Natural and synthetic biodegradable nanofibers are extensively used for biomedical applications and tissue engineering. Biocompatibility and a well-established safety profile for polycaprolactone (PCL) and collagen represent a favorable matrix for preparing a dermal substitute for engineering skin. Collagen synthesized by fibroblasts is a good surface active agent and demonstrates its ability to penetrate a lipid-free interface. During granulation tissue formation, fibronectin provides a temporary substratum for migration and proliferation of cells and provides a template for collagen deposition, which increases stiffness and tensile strength of this healing tissues. The objective of this study was to fabricate nanofiber matrices from novel biodegradable PCL and collagen to mimic natural extracellular matrix (ECM) and to examine the cell behavior, cell attachment, and interaction between cells and nanofiber matrices. Collagen nanofiber matrices show a significant (p < 0.001) level of fibroblast proliferation and increase up to 54% compared with control tissue culture plate (TCP) after 72 h. The present investigation shows that PCL-coated collagen matrices are suitable for fibroblast growth, proliferation, and migration inside the matrices. This novel biodegradable PCL and collagen nanofiber matrices support the attachment and proliferation of human dermal fibroblasts and might have potential in tissue engineering as a dermal substitute for skin regeneration.