Skin wound repair by biomaterials has been extensively investigated. However, there have been relatively few reports on the use of electrospun nanofibers as novel biodegradable materials in skin defect repair. In this context, we evaluated electrospun scaffolds of collagen/chitosan (CL/CS) nanofibers at an 80: 20 ratio and silk fibroin/chitosan (SF/CS) nanofibers at a 50: 50 ratio. To test the biocompatibility of these two novel nanofiber scaffolds, we cocultured the materials with cells in vitro and applied the scaffolds in an animal skin defect repair model in vivo. The morphology of the electrospun nanofiber scaffolds was observed by field emission scanning electron microscopy. The proliferation of Sprague-Dawley (SD) rat skin fibroblasts (FBs) and keratinocytes (KCs) seeded on tissue culture plastic and CL/CS and SF/CS scaffolds was studied using methylthiazol tetrazolium assays. The morphology of cells on the nanofiber scaffolds was investigated by scanning electron microscopy. In vivo, the biocompatibility of the compound nanomaterials with the surface of the wounded rat skin was assessed by conducting macroscopic observations and hematoxylin and eosin staining of wound tissue samples. The results showed that the FBs and KCs isolated from SD rat skin proliferated well on and within the nanofiber scaffolds. Compared with the control group (gauze dressing), the CL/CS and SF/CS scaffolds had good biocompatibility and promoted wound healing. Therefore, CL/CS and SF/CS nanofiber scaffolds may be good candidates for skin tissue engineering applications.