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
Submitted: 28/06/2019
Accepted: 30/08/2019