Michala Rampichová,1,2 Matej Buzgo,1 Andrea MíÄková,1,2 Karolína Vocetková,2 Vera Sovková,2 Vera Lukášová,2 Eva Filová,2 Franco Rustichelli,2 Evžen Amler1,2 1Indoor Environmental Quality, University Center for Energy Efficient Buildings, Czech Technical University in Prague, Buštehrad, 2Laboratory of Tissue Engineering, Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic Abstract: Bone and cartilage are tissues of a three-dimensional (3D) nature. Therefore, scaffolds for their regeneration should support cell infiltration and growth in all 3 dimensions. To fulfill such a requirement, the materials should possess large, open pores. Centrifugal spinning is a simple method for producing 3D fibrous scaffolds with large and interconnected pores. However, the process of bone regeneration is rather complex and requires additional stimulation by active molecules. In the current study, we introduced a simple composite scaffold based on platelet adhesion to poly-ε-caprolactone 3D fibers. Platelets were used as a natural source of growth factors and cytokines active in the tissue repair process. By immobilization in the fibrous scaffolds, their bioavailability was prolonged. The biological evaluation of the proposed system in the MG-63 model showed improved metabolic activity, proliferation and alkaline phosphatase activity in comparison to nonfunctionalized fibrous scaffold. In addition, the response of cells was dose dependent with improved biocompatibility with increasing platelet concentration. The results demonstrated the suitability of the system for bone tissue. Keywords: centrifugal spinning, 3D scaffold, platelets, growth factors, cytokines, PCL