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Cotton is cheap, easily available and widely used as surgical material. Therefore, cotton would be good raw material to design a scaffold for tissue engineering applications. In this work, the gelatin-coated microfibrous paper scaffold was fabricated successfully by a papermaking process. Microfibers-based scaffold could overcome the limitations of nanofibrous material-based scaffold for tissue engineering application. The physicochemical and mechanical properties of the scaffolds were characterized. The results revealed that the gelatin contributed to the enhanced cell attachment and proliferation over the scaffolds, whereas cellulose as a strong backbone in the scaffold to support it for keeping its appearance. Their tensile strength and water absorption capacity were improved, but pore size and porosity were decreased after incorporation of gelatin. Hence, results suggested that fabricated scaffolds have huge prospective as a bioactive, well-designed and economical scaffold stand for tissue engineering application.