The synthesis of carbon dots (CDs) is gaining wide-ranging interest due to their broad applicability, owing to their small size and luminescence. CDs were prepared from charcoal via a one-step process using laser ablation in liquid without the use of reagents. The adopted method was based on the use of a commercially available continuous wave (CW) laser diode emitting a 450 nm wavelength and, for the liquid, a phosphate-buffered saline (PBS) solution, routinely used in the biological field. Photoluminescence analysis revealed fluorescence, at 480 nm, increasing with laser irradiation time. The atomic force microscopy (AFM) of the CDs revealed an average sphere shape with a size of about 10 nm. Biodegradable polycaprolactone (PCL), typically adopted in biomedicine applications, was used as a matrix to show the preserved luminescence, ideal for the non-invasive monitoring of implanted scaffolds in tissue engineering.