Diyabetik Ayak Yaraları Üzerine İmmunohistokimyasal Bir Çalışma; MMP-2 ve TNF- α Ekspresyonlarının İncelenmesi
Öz
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
Kaynakça
- Rodrigues J, Mitta N. Diabetic Foot and Gangrene. In Tech Open 2011; 29: 1–25.
- Williams DT, Hilton JR, Harding KG. Diagnosing foot infection in diabetes. Clin Infect Dis 2004; 39: 83–6.
- Berlanga J, Valdéz C, Savigne W. Cellular and molecular insights into the wound healing mechanism in diabetes, Biotecnol Apl 2010; 27: 255–61.
- Acosta JB, del Barco DG, Vera DC et al. The pro-inflammatory environment in recalcitrant diabetic foot wounds. Int Wound J 2008; 5(4): 530-9.
- Xu F, Zhang C, Graves DT. Abnormal cell responses and role of TNF-α in impaired diabetic wound healing. BioMed Res Int 2013; 754802.
- Aggarwal BB. Signalling pathways of the TNF superfamily: a double-edged sword, Nat Rev Immunol 2003; 3(9): 745–56.
- Wetzler C, Kampfer H, Stallmeyer B, Pfeilschifter J, Frank S. Large and sustained induction of chemokines during impaired wound healing in the genetically diabetic mouse: prolonged persistence of neutrophils and macrophages during the late phase of repair, J Invest Dermatol 2000;115: 245–253.
- Nagase H, Woessner JF. Matrix metalloproteinases, J Biol Chem 1999; 274(31): 21491-4.
- Overall CM, Lopez-Otin C. Strategies for MMP inhibition in cancer: innovations for the post-trial era, Nat Rev Cancer, 2002; 2: 657–72.
- Hu J, Van den Steen PE, Sang QX, Opdenakker G. Matrix metalloproteinase inhibitors as therapy for inflammatory and vascular diseases. Nat Rev Drug Discov 2007; 6(6):480-98.
- Sawicki G. Intracellular Regulation of Matrix Metalloproteinase-2 Activity: New Strategies in Treatment and Protection of Heart Subjected to Oxidative Stress. Scientifica 2013; ID 130451.
- Tellechea A, Le Veves A, Carvalho E. Inflammatory and angiogenic abnormalities in diabetic wound healing: role of neuropeptides and therapeutic perspectives, Open Circ Vasc J 2010; 3: 43–55.
- Leung PC. Diabetic foot ulcers, a comprehensive review. Surgeon 2007; 5(4): 219-31.
- Sibbald RG, Woo KY. The biology of chronic foot ulcers in persons with diabetes, Diabetes Metab Res Rev.2008; 24(1): 25–30.
- Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic-Canic M. Growth factors and cytokines in wound healing, Wound Repair Regen 2008; 16(5): 585-601.
- Borst SE. The role of TNF-alpha in insulin resistance. Endocrine 2004; 23(2-3): 177-82.
- Rapala K, Laato M, Niinikoski J, et al. Tumor necrosis factor alpha inhibits wound healing in the rat. Eur Surg Res 1991;23: 261–8.
- Löffek S, Schilling O, Franzke CW. Biological role of matrix metalloproteinases: A critical balance. Eur Respir J 2011; 38: 191-208.
Öz
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.
Anahtar Kelimeler
Diabetic foot MMP-2 TNF- α histopathology immunohistochemistry
Kaynakça
- Rodrigues J, Mitta N. Diabetic Foot and Gangrene. In Tech Open 2011; 29: 1–25.
- Williams DT, Hilton JR, Harding KG. Diagnosing foot infection in diabetes. Clin Infect Dis 2004; 39: 83–6.
- Berlanga J, Valdéz C, Savigne W. Cellular and molecular insights into the wound healing mechanism in diabetes, Biotecnol Apl 2010; 27: 255–61.
- Acosta JB, del Barco DG, Vera DC et al. The pro-inflammatory environment in recalcitrant diabetic foot wounds. Int Wound J 2008; 5(4): 530-9.
- Xu F, Zhang C, Graves DT. Abnormal cell responses and role of TNF-α in impaired diabetic wound healing. BioMed Res Int 2013; 754802.
- Aggarwal BB. Signalling pathways of the TNF superfamily: a double-edged sword, Nat Rev Immunol 2003; 3(9): 745–56.
- Wetzler C, Kampfer H, Stallmeyer B, Pfeilschifter J, Frank S. Large and sustained induction of chemokines during impaired wound healing in the genetically diabetic mouse: prolonged persistence of neutrophils and macrophages during the late phase of repair, J Invest Dermatol 2000;115: 245–253.
- Nagase H, Woessner JF. Matrix metalloproteinases, J Biol Chem 1999; 274(31): 21491-4.
- Overall CM, Lopez-Otin C. Strategies for MMP inhibition in cancer: innovations for the post-trial era, Nat Rev Cancer, 2002; 2: 657–72.
- Hu J, Van den Steen PE, Sang QX, Opdenakker G. Matrix metalloproteinase inhibitors as therapy for inflammatory and vascular diseases. Nat Rev Drug Discov 2007; 6(6):480-98.
- Sawicki G. Intracellular Regulation of Matrix Metalloproteinase-2 Activity: New Strategies in Treatment and Protection of Heart Subjected to Oxidative Stress. Scientifica 2013; ID 130451.
- Tellechea A, Le Veves A, Carvalho E. Inflammatory and angiogenic abnormalities in diabetic wound healing: role of neuropeptides and therapeutic perspectives, Open Circ Vasc J 2010; 3: 43–55.
- Leung PC. Diabetic foot ulcers, a comprehensive review. Surgeon 2007; 5(4): 219-31.
- Sibbald RG, Woo KY. The biology of chronic foot ulcers in persons with diabetes, Diabetes Metab Res Rev.2008; 24(1): 25–30.
- Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic-Canic M. Growth factors and cytokines in wound healing, Wound Repair Regen 2008; 16(5): 585-601.
- Borst SE. The role of TNF-alpha in insulin resistance. Endocrine 2004; 23(2-3): 177-82.
- Rapala K, Laato M, Niinikoski J, et al. Tumor necrosis factor alpha inhibits wound healing in the rat. Eur Surg Res 1991;23: 261–8.
- Löffek S, Schilling O, Franzke CW. Biological role of matrix metalloproteinases: A critical balance. Eur Respir J 2011; 38: 191-208.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Klinik Tıp Bilimleri |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 29 Ağustos 2019 |
| Gönderilme Tarihi | 15 Mart 2019 |
| Kabul Tarihi | 10 Mayıs 2019 |
| Yayımlandığı Sayı | Yıl 2019 Cilt: 16 Sayı: 2 |
Harran Üniversitesi Tıp Fakültesi Dergisi / Journal of Harran University Medical Faculty