[Treatment of deep chondral defects of the knee using autologous chondrocytes cultured on a support--preparation of the cartilage graft].
OBJECTIVE
Chondral and osteochondral defects represent an important topic in the knee surgery. The knee traumas are often followed by premature development of osteoarthritis due to limited reparative processes in the cartilage. Today's diagnostic possibilities and progress in arthroscopic techniques promote the early diagnostics and exact classification of osteochondral defects. Authors present therapeutic ways for treatment of cartilage injuries and adjacent part of the subchondral bone. New possible method for treatment of deep chondral defects on the weight-bearing area of the knee is the application of autologous cultivated chondrocytes in Tissucol (Baxter, Austria).


METHODS
Chondrocyte retrieval and in vitro cultivation is today a commonly known method. The goal of the knee surgery is to find a suitable three-dimensional bearer for chondrocyte culture. After serial laboratory tests we decided to use the tissue fibrin glue Tissucol (Baxter, Austria) as a chondrocyte carrier. Very good cell viability and capability of cell migration and outgrow in Tissucol were documented. Authors describe a new chondrograft type, its characteristics. They focused on quality control and documentation of key parameters necessary for graft use in humans. Following methods for quality control were used: Sterility control Determination of cells number Determination of cell viability by trypan blue staining. Viability between 90-95% was demonstrated. Determination of proliferative activity was tested using growth quality control. Migration and outgrowth of chondrocytes was documented. The morphological characteristics were studied by: Light microscopy (haematoxylin-eosin staining). Electron microscopy (transmission and scanning). A good stage of cellular organelles and production of extracellular matrix with protocollagenic fibrils were found in all cases. Immuno-fluorescence microscopy (using monoclonal antibodies against vimentin).


RESULTS
Preclinical testing on pigs and cadaverous was provided following laboratory investigations. Testing on cadaverous demonstrated a chondrograft ability to fill up completely the chondral defect to renovate the anatomic surface of the joint cartilage. Tests on pigs described hyaline-like cartilage. Publication contains detailed description of chondrograft preparation to allow reproducibility of described technique. Chondrograft transplantation consists of following phases: Arthroscopic diagnostic of defect. Arthroscopic sampling of cartilage for in vitro cultivation. Transport of samples to tissue bank. Enzymatic isolation, in vitro cultivation of chondrocytes. Graft formation (combination of fibrin glue Tissucol with chodrocytes), quality control. Transplantation (chondrograft agglutination into defect).


CONCLUSIONS
The goal of the nowadays tissue engineering is to find an optimal three-dimensional carrier for autologous chondrocyte culture. A new type of carrier based on hyaluronic acid--Hyalograft C was presented by Marcani et al in 2001 year. Published results from the first 20 clinical cases using Hyalograft C do not report any method complications. Our chondrograft technique represents an alternative option when compared to method described by Marcani.


CONCLUSIONS
Based on the successful preclinical outcomes observed in this study, authors conclude that autologous chondrocyte transplantation in Tissucol offers an important treatment option for large full thickness chondral lesions located on the weight-bearing surfaces of the femur.
Visna
P
P
Urazová nemocnice, Brno.
Pasa
L
L
Adler
J
J
Folvarský
J
J
Horký
D
D
D002358:Cartilage, Articular / Q000166:cytology / Q000601:surgery*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000637:transplantation*; D015718:Fibrin Tissue Adhesive; D006801:Humans; D007719:Knee Joint / Q000601:surgery*; D023822:Tissue Engineering / Q000379:methods*
D015718:Fibrin Tissue Adhesive