Effect of femtosecond laser interaction with human fibroblasts: a preliminary study

In in vitro methods and cell culture models, femtosecond (fs) laser interaction has been employed to assess its effect on the proliferation and morphology of human skin fibroblasts. We cultured a primary human skin fibroblast cell line on a glass plate, passages 17-23. The cells were irradiated with a 90-fs laser at a wavelength of 800 nm and a repetition rate of 82 MHz. The target received an average power of 320 mW for 5, 20, and 100 s, corresponding to the radiation exposures of 22.6, 90.6, and 452.9 J/cm², respectively. Using a laser scanning microscopy technique, the photon densities were measured to be 6.4 × 10¹⁸, 2.6 × 10¹⁹, and 1.3 × 10²⁰ photons/cm² in a spot area of 0.07 cm²; the recorded spectra were obtained from the laser interaction after 0.00, 1.00, 25.00, and 45.00 h. The cell count and morphological changes showed that the cultured cells were affected by laser irradiation under photon stress; some fibroblasts were killed, while others were injured and survived. We discovered evidence of the formation of several coenzyme compounds, such as flavin (500-600 nm), lipopigments (600-750 nm), and porphyrin (500-700 nm). This study is motivated by the future development of a novel, ultra-short fs laser system and the need to develop a basic in vitro understanding of photon-human cell interaction. The cell proliferation indicated that cells are partly killed or wounded. The exposure of fibroblasts to fs laser fluence up to 450 J/cm² accelerates cell growth of the viable residual cell.