Evaluation of in vitro toxicity of Rumalaya liniment using mouse embryonic fibroblasts and human keratinocytes

Sandeep Ravi Varma; Ashok Godavarthi; Satyakumar Vidyashankar; Krishna S. Nandakumar; Pralhad Sadashiv Patki

doi:10.22377/ijgp.v5i1.165

Evaluation of in vitro toxicity of Rumalaya liniment using mouse embryonic fibroblasts and human keratinocytes

Sandeep Ravi Varma, Ashok Godavarthi, Satyakumar Vidyashankar, Krishna S. Nandakumar, Pralhad Sadashiv Patki

Abstract

The skin irritation potential of topical formulations is investigated prior to human exposure to identify the chemicals which might
induce 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 Î¼g/ml and was nontoxic at further lower concentrations, whereas the toxicity of CS was above 59% even at 400 Î¼g/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

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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.

Oâ€™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.

DOI: https://doi.org/10.22377/ijgp.v5i1.165

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