Nanotechnology is one of the most popular branches of science in the current decade where nanofiber scaffolds (NS) attain principal attraction. NS are composed of several nanometer to micrometer diameter fibers similar in nature to collagen, keratin and elastin fibers of the extracellular matrix (ECM) and helps in cell attachment, proliferation and differentiation. NS with desired features can be fabricated using various chemical, physical and biological methods such as electrospinning, phase separation, self-assembly, templating etc. Most often NS are fabricated using ECM derived compounds, naturally occurring polysaccharides such as chitosan, pullulan, dextran and many other synthetic compounds. Like ECM in natural niche NS interacts with cells and hence holds wide applications in Tissue engineering (TE)/ regenerative medicine where they are used as replacement for nerve, blood vessels, skin, muscle, bone and cartilage. They have also established applications in wound healing, drug testing and drug/ growth factor/ gene delivery. Application of cell free scaffolds for the culture of stem cells isolated from individual persons could pave way towards personalised therapy using TE. Cancer cells cultured in 3D microenvironment created with NS were observed to enhance the epithelial to mesenchymal transition. The present review principally focuses on various fabrication techniques and the application of various natural and synthetic compounds in tissue engineering.