Kompozit doku allotransplantasyonu yapılan sıçanlarda, yağ doku kaynaklı mezenekimal kök hücrelerin ilk 24 saatte meydana gelen sistemik yanıta etkisi
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2015-01-19
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Selçuk Üniversitesi Tıp Fakültesi
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info:eu-repo/semantics/openAccess
Özet
AMAÇ: Kompozit doku nakilleri eş zamanlı birçok farklı doku türünün rekonstrüksiyonu açısından avantajlıdır. Ancak bu işlemin önündeki en büyük sorun tüm doku çeşitlerine karşı ayrı ayrı oluşan sistemik ve immün yanıttır. Biz bu çalışmamızda kompozit doku nakillerine bağlı ilk 24 saatte meydana gelen sistemik yanıtı ve oluşan sistemik yanıta yağ doku kaynaklı mezenkimal kök hücrenin (MKH) etkisini gözlemlemeyi amaçladık. YÖNTEM: Çalışmada 29 adet Wistar Albino (WA) , 29 adet Sprague Dawley (SD) türü sıçan kullanılmıştır. 24 adet WA sıçan 12'şerli 2 ayrı gruba ayrılmıştır. ( Grup I, Grup III), 24 adet SD sıçan 12'şerli 2 ayrı gruba ayrılmıştır. ( Grup II, Grup IV) . Kalan 5 adet WA ve 5 adet SD türü sıçan MKH eldesi ve bazal karaciğer apoptoz değerlerinin ölçülmesinde kullanılmıştır. Bu gruplardan elde edilen MKH süspansiyonları kendi türlerinden sıçanlara enjekte edilmek üzere -80 ºC de saklanmış ve enjeksiyondan 40 dakika önce uygun koşullarda eritilmiştir. Grup I ile Grup II arasında, Grup III ile Grup IV arasında çapraz sol arka ekstremite transferi planlanmıştır. Transfer öncesi tüm sıçanlardan kuyruk kanı alınarak bazal AST, ALT, Üre ve Kreatinin değerleri ölçülmüştür. Transfer esnasında anastomoz açılıp ekstremite dolaşımı gözlenen Grup III ve Grup IV sıçanlara kendi t ürlerinden elde edilen adipoz kaynaklı MKH içeren süspansiyon intraperitoneal olarak enjekte edilmiştir. Grup I ve Grup II sıçanlara aynı miktar serum fizyolojik intraperitoneal olarak enjekte edilmiştir. Transfer işleminden 24 saat sonra bacak dolaşımı yeterli olan sıçanlar sakrifiye edilerek hem kan hemde karaciğer doku örneği alınmıştır. Elde edilen veriler hem grup içinde hemde gruplar arasında istatistiksel olarak analiz edilmiştir. BULGULAR: İskemi süreleri ile 24 saat sonunda kandan ölçülen AST, ALT, Üre, Kreatinin değerleri ve karaciğer dokusundan akım sitometrisi ile elde edilen ölü hücre, canlı hücre, erken ve geç apoptotik hücre oranları arasında korelasyon olmadığı görülmüştür. Kompozit doku nakli sonrası tüm gruplarda AST değerlerinde artış olmasına rağmen bu artış sadece 1 grupta anlamlı bulunmuştur. Kreatinin ve ALT değerlerinde tüm gruplarda anlamlı oranda azalma gözlenmiştir. Üre değerleri MKH enjeksiyonu yapılmayan gruplarda azaldığı halde enjeksiyon yapılan gruplarda artmış olarak gözlendi. Akım sitometrisi incelemesinde MKH enjeksiyonu yapılan gruplarda canlı hücre, erken ve geç apoptotik hücre oranları arasında fark gözlenmedi. Sadece 1 grupta MKH enjeksiyonu sonrası ölü hücre oranında azalma olduğu gözlendi. MKH eldesi için kullanılan sıçanlardan alınan karaciğer örneklerinin akım sitometrik incelemesi sonuçları, cerrahi işlem yapılıp MKH enjeksiyonu yapılmayan ve 24 saat sonunda sakrifiye edilerek karaciğer örnekleri alınan sıçanlar ile karşılaştırıldığında ölü hücre, canlı hücre, erken ve geç apoptotik hücre oranları açısından anlamlı fark gözlenmemiştir. SONUÇ: Kompozit doku nakilleri sonrası ilk 24 saatte meydana gelen sistemik yanıtın izlenmesinde AST, ALT, Üre ve Kreatinin değerlerinin anlamlı bulgu vermediği, iskemi süresinin karaciğer ve böbrek fonksiyon testleri ile korelasyon göstermediği, AST ve ALT değerlerinin karaciğerde mevcut apoptoz ve ölü hücre miktarı ile korelasyon göstermediği bu nedenle takip açısından yeterli olmadığı, MKH' lerin kompozit doku nakli yapılan sıçanlarda ilk 24 saatte meydana gelen sistemik yanıta belirgin olumlu etkisinin olmadığı hatta bazı parametrelerde olumsuz etki oluşturabileceği sonucuna varılmıştır.
OBJECTIVE: Composit tissue transplantations are advantageous for reconstructions of many tissue. However the biggest problem of this process is the sistemic and immune response against all tissue types seperately. In our study we aimed to observe sistemic response in the first 24 hours due to vascularised composit tissue allotransplantations (VCA) and the effect of adipose derived mesenchimal stem cell to the systemic response. METHOD: 29 Wistar Albino and 29 Sprague Dawley rats were used in the study. Twentyfour WA rats were seperated to 2 groups with 12 rats.(group 1,3) 24 SD rats were seperated to 2 groups with 12 rats (group 2,4) Remaining 5 WA and 5 SD rats were used for obtaining Mesenchimal Stem Cell (MSC) and for measure the basal liver apoptosis values. These MSC suspensions were preserved in -80 degrees in order to inject the same species and melted 40 minutes before the injection in favourable conditions. Between group 1 and 2 and between group 3 and 4 the cross left hind extremity transfer was planned. Before transfer, all rats' basal AST, ALT, BUN, Creatinin values were measured by taking the tail blood. The suspension including the adipose derived MSC which is obtained from the same species, was injected intraperitoneally to group 3 and 4 rats that had been observed the extremity circulation by opening the anastomosis during the transfer. Same amount of serum physiologic was injected intraperitoneally to the group 1 and 2. Twentyfour hours after the transfer, the rats which had the enough leg circulation were sacrificed and both blood and liver tissue samples were obtained. The obtained results were statistically analysed both in the group and between the groups. RESULTS: We didn't find any correlation between ischemia time and AST, ALT, BUN and Creatinin values which were measured 24 hours after VCA. Alike any correlation with flow cytometry results. Despite the increase in AST values after composite tissue transplantation, this increase was significant in only one group. Creatinin and ALT values were significantly decreased in all groups. BUN values were decreased in the groups which wasn't injected MSC, however it is observed as increased in the groups which was injected MSC. Significant difference wasn't observed between the dead cell, live cell, early and late apoptotic cell in the groups that was injected MSC in the flow cytometry. Only in one group, it is observed that the dead cell quantitiy was decreased after the MSC injection. The flow cytometric results of liver samples of the rats which was utilized for obtaining the MSC weren't differs from the rats which were operated but not get the MSC injection and sacrificed after 24 hours from the point of dead cell, live cell, early and late apoptotic cell. CONCLUSION: It is concluded that AST, ALT, BUN and Creatinin values aren't give significant result for monitoring the systemic response in the first 24 hours after the composite tissue transplantations. Ischemia time doesn't corelate with liver and kidney function tests; AST and ALT values don't corelate with the amount of liver apoptosis and dead cell and so not enough for monitoring; MSC has no significantly positive effect on the systemic response in the first 24 hours of the composite tissue transplantation, even has negative effects on some parameters.
OBJECTIVE: Composit tissue transplantations are advantageous for reconstructions of many tissue. However the biggest problem of this process is the sistemic and immune response against all tissue types seperately. In our study we aimed to observe sistemic response in the first 24 hours due to vascularised composit tissue allotransplantations (VCA) and the effect of adipose derived mesenchimal stem cell to the systemic response. METHOD: 29 Wistar Albino and 29 Sprague Dawley rats were used in the study. Twentyfour WA rats were seperated to 2 groups with 12 rats.(group 1,3) 24 SD rats were seperated to 2 groups with 12 rats (group 2,4) Remaining 5 WA and 5 SD rats were used for obtaining Mesenchimal Stem Cell (MSC) and for measure the basal liver apoptosis values. These MSC suspensions were preserved in -80 degrees in order to inject the same species and melted 40 minutes before the injection in favourable conditions. Between group 1 and 2 and between group 3 and 4 the cross left hind extremity transfer was planned. Before transfer, all rats' basal AST, ALT, BUN, Creatinin values were measured by taking the tail blood. The suspension including the adipose derived MSC which is obtained from the same species, was injected intraperitoneally to group 3 and 4 rats that had been observed the extremity circulation by opening the anastomosis during the transfer. Same amount of serum physiologic was injected intraperitoneally to the group 1 and 2. Twentyfour hours after the transfer, the rats which had the enough leg circulation were sacrificed and both blood and liver tissue samples were obtained. The obtained results were statistically analysed both in the group and between the groups. RESULTS: We didn't find any correlation between ischemia time and AST, ALT, BUN and Creatinin values which were measured 24 hours after VCA. Alike any correlation with flow cytometry results. Despite the increase in AST values after composite tissue transplantation, this increase was significant in only one group. Creatinin and ALT values were significantly decreased in all groups. BUN values were decreased in the groups which wasn't injected MSC, however it is observed as increased in the groups which was injected MSC. Significant difference wasn't observed between the dead cell, live cell, early and late apoptotic cell in the groups that was injected MSC in the flow cytometry. Only in one group, it is observed that the dead cell quantitiy was decreased after the MSC injection. The flow cytometric results of liver samples of the rats which was utilized for obtaining the MSC weren't differs from the rats which were operated but not get the MSC injection and sacrificed after 24 hours from the point of dead cell, live cell, early and late apoptotic cell. CONCLUSION: It is concluded that AST, ALT, BUN and Creatinin values aren't give significant result for monitoring the systemic response in the first 24 hours after the composite tissue transplantations. Ischemia time doesn't corelate with liver and kidney function tests; AST and ALT values don't corelate with the amount of liver apoptosis and dead cell and so not enough for monitoring; MSC has no significantly positive effect on the systemic response in the first 24 hours of the composite tissue transplantation, even has negative effects on some parameters.
Açıklama
Anahtar Kelimeler
Yağ doku, Kök hücre, Kompozit doku nakli, Sistemik yanıt, Adipose tissue, Stem cell, Composite tissue allotransplantation, Systemic response
Kaynak
WoS Q Değeri
Scopus Q Değeri
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Sayı
Künye
Akatekin, A. (2015). Kompozit doku allotransplantasyonu yapılan sıçanlarda, yağ doku kaynaklı mezenekimal kök hücrelerin ilk 24 saatte meydana gelen sistemik yanıta etkisi. Selçuk Üniversitesi, Yayımlanmış uzmanlık tezi, Konya.
