DEVELOPMENT OF LIPOSOME AND NANOPARTICLE FORMULATION CONTAINING DOXYCYCLINE AND SODIUM TAUROCHOLATE AND TRANSPORT PROPERTIES THROUGH CACO-2 CELL LINES

Purpose: Evaluation of liposome and nanoparticle formulations containing doxycycline and investigation of transport properties of these formulations on Caco-2 cell line that used as a model. Material and Methods: Doxycycline and sodium taurocholate (NaTC) liposomes were prepared using dry film hydration method using cholesterol and dipalmytoilphosphatidiylcholine (DPPC). Nanoparticles were prepared using emulsion polymerisation. Eudragit RS 100 was used as a polymer.MTT (-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) test was performed for studying doxycycline and NaTC (increased paracellular absorbtion due to opening of tight junctions between epithelial cells) effect on the viability of Caco-2 cells.For matrix metalloproteinase-2 (MMP-2) determination, approximately 5x105 Caco-2 cells were seeded to 35 mm wells1. They were incubated for two days and washed with DMEM. Studied groups were incubated with 1 mL DMEM for 18 h. After cells were centrifugated MMP-2 was determined from the upper clear part of the kit. Results: According to the MTT test results, doxycycline and NaTC concentration did not exceed to 10 µg/mL and 1.875 mM for 24 hours of transport period. The cumulative amount of doxycycline and NaTC liposome and nanoparticle formulations transported through apical to basolateral side of Caco-2 cells was found to be 68.4% and 53.4% (n=3) for doxycycline respectively. Apparent permeability coefficients (k) were calculated as 5.83±0.07, 2.13±0.09 (n=3) for doxycycline and NaTC liposomes and nanoparticles respectively. Doxycycline is a type of tetracycline antibiotic that plays a key role in the growth and spread of the tumor as it inhibits the MMP-2 enzyme and produces an antitumor effect. Based on the secretion of MMP-2 from cancer cells, two formulations containing MMP inhibitor, doxycycline, have been tested and it was found that the growth of the cancer cells was inhibited by developed doxycycline formulations. Inhibition percentage 95.1% and 98.4 % were seen with liposome and nanoparticle formulations respectively. Discussion: It was concluded that doxycycline permeability coefficient and % cumulative amount of doxycycline were found to be higher for liposome formulation compared to nanoparticles. MMP-2 was found to be inhibited more with doxycycline when liposome formulation was used. This suggests that liposome formulation may be more effective and it can be used for the treatment of cancer cells.