Use of a novel joint-simulating culture system to grow organized ex-vivo three-dimensional cartilage-like constructs from embryonic epiphyseal cells.

A method for growth and maintenance of vital cartilaginous tissue is necessary for cartilage repair by in-vitro produced biologic implants. A previously tested perfusion system simulating joint activity was used. Whole epiphyses collected from thirty 11-day-old chick embryos were divided into two groups. One group was grown in a tissue culture dish for 10 days. The other group was placed in a perfusion system termed a joint-simulating device (JSD). After a period of 10 days, histology and immunohistochemistry were performed on five epiphyses from each group. Histologically, epiphyses grown in the device coalesced into a homogenous three-dimensional mass. The bridging tissue between individual epiphyses was highly cellular (PCNA staining positive) and was composed of mesenchymal stem cells as shown by expression of FGF receptor 3. No such tissue formed between epiphyses in the tissue culture dish and the epiphyseal cores were shown to be necrotic. The rest of the epiphyses were evaluated for radioactive sulfate incorporation into glycosaminoglycans (GAGs). A tenfold increase in sulfate incorporation occurred in epiphyses grown within the JSD as compared to the traditional culture method. In conclusion, embryonic epiphyses could be a suitable source for the ex-vivo growth of tissue-engineered cartilage constructs that might later be used as an in-vivo cartilage implant. The joint simulating device effectively maintains cartilage viability and bioactivity for as long as 10 days.