Phototherapy stimulates metabolic processes in healing wounds. Despite worldwide interest, phototherapy is not firmly established or practiced in South Africa. This study aimed to determine which dose and wavelength would better induce healing in vitro. Diabetic-induced wounded fibroblasts were irradiated with 5 or 16 J/cm(2) at 632.8, 830, or 1,064 nm. Cellular morphology, viability (Trypan blue and apoptosis), and proliferation (basic fibroblast growth factor) were then determined. Cells irradiated with 5 J/cm(2) at 632.8 nm showed complete wound closure and an increase in viability and basic fibroblast growth factor (bFGF) expression. Cells irradiated at 830 nm showed incomplete wound closure and an increase in bFGF expression. Cells irradiated at 1,064 nm showed incomplete closure and increased apoptosis. All cells irradiated with 16 J/cm(2) at all three wavelengths showed incomplete wound closure, increased apoptosis, and decreased bFGF expression. This study showed that diabetic-wounded cells respond in a dose- and a wavelength-dependent manner to laser light. Cells responded the best when irradiated with a fluence of 5 J/cm(2) at a wavelength of 632.8 nm.