Hydrogels are suitable for osteochondral defect regeneration as they mimic the viscoelastic environment of cartilage. However, their biomechanical properties are not sufficient to withstand high mechanical forces. Therefore, we have prepared electrospun poly-ε-caprolactone-chitosan (PCL-chit) and poly(ethylene oxide)-chitosan (PEO-chit) nanofibers, and FTIR analysis confirmed successful blending of chitosan with other polymers. The biocompatibility of PCL-chit and PEO-chit scaffolds was tested; fibrochondrocytes and chondrocytes seeded on PCL-chit showed superior metabolic activity. The PCL-chit nanofibers were cryogenically grinded into microparticles (mean size of about 500 µm) and further modified by polyethylene glycol-biotin in order to bind the anti-CD44 antibody, a glycoprotein interacting with hyaluronic acid (PCL-chit-PEGb-antiCD44). The PCL-chit or PCL-chit-PEGb-antiCD44 microparticles were mixed with a composite gel (collagen/fibrin/platelet rich plasma) to improve its biomechanical properties. The storage modulus was higher in the composite gel with microparticles compared to fibrin. The Eloss of the composite gel and fibrin was higher than that of the composite gel with microparticles. The composite gel either with or without microparticles was further tested in vivo in a model of osteochondral defects in rabbits. PCL-chit-PEGb-antiCD44 significantly enhanced osteogenic regeneration, mainly by desmogenous ossification, but decreased chondrogenic differentiation in the defects. PCL-chit-PEGb showed a more homogeneous distribution of hyaline cartilage and enhanced hyaline cartilage differentiation.

Department of Organic Chemistry Faculty of Science Palacky University 17 listopadu 12 771 46 Olomouc Czech Republic

Department of Tissue Engineering Institute of Experimental Medicine of the Czech Academy of Science Videnska 1083 142 20 Prague 4 Czech Republic

Faculty of Textile Engineering Technical University of Liberec Studentská 2 461 17 Liberec Czech Republic

Hospital of Rudolfa and Stefanie a s Máchova 400 256 30 Benešov Czech Republic

Institute of Biophysics 2nd Faculty of Medicine Charles University 5 Uvalu 84 150 06 Prague 5 Czech Republic

Institute of Histology and Embryology and Biomedical Center Faculty of Medicine in Pilsen Charles University Prague Husova 3 305 06 Pilsen Czech Republic

Student Science s r o Národních Hrdinů 279 Dolní Počernice 190 12 Prague Czech Republic

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