Perikondrium etkisiyle yağ dokusu kaynaklı yetişkin mezenşimal kök hücrelerden kıkırdak elde edilmesi: (Tavşanlar üzerinde deneysel çalışma)

Abstract

Belirli boyutun üzerindeki kıkırdak defektlerinin onarımı zordur. Bu defektlerin onarımında en sık kullanılan yöntem otolog kıkırdak greftleridir. Donör alan kısıtlaması bu yöntemin en önemli dezavantajıdır. Bu Çalışmada yetişkin yağ dokusu kökenli mezenşimal kök hücrelerden perikondriumun salgıladığı büyüme faktörlerinin etkisiyle kıkırdak elde edilebilirliği ve kıkırdak defektlerinin bu yolla onarımı araştırıldı. Bu amaçla tavşan kulağında perikondrium korunarak 2x2 cm kıkırdak defekti oluşturuldu, aynı tavşandan cilt altı yağ dokusu alınarak SVF elde edildi. ADAS hücreleri DiI boyası ile işaretlenerek üç farklı grupta deneye tabi tutuldu. Kıkırdak defektlerine Grup 1 de ADAS tan zengin SVF, Grup 2 de kültüre edilerek çoğaltılmış ADAS hücreleri ve Grup 3 te kültüre edilerek çoğaltılmış ve kartilojeneze yönlendirilmiş ADAS hücreleri verildi. 40 gün sonra tavşanlar sakrifiye edilerek kıkırdak defektlerinde oluşan onarım dokusu makroskobik ve mikroskobik olarak değerlendirildi. Sonuçlar Student t-testi ile istatistiksel analize tabii tutuldu(p 2 defektlerin tam ve tama yakın onarıldığı saptandı. Fakat oluşan kıkırdağın kalitesinin orijinal kulak elastik kıkırdak kalitesinden düşük olduğu görüldü. Repair of large cartilage defects is mostly problematic. For this, autologous cartilage grafts are most preferred one among the reconstructive alternatives. The most important disadvantage of this method is limited donor area. In this study, it was investigated the possibility of regeneration of new cartilage tissue creating by adult adipose derived mesenchymal stem cells to be stimulated by perichondrium which secretes growth factors. For this purpose, three different groups (Group 1, 2 and 3) were designed by using both ears of the rabbits. The 2x2 cm cartilage defects were created leaving perichondrium intact in the rabbit's ears. The defects of the right ears were used as experimental side and the left ones were used as control side in the same animal. Stromal vascular fraction (SVF) was obtained by taking subcutaneous fat tissue from the same animal. Adipose derived adult stemcell (ADAS) were put to assay in three different groups by marking DiI stain. ADAS rich SVF was applied into the cartilage defect in group 1, replicated ADAS in culture into the cartilage defect in group 2 and replicated and redirected in culture ADAS into the cartilage defect in group 3. After follow-up period, the rabbits have sacrificed after 40 days and the repaired tissue in the cartilage defects were evaluated by macroscopic and microscopic methods. The results were statistically analyzed with the Student t- test (p value was 0.05). As a result, although the experimental defects in the group 1 have showed cartilage-like tissue, no important statistically difference was found with the result of the controls. The group 2 and 3 have showed complete or near-complete cartilage formation in the defects by application ADAS. 3 In conclusionæ the replicated ADAS used in the group 2 and the replicated and redirected ADAS used in the group 3 have produced complete or near-complete cartilage formation in the defects. But, the quality of the new-formed cartilage has showed lower than the original elastic cartilage of the ear.