Lazerio Pharos ultravioletinės spinduliuotės genotoksinio poveikio žmogaus akies ragenos epitelio pirminėms ląstelėms in vitro tyrimas neplanuotos DNR sintezės metodu / ; The investigation of genotoxic effect of laser pharos ultraviolet radiation on primary human corneal epithelial cells in vitro using unscheduled dna synthesis (uds).

The Investigation of Genotoxic Effect of Laser Pharos Ultraviolet Radiation on Primary Human Corneal Epithelial Cells In Vitro Using Unscheduled DNA Synthesis (UDS) According to many different indicators, the femtosecond UV radiation laser Pharos, which was developed by scientists of UAB MGF “Šviesos konversija”, is considered as a new and advanced tool in laser microsurgery of the eye. However, in order for this laser to be used in ophthalmology, it is important to examine not only the characteristics of the interaction between the tissue and the radiation it emits, but also to assess the genetic risk of laser radiation. When genetic material is damaged, DNA repair enzymes are activated, and cellular DNA synthesis occurs. This process is called unscheduled DNA synthesis (UDS). Unscheduled DNA synthesis can be used to measure the extent of DNA damage in vitro and in vivo. Since there is no possibility to investigate the genotoxicity of laser radiation on human corneal epithelial cells in vivo, it is necessary to explore the effects of radiation on these cells in vitro. The purpose of this thesis was to evaluate the genotoxic effects of laser Pharos 206 nm radiation on human primary corneal epithelium cells (HPCE) in vitro using an unscheduled DNA synthesis method. To achieve this goal, a method was developed for affecting the corneal epithelial primary cells by laser and lamp UV radiation by irradiating the primary cell monolayer on 0.1% gelatin-coated microplates. Also, the Click-iT® EdU Imaging Kits protocol was adapted and modified. This study revealed that the radiation effects on cells were dependent on the wavelength used – 257 nm laser radiation caused the most DNA reparation in irradiated cells, followed by 254 nm constant wave radiation, while 206 nm waves were the least genotoxic. Also, it is important to say that in all cases, the intensity of the reparation increased as the dose of the exposure increased. Thus, the results of the research have shown that, although Pharos femtosecond 206 nm radiation ...