Krzysztof MARYCZ, Agnieszka SMIESZEK, NEZİR YAŞAR TOKER, Jakup NİCPON

The morphology and proliferation rate of canine and equine adipose derived mesenchymal stem cells cultured with flunixin meglumine-in vitro

Hayvanlarda özellikle kök hücrelerinin kullanıldığı rejeneratif (yenilenebilir) tıp hızla gelişim gösteren bir alandır. Günümüzde hem büyük hem de küçük hayvanların ortopedik hastalıklarında kök hücre tedavisinden faydalanılmaktadır. Klinik pratikte rutin olarak kullanılan non-steroidal anti-inflamatuar ilaçlar, adipoz kökenli mezenşimal kök hücresi tedavisine de dahil edilmektedir. Bu çalışmada köpek ve atlardan elde edilmiş adipoz kökenli mezenşimal kök hücrelerinin, yaygın kullanılan bir non-steroidal anti-inflamatuar ilaç (flunixin meglumine) ile kültüre edilmesinin, hücre morfolojisi ve proliferasyonu üzerine etkileri araştırıldı. Hücrelerin morfolojik takibinde epi- fuloresans mikroskop ve taramalı elektron mikroskobu (SEM) kullanıldı. Hücrelerin mikrovezikül sekresyonları da SEM aracılığıyla izlendi. Adipoz kökenli mezenşimal kök hücrelerinin proliferasyon aktiviteleri resazurin-temelli bir test yardımıyla değerlendirildi. Araştırma sonuçlarına göre fluniksin megluminin en düşük konsantrasyonlarının (0.01 mg/ml) köpeklerden elde edilen adipoz kökenli mezenşimal kök hücrelerinde proliferasyonu uyardığı; ancak aynı konsantrasyonun at kökenli hücrelerde proliferasyonu belirgin düzeyde yavaşlattığı gözlendi. Enteresan bir sonuç olarak 0.01 mg/ml konsantrasyondaki flunixin megluminein, incelenen kök hücre populasyonunun morfolojilerine etki etmediği belirlendi. Çok aşamalı bu araştırmadan elde edilen bulgular ışığında, köpeklerden elde edilen adipoz kökenli mezenşimal kök hücrelerinin ortopedik tedavilerde kullanımı sırasında, flunixin megluminein en düşük konsantrasyonlarda kullanılmasının yararlı olacağı sonucuna ulaşıldı.

Flunixin Meglumine İle in vitro Kültüre Edilen Köpek ve At Yağ Kökenli Mezenkimal Kök Hücrelerinin Morfolojisi ve Çoğalma Hızı

The regenerative medicine in animals is a rapidly growing field, especially when therapies with stem cells are applied. Currently, stem cells are used for treatment of orthopedic diseases occurring both in small and large animals. Non-steroidal anti-inflammatory drugs (NSAIDs) are routinely used and are often accompanied with adipose derived mesenchymal stem cells (ADMSCs) therapy. Therefore, it is reasonable to monitor morphology of cells and the proliferation status of canine and equine ADMSCs cultured with NSAIDs.focused on the analysis of the above mentioned parameters. Morphology of investigated cells was monitored using an epifluorescence microscope and scanning electron microscope (SEM). Moreover, SEM analysis was carried out for determination of microvesicles secretion. Proliferation activity of AdMSCs was evaluated with a resazurin-based test. Our research showed that the lowest concentration of Flunixin meglumine (0.01 mg/ml) had a stimulating effect on canine AdMSCs proliferation, while the same concentration significantly slowed down equine stem cell growth. Interestingly, the 0.01 mg/ml concentration of Flunixin meglumine did not effect morphologythe investigated stem cells population. Thus, results obtained from multilevel research allow us to conclude that the lowest concentration of Flunixin meglumine may be accompanied with canine adipose derived mesenchymal stem cells in orthopedic treatment.

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