Fig. 3

Tumour treatment with MTX-MPs in vivo. A–D Methotrexate-loaded MPs in the treatment of LLC subcutaneous tumour model. C57 mice were inoculated with 1 × 10⁵ Lewis cells subcutaneously on Day 0. On Day 7, 10 µg SMPs and 10 µg LMPs were injected intravenously, once every other day, for a total of 6 times, and the tumour volume was measured every other day. 6 mice were dissected and the tumour weights were measured on Day 22, and the survival period of the remaining 6 mice was counted. A The volume and B the weight of the mice subcutaneous tumour. C Pictures of tumours after MPs treatment. D Survival curve of mice (n = 6). E–H Methotrexate-loaded MPs in the treatment of A549 subcutaneous tumour model. BALB/c Nude mice were inoculated with 5 × 10⁵ A549 cells subcutaneously on Day 0. On Day 7, 10 µg SMPs and 10 µg LMPs were injected intravenously, once every other day, for a total of 6 times, and the tumour volume was measured every other day. 6 mice were dissected and the tumour weights were measured on Day 20, and the survival period of the remaining 6 mice was counted. E The volume and F the weight of the mice subcutaneous tumour. G Pictures of tumours after MPs treatment. H Survival curve of mice (n = 6). Statistical analysis was performed by Student’s t-test in Graphpad Prism 8 software (Graphpad Software Inc., USA), n = 6 in each group, and the values were reported as mean ± standard error of mean (SEM), *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001. I–L In vivo safety of SMPs.Healthy BALB/c mice were injected intravenously with SMPs, once every other day, 4 times in total. Blood was collected 48 h after the last injection to detect the biochemical indicators of the mice. I Body weight of mice. J Alanine transaminase (ALT). K Aspartate aminotransferase (AST). L Creatinine. N = 5 in each group, and the values were reported as mean ± SEM