Th THC or CBD. In addition, as the combined administration of submaximal doses of THC and CBD (1:1) has been shown to reduce the growth of glioma xenografts [18], animal were also treated with a mixture of THC and CBD MPs.(1:1 w:w) In the same experiment, another set of tumours was treated daily with a single peritumoral injection of a solution containing vehicle, THC, CBD or a mixture of THC and CBD (1:1). As shown in Figure 2 administration every five days of cannabinoid-loaded microparticles (THC, CBD or THC + CBD) reduced tumor growth at the same extent than daily treatment with THC, CBD or THC + CBD in solution (Figure 2A ). A similar effect was observed when the weight of the tumors on the last day of the treatment was analyzed (Figure 3A and 3B). To note, animals treated with cannabinoids in solution and with cannabinoid-loaded MPs received approximately the same amount of cannabinoids along the treatment (Table 2). Thus, we found that 59 of the initial amount of CBD present in CBDloaded MPs (5.360.22 mg CBD/100 mg of MPs of the initial 8.9360.13 mg CBD/100 mg MPs) and 58 of THC present in THC-loaded MPs (4.7360.13 mg THC/100 mg MPs of the initial 8.2160.07 mg THC/100 mg MPs) were still present in the MPs remaining at the site of injection at the end of the experiment (Table 2). Taken together, these observations support that administration of cannabinoid-loaded MPs every five days reduces tumor growth with the same efficiency than a daily injection ofTerminal deoxyribonucleotidyl transferase ediated dUTP nick end labelingTerminal deoxyribonucleotidyl transferase ediated dUTP nick end labeling (TUNEL) was done using the in situ cell death detection kit (Roche).StatisticsStatistical Calyculin A analysis for tumor volume data were performed by ANOVA with a post hoc analysis by the Student-Neuman-Keuls test.Results Preparation and characterization of cannabinoid-loaded microparticlesIn order to evaluate the potential anticancer efficacy of microencapsulated cannabinoids, we prepared biodegradable polymeric poly-e-caprolactone (PCL) microparticles (MPs) containing THC or CBD by using the oil-in-water emulsion solvent evaporation technique. Microparticles prepared by this procedure were spherical, showed a smooth surface (Figure 1A) and had an average size of 50 mm (Figure 1B). The encapsulation efficiencies of CBD and THC into PCL MPs were 99.0965.14 ML240 biological activity andCannabinoid Microparticles Inhibit Tumor GrowthFigure 2. Cannabinoid-loaded microparticles reduce the growth of U87MG cell-derived tumour xenografts. (A) Effect of the local administration of placebo MPs, THC-loaded MP (75 mg of MP containing approximately 6.15 mg of THC per administration, one administration every 5 days), CBD-loaded MP (75 mg of MP containing approximately 6.7 mg of CBD per administration, one administration every 5 days), a mixture (1:1 w:w) of THC- and CBD-loaded MP (37.5 mg of THC-loaded MP and 37.5 mg of CBD-loaded MP per administration, one administration every 5 days), THC (15 mg/kg/day corresponding to 0.5 mg THC per day), CBD (15 mg/kg/day corresponding to 0.5 mg THC per day) or THC + CBD (7.5 mg/kg/day of THC and 7.5 mg/kg/day CBD corresponding to 0.25 mg of THC and 0.25 mg of CBD per day) on the growth of U87MG cell-derived tumor xenografts. No significant differences were found between tumours treated with vehicle in solution or placebo MPs and these data were represented together. For the sake of clarity, comparisons between the effect of THC-loaded MPs and THC.Th THC or CBD. In addition, as the combined administration of submaximal doses of THC and CBD (1:1) has been shown to reduce the growth of glioma xenografts [18], animal were also treated with a mixture of THC and CBD MPs.(1:1 w:w) In the same experiment, another set of tumours was treated daily with a single peritumoral injection of a solution containing vehicle, THC, CBD or a mixture of THC and CBD (1:1). As shown in Figure 2 administration every five days of cannabinoid-loaded microparticles (THC, CBD or THC + CBD) reduced tumor growth at the same extent than daily treatment with THC, CBD or THC + CBD in solution (Figure 2A ). A similar effect was observed when the weight of the tumors on the last day of the treatment was analyzed (Figure 3A and 3B). To note, animals treated with cannabinoids in solution and with cannabinoid-loaded MPs received approximately the same amount of cannabinoids along the treatment (Table 2). Thus, we found that 59 of the initial amount of CBD present in CBDloaded MPs (5.360.22 mg CBD/100 mg of MPs of the initial 8.9360.13 mg CBD/100 mg MPs) and 58 of THC present in THC-loaded MPs (4.7360.13 mg THC/100 mg MPs of the initial 8.2160.07 mg THC/100 mg MPs) were still present in the MPs remaining at the site of injection at the end of the experiment (Table 2). Taken together, these observations support that administration of cannabinoid-loaded MPs every five days reduces tumor growth with the same efficiency than a daily injection ofTerminal deoxyribonucleotidyl transferase ediated dUTP nick end labelingTerminal deoxyribonucleotidyl transferase ediated dUTP nick end labeling (TUNEL) was done using the in situ cell death detection kit (Roche).StatisticsStatistical analysis for tumor volume data were performed by ANOVA with a post hoc analysis by the Student-Neuman-Keuls test.Results Preparation and characterization of cannabinoid-loaded microparticlesIn order to evaluate the potential anticancer efficacy of microencapsulated cannabinoids, we prepared biodegradable polymeric poly-e-caprolactone (PCL) microparticles (MPs) containing THC or CBD by using the oil-in-water emulsion solvent evaporation technique. Microparticles prepared by this procedure were spherical, showed a smooth surface (Figure 1A) and had an average size of 50 mm (Figure 1B). The encapsulation efficiencies of CBD and THC into PCL MPs were 99.0965.14 andCannabinoid Microparticles Inhibit Tumor GrowthFigure 2. Cannabinoid-loaded microparticles reduce the growth of U87MG cell-derived tumour xenografts. (A) Effect of the local administration of placebo MPs, THC-loaded MP (75 mg of MP containing approximately 6.15 mg of THC per administration, one administration every 5 days), CBD-loaded MP (75 mg of MP containing approximately 6.7 mg of CBD per administration, one administration every 5 days), a mixture (1:1 w:w) of THC- and CBD-loaded MP (37.5 mg of THC-loaded MP and 37.5 mg of CBD-loaded MP per administration, one administration every 5 days), THC (15 mg/kg/day corresponding to 0.5 mg THC per day), CBD (15 mg/kg/day corresponding to 0.5 mg THC per day) or THC + CBD (7.5 mg/kg/day of THC and 7.5 mg/kg/day CBD corresponding to 0.25 mg of THC and 0.25 mg of CBD per day) on the growth of U87MG cell-derived tumor xenografts. No significant differences were found between tumours treated with vehicle in solution or placebo MPs and these data were represented together. For the sake of clarity, comparisons between the effect of THC-loaded MPs and THC.
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