The influence of environmental conditions on the thermo-physiological comfort of men's socks was investigated using an objective assessment of comfort under dynamic conditions. Men's socks made in a 3:1 rib construction from three types of basic yarns (77%) were used: bamboo, cotton, and a cotton/polyester blend. The remaining composition of the socks is identical: filament polyamide yarn (22%) and wrapped rubber thread (1%) for the welt. The influence of climate factors artificially created in the climate chamber was analyzed by measuring three physiological parameters: skin temperature, relative humidity of the skin, and microclimate in the space between the sock and the shoe on both feet. The research results show that the raw material composition of the base yarn used in the manufacture of men's socks has a significant impact on thermo-physiological comfort when wearing socks. Under artificially created conditions in the climate chamber, an ambient temperature of 20 °C and relative humidity of 50% (autumn and spring conditions), socks with basic yarn made of a cotton/polyester blend are preferred, then with a base yarn made of cotton, and finally socks with a base yarn made of bamboo. The data from the study provide valuable information for the clothing industry in designing and defining the choice of base yarn to produce socks, depending on the conditions of use.