Diyabetik Ayak Yaraları Üzerine İmmunohistokimyasal Bir Çalışma; MMP-2 ve TNF- α Ekspresyonlarının İncelenmesi

Amaç: Diyabetik ayak, diyabetin önemli ve uzun süreli komplikasyonlarından biridir. Bilindiği üzere diyabetik bireylerde yara iyileşmesi yavaş olmaktadır ve bu duruma bakteriyel invazyonun eklenmesi sonucu uzun süreli inflamasyon eşliğinde iyileşmeyen diyabetik ayak yaraları ortaya çıkmaktadır. Söz konusu çalışmanın amacı, diyabetik ayak yara dokusunda proinflamatuvar sitokinlerden TNF-α ve kollajenin parçalanmasında rol oynayarak dokunun yeniden şekillenmesini sağlayan matriks metaloprotein MMP-2 ekspresyonunu immunohistokimyasal yöntemlerle tespit etmektir.Materyal ve metod: Bu çalışmaya 30 erkek ve 30 kadın olmak üzere, diyabetik ayak tanısı almış, ve ayaklarında açık yara bulunan 60 birey dahil olmuştur. Çalışmaya alınacak ayak, izotonik çözelti ile yıkandıktan sonra yaralar kesilip çıkarılmış ve dokular %10’luk formaldehit solüsyonunda tespit edilmiştir. Rutin histolojik takip sonrası kesitler parafine gömülmüş ve yarı-ince kesitleri alınarak histopatolojik incelemeleri yapılmıştır. İmmunohistokimyasal analiz için, doku örnekleri, MMP-2 ve TNF-α primer antikorları ile boyanarak mikroskop altında incelenmiştir. Bulgular: Çalışmamızın sonuçlarına göre diyabetik ayak yara dokusunda, ligamenter dokunun içinde lökositler, lenfositler ve monositlerin yoğun olduğu izlenmiştir. Kollajen liflerde dejenerasyon ve kan damarlarında dilatasyon, konjesyon ve ödem görülmüştür. İnflamatuvar hücrelerde ve nekroze olan alanlarda TNF-α ekspresyonunda artış izlenmiştir. Damar çevresinde görülen yoğun inflamasyonunun arasında, dejenere kollajen lif ve fibroblast hücreleri ve ekstrasellüler matrikste MMP-2 ekspresyonu pozitif olarak gözlenmiştir.Sonuç: Diyabetik ayak yarası tedavisinde MMP ekspresyonu yönünde düzenleme yapılarak, her geçen gün genişleyen diyabetik popülasyonda iyileşmeyen ayak yaralarına karşı bir yaklaşım geliştirilebilir düşüncesindeyiz.

Anahtar Kelimeler:

Diyabetik ayak, MMP-2, TNF- α, histopatoloji, immunohistokimya

An immunohistochemical study on diabetic foot wounds; examination of MMP-2 and TNF- α expressions

Objectives: Diabetic foot is one of the major and long-term complications of diabetes. As known, wound healing is slow in diabetic individuals and as a result of inclusion of bacterial invasion to this condition, non-healing diabetic foot wounds occur with prolonged inflammation. The aim of this study was to determine the expressions of TNF-α, a proinflammatory cytokine and matrix metalloprotein MMP-2, involved in tissue remodeling via collagen degradation by immunohistochemical methods in diabetic foot wound tissues.Materials and Methods: This study included 30 males and 30 females, a total of 60 patients diagnosed with diabetic foot having open wounds. After washing the foot with isotonic solution, the hole wounds were cut and tissues were fixed in 10% formaldehyde solution. After routine histological follow-up, the sections were embedded in paraffin and semi-thin sections were cut and histopathological examinations were performed. For immunohistochemical analysis, tissue samples were stained with MMP-2 and TNF-α primary antibodies and examined under a microscope.Results: In our study, leukocytes, lymphocytes and monocytes were observed in the diabetic foot wound tissue. Degeneration of collagen fibers and dilatation of blood vessels, congestion and edema were observed. TNF-α expression was increased in inflammatory cells and necrosis areas. MMP-2 expression was found to be positive in degenerated collagen fibers, fibroblasts and extracellular matrix among the intense inflammation around the vein. Conclusion: Regarding the MMP expression in diabetic foot wound treatment, we think that an approach to the healing of non-healing foot wounds may be developed in the diabetic population.

Keywords:

Diabetic foot, MMP-2, TNF- α, histopathology, immunohistochemistry,

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