In the Anthropocene era, lake ecosystems are increasingly subjected to significant human-induced pressures, leading to declines in both biodiversity and habitat quality. However, restoration initiatives offer promising avenues for enhancing the resilience of freshwater environments. This research investigated a range of established and novel methods aimed at promoting the growth of the macrophyte Vallisneria spiralis in the littoral zone of Lake Como, a southern alpine lake in Italy. To conduct this study, samples of Vallisneria spiralis were collected and placed in tanks containing four different types of 3D-printed biodegradable substrates. The optimal conditions for the growth of this species were identified as follows: a temperature range of 25 to 27 °C, the continuous operation of a circulation pump equipped with a filter, the presence of a fertile substrate, and light cycles comprising 6 h of peak illumination followed by 6 h of darkness. Remarkably, the plants exhibited a growth rate of 4 mm per day, increasing from an initial count of 12 specimens to 400 within four months, with a total of over 700 plants by the end of the study. Among the substrates tested, the patch substrate was found to be the most effective. After their introduction into the natural environment, the survival rate of plants established on stable substrates in contact with the lakebed reached an impressive 85.7%. This research represents a pioneering step in demonstrating that Vallisneria spiralis may serve as a viable option for restoration projects in coastal lake habitats, particularly when employing biodegradable substrates.