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Partially degradable friction-welded pure iron-stainless steel 316L bone pin

Nasution AK, Murni NS, Sing NB, Idris MH, Hermawan H

J Biomed Mater Res B Appl Biomater, 2015 Jan;103(1):31-8.
PMID: 24757071 DOI: 10.1002/jbm.b.33174

This article describes the development of a partially degradable metal bone pin, proposed to minimize the occurrence of bone refracture by avoiding the creation of holes in the bone after pin removal procedure. The pin was made by friction welding and composed of two parts: the degradable part that remains in the bone and the nondegradable part that will be removed as usual. Rods of stainless steel 316L (nondegradable) and pure iron (degradable) were friction welded at the optimum parameters: forging pressure = 33.2 kPa, friction time = 25 s, burn-off length = 15 mm, and heat input = 4.58 J/s. The optimum tensile strength and elongation was registered at 666 MPa and 13%, respectively. A spiral defect formation was identified as the cause for the ductile fracture of the weld joint. A 40-µm wide intermetallic zone was identified along the fusion line having a distinct composition of Cr, Ni, and Mo. The corrosion rate of the pin gradually decreased from the undeformed zone of pure iron to the undeformed zone of stainless steel 316L. All metallurgical zones of the pin showed no toxic effect toward normal human osteoblast cells, confirming the ppb level of released Cr and Ni detected in the cell media were tolerable.

Matched MeSH terms: Cell Line
Fulltext Evaluation of Hydra HALT-1 as a toxin moiety for recombinant immunotoxin

Jiemy WF, Hiew LF, Sha HX, In LLA, Hwang JS

BMC Biotechnol, 2020 Jun 17;20(1):31.
PMID: 32552895 DOI: 10.1186/s12896-020-00628-9

BACKGROUND: Immunotoxin is a hybrid protein consisting of a toxin moiety that is linked to a targeting moiety for the purpose of specific elimination of target cells. Toxins used in traditional immunotoxins are practically difficult to be produced in large amount, have poor tissue penetration and a complex internalization process. We hypothesized that the smaller HALT-1, a cytolysin derived from Hydra magnipapillata, can be used as the toxin moiety in construction of a recombinant immunotoxin.
RESULTS: In this study, pro-inflammatory macrophage was selected as the target cell due to its major roles in numerous inflammatory and autoimmune disorders. We aimed to construct macrophage-targeted recombinant immunotoxins by combining HALT-1 with anti-CD64-scFv in two orientations, and to assess whether their cytotoxic activity and binding capability could be preserved upon molecular fusion. The recombinant immunotoxins, HALT-1-scFv and scFv-HALT-1, were successfully constructed and expressed in Escherichia coli (E. coli). Our data showed that HALT-1 still exhibited significant cytotoxicity against CD64+ and CD64- cell lines upon fusion with anti-CD64 scFv, although it had half cytotoxic activity as compared to HALT-1 alone. As positioning HALT-1 at N- or C-terminus did not affect its potency, the two constructs demonstrated comparable cytotoxic activities with IC50 lower in CD64+ cell line than in CD64- cell line. In contrast, the location of targeting moieties anti-CD64 scFv at C-terminal end was crucial in maintaining the scFv binding capability.
CONCLUSIONS: HALT-1 could be fused with anti-CD64-scFv via a fsexible polypeptide linker. Upon the successful production of this recombinant HALT-1 scFv fusion protein, HALT-1 was proven effective for killing two human cell lines. Hence, this preliminary study strongly suggested that HALT-1 holds potential as the toxin moiety in therapeutic cell targeting.

Matched MeSH terms: Cell Line
Biogenic Synthesis, Characterization and Evaluation of Silver Nanoparticles from Aspergillus niger JX556221 Against Human Colon Cancer Cell Line HT-29

Chengzheng W, Jiazhi W, Shuangjiang C, Swamy MK, Sinniah UR, Akhtar MS, et al.

J Nanosci Nanotechnol, 2018 May 01;18(5):3673-3681.
PMID: 29442882 DOI: 10.1166/jnn.2018.15364

Nanobiotechnology has emerged as a promising technology to develop new therapeutically active nanomaterials. The present study was aimed to biosynthesize AgNPs extracellularly using Aspergillus niger JX556221 fungal extract and to evaluate their anticancer potential against colon cancer cell line, HT-29. UV-visible spectral characterization of the synthesized AgNPs showed higher absorption peak at 440 nm wavelength. Transmission Electron Microscopy (TEM) analysis revealed the monodispersed nature of synthesized AgNPs occurring in spherical shape with a size in the range of 20-25 nm. Further, characterization using Energy Dispersive Spectroscopy (EDX) confirmed the face-centred cubic crystalline structure of metallic AgNPs. FTIR data revealed the occurrence of various phytochemicals in the cell free fungal extract which substantiated the fungal extract mediated AgNPs synthesis. The cytotoxic effect of AgNPs was studied by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results evidenced the cytotoxic effect of AgNPs on HT-29 cell lines in a dose dependent manner. The highest activity was found at 100 μg/ml concentration after 24 h of incubation. Use of propidium iodide staining examination method confirmed the cytotoxic effect of AgNPs through inducing cell apoptosis. AgNPs cytotoxicity was found to be through elevating reactive oxygen species (ROS), and caspase-3 activation resulting in induced apoptosis. Therefore, this research finding provides an insight towards the development of novel anticancer agents using biological sources.

Matched MeSH terms: Cell Line
In vitro evaluation of cytotoxic properties of 5-Aminolevulinic acid (5-ALA) on bladder cancer cells

Fahmy UA, Fahmy O

Photodiagnosis Photodyn Ther, 2020 Jun;30:101714.
PMID: 32165337 DOI: 10.1016/j.pdpdt.2020.101714

INTRODUCTION: 5-Aminolevulinic (5-ALA) may be used as a photo diagnostic agent in bladder cancer. The aim of this study was to investigate the cytotoxic effect of 5-ALA on bladder cancer cells.
MATERIALS AND METHODS: T24 cells treated with various concentrations of mitomycin (MC), 5-ALA and an MC/5-ALA mixture were evaluated to determine the role of 5-ALA on MC cytotoxicity. Cell cycle analysis was conducted, and apoptosis was analyzed by flow cytometry. Caspase 3 enzyme and reactive oxygen species were measured.
RESULTS: Our initial studies exploring the impact of combination therapy on cell viability demonstrated improved cytotoxic effects on T24 and RT cells with relatively low doses of 5-ALA/MC in conjunction with MC alone. Indicated no significant difference between the IC50 of MC and MC/5-ALA in T24 cells, while IC50 value was decreased by 25 % in RT4 cells in 5-ALA/MC in comparing with MC alone. However, examination of cell cycle phase arrests by flow cytometry revealed significant PreG1 apoptosis and cell growth arrest in G2/M in T24 cells treated with the MC/5-ALA mixture compared with MC treatment. In addition, caspase 3 enzyme was increased by 1.15-fold in T24 cells treated with MC/5-ALA in comparison with MC alone.
CONCLUSION: These results suggest that 5-ALA might possess anti-cancer properties and is not only a photo diagnostic element.

Matched MeSH terms: Cell Line, Tumor
Fulltext The effect of DMEM and DMEM:F12 culture media on the growth of SH-SY5Y cells

Aishah Mohammed Izham, Min, Jasmine Chia Siew, Vidyadaran, Sharmili, Mohd Roslan Sulaiman, Hemabarathy, Bharatham B., Perimal, Enoch Kumar

Life Sciences, Medicine and Biomedicine, 2018;2(3):6-0.
MyJurnal

The human neuroblastoma cell line, SH-SY5Y cells, derived from the parental SK-N-SH cell line, is commonly used as an in vitro model for neuroscience and neurobiology research. Since SH-SY5Y cells are widely cultured for research, several different culture media have been used to optimize the growth of the cells, including Eagle's Minimum Essential Medium (EMEM), Dulbecco’s modified Eagle’s medium (DMEM) and other recently developed culture media. SH-SY5Y cells has the ability to reach confluency in culture flasks ranges from 5 days to 15 days, depending on the culture media used. Hence, the optimization of the culture media is crucial to achieve the fastest growth rate for the cells. The objective of the study is to evaluate the culture media for the proliferation of SH-SY5Y cells. We compared the growth rate of SH-SY5Y cells cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 15% heat-inactivated fetal bovine serum (hiFBS), Dulbecco’s modified Eagle’s medium: Nutrient mixture F-12 (DMEM:F12) + supplemented with 15% hiFBS and DMEM:F12 supplemented with 10% hiFBS. In DMEM:F12 supplemented with 15% hiFBS, cells grew up to 6.67E+05 cells. In DMEM:F12 supplemented with 10% hiFBS, cells grew up to 5.28E+05 cells. In DMEM supplemented with 15% hiFBS, the cells grew up to 4.76E+05 cells. There was a significant difference between culture media DMEM:F12 supplemented with 15% hiFBS as compared to DMEM:F12 supplemented with 10%hiFBS and DMEM supplemented with 15% hiFBS (p0.05). We found that DMEM:F12 supplemented with 15% hiFBS could serve as an optimized culture media for high proliferation rate of SH-SY5Y cells.

Matched MeSH terms: Cell Line
Single Agent and Synergistic Activity of Maritoclax with ABT-263 in Nasopharyngeal Carcinoma (NPC) Cell Lines

Xiang BLS, Kwok-Wai L, Soo-Beng AK, Mohana-Kumaran N

Trop Life Sci Res, 2020 Oct;31(3):1-13.
PMID: 33214852 DOI: 10.21315/tlsr2020.31.3.1

The BCL-2 anti-apoptotic proteins are over-expressed in many cancers and hence are attractive therapeutic targets. In this study, we tested the sensitivity of two Nasopharyngeal Carcinoma (NPC) cell lines HK1 and C666-1 to Maritoclax, which is reported to repress anti-apoptotic protein MCL-1 and BH3 mimetic ABT-263, which selectively inhibits anti-apoptotic proteins BCL-2, BCL-XL and BCL-w. We investigated the sensitisation of the NPC cell lines to these drugs using the SYBR Green I assay and 3D NPC spheroids. We report that Maritoclax repressed anti-apoptotic proteins MCL-1, BCL-2, and BCL-XL in a dose- and time-dependent manner and displayed a single agent activity in inhibiting cell proliferation of the NPC cell lines. Moreover, combination of Maritoclax and ABT-263 exhibited synergistic antiproliferative effect in the HK1 cells. Similar results were obtained in the 3D spheroids generated from the HK1 cells. More notably, 3D HK1 spheroids either treated with single agent Maritoclax or combination with ABT-263, over 10 days, did not develop resistance to the treatment rapidly. Collectively, the findings illustrate that Maritoclax as a single agent or combination with BH3 mimetics could be potentially useful as treatment strategies for the management of NPC.

Matched MeSH terms: Cell Line
Fulltext A Study of the in vitro cytotoxic activity of Gelsemium elegans using human ovarian and breast cancer cell lines

Abdul Wahab K, Ahmad FB, Din LB, Cheah SH, Mock SL

Trop Biomed, 2004 Dec;21(2):139-44.
PMID: 16493406 MyJurnal

The crude methanol extracts of Gelsemium elegans leaves were assessed for their cytotoxic activity using the microculture 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cellular viability. This study utilized two different types of human cancer cell lines, CaOV-3 (human ovarian cancer cells) and MDA-MB-231 (human estrogen receptor negative breast cancer cells), allowing for comparison of toxicity of G. elegans against these two cancer cells lines. Our results showed that the methanol extract of G. elegans exhibited high cytotoxicity against the human ovarian cancer cell line CaOV-3 with an IC50 value of 5microg/ml after 96 h incubation. However, G. elegans displayed discernibly less toxicity against the MDA-MB-231 cells with an IC50 value 40microg/ml after 96 h incubation and this effect was dose- and time-dependent, up to 72h and 20-30 microg/ml. In conclusion, our results demonstrated that G. elegans is potently cytotoxic against the human ovarian cancer cell line CaOV-3 and to a lesser extend towards the human breast carcinoma cancer MDA-MB-231 cells, suggesting that the extract is selective towards CaOV-3 cells and may have a chemotherapeutic role for ovarian cancer treatment in the future.

Matched MeSH terms: Cell Line
Phenolic constituents and anticancer properties of Morus alba (white mulberry) leaves

Chan EWC, Wong SK, Tangah J, Inoue T, Chan HT

J Integr Med, 2020 May;18(3):189-195.
PMID: 32115383 DOI: 10.1016/j.joim.2020.02.006

Flavonoids are by far the most dominant class of phenolic compounds isolated from Morus alba leaves (MAL). Other classes of compounds are benzofurans, phenolic acids, alkaloids, coumarins, chalcones and stilbenes. Major flavonoids are kuwanons, moracinflavans, moragrols and morkotins. Other major compounds include moracins (benzofurans), caffeoylquinic acids (phenolic acids) and morachalcones (chalcones). Research on the anticancer properties of MAL entailed in vitro and in vivo cytotoxicity of extracts or isolated compounds. Flavonoids, benzofurans, chalcones and alkaloids are classes of compounds from MAL that have been found to be cytotoxic towards human cancer cell lines. Further studies on the phytochemistry and anticancer of MAL are suggested. Sources of information were PubMed, PubMed Central, ScienceDirect, Google, Google Scholar, J-Stage, PubChem and China National Knowledge Infrastructure.

Matched MeSH terms: Cell Line, Tumor
Recent advances in TPGS-based nanoparticles of docetaxel for improved chemotherapy

Choudhury H, Gorain B, Pandey M, Kumbhar SA, Tekade RK, Iyer AK, et al.

Int J Pharm, 2017 Aug 30;529(1-2):506-522.
PMID: 28711640 DOI: 10.1016/j.ijpharm.2017.07.018

Docetaxel (DTX) is one of the important antitumor drugs, being used in several common chemotherapies to control leading cancer types. Severe toxicities of the DTX are prominent due to sudden parenteral exposure of desired loading dose to maintain the therapeutic concentration. Field of nanotechnology is leading to resist sudden systemic exposure of DTX with more specific delivery to the site of cancer. Further nanometric size range of the formulation aid for prolonged circulation, thereby extensive exposure results better efficacy. In this article, we extensively reviewed the therapeutic benefit of incorporating d-α-tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS, or simply TPGS) in the nanoparticle (NP) formulation of DTX for improved delivery, tumor control and tolerability. TPGS is well accepted nonionic-ampiphilic polymer which has been identified in the role of emulsifier, stabilizer, penetration enhancer, solubilizer and in protection in micelle. Simultaneously, P-glycoprotein inhibitory activity of TPGS in the multidrug resistant (MDR) cancer cells along with its apoptotic potential are the added advantage of TPGS to be incorporated in nano-chemotherapeutics. Thus, it could be concluded that TPGS based nanoparticulate application is an advanced approach to improve therapeutic efficacy of chemotherapeutic agents by better internalization and sustained retention of the NPs.

Matched MeSH terms: Cell Line, Tumor
Fulltext Crocodylus porosus Gut Bacteria: A Possible Source of Novel Metabolites

Khan NA, Soopramanien M, Maciver SK, Anuar TS, Sagathevan K, Siddiqui R

Molecules, 2021 Aug 18;26(16).
PMID: 34443585 DOI: 10.3390/molecules26164999

Crocodiles are remarkable animals that have the ability to endure extremely harsh conditions and can survive up to a 100 years while being exposed to noxious agents that are detrimental to Homo sapiens. Besides their immunity, we postulate that the microbial gut flora of crocodiles may produce substances with protective effects. In this study, we isolated and characterized selected bacteria colonizing the gastrointestinal tract of Crocodylusporosus and demonstrated their inhibitory effects against three different cancerous cell lineages. Using liquid chromatography-mass spectrometry, several molecules were identified. For the first time, we report partial analyses of crocodile's gut bacterial molecules.

Matched MeSH terms: Cell Line, Tumor
In vitro adhesion and invasion by Cladosporium sphaerospermum in human bronchial epithelial cells (BEAS-2B) and human pulmonary alveolar epithelial cells (HPAEpiC)

Lo SG, Wong SF, Mak JW, Choo KK, Ng KP

Trop Biomed, 2019 Dec 01;36(4):958-971.
PMID: 33597466

Cladosporium spores are ubiquitous in indoor and outdoor environment and may potentially trigger allergic responses upon inhalation. To date, there is limited investigation on the fate of Cladosporium spores after being inhaled into the respiratory tract. This study was conducted to investigate the interaction of Cladosporium sphaerospermum with Human Bronchial Epithelial Cells (BEAS-2B) and Human Pulmonary Alveolar Epithelial Cells (HPAEpiC). C. sphaerospermum conidia were harvested and co-cultured with BEAS-2B or HPAEpiC cells for 72 hours. At each time point (30 minutes, 2, 4, 24, 48 and 72 hours), adherence and invasion of the cells by C. sphaerospermum conidia (and hyphae) were investigated by immunofluorescence staining. This study demonstrated the adherence and internalization of C. sphaerospermum conidia within these epithelial cells. In addition, the conidia were able to germinate and invade the epithelial cells. The ability of the fungal conidia to adhere, internalize, germinate and invade both the bronchial and alveolar epithelial cells of the respiratory tract in vitro might contribute to the understanding of the pathogenesis of Cladosporium in respiratory infection and allergy in vivo.

Matched MeSH terms: Cell Line
Fulltext Interleukin 6 and Interleukin 17a enhance proliferation and differentiation of murine osteoblast and human foetal osteoblast cell lines

Shaminea S, Kannan TP, Norazmi MN, Nurul AA

International Medical Journal Malaysia, 2014;13(2):35-40.
MyJurnal

Introduction: Cytokines have been gaining great focus due to their role in enhancing osseointegration as well as their potential in bone reconstruction. Osseointegration often faces complications in its compatibility with the implant due to rejection by the recipients own immune system. Therefore, extensive studies are being carried out to enhance osteoblast development to minimize such complication. The aim of this study was to determine the effect of different concentrations of Interleukin 6 (IL-6) and Interleukin 17a (IL-17A) in the proliferation and differentiation of murine and human osteoblasts.

Methods: Various concentrations (5, 10, 25 and 50 ng/ml) of rIL-6 and rIL-17A were tested on both murine osteoblast (MC3T3-E1) and human feotal osteoblast (hFOB) cell lines using [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] (MTS) and alkaline phosphatise (ALP) assays. MTS was carried out at 24, 48, 72, 96 and 120 hours while ALP assay was done on day 1, 3, 7, 10 and 14.

Results: MC3T3-E1 cells showed steadier proliferation and differentiation compared to hFOB. Both cell lines expressed responses in dose-dependent manner. The concentration of 10ng for IL-6 and IL-17A in the case of MC3T3-E1 cell line was found to be the most suitable for further studies.

Conclusion: IL-6 and IL-17A enhance proliferation and ALP activity of both MC3T3-E1 and hFOB cell lines.

Matched MeSH terms: Cell Line
Fulltext Hypoglycaemic properties of Malaysian cocoa (Theobroma cacao) polyphenols-rich extract

Ruzaidi, A., Abbe Maleyki, Amin, I., Nawalyah, A.G., Muhajir, H., Pauliena, M.B.S.M., et al.

International Food Research Journal, 2008;15(3):305-312.
MyJurnal

The objective of the study was to investigate the hypoglycaemic properties of Malaysian cocoa (Theobroma cacao) polyphenols extract in-vivo and insulin sensitivity in-vitro. Cocoa extract (CE) (containing 190 - 286 mg total polyphenol per gram extract) was prepared from fermented and roasted (140°C, 20 min) beans by extracting with 80% ethanol in the ratio of 1 to 10. For the in-vivo study, the CE was administered in three dosages (1%, 2%, and 3%) to groups of normal and diabetic rats for a period of 4 weeks by forcefeeding. Results showed that dosages of 1% and 3% CE significantly reduced (p < 0.05) plasma glucose levels in the diabetic rats. An in-vitro study (BRIN-BD11 cell lines) was used to evaluate the effect of CE on insulinsensitivity. The results demonstrated that CE at a concentration of 0.1 mg/ml significantly increased (p < 0.05) insulin level compared to the control. The results of this study showed that Malaysian cocoa polyphenol extract have the potential of being an insulin-mimetic agent. Further studies are on-going to elucidate the underlying mechanisms of polyphenols present in CE that contribute to the reduction of plasma glucose levels and insulin mimicking activity.

Matched MeSH terms: Cell Line
Fulltext A novel 4-arm DNA/RNA Nanoconstruct triggering Rapid Apoptosis of Triple Negative Breast Cancer Cells within 24 hours

Tung J, Tew LS, Hsu YM, Khung YL

Sci Rep, 2017 04 11;7(1):793.
PMID: 28400564 DOI: 10.1038/s41598-017-00912-3

Measuring at ~30 nm, a fully customizable holliday junction DNA nanoconstruct, was designed to simultaneously carry three unmodified SiRNA strands for apoptosis gene knockout in cancer cells without any assistance from commercial transfection kits. In brief, a holliday junction structure was intelligently designed to present one arm with a cell targeting aptamer (AS1411) while the remaining three arms to carry different SiRNA strands by means of DNA/RNA duplex for inducing apoptosis in cancer cells. By carrying the three SiRNA strands (AKT, MDM2 and Survivin) into triple negative breast MDA-MB-231 cancer cells, cell number had reduced by up to ~82% within 24 hours solely from one single administration of 32 picomoles. In the immunoblotting studies, up-elevation of phosphorylated p53 was observed for more than 8 hours while the three genes of interest were suppressed by nearly half by the 4-hour mark upon administration. Furthermore, we were able to demonstrate high cell selectivity of the nanoconstruct and did not exhibit usual morphological stress induced from liposomal-based transfection agents. To the best of the authors' knowledge, this system represents the first of its kind in current literature utilizing a short and highly customizable holliday DNA junction to carry SiRNA for apoptosis studies.

Matched MeSH terms: Cell Line, Tumor
Fulltext In vitro cytotoxicity evaluation of dental porcelain using human cell lines

Hazem Yousef Abu Sharbeh, Kannan, Thirumulu Ponnuraj, Raja Azman Raja Awang, Adam Husein

Archives of Orofacial Sciences, 2012;7(2):68-74.
MyJurnal

The in vitro cytotoxic potential of locally produced dental porcelain was evaluated in this study. The cellular response of human osteoblast and fibroblast cell lines were assessed using MTT assay by incubating with the fluid extract of dental porcelain powder and dental porcelain discs (direct test). Aging process was carried out by submerging the discs into 3% bovine serum albumin (BSA) solution. Tests on extracts showed that dental porcelain was significantly different from the control at a concentration of 250 mg/ml. Direct test showed that dental porcelain after aging was not significantly different from the control with a mean (SD) of 89.2 (13.4)%, whereas, it was significantly different from the control before conditioning of BSA with a mean (SD) of 88.5 (12.1)%. However, the dental porcelain caused mild suppression of succinate dehydrogenase activity (

Matched MeSH terms: Cell Line
Fulltext Optimization of cell density and LPS concentration for the evaluation of nitric oxide production on BV-2 cells in a griess assay

Nasim Karim Hosseini, Jose, Shinsmon, Vidyadaran, S., Syafinaz Amin Nordin

Malaysian Journal of Medicine and Health Sciences, 2014;10(2):1-8.
MyJurnal

Introduction: Production of nitric oxide (NO) is one of the main responses elicited by a variety of
immune cells such as macrophages (e.g. microglia, resident macrophages of brain), during inflammation. Evaluation of NO levels in the inflammatory milieu is considered important to the understanding of the intensity of an immune response; and has been performed using different methods including the Griess assay. To assay NO in culture, an appropriate number of cells are stimulated into an inflammatory phenotype. Common stimuli include lipopolysaccharide (LPS), IFN-γ and TNF-α. However, overt stimulation could cause cell cytotoxicity therefore an ideal concentration of LPS should be used. Objective: To set-up a model of BV-2 cell activation that allows the assay of detectable levels of NO. Optimization of BV-2 microglia cell density and LPS concentrations after stimulation by bacterial lipopolysaccharide (LPS) for the Griess assay is demonstrated in this study. Methods: BV-2 microglia were cultured at different cell densities, and treated with LPS at three concentrations (1, 5, 10 μg/ml). NO production in culture supernatants were then measured at 18, 24, 48 and 72 hours. Moreover, methyl tetrazolium assay (MTT) was also performed to ensure that NO measurement is performed at no-cytotoxic concentrations of LPS. Results and Conclusions: NO production follows a temporal pattern. The density of 25000 cells/ well was the ideal seeding density for NO evaluation in BV-2 cells. BV-2 stimulation by LPS is dose dependent, and NO levels are increased proportional to the LPS concentration up to 1.0μg/ml, whereas the higher LPS concentrations are associated with decreased cell viability may be caused by the high toxic levels of LPS or NO. Although Griess assay has been commonly used by the scientists, however, optimization of its parameters on BV-2 cells will be useful for the experiments which will be performed on this particular cell line. The optimized pattern of Griess assay on BV-2 cells was achieved in this study, hence easier and more practical for the future scientists to perform Griess assay on BV-2 cells.

Matched MeSH terms: Cell Line
Fulltext Optimization and validation of a cell-based tyrosinase assay for screening of tyrosinase inhibitors

Chan, Y.Y., Kim, K.H., Cheah, S.H.

JUMMEC, 2011;14(2):1-4.
MyJurnal

Tyrosinase is a key enzyme that catalyzes melanogenesis in human skin. It oxidizes tyrosine to L-3,4-dihydroxyphenylalanine (L-DOPA) and subsequently to dopachrome, which further polymerizes to melanin pigments. Therefore finding an effective tyrosinase inhibitor, either from synthetic or natural sources, is not only useful as skin whitening agents in cosmetic application, but also beneficial in treating melanin-related disorders. The present study reports of the optimized and validated results of a cell-based tyrosinase assay using B16F10 murine melanoma cell line, which produces melanin pigments and has been used extensively in antimelanogenesis studies. The optimization studies involved 3 parameters (1) optimal seeding cell number per well for total protein extraction; (2) optimal dopachrome formation from enzymatic reaction between total protein (tyrosinase source) and L-DOPA (substrate); and (3) optimal incubation period after the addition of substrate. The present study demonstrates that using seeding cell number of 2 × 105 cells/well, total protein of 40 μg, L-Dopa of 5 mM,and at an incubation period of 1 hour at 37°C provided the optimal response on cultured melanoma cells. Kojic acid, a standard tyrosinase inhibitor, was used as a positive control in the optimized cell-based tyrosinase assay to validate the usefulness of the assay. CONCLUSION: The use of the mentioned protocol is sensitive to determine changes in melanoma cells as the result of tyrosine inhibitors.

Matched MeSH terms: Cell Line
Fulltext Gamma-tocotrienol Alters Protein Expression of HepG2 Cell Line

Farahani, A.S.R., Zakiah, J., Abdul Rahman, M., Karsani, S.A., Wan, Ngah Wz

Medicine & Health, 2008;3(2):256-262.
MyJurnal

Gamma-tocotrienol (GTT) has been shown to exhibit significant antitumor activity in a variety of tumor cells. Previous findings have demonstrated that GTT had antiprolifera-tive effects on a liver cancer cell line (HepG2) with an IC50 value of 170μM. In this study, two dimensional gel electrophoresis (2DE) was used to determine changes in protein expression in HepG2 cell line following treatment with GTT. The ultimate aim is to identify the possible molecular mechanisms involved in GTT antitumor activity. This study is focused on obtaining a 2DE protein profile for HepG2 cell line with and without
GTT treatment. In the preliminary analysis of the resulting 2DE profiles, 18 protein spots were found to be differentially expressed in cells treated with GTT. This observa-tion is confirmed by extending the analysis to a larger sample size. By studying the effects of GTT treatment on differential protein expression in HepG2 cells the underly-ing mechanisms involved in the antitumor activity of GTT may be elucidated.

Matched MeSH terms: Cell Line
Limonoids and triterpenoid from fruit of Swietenia macrophylla

Sun YP, Zhu LL, Liu JS, Yu Y, Zhou ZY, Wang G, et al.

Fitoterapia, 2018 Mar;125:141-146.
PMID: 29325928 DOI: 10.1016/j.fitote.2018.01.004

Five new limonoids, swieteliacates A-E (1-5) and a tirucallane-type triterpenoid, swietesenin (6), together with four known compounds (7-10) were isolated from fruit of Swietenia macrophylla. Their structures were determined by spectroscopic analyses. The new compounds were tested in vitro for their cytotoxic effects against five human cancer cell lines. Compound 2 exhibited moderate cytotoxic activities against SW480 and HL-60 cancer cell lines with IC50 values of 30.6 and 32.9μM, respectively.

Matched MeSH terms: Cell Line, Tumor
Fulltext Design, synthesis, antimicrobial and cytotoxicity study on human colorectal carcinoma cell line of new 4,4'-(1,4-phenylene)bis(pyrimidin-2-amine) derivatives

Kumar S, Lim SM, Ramasamy K, Mani V, Shah SAA, Narasimhan B

Chem Cent J, 2018 Jun 25;12(1):73.
PMID: 29938365 DOI: 10.1186/s13065-018-0440-3

BACKGROUND: Pyrimidine molecules attracted organic chemists very much due to their biological and chemotherapeutic importance. Their related fused heterocycles are of interest as potential bioactive molecules so, we have designed and prepared a new class of 4,4'-(1,4-phenylene)bis(pyrimidin-2-amine) molecules and screened for their in vitro antibacterial, antifungal and cytotoxicity studies.
RESULTS: The structures of synthesized bis-pyrimidine molecules were confirmed by physicochemical and spectral means. The synthesized compounds were further evaluated for their in vitro biological potentials i.e. antimicrobial activity using tube dilution method and anticancer activity against human colorectal carcinoma (HCT116) cancer cell line by Sulforhodamine B assay.
CONCLUSIONS: The biological study demonstrated that compounds s7, s8, s11, s14, s16, s17 and s18 have shown more promising antimicrobial activity with best MIC values than the cefadroxil (antibacterial) and fluconazole (antifungal) and compound s3 found to have better anticancer activity against human colorectal carcinoma (HCT116) cancer cell line.

Matched MeSH terms: Cell Line

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