In this study, to combine the biological properties of lanolin and the advantages of electrospun nanofiber mats for developing a potent topical wound dressing/nursing pads that includes lanolin, lanolin incorporated electrospun cellulose acetate (CA), polyethylene oxide (PEO), polyethylene oxide/chitosan (PEO/chitosan) and thermoplastic polyurethane (TPU) nanofibers were developed and characterized. Surface tension, viscosity and conductivity measurements of each polymer solution were carried out in order to characterize the spinning solutions. Lanolin addition did not affect the conductivity values but increase the viscosity of each spinning solution. According to the SEM images, smooth nanofiber surfaces were produced which could be appropriate for a top layer of a nursing pad. Slightly attached, thicker nanofibers could be seen from the TPU/Lanolin images. Similar to TPU nanofibers, CA nanofibers were smoother and thinner than CA/Lanolin nanofibers. Lanolin effected the uniformity of the PEO nanofibers, but it could be said that homogeneous distribution of lanolin was achieved. Characterization of these nanofiber membranes were also carried out with FTIR, DSC, swelling and weight loss analysis. Since TPU nanofibers were hydrophobic and the addition of lanolin made the nanofiber more hydrophobic, they did not show high swelling ratios. CA nanofibers swelled 610% due to their relatively hydrophilic nature and bulky structure. PEO nanofibers were totally dissolved in the water, a very thin layer remained for PEO-Chitosan/lanolin nanofibers. So, using electrospun CA and TPU nanofiber membranes for a nursing pad upper layer could be suitable. CA/lanolin nanofibers showed better hydrophilicity and swelling when compared to TPU nanofibers. On the other hand, TPU nanofibers are more elastic and durable with lower swelling.