Characterization of black cumin (Nigella sativa L.) derived lipid nanoparticle as a novel nanovector Marselina Irasonia Tan, Azzania Fibriania, Adelina Khristiani Rahayu
School of Life Sciences and Technology, Institut Teknologi Bandung
Abstract
Liposomes as non-viral vectors is one of method to deliver water-soluble drug or gene theraphy. However, commercial liposomes that are currently available are also quite expensive, so that innovation of commercial liposome substitute materials that have the same capabilities with an economical price and has an abundance of materials for production is very necessary, by using nanoparticles derived from plants or called lipid nanoparticles. Lipid nanoparticles derived from plant have a lipid bilayer structure similar to that of conventional liposomes. When interacting with water, polar lipids gather and form colloidal particles so that they can encapsulate water-soluble drug or gene. Therefore, the aim of this study was to isolate black cumin derived lipid nanoparticles (Nigella sativa), so that they can be used as nanovector.
Isolation of black cumin derived lipid nanoparticles was carried out by gradient sucrose-centrifugation. The isolated lipid nanoparticles were centrifuged at a speed of 11,000 rpm, 14,000 rpm, gradient centrifugation and using amicon. The morphology of black cumin derived lipid nanoparticles and DOTAP was observed using TEM. Cytotoxicity assay was carried out using MTT assay. Internalization of black cumin and DOTAP were observed using confocal microscope.
The results showed that black cumin derived lipid nanoparticles were successfully isolated using the sucrose gradient method (the amount of sucrose used was 0.5mL, with a sucrose concentration of 90% - 10%). Black cumin derived exosome-like nanoparticles that were successfully centrifuged and collected had diameters varying between 100 nm - 200 nm, spherical in shape, and had a negative surface charge (-). The characteristics possessed by black cumin derived lipid nanoparticles are the same as those of DOTAP (commercial liposomes), which are round in shape. Based on the results of the cytotoxicity assay using MTT, the MCF-7 cell line after treatment with DOTAP (128ug/mL) still had a percentage of cell viability of up to 59% when compared to cell control. The MCF-7 cell line after treatment with lipid nanoparticles (320ug/mL) also still had a percentage of cell viability of up to 94.55% when compared to control. This indicated that black cumin derived lipid nanoparticles did not have anti-cancer activity against the MCF-7 cell line (non-toxic), similar to DOTAP. Black cumin derived lipid nanoparticles successfully internalized by MCF-7 at 45 mins after treatment. Those result showed that lipid nanoparticle has potential as novel nanovector in drug delivery systems.
Keywords: lipid nanoparticle, black cumin, nanovector, drug delivery system, breast cancer
