Testing NP blood circulation time in a mouse model
Purpose: The use of drug-containing nanoparticles (NP) is a promising growing field of research aimed at the treatment of cancer and bacterial infections. The main purpose of the planned experiments in the mouse is to test the circulation time in the blood of different nanoparticle formulations produced in our lab or obtained from three international collaborating groups. Understanding this parameter is of paramount importance to design better NP aimed at the treatment of tuberculosis and cancer. Our previous research on the blood circulation time in zebrafish embryos using those NP suggests that this animal model has great predictive value for these parameters in rodent models. We therefore want to test whether this is unequivocally the case aiming to establish the zebrafish embryo model as a valid replacement for the mouse model for such experiments.
Distress: The level of distress for the animals would be mild. They will receive a single intravenous injection of different types of NP. Only one of these nanocarrier formulations is a commercially available drug called, Caelyx (liposome-based drug containing doxorubicin; Caelyx is called Doxil in US), which will be given at half the concentration known from the literature to have an effect against cancer in mouse. The lowered concentration is given to avoid distress in the mice as we only intend to study the NP circulation time rather than the drug efficacy. The blood samples necessary for our analysis will be collected, under terminal anesthesia, from the heart. Organs will be harvested post mortem for NP biodistribution studies.
Expected benefit: The circulation times and biodistribution of NP aimed at the treatment of tuberculosis and cancer are fundamental parameters for knowing the potential of each formulation in treating the disease. We would therefore obtain important information, which will be used to select, among the tested NP, the most successful ones for further tests in zebrafish and mice models of tuberculosis and cancer. In addition, these experiments are intended to establish the zebrafish embryo as a valid model for testing NP blood circulation times with the potential to replace experiments in mice. This would improve animal welfare, cutting the costs of research because the zebrafish is a cheaper and an experimentally faster model. Furthermore, experiments using the zebrafish embryo model require much less amounts of NP and drugs, which constitute a high cost-factor.
Number of animals and what kind, How to adhere to 3R: We are planning to use 240 BALB/cAnNRj mice with 80 mice (2 extra per group) as backup if needed. The number of animals was determined in compliance with the 3R standards using thorough statistical analysis (Power analysis). Importantly, if our hypothesis that the zebrafish embryo results could match those obtained in the mouse we would be able to: Reduce and possibly even Replace the use of mice to study nanoparticle blood circulation times. We would also be able to Refine the method as the experiments in the zebrafish would be carried out in animals which are in early phase of development, thereby minimizing the pain that an adult mice would suffer for the same experiment.
Distress: The level of distress for the animals would be mild. They will receive a single intravenous injection of different types of NP. Only one of these nanocarrier formulations is a commercially available drug called, Caelyx (liposome-based drug containing doxorubicin; Caelyx is called Doxil in US), which will be given at half the concentration known from the literature to have an effect against cancer in mouse. The lowered concentration is given to avoid distress in the mice as we only intend to study the NP circulation time rather than the drug efficacy. The blood samples necessary for our analysis will be collected, under terminal anesthesia, from the heart. Organs will be harvested post mortem for NP biodistribution studies.
Expected benefit: The circulation times and biodistribution of NP aimed at the treatment of tuberculosis and cancer are fundamental parameters for knowing the potential of each formulation in treating the disease. We would therefore obtain important information, which will be used to select, among the tested NP, the most successful ones for further tests in zebrafish and mice models of tuberculosis and cancer. In addition, these experiments are intended to establish the zebrafish embryo as a valid model for testing NP blood circulation times with the potential to replace experiments in mice. This would improve animal welfare, cutting the costs of research because the zebrafish is a cheaper and an experimentally faster model. Furthermore, experiments using the zebrafish embryo model require much less amounts of NP and drugs, which constitute a high cost-factor.
Number of animals and what kind, How to adhere to 3R: We are planning to use 240 BALB/cAnNRj mice with 80 mice (2 extra per group) as backup if needed. The number of animals was determined in compliance with the 3R standards using thorough statistical analysis (Power analysis). Importantly, if our hypothesis that the zebrafish embryo results could match those obtained in the mouse we would be able to: Reduce and possibly even Replace the use of mice to study nanoparticle blood circulation times. We would also be able to Refine the method as the experiments in the zebrafish would be carried out in animals which are in early phase of development, thereby minimizing the pain that an adult mice would suffer for the same experiment.