Effectiveness of nanoencapsulated methotrexate against osteosarcoma cells: in vitro cytotoxicity under dynamic conditions.

Cancer is a leading cause of mortality in the world, with osteosarcoma being one of the most common types among children between 1 and 14 years old. Current treatments including preoperative chemotherapy, surgery and postoperative chemotherapy produce several side effects with limited effectiveness. The use of lipid nanoparticles as biodegradable shells for controlled drug delivery shows promise as a more effective and targeted tumor treatment. However, in vitro validation of these vehicles is limited due to fluid stagnation in current techniques, in which nanoparticles sediment onto the bottom of the wells killing the cells by asphyxiation. In the current series of experiments, results obtained with methotrexate-lipid nanoparticles under dynamic assay conditions are presented as a promising alternative to current free drug based therapies. Effects on the viability of the U-2 OS osteosarcoma cell line of recirculation of cell media, free methotrexate and blank and methotrexate containing lipid nanoparticles in a 11 µM concentration were successfully assessed. In addition, several designs for the microfluidic platform used were simulated using COMSOL-Multiphysics, optimized devices were fabricated using soft-lithography and simulated parameters were experimentally validated. Nanoparticles did not sediment to the bottom of the platform, demonstrating the effectiveness of the proposed system. Moreover, encapsulated methotrexate was the most effective treatment, as after 72 h the cell population was reduced nearly 40% while under free methotrexate circulation the cell population doubled. Overall, these results indicate that methotrexate-lipid nanoparticles are a promising targeted therapy for osteosarcoma treatment.

Immune response to two hours of rowing in elite female rowers.

The influence of carbohydrate (C) versus placebo (P) beverage consumption on the immune and hormonal responses to normal rowing training sessions was measured in 15 elite female rowers residing at the U.S. Olympic Training Center. In a randomized, counterbalanced design, the athletes received C or P beverages (double-blind) before, during, and after two 2-hour bouts of rowing (one day apart). Blood samples were collected before, and 5-10 minutes and 1.5 hours after rowing. Metabolic measures indicated that training was performed at moderate intensities, with some high intensity intervals interspersed throughout the sessions (mean oxygen uptake of 2,307+/-169 m x min(-1), 57% of VO2max). Glucose and insulin were significantly lower after two hours of rowing with ingestion of P compared to C. The patterns of change in cortisol, growth hormone, epinephrine, and norepinephrine did not differ between C and P rowing trials. Blood neutrophil cell counts and the neutrophililymphocyte ratio were significantly higher following P versus C rowing sessions. The patterns of change in blood lymphocyte and lymphocyte subset counts, and lymphocyte proliferative responses did not differ between P and C trials, except for a slight difference in NK cell counts and activity. In summary, minimal changes in blood hormonal and immune measures were found following two-hour bouts of training in elite female rowers. Carbohydrate compared to placebo ingestion attenuated the moderate rise in blood neutrophil counts, but had slight or no effects on other immune parameters.