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The aim of this study was to develop biodegradable medical nonwoven fabrics with added value, focusing on environmentally friendly natural substances for use in wound care. Propolis, a natural antibiotic, with confirmed antimicrobial and anti-inflammatory action was used. It was used in two formulations: ethanol extract of propolis and powder of native propolis. In order to obtain an environmentally friendly product, no chemical binder was used. Three modified procedures of applying propolis by immersing and scattering were used. Antimicrobial effect of the performed treatment was tested according to standard EN ISO 20645:2004 on three types of bacteria: Gram-positive Staphylococcus aureus (ATCC 6538), Gram-negative Escherichia coli (ATCC 11229) and Klebsiella pneumonia (ATCC4352). In this study, the microbial barrier properties of textiles for two bacterial endospores were investigated, with the aim of determining the microorganism mechanical permeability of textile. Fourier-transform infrared–attenuated total reflection was used to investigate the structural changes of Tencel nonwoven fabrics/propolis samples. On the basis of the results it can be concluded that the application of bee products, in this case of propolis in both formulations, on Tencel nonwoven fabrics provides satisfactory antimicrobial effects and an effective microbial barrier. Obtained results show that treatments 1 and 3 provide better antibacterial properties, indicating the further development potential of these treatments. AATCC Test Method 79, Absorbency of Textiles, was used to test absorption. The results show an increase in the absorption time of finished samples. The absorption time of finished samples can be reduced if they are stacked in a layered structure.