, 1995]. Intermediate-frequency EFs (>10 KHz to MHz) were mostly considered as having no biological effect [Elson, 1995] and, hence, medical application, though several non- thermal cellular effects have been observed [Zimmerman et al, 1981; Holzapfel et al, 1982; Pawlowski et al, 1993]. Recently, in vitro and in vivo studies on effects of alternating EFsas anticancer, have been documented [Kirson et al, 2004; Kirson et al, 2007; Zimmerman et al, 2012; Barbault et al, 2009]. These studies showed that anticancer effects were achieved at specific (for the cancer cell type) modulation frequencies and demonstrated proliferative inhibition and mitotic spindle disruption following exposure to alternating electric fields [Kirson et al, 2007; Zimmerman et al 2013].Designing a therapy that specifically recognizeand kills cancer cells without affecting normal cells, has been a recentobjective in theranostics [Kievit and Zhang, 2011]. Thus, it is important to observe the effect of EFs on normal cells. In this study, we evaluated effect of EFs on normal fibroblast cells (BJ). 2. Materials And Methods Cell Culture BJ cells [ATCC®CRL-2522] were provided by Biomolecular and Biomedical Research Center, Aretha Medika Utama. Cells were grown in α-Minimum Essential Medium Eagle(α-MEM) [Biowest L0475], 10% Fetal Bovine Serum (FBS) [Biowest S181H], 1% Pennicilin Streptomycin [Biowest L0022], and maintainedat 37°C in humidified atmosphere and 5% CO 2 until the cells were 80- 90% confluence. Growth medium was removed and washed with phosphate buffer saline (PBS) [Gibco 14200075]. Cells were then added with trypsin-EDTA [Biowest L0931-500], incubated at 37 o C for 3 min. Tripsynization was stopped by adding growth medium in equal volume. Cells were suspended and replaced into tube, centrifuged at 500 xgfor 4 min. Supernatant was removed and pellet were resuspendedwith 4-5 mL growth medium. Cell suspension was aliquoted into T-flask containing growth medium with density of8000 cell/cm 2 .Medium was replaced every two days. Cells were incubated at 37 o C, 5% CO 2 . Treatments Cells were washed with 1 mL 1X PBS twice.Cell were then added with 1 mL trypsin EDTA, incubated for 3 min at37 o C, 5% CO 2 .Cells were removed to tube containing 5 mL culture medium, and centrifuged at 500 g for 5 min.Supernatant was removed, cells were resuspended with 1 mL culture medium.Cells were plated into 96 wellswith 5000 cells eachwellin 100 μl medium. . Cells were incubated for 2, 4, and 6 days at 37 o C, 5% CO 2 with frequency of50 KHz, 100 KHz, 150 KHz, 200 KHz, 300 KHz.Cells number and viability were measured with 20μL (3-(4,5-dimethylthiazol- 2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium (MTS) [Promega, Madison, WI, USA],and incubated at 37 o C for 3 hours.Absorbance was measured usingmicroplate reader (MultiSkan-Go)at 490 nm wavelength. 3. Results Cell number and viability were evaluated by MTS assay. MTS assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays (Malich et al., 1997, Widowati et al., 2013). Absorbance was measured using microplate reader at 490 nm wavelength. Cells number of BJ cells can be seen in Table 1. Paper ID: ART20174172 DOI: 10.21275/ART20174172 626