Storing live embryonic and adult human cartilage grafts for transplantation using a joint simulating device.
OBJECTIVE
Cartilage transplantation as a means to replace damaged articular surfaces is of interest. A major obstacle is the long-term preservation of cartilage grafts. The commonly used technique of freezing the grafts inevitably leads to cellular death. The current study compares the technique to an innovative approach using a pulsed-pressure perfusion system termed a joint simulating device (JSD), intended to simulate intra-articular mechanical forces.


METHODS
Human articular cartilage explants were harvested from both embryonic epiphyseal tissue and femoral heads of elderly women (over 70 years of age) undergoing a partial joint replacement (hemi-arthroplasty) and were divided in two groups: half of the samples were incubated in the JSD while the remaining half were grown in static culture within tissue culture plates. After 10 days all samples were evaluated for: (a) cell vitality as assessed by image analysis and XTT assay; (b) biosynthetic activity as expressed by radioactive sulfate incorporation into glycosaminoglycans (GAG's); and (c) proteoglycan content as assessed by alcian blue staining intensity.


RESULTS
A 10-fold increase in sulfate incorporation in samples held in the JSD compared to the static culture group was observed in embryonic cartilage. In adult cartilage culture in the JSD elevated sulfate incorporation by threefold as compared to static culture. Central necrosis was observed in specimens grown in the static culture plates, while it did not occur in the samples held in the JSD. Cell vitality as assessed by XTT assay was significantly better in the JSD group as compared to static culture. The difference was more pronounced in the embryonic specimens as compared to adult cartilage. The specimens cultured within the JSD retained proteoglycans significantly better than those cultured in static culture.


CONCLUSIONS
Maintenance of cartilage specimens in a JSD was highly effective in keeping the vitality of cartilage explants in vitro over a 10-day period. A possible future application may be a long-term preservation of chondral grafts, without freezing. Avoidance of freezing of cartilage grafts, might prevent the cartilage degeneration often observed in frozen osteochondral grafts.
Cohen
I
I
The Orthopaedic Division, Sheba Medical Center, Tel-Hashomer, Israel.
Robinson
D
D
Cohen
N
N
Nevo
Z
Z
D000368:Aged; D002358:Cartilage, Articular / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism / Q000637:transplantation; D002470:Cell Survival; D003470:Culture Media; D004838:Epiphyses / Q000196:embryology; D004867:Equipment Design; D005260:Female; D005270:Femur Head; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009038:Motion; D009924:Organ Culture Techniques / Q000295:instrumentation*; D009926:Organ Preservation / Q000295:instrumentation*; D010477:Perfusion / Q000295:instrumentation*; D011312:Pressure; D011509:Proteoglycans / Q000096:biosynthesis; D013194:Staining and Labeling; D013431:Sulfates / Q000378:metabolism; D009927:Tissue and Organ Procurement / Q000379:methods*
D003470:Culture Media; D006025:Glycosaminoglycans; D011509:Proteoglycans; D013431:Sulfates