Evaluation of in vitro toxicity of Rumalaya liniment using mouse embryonic fibroblasts and human keratinocytes
DOI:
https://doi.org/10.22377/ijgp.v5i1.165Abstract
The skin irritation potential of topical formulations is investigated prior to human exposure to identify the chemicals which mightinduce adverse skin reactions. Rumalaya liniment (RL) a novel formulation using natural oils and plant extracts is used for reducing inflammations associated with musculoskeletal disorders. Preclinical studies on RL are needed prior to skin application. The aim of the study was to evaluate the possible cytotoxic effects of RL and a commercial sample (CS) on mouse embryo fibroblasts and human keratinocytes using neutral red uptake, (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and resazurin assay. The CTC50 values obtained for RL was significantly higher than that of CS, which revealed that RL is less toxic to CS. RL was less toxic (<17%) on both cell lines at 400 μg/ml and was nontoxic at further lower concentrations, whereas the toxicity of CS was above 59% even at 400 μg/ml. It was observed from the present study that by using three different assay methods and two different
cell lines, the toxicity of RL was significantly lower than that observed with CS. From the study, it could be concluded that RL could be safer to skin due to their low cytotoxicity as compared with CS.
Key words: Cell based assay, cell lines, cytotoxicity, herbal formulation, skin irritation
Downloads
References
Vinardell MP, Benavides T, Mitjans M, Infante MR, Clapes
P, Clothier R. Comparative evaluation of cytotoxicity and
phototoxicity of mono and diacylglycerol amino acid-based
surfactants. Food Chem Toxicol 2008;46:3837-41.
Zanatta CF, Ugartondo V, Mitjans M, Rocha-Filho P, Vinardell MP.
Low cytotoxicity of creams and lotions formulated with Buriti oil
(Mauritia flexuosa) assessed by the neutral release test. Food Chem
Toxicol 2008;46:2776-81.
Nemova EP, Smolnikova NM, Tanirbergenov TB, Durnev AD.
Studies on reproductive toxicity of Rumalaya. Pharm Chem J
;41:399-402.
Sanchez L, Mitjans M, Infante MR, Vinardell MP. Assessment of
the potential skin irritation of lysine-derivative anionic surfactants
using mouse fibroblasts and human keratinocytes as an alternative
to animal testing. Pharm Res 2004;21:1637-41.
Moreno JJ. Arachidonic acid release and prostaglandin E-2
synthesis as irritant index of surfactants in 3T6 fibroblast cultures.
Toxicology 2000;143:275-82.
Korting HC, Herzinger T, Hartinger A, Kerscher M, Angerpointner
T, Maibach HI. Discrimination of the irritancy potential of
surfactants in vitro by two cytotoxicity assays using normal human
keratinocytes, HaCaT cells and 3T3 mouse fibroblast: Correlation
with in vivo data from a soap chamber assay. J Dermatol Sci
;7:119-29.
Zuang V, Balls M, Botham PA, Coquette A, Corsini E, Curren RD,
et al. Follow-up to the ECVAM prevalidation study on in vitro tests
for acute skin irritation: The European centre for the validation of
alternative methods skin irritation task force report 2. Altern Lab
Anim 2002;30:109-29.
Spielmann H, Balls M, Dupuis J, Pape WJ, Pechovitch G, de
Silva O, et al. The international EU/COLIPA in vitro phototoxicity
validation study results of phase II (blind trial), Part 1: The 3T3
NRU phototoxicity test. Toxicol In Vitro 1998;12:305-27.
Wilhelm KP, Boettjer B, Siegers CP. Quantitative assessment of
primary skin irritants in vitro in a cytotoxicity model: Comparison
with in vivo human irritation tests. Br J Dermatol 2001;145:709-15.
Borenfreund E, Puerner JA. Toxicity determined in vitro by
morphological alterations and neutral red absorption. Toxicol
Lett 1985;24:119-24.
Ridell RJ, Clothier RH, Ball M. An evaluation of three in vitro
cytotoxicity assays. Food Chem Toxicol 1986;24:469-71.
Mosmann T. Rapid colorimetric assay for cellular growth and
survival: Application to proliferation and cytotoxicity assays. J
Immunol Met 1983;65:55-63.
O’Brien J, Wilson I, Orton T, Pognan F. Investigation of the alamar
blue (resazurin) fluorescent dye for the assessment of mammalian
cell cytotoxicity. Eur J Biochem 2000;267:5421-6.
Clothier R, Starzec G, Pradel L, Baxter V, Jones M, Cox H, et al.
The prediction of human skin responses by using the combined
in vitro fluorescein leakage/Alamar Blue (resazurin) assay. ATLA
Altern Lab Anim 2002;30:493-504.
Osborne R, Perkins MA. An approach for development of
alternative test methods based on mechanisms of skin irritation.
Food Chem Toxicol 1994;32:133-42.
Suman G, Jamil K. Application of human lymphocytes for
evaluating toxicity of anti-cancer drugs. Int J Pharm 2006;2:374-81.
Clothier RH, Beed M, Samson R, Ward R. An in vitro approach
to the evaluation of repeat exposure in the prediction of toxicity.
Toxicol In Vitro 1997;11:679-82.
Lee JK, Kim DB, Kim JI, Kim PY. In vitro cytotoxicity tests on
cultured human skin fibroblasts to predict skin irritation potential
of surfactants. Toxicol In Vitro 2000;14:345-9.
Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham
A, Fusenig NE. Normal keratinisation in a spontaneously
immortalized aneuploid human keratinocyte cell line. J Cell Biol
;106:761-71.
