Research is being carried out worldwide on the development of multilayer wound dressing systems with each layer satisfying the requirements of an ideal wound dressing system for achieving better wound healing performance. In this paper, mats were prepared from Eri silk fibroin (ESF) and its blend with poly-L-lactic acid (PLA) polymer by the electrospinning process and evaluated for the properties required for the skin contact layer of wound dressing systems. Tetracycline hydrochloride (TCH) was added to the polymer solution and was spun into a mat. The ESF fibres had a mean diameter of 320 nm and ESF, PLA blend (ESF–PLA) fibres had 502 nm. The phosphate buffer saline (PBS) uptake was less for the ESF–PLA mat compared to that of the 100% ESF mat. The contact angle with water was 23° for the ESF mat and 79° for the ESF–PLA mat. The vapour transmission rate was lower for the ESF–PLA mat compared to that of the 100% ESF mat, however, they were found to be lower than the level at which dehydration would occur. TCH loaded mats showed an effective zone of inhibition against gram positive and gram negative bacteria. A cytotoxicity study on neuroblast cells showed that the ESF mat did not impair the growth of cells. The in vivo study on a rat model showed that the wound healing performance of the mats developed from 100% ESF and ESF–PLA were better compared to the conventional open wound and gauze cloth wound dressing systems.