Metilkobalamin ve takrolimus içeren Polikaprolakton (PCL) nanofiberlerin periferik sinir rejenerasyonuna etkilerinin karşılaştırılması
Yükleniyor...
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Necmettin Erbakan Üniversitesi Meram Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Periferik sinir yaralanmaları sık görülen; etkilenen hasta grubunda, duyusal ve motor fonksiyonlarda kayba neden olabilen bir durumdur. Kusursuz yapılan cerrahiler sonrasında bile motor ve duyusal fonksiyonlarda tatmin edici sonuçlara ulaşılamayabilir. Periferik sinir yaralanmalarının tam olarak iyileşmemesi genellikle etkilenen uzuvlarda uyuşma, kronik ağrı , duyusal ve/veya motor fonksiyonlarda azalma gibi birçok olumsuz sonuca yol açmakta ve bu nedenlerle periferik sinir yaralanmaları dünya çapında bir klinik sorun teşkil etmektedir. Tam anlamıyla başarılı olarak uygulanabilecek farmakolojik ajanın bulunması için güncel araştırmalar bu yöne doğru evrilmiştir. Bu araştırmalardan bir tanesi de nanoteknoloji ve bu yöntemle üretilen nanomateryallerdir. Çalışmamızda periferik sinir rejenerasyonuna farklı mekanizmalar üzerinden katkı yapan metilkobalamin ve takrolimus içeren nanofiber sinir sargılarının etkilerinin karşılaştırılaştırılması amaçlanmıştır.
Her grupta 8’er adet sıçan olacak şekilde 6 grup oluşturularak işlemler gerçekleştirildi. Tüm gruplarda anestezi altında sıçanların sol siyatik siniri dorsal yaklaşım ile eksplore edildi ve Grup 1 hariç diğer tüm gruplarda sol siyatik trifurkasyonunun 1 cm proximalinde mikroskop altında mikromakas ile tam kat düz kesi yapıldı. Kesi sonrası aynı seansta siyatik sinir 9/0 nylon dikişler ile epinöral onarım yapılarak koaptasyon sağlandı. Grup 2 için sadece epinöral onarım yapıldı. Grup 3 için epinöral onarım hattı etrafına elektro-eğirme yöntemi ile üretilen PCL sinir sargısı sarıldı. Grup 4 için epinöral onarım hattı etrafına içerisinde Metilkobalamin bulunan, Grup 5 için Takrolimus bulunduran, Grup 6 için hem Metilkobalamin hem de Takrolimus bulunduran PCL nanofiber sinir sargıları sarıldı. Sıçanlar 8 hafta süre ile takip edildi. Sıçanlar sakrifiye edilmeden önce fonksiyonel değerlendirme için yürüme testi ve elektrofizyolojik değerlendirmeler yapıldı. Sakrifikasyon anestezi altında gerçekleştirildikten sonra sol siyatik sinirden örnek alınarak histopatolojik inceleme yapıldı. Bilateral gastroknemius kasları origo-insersiyolarından ayrılarak hassas laboratuvar terazisi ağırlıkları ölçüldü ve gastroknemius kas ağırlık indeksi hesaplandı.
v
8. hafta sonunda yapılan değerlendirmelerde siyatik sinirde tam kat kesi yapılarak cerrahi işlem uygulanan gruplar arasında en iyi SFI değeri Grup 4’te bulundu. Siyatik sinirde tam kat kesi yapılarak cerrahi işlem uygulanan gruplar arasında sham grubuna en yakın, en iyi gastroknemius kas ağırlık indeksi değerleri Grup 4 ve 5 ’te bulundu. Grup 4 ve 5 arasında istatiksel olarak anlamlı fark saptanmadı (p=0.600). Grup 1’den Grup 6’ya sırasıyla ortalama latans süreleri; 1,42±0,13 , 2,15±0,47 , 1,83±0,22 ,1,63±0,22 , 1,74±0,25 , 1,82±0,16 ms olarak bulundu. Grup 1’den Grup 6’ya sırasıyla ortalama amplitüd değerleri; 11,91±2,30 , 3,59±0,76 , 4,86±3,71 , 9,27±2,41 , 7,75±2,51 , 5,62±3,37 mV olarak bulundu. Histopatolojik değerlendirmede ortalama fibrozis değeri Grup 5’te en düşük olarak bulundu. İnflamasyona bakıldığında Grup 5 ve 6’da diğer gruplara göre daha az inflamasyon görüldü. Grup 4 ve 5’te aksonal dejenerasyon derecesi %50’den fazla olan sıçan görülmedi. Ortalama vaskülarizasyon değerinin en yüksek Grup 3’te olduğu ve Grup 4,5 ve 6’nın da benzer vaskülarizasyon değerlerine sahip olduğu görüldü. Gruplar mm2 ‘deki aksonal yoğunluk açısından karşılaştırıldığında en fazla ortalama akson sayısının Grup 5’te olduğu görüldü. Fasiküler organizasyon bozukluğunun siyatik sinire tam kat kesi yapılarak cerrahi işlem uygulanan gruplar arasında en az Grup 4’te olduğu görüldü. Grup 4’te %50 fazla fasiküler organizasyon bozukluğu bulunan sıçan görülmedi.
Sonuç olarak hem metilkobalamin hem takrolimus içeren nanofiber sinir sargılarının ayrı ayrı kullanıldığında fonksiyonel ve histopatolojik olarak sinir rejenerasyonuna olumlu katkıda bulunduğu görüldü. İki ajan fonksiyonel ve histopatolojik olarak karşılaştırıldığında sinir rejenerasyonu açısından birbirlerine belirgin üstünlüğü olmadığı görüldü. Metilkobalamin ve takrolimusun birlikte kullanımının sinir rejenerasyonu üzerinde sinerjistik bir etki yaratmadığı ve sinir iyileşmesi için olumlu katkıları bulunmasına karşın istatistiksel olarak anlamlı fark oluşturmadığı gözlemlendi.
Peripheral nerve injuries are common and can cause loss of sensory and motor functions in the affected patient group. Even after successful surgeries, satisfactory results in motor and sensory functions may not be achieved. Incomplete healing of peripheral nerve injuries often leads to many adverse consequences such as numbness, chronic pain, and decreased sensory and/or motor function in the affected limbs, and for these reasons, peripheral nerve injuries constitute a worldwide clinical problem. Current research has evolved in this direction in order to find a pharmacological agent that can be successfully administered. One of these researches is nanotechnology and nanomaterials produced with this method. In our study, we aimed to compare the effects of nanofiber nerve wraps containing methylcobalamin and tacrolimus, which contribute to peripheral nerve regeneration through different mechanisms. Six groups were constituted with 8 rats in each group and the studies were performed. The left sciatic nerve of rats was explored by dorsal approach under anesthesia in all groups. In all groups except Group 1, a full-thickness straight transection was performed with micro scissors 1 cm proximal to the left sciatic trifurcation under the microscope. After the transection, epineural repair of the sciatic nerve was performed with 9/0 nylon sutures in the same session and coaptation was achieved. For Group 2, only epineural repair was performed. For group 3, PCL nerve wrap produced by electro-spinning method was wrapped around the epineural repair site. PCL nanofiber nerve wraps containing Methylcobalamin for Group 4, Tacrolimus for Group 5 and both Methylcobalamin and Tacrolimus for Group 6 were wrapped around the epineural repair site. The rats were followed up for 8 weeks. Before the rats were sacrificed, walking track test and electrophysiologic evaluations were performed for functional assessment. After sacrification under anesthesia, the left sciatic nerve was sampled for histopathological examination. Bilateral gastrocnemius muscles were separated from their origo-insertions and weights were measured on a sensitive laboratory scale and gastrocnemius muscle weight index was calculated. vii At the end of the 8th week, the best SFI value was found in Group 4 among the groups that underwent surgical procedure with a full-thickness transection of the sciatic nerve. The highest and best gastrocnemius muscle weight index values were found in Groups 4 and 5, which were closest to the sham group among the groups that underwent surgical procedure by making a full-thickness transection in the sciatic nerve. There was no statistically significant difference between Groups 4 and 5 (p=0.600). The mean latency periods from Group 1 to Group 6 were 1.42±0.13, 2.15±0.47, 1.83±0.22, 1.63±0.22, 1.74±0.25, 1.82±0.16 ms, respectively. The mean amplitude values from Group 1 to Group 6 were 11.91±2.30, 3.59±0.76, 4.86±3.71, 9.27±2.41, 7.75±2.51, 5.62±3.37 mV, respectively. In histopathological assessment, the mean fibrosis value was found to be the lowest in Group 5. When inflammation was evaluated, less inflammation was observed in Groups 5 and 6 compared to the other groups. In Groups 4 and 5, there were no rats with more than 50% axonal degeneration. Group 3 had the highest mean vascularization value and Groups 4, 5 and 6 had similar vascularization values. When the groups were compared in terms of axonal density per mm2 , the highest mean axon number was seen in Group 5. Fascicular disorganisation was found to be the least in Group 4 of the groups that underwent surgery with full-thickness transection of the sciatic nerve. In group 4, there were no rats with more than 50% fascicular disorganisation. In conclusion, nanofiber nerve wraps containing either methylcobalamin or tacrolimus contributed favorably to nerve regeneration functionally and histopathologically when used separately. When the two agents were compared functionally and histopathologically, no significant superiority was observed in terms of nerve regeneration. It was observed that the combined use of methylcobalamin and tacrolimus did not have a synergistic effect on nerve regeneration and did not result in a statistically significant difference, although there were positive contributions to nerve regeneration.
Peripheral nerve injuries are common and can cause loss of sensory and motor functions in the affected patient group. Even after successful surgeries, satisfactory results in motor and sensory functions may not be achieved. Incomplete healing of peripheral nerve injuries often leads to many adverse consequences such as numbness, chronic pain, and decreased sensory and/or motor function in the affected limbs, and for these reasons, peripheral nerve injuries constitute a worldwide clinical problem. Current research has evolved in this direction in order to find a pharmacological agent that can be successfully administered. One of these researches is nanotechnology and nanomaterials produced with this method. In our study, we aimed to compare the effects of nanofiber nerve wraps containing methylcobalamin and tacrolimus, which contribute to peripheral nerve regeneration through different mechanisms. Six groups were constituted with 8 rats in each group and the studies were performed. The left sciatic nerve of rats was explored by dorsal approach under anesthesia in all groups. In all groups except Group 1, a full-thickness straight transection was performed with micro scissors 1 cm proximal to the left sciatic trifurcation under the microscope. After the transection, epineural repair of the sciatic nerve was performed with 9/0 nylon sutures in the same session and coaptation was achieved. For Group 2, only epineural repair was performed. For group 3, PCL nerve wrap produced by electro-spinning method was wrapped around the epineural repair site. PCL nanofiber nerve wraps containing Methylcobalamin for Group 4, Tacrolimus for Group 5 and both Methylcobalamin and Tacrolimus for Group 6 were wrapped around the epineural repair site. The rats were followed up for 8 weeks. Before the rats were sacrificed, walking track test and electrophysiologic evaluations were performed for functional assessment. After sacrification under anesthesia, the left sciatic nerve was sampled for histopathological examination. Bilateral gastrocnemius muscles were separated from their origo-insertions and weights were measured on a sensitive laboratory scale and gastrocnemius muscle weight index was calculated. vii At the end of the 8th week, the best SFI value was found in Group 4 among the groups that underwent surgical procedure with a full-thickness transection of the sciatic nerve. The highest and best gastrocnemius muscle weight index values were found in Groups 4 and 5, which were closest to the sham group among the groups that underwent surgical procedure by making a full-thickness transection in the sciatic nerve. There was no statistically significant difference between Groups 4 and 5 (p=0.600). The mean latency periods from Group 1 to Group 6 were 1.42±0.13, 2.15±0.47, 1.83±0.22, 1.63±0.22, 1.74±0.25, 1.82±0.16 ms, respectively. The mean amplitude values from Group 1 to Group 6 were 11.91±2.30, 3.59±0.76, 4.86±3.71, 9.27±2.41, 7.75±2.51, 5.62±3.37 mV, respectively. In histopathological assessment, the mean fibrosis value was found to be the lowest in Group 5. When inflammation was evaluated, less inflammation was observed in Groups 5 and 6 compared to the other groups. In Groups 4 and 5, there were no rats with more than 50% axonal degeneration. Group 3 had the highest mean vascularization value and Groups 4, 5 and 6 had similar vascularization values. When the groups were compared in terms of axonal density per mm2 , the highest mean axon number was seen in Group 5. Fascicular disorganisation was found to be the least in Group 4 of the groups that underwent surgery with full-thickness transection of the sciatic nerve. In group 4, there were no rats with more than 50% fascicular disorganisation. In conclusion, nanofiber nerve wraps containing either methylcobalamin or tacrolimus contributed favorably to nerve regeneration functionally and histopathologically when used separately. When the two agents were compared functionally and histopathologically, no significant superiority was observed in terms of nerve regeneration. It was observed that the combined use of methylcobalamin and tacrolimus did not have a synergistic effect on nerve regeneration and did not result in a statistically significant difference, although there were positive contributions to nerve regeneration.
Açıklama
Anahtar Kelimeler
Metilkobalamin, Nanofiber, Periferik sinir rejenerasyonu polikaprolakton, Sinir sargısı, Takrolimus, Methylcobalamin, Nanofiber, Nerve wrap, Peripheral nerve regeneration, Polycaprolactone, Tacrolimus
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Tekecik, M. (2023). Metilkobalamin ve takrolimus içeren Polikaprolakton (PCL) nanofiberlerin periferik sinir rejenerasyonuna etkilerinin karşılaştırılması. (Yayınlanmamış tıpta uzmanlık tezi) Necmettin Erbakan Üniversitesi, Meram Tıp Fakültesi Cerrahi Tıp Bilimleri Bölümü Plastik Cerrahi Anabilim Dalı, Konya.
