Theoretical and Experimental Assay of Shock Experienced by Yeast Cells during Laser Bioprinting

Int J Mol Sci. 2022 Aug 29;23(17):9823. doi: 10.3390/ijms23179823.

Abstract

Laser-induced forward transfer (LIFT) is a useful technique for bioprinting using gel-embedded cells. However, little is known about the stresses experienced by cells during LIFT. This paper theoretically and experimentally explores the levels of laser pulse irradiation and pulsed heating experienced by yeast cells during LIFT. It has been found that only 5% of the cells in the gel layer adjacent to the absorbing Ti film should be significantly heated for fractions of microseconds, which was confirmed by the fact that a corresponding population of cells died during LIFT. This was accompanied by the near-complete dimming of intracellular green fluorescent protein, also observed in response to heat shock. It is shown that microorganisms in the gel layer experience laser irradiation with an energy density of ~0.1-6 J/cm2. This level of irradiation had no effect on yeast on its own. We conclude that in a wide range of laser fluences, bioprinting kills only a minority of the cell population. Importantly, we detected a previously unobserved change in membrane permeability in viable cells. Our data provide a wider perspective on the effects of LIFT-based bioprinting on living organisms and might provide new uses for the procedure based on its effects on cell permeability.

Keywords: cell death; laser bioprinting; laser-induced forward transfer (LIFT); membrane perturbation; yeast.

MeSH terms

  • Bioprinting* / methods
  • Cell Count
  • Lasers
  • Light
  • Saccharomyces cerevisiae

Grants and funding

This work was supported by the Russian Science Foundation (Grant No. 20-14-00286) in part of improving the technology of microbial systems engineering, and partly supported by the Ministry of Science and Higher Education (within the State assignment FSRC «Crystallography and Photonics» RAS in part of «development of laser technologies») as well in the base funding of the work of AIA (within the State assignment of the FRC of Biotechnology of the RAS).