Skin injuries are very common in clinical practice and existing tissue engineering approaches focus on skin regeneration procedures using polymeric scaffolds. Skin regeneration is an important field of tissue engineering. in large burns, present treatments are insufficient in preventing scar formation. Electrospinning (ES) is a simple and effective method for preparing nanofibers and nano scaffold with diameters ranging from 5 to 500 nm, 10 2 to 10 4 times smaller than those prepared by the traditional methods of solution or melt spinning. The aim of this thesis was to fabricate electrospun nanoscaffolds from mixture of chitosan (CHI) (75-85% deacetylated, low molecular weight) and collagen type I (COL) extraction from rat tail tendon. Collagen is a poly protein and has good properties such as having a biological origin, non-immunogenicity, excellent biocompatibility and biodegradability and it provides a favourable surface for cellular attachment. Chitosan can modify the properties of collagen and improve mechanical properties of collagen. Collagen and chitosan could not form nanofibers during ES due to their large net charges. A synthetic polymer (PEO) was therefore added with an effort to decrease the conductivity of the solution. The effects of honey on wound healing have been reported in both acute (burns, lacerations) and chronic wounds (venous legulcers, pressure ulcers). In this study, we were used honey in solution of polymers , use of honey showed powerful antibacterial efficacy, farthermore it improved and facilitated electrospining process. The aim of this thesis was to fabricate CHI-COL-PEO nanofiber via electrospinning method. First, Acidic solution (10% acetic acid) of type I collagen, chitosan, and PEO were prepared with the total polymer and PEO concentrations fixed at 3% (w/w). The collagen/chitosan mass ratios were varied between 80/20,60/40,50/50. The solutions were analyzed for conductivity and viscosity. Nanofibers were collected on a collector by connecting a high voltage power supply to a steel needle fitted on a glass syringe that was filled with the polymer solution. A syringe pump was used for delivering the polymer solution at a rate 0.8 ml/h. Electrospining was carried out at voltage from 12 to 21 kV and with a needle to collector gap distance of 10 to18 cm. To reduce the solubility in water, electrospun membrane was crosslinked with glutaraldehyde vapor which would crosslink amino groups in collagen and chitosan, and the dried for 2 d in an oven at 37 ? C. The electrospun Nanofibers were coated with gold and analyzed for fiber diameters by scanning electron microscopy (SEM). The diameter distribution of the electrospun fibers were calculated by measuring at least 20 fibers at random. Then human fibroblast cells were maintained in humidified environment(37 ? C, 5% CO2) and cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum(FBS) and1% antibiotic agent. For in vitro cell culture study, scaffolds electrospun on cover slip. efore cell seeding, scaffolds washed repeatedly with followed by overnight incubation in cell culture medium. Fibroblasts were seeded onto the scaffold at a density of 10000 cells/scaffold. The samples were incubated at 37 ? C, and culture medium was replaced for every 24 h. The results of analysis demonstrated that these nanofibre have high swelling ratio , because of high porosity. The consequence of MTS assay showed the increasing of cells and good cell behaviours. Results of this study showed that nanoscaffols of Chi/Col/PEO with honey would be a potential condidate for burn treatment. Keywords: electrospining, chitosan, collagen, honey.