The versatility of spacer fabrics includes the ability to knit two entirely different fabrics having different properties and connect them to form a unique structure. Their characteristics like excellent compression elasticity and cushioning, high breathability and air permeability, high thermal insulation and temperature regulation, surface resistance make them concept of study for footwear area. This paper aims to investigate the physical and thermal comfort attributes of flat knitted spacer and interlock fabrics designed for shoe linings. Cotton, Cotton/Bamboo, Cotton/Type A blended yarn and Cotton/Type B blended yarn (20/1 Nm) were used for the production of interlock and flat knitted spacer fabrics. In the production of spacer fabrics, polyamide (20/1 Nm) yarn was used as spacer yarn. Characteristics like fabric extensibility, friction coefficient, water vapour permeability, air permeability and thermal comfort properties of the fabrics were investigated, in order to select the most suitable materials and fabric structures for footwear linings.