In-Vitro Screening of Etoposide and Its Thermosensitive Hydrogel in Lung Cancer (L132) Cell Lines

Abstract

Introduction: Cancer causes genetic abnormality (mutation) in healthy cells which leads overgrowth of abnormal cells leads to the development of tumors. In lung cancer, abnormal cell overgrowth occurs, which are vital organ for breathing. Sustained release drug delivery system is comparatively better than the conventional drug therapy because the conventional methods for delivery fail to achieve desire therapeutic concentrations and effectiveness of drugs, despite reaching toxicity, hydrogel overcomes the shortcomings of the conventional delivery. Thermosensitive hydrogel (Poloxamer 407) of etoposide are excellent candidates for long term depot formations in our body. Etoposide (DNA Topoisomerase-II inhibitor) is one of the most commonly used drugs in the chemotherapy of cancer.

Objective: To carried out in-vitro screening of etoposide and its thermosensitive hydrogel in lung cancer (L132) cell lines.

Methods: This study was carried out to evaluate cell viability & proliferation activity on Lung cancer cell line (L132) by etoposide hydrogels & other chemotherapeutic drugs. Fresh L132 cell lines were observed under microscope for any contamination. Cell lines were incubated for 24 hours in a CO2 incubator (Heraeus Hera cell) at 37 °C and 5% CO2. In vitro cell viability assays with cell lines are mainly used for drug screening. For MTT Assay, 100 µl of DMEM media was placed in 96-well plates. 98 µl L132 cell line was then added into the DMEM media and incubated for 24 hours at 37°C and 5% CO2 in a CO2 incubator. The average value were determined from reading & proliferation indices & percentage proliferation were calculated.

Results: Anticancer activity of  hydrogels were compared with different established chemotherapeutic drugs by MTT method and etoposide hydrogel 2 showed maximum anticancer activity when compared to all the established chemotherapeutic drug including standard etoposide with a percentage of inhibition of cancerous cells of 147.09% (0.63±0.007).  The standard etoposide showed 125% percent inhibition of cancerous cells (0.536±0.011).  Etoposide hydrogel 1 showed 117.8% inhibition of lung cancer cells (0.502±0.014).  Anticancer activity of etoposide hydrogel 1 was found to be statistically very significant when compared to etoposide hydrogel 2, Dacarbazine, Ifosfamide with a p value of <0.001 and significant when compared to normal control with a p value of <0.05.

Conclusion: Etoposide loaded Thermosensitive hydrogel could be effectively used for continuous release of drug. The present study will not only approach the cancerous cells but could be able to protect normal cell cellularity & morphology.