Dimethyl sulfoxide toxic and stimulation effects in retinal pigment epithelial cells

Girish K. Srivastava1,2 , Antonio Dueñas-Laita1,3, J Carlos Pastor1,2,4

1. Instituto Universitario de Oftalmobiología Aplicada (IOBA), Universidad de Valladolid, Valladolid, Spin
2. Centro en Red de Medicina Regenerativa y Terapia Celular, Junta de Castilla y Leon, Valladolid, Spin
3. Departamento de Toxicología,Universidad de Valladolid, Valladolid, Spin
4. Departamento de Oftalmologia, Hospital Clinico Universitario of Valladolid, Valladolid, Spain

Abstract

Dimethyl sulfoxide (DMSO) is used for drug formulations as a active pharmaceutical ingredients (API) or vehicle of an API. Eye is considered as a relatively isolated organ, protected by barriers similar to the blood-brain one. Thus many systemically administered drugs are not able to reach adequate intraocular levels because they are unable to cross these blood-ocular barriers. Direct injection of drugs into the vitreous cavity has become very popular in recent years. Therefore, it is crucial to establish the level of toxicity and safety of the drugs, since the intraocular tissues are especially sensitive to certain situations that would be perfectly tolerated by other routes of administration. This study has been focused on evaluating the effect on RPE cells directly exposed to DMSO for short to long periods by analyzing cell morphology, cell culture´s confluence level, cell proliferation rate and viability. ARPE-19 cell cultures were exposed to 10%, 5%, 2%, 1%, 0.50%, 0.20% and 0.10% DMSO concentrations and grown for 1, 4 and 14 days. Microscopic observations were performed at 3 hours and 4 days to note any changes in cell morphology and cell culture confluence level. MTT assay was performed to assess cell viability / toxicity and proliferation. The results showed that cells exposed to 10% to 0.50% DMSO were affected for cell morphology and viability / proliferation or only morphology or level of cell culture confluence, even showing dead cell debris as they grew from 1 to 14 days. However, they retained cell morphology, viability and confluent level at concentrations of 0.20% to 0.01% DMSO. But even at lower concentrations, 0.05% to 0.01%, it was noted significant cell proliferations. This study provides data for DMSO effect assessments in RPE cell culture in in vitro conditions for measuring cell morphology, viability / toxicity and proliferation, which could be useful in drug formulations. However, the study results need to be evaluated by animal experimentations, then follow-up by clinical studies as well before use.    

Keywords: Cell toxicity, cell stimulation, Dimethyl sulfoxide, DMSO, RPE cells

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Publication
Ms. Ref. No.: A0201007     Reviewer   Author
Submitted: 28/06/2019
Accepted: 30/08/2019
Published: 15/02/2020

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