Nano-phytoremediation is a novel green technique to remove toxic pollutants from the environment. In vitro regenerated Ceratophyllum demersum (L.) plants were exposed to different concentrations of chromium (Cr) and exposure times in the presence of titania nanoparticles (TiONPs). Response surface methodology was used for multiple statistical analyses like regression analysis and optimizing plots. The supplementation of NPs significantly impacted Cr in water and Cr removal (%), whereas NP × exposure time (T) statistically regulated all output parameters. The Firefly metaheuristic algorithm and the random forest (Firefly-RF) machine learning algorithms were coalesced to optimize hyperparameters, aiming to achieve the highest level of accuracy in predicted models. The R scores were recorded as 0.956 for Cr in water, 0.987 for Cr in the plant, 0.992 for bioconcentration factor (BCF), and 0.957 for Cr removal through the Firefly-RF model. The findings illustrated superior prediction performance from the random forest models when compared to the response surface methodology. The conclusion is drawn that metal-based nanoparticles (NPs) can effectively be utilized for nano-phytoremediation of heavy metals. This study has uncovered a promising outlook for the utilization of nanoparticles in nano-phytoremediation. This study is expected to pave the way for future research on the topic, facilitating further exploration of various nanoparticles and a thorough evaluation of their potential in aquatic ecosystems.