Investigating the potential effects of 26 GHz 5G microwaves on UV-induced CPD levels in HaCaT keratinocytes in vitro
The 5G, the fifth generation of wireless cellular network technology, promises faster speeds, higher data transfer, and lower latency. It is being deployed globally, making it crucial to understand how exposure to 5G networks affects human tissues. In Europe, the 26 GHz frequency band has been approved for 5G telecommunications. While radiofrequency used in 4G networks has been extensively studied, millimeter-wavelength microwaves like those in the 26 GHz range remain less researched. As frequency increases, penetration into human tissue decreases, therefore 26 GHz microwaves mainly absorbed by the skin. Ultraviolet (UV) radiation, part of the electromagnetic spectrum, includes UVA (315–400 nm), UVB (280–315 nm), and UVC (100–280 nm) ranges. While UVA and part of the UVB range reach the Earth's surface, UVC and 95% of UVB are absorbed by the ozone layer. Humans are also exposed to artificial UVC radiation from sterilizing lamps. The genetic material suffers significant damage during the absorption of UV radiation, especially in the UVC and UVB ranges, causing cross-links between constituent nucleic acids. The absorption of UV can result in the formation of intrastrand cyclobutane-pyrimidine dimers in DNA, which can lead to mutations or cell death. Therefore, solar UV radiation has high carcinogenic activity and gives rise to a long-term risk of skin cancer. As a result, solar UV radiation is carcinogenic, classified by the World Health Organization’s IARC as a Group 1 carcinogen to humans. Both 26 GHz microwaves and solar UV radiation are absorbed in the upper layers of the skin, making it essential to study their combined effects. HaCaT cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with fetal bovine serum (FBS) and antibiotics, maintained at 37°C with 5% CO2. Cells were seeded into 96- well plates or 35 mm Petri dishes before exposure to UV and microwave radiation. UV exposure was conducted using a SOL-500 solar simulator equipped with a filter to transmit UVB and UVA wavelengths. The UV dose delivered was controlled by adjusting exposure time and lamp distance, with an effective dose of 40 J/m². For 26 GHz microwave exposure, the cells were irradiated using a horn antenna, with an average power density of 5–10 W/m². A software-defined radio (SDR) system generated a 5G signal, up-converted to the 26 GHz band, amplified, and transmitted to the cells. The exposure system was placed in a CO2 incubator, and the temperature was carefully monitored to stay below 37°C. DNA damage was assessed using the T4 enzyme-modified single-cell gel electrophoresis (comet) assay, which detects DNA strand breaks and UV-induced pyrimidine dimers. After irradiation, cells were harvested, mixed with 1% low-melting point agarose, dropped onto agarose pre-coated slides and covered with coverslips. Cells were lysed in lysis solution overnight at 4°C, then treated with a 1:1000 dilution of FPG enzyme (New England Biolabs, USA) and incubated at 37°C for 30 minutes. After alkaline treatment (pH>13) and electrophoresis, the slides were stained and analyzed using Comet Assay IV software (Instem, UK). The aim of this study was to assess the combined effects of 26 GHz microwave and UV radiation exposure on CPD formation in keratinocytes. The findings will provide insight into the potential risks of combined exposure to these non-ionizing radiations, which could have significant implications for human health as 5G technology becomes more widespread.
Read more