An ovine in vitro model for chondrocyte-based scaffold-assisted cartilage grafts

Journal of Orthopaedic Surgery and Research

Christian Kaps¹  Raimund W Kinne³  Marcus Stoffel⁴  David Pretzel³  Undine Freymann¹  Bei Zhou⁴  Katja Neumann¹  Michaela Endres² 

[1] TransTissue Technologies GmbH, Charitéplatz 1, Berlin, 10117, Germany;Tissue Engineering Laboratory, Department of Rheumatology and Clinical Immunology, Charité-Universitätsmedizin Berlin, Charitéplatz 1, Berlin, 10117, Germany;Experimental Rheumatology Unit, Department of Orthopedics, University Hospital Jena, Waldkrankenhaus, Rudolf Elle, Klosterlausnitzer Str. 81, Eisenberg, 07607, Germany;Institute of General Mechanics, RWTH Aachen, Templergraben 64, Aachen, 52062, Germany

关键词: Biomechanics;    Differentiation;    Regenerative medicine;    Tissue engineering;    Polyglycolic acid;    Fibrin;    Scaffolds;    In vitro model;    Chondrocytes;    Cartilage grafts;   

Others  :  817841
DOI  :  10.1186/1749-799X-7-37

 received in 2011-08-09, accepted in 2012-11-06,  发布年份 2012

【 摘 要 】

Background

Scaffold-assisted autologous chondrocyte implantation is an effective clinical procedure for cartilage repair. From the regulatory point of view, the ovine model is one of the suggested large animal models for pre-clinical studies. The aim of our study was to evaluate the in vitro re-differentiation capacity of expanded ovine chondrocytes in biomechanically characterized polyglycolic acid (PGA)/fibrin biomaterials for scaffold-assisted cartilage repair.

Methods

Ovine chondrocytes harvested from adult articular cartilage were expanded in monolayer and re-assembled three-dimensionally in PGA-fibrin scaffolds. De- and re-differentiation of ovine chondrocytes in PGA-fibrin scaffolds was assessed by histological and immuno-histochemical staining as well as by real-time gene expression analysis of typical cartilage marker molecules and the matrix-remodelling enzymes matrix metalloproteinases (MMP) -1, -2 and −13 as well as their inhibitors. PGA scaffolds characteristics including degradation and stiffness were analysed by electron microscopy and biomechanical testing.

Results

Histological, immuno-histochemical and gene expression analysis showed that dedifferentiated chondrocytes re-differentiate in PGA-fibrin scaffolds and form a cartilaginous matrix. Re-differentiation was accompanied by the induction of type II collagen and aggrecan, while MMP expression decreased in prolonged tissue culture. Electron microscopy and biomechanical tests revealed that the non-woven PGA scaffold shows a textile structure with high tensile strength of 3.6 N/mm² and a stiffness of up to 0.44 N/mm², when combined with gel-like fibrin.

Conclusion

These data suggest that PGA-fibrin is suited as a mechanically stable support structure for scaffold-assisted chondrocyte grafts, initiating chondrogenic re-differentiation of expanded chondrocytes.

【 授权许可】

   
2012 Endres et al.; licensee BioMed Central Ltd.

【 图 表 】

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