Safety evaluation of a ketamine-dodecyl maltoside combination using angiogenesis and embryonic development models

Ketamine (KET) exhibits potential anticancer activity, with enhanced cytotoxicity against melanoma cells when combined with the permeation enhancer dodecyl maltoside (DDM). To support clinical translation, this study evaluated the embryotoxicity, effects on angiogenesis, and cytocompatibility of KET, DDM, and their combination (KET + DDM) using a chicken embryo model. The chorioallantoic membrane (CAM) assay was used to assess angiogenesis following treatment with KET (1000 µM), DDM (19.6 µM), or KET + DDM in fertilized eggs. Embryonic survival and morphology were monitored for five days. Quantitative PCR analysis of heart, lung, kidney, and brain tissues evaluated apoptosis-related genes (Caspase 3, Caspase 8, Caspase 9, BAX) and VEGF expression. Cytocompatibility was examined in primary embryonic fibroblasts (EFBs) using AlamarBlue assays and morphological assessment. The results showed no significant differences in vascular density, vessel length, or branching in the CAM assay across all treatments. Embryos treated with KET or KET + DDM showed normal survival and morphology, while DDM alone reduced viability. Apoptotic and angiogenic gene expression remained unchanged in major organs. In vitro, KET and KET + DDM did not reduce EFB viability or alter morphology. Overall, KET and KET + DDM did not produce detectable adverse effects in embryonic and cellular models, preserving angiogenesis and development. Notably, the KET + DDM combination showed no detectable adverse effects, supporting the preliminary safety of this combination for further anticancer investigation.