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Introduction
Cartilage repair has been a challenge to orthopedic surgeons. The conventional techniques usually lead to a symptom improve only, and the cartilage lesion persists. The surgical techniques such as drilling, abrasion and microfracture are based on filling the cartilage with mesenchymal cells, this results in filling the cartilage lesions with a repair tissue, but most times this tissue is fibrous. In order to achieve the cartilage regeneration, that fills the lesion with a hyaline tissue, the autologous chondrocyte transplantation was developed. This technique consists of applying cell culture to proliferate the patient's chondrocytes. These cultured cells are afterwards transplanted into the patient, at the cartilage lesion site, and the cells are able to regenerate the damaged tissue. The culture systems to proliferate the cells have been improved in order to achieve higher rates of cellular proliferation and safety culture conditions. During the implantation of this biological technique in a Brazilian laboratory we tested the chondrocyte culture conditions. The parameters analyzed were cellular viability, proliferation, morphology and the absence contaminants at the cultures.
Methods
Articular cartilage was obtained during arthroscopy procedure. The eleven cartilage samples were transported to the laboratory and submitted to enzymatic digestion to achieve chondrocyte isolation. The cells were then cultured in HAMF12:DME medium with 10% fetal calf serum and 1% penicillin/streptomycin up to 40 days. Cellular counting was carried on with Neubauer chamber and the viability determination was made with Tripan Blue exclusion method. For the morphological observations we used a Nikon TS100 inverted microscope.
Results and Discussion
The cellular viability was determined just after the isolation and the numbers were superior to 90%. The average culture period was 31 days, to obtain an 23 time increase in cultured cell number. The cells showed typical morphology, flattened and fibroblast-like. Cellular divisions were often observed, resulting in cell groups. The microbiological testing of the cultures showed the absence of contaminants.
Conclusion
With the presented results we concluded that the procedure tested to cultivate chondrocytes was able to maintain the cell viability and achieve the proliferation rates necessary to autologous transplantation procedure.
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