Ulus Travma Acil Cerrahi Derg. Baskýdaki Makaleler: UTD-37711

Diyabetik Sýçanlarda Deri Yarasýnda Kombine Hücresel Tedaviye Probiyotiklerin Katkýsý

Emine Müge Karakayalý¹, Mehmet Ýbrahim Tuðlu^2
1Týbbi Mikrobiyoloji Anabilim Dalý, Ýzmir Demokrasi Üniversitesi Týp Fakültesi, Ýzmir
²Histoloji Ve Embriyoloji Anabilim Dalý, Celal Bayar Üniversitesi Týp Fakültesi, Manisa

Amaç: Diyabetik (DB) yara bakýmý, kronik ve iyileþmeyen, enfeksiyona yatkýn ve ampütasyona yol açan ülserler nedeniyle hala çok önemli bir sorundur. Mezenkimal Kök Hücreler (MKH), Trombositten Zengin Plazma (TZP) ve Stromal Vasküler Fraksiyon (SVF), son yýllarda yaygýn olarak kullanýlan Hücresel Terapi (HT) ürünleridir. Tek baþlarýna ve ikili kombinasyonlarda kullanýmlarý üzerine çalýþmalar olmasýna raðmen, üçlü kombinasyon olarak Kombine Hücresel Terapi (KHT) çalýþýlmamýþtýr. Probiyotikler (PB), baðýrsakta yaptýklarý mikrobiyata deðiþiklikler ile iyileþmeyi hýzlandýrýr. Bu çalýþmada, KHT ile oluþan DB yara iyileþmesinde PB katkýsýnýn incelenmesi amaçlandý.
Gereç ve Yöntem: DB model, eriþkin Albino sýçanlarda 40 mg/kg STZ i.p. uygulanmasýyla oluþturuldu. 1x1 cm2 tam kat kutanöz yara yapýldý. Denekler, DB, DB+PB, DB+CCT ve DB+ CCT+PB olmak üzere her biri 6 yetiþkin içeren gruplara bölündü. CCT olarak, 100 µL 1x106 MSC, 100 µL SVF ve 100 µL PRP içeren toplam 300 µL solüsyonun her biri 30 µL olacak þekilde yara kenarlarýna uygulanmasý ile yapýldý. PB, yaklaþýk 200 mg/günlük dozlarda oral yoldan uygulandý. Histokimyasal analizler HE ve MT ile gerçekleþtirildi. Ýmmünohistokimyasal analizleri için eNOS, Kaspaz 3, IL10, VEGF ve Kolajen I kullanýldý. PB için taksonomik yapý fekal metagenomik analizi ile gösterildi.
Bulgular: DB yaranýn CCT ile daha etkin ve hýzlý kapandýðý görüldü. PB ile bu sürecin hýzlandýrýldýðý ve daha etkin iyileþme saðlandýðý saptandý. Ayrýca KHT ile yüksek eNOS, Kaspaz 3 ve IL-10 ekspresyonlarýnýn azaltýldýðý ayrýca düþük VEGF ve Kollajen I tersine çevrildiði gözlendi. PB uygulamasýnýn KHT ile oluþan bu olumlu etkilerine anlamlý bir katký saðladýðý gösterildi. PB katkýsý ile KHT oksidatif stresi, apoptozu ve inflamasyonu önleyip vaskülarizasyon ve kollajen organizasyonunu artýrarak yara iyileþmesini önemli ölçüde geliþtirdi.
Sonuç: DB yara tedavisinde KHT ile PB katkýsýnýn önemli olduðu ve hastanýn kötü yaþam kalitesini düzeltebileceði düþünüldü.
Anahtar kelimeler: Probiyotikler, Diyabetik Cilt, Yara Ýyileþmesi, Kombine Hücre Tedavisi.

Anahtar Kelimeler: Rat, Probiotics, Diabetic Skin, Wound Healing, Cell Therapy

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The Contribution of probiotics to the cellular therapy on skin wound in diabetic rats

Emine Müge Karakayalý¹, Mehmet Ýbrahim Tuðlu^2
1Department Of Medical Microbiology, Izmir Democracy University Faculty Of Medicine, Ýzmir, Turkey
²Department Of Histology And Embryology, Manisa Celal Bayar University Faculty Of Medicine, Manisa, Turkey

ABSTRACT
Background: Diabetes-related wound care is still a major issue because of the chronic and unhealed ulcers that are prone to infection and ultimately to amputation. In recent years, cellular therapy (CT) products such as mesenchimal stem cells (MSC), platelet-rich plasma (PRP) and stromal vascular fraction (SVF) have been widely used. A Combined Cellular Therapy (CCT) has not been explored as a triple combination, despite investigations on its usage both alone and in dual combinations. Probiotics (PB) accelerated healing by causing changes in the intestine mictobiata. This study aims to examine the role of PB in the effects of CCT on diabetic wound healing.
Methods: A 1×1 cm2 full-thickness cutaneous wound was made after administering 40 mg/kg STZ i.p. to create a diabetic (DB) animal model. They were divided into four groups: DB, DB+PB, DB+CCT and DB+ CCT+PB, each with five (6) adult Albino rats. The wound edges were treated with a total of 300 µL a solution including 30 µL of each of 100 uL 1×106 MSC, 100 µL SVF, and 100 uL PRP as CCT. PB was administered orally in doses of about 200 mg daily. Histochemical analyses were performed using HE and MT. Immunohistochemical analyses were done by eNOS, Caspase 3, IL10, VEGF and Collagen I. The intestinal microbiome study revealed by metagenomic analyses for taxonomic structure.
Results: CCT provided more effective and faster healing in DB animals. It was further discovered that PB accelerated this process, resulting in greater improvement. Furthermore, CCT was observed to reverse high eNOS, Caspase 3 and IL-10 expressions as well as low VEGF and Collagen I. Additionally, PB therapy was shown to contribute significantly to these positive effects of CCT. CCT in combination with PB significantly improved wound healing by preventing oxidative stress, apoptosis and inflammation with improvement of vascularization and collagene organization.
Conclusions: PB support was considered important for diabetic wound healing. It was suggested that this contribution could improve the patient’s poor quality of life.
Key words: Probiotics, Diabetic Skin, Wound Healing, Cell Therapy.

Keywords: Rat, Probiotics, Diabetic Skin, Wound Healing, Cell Therapy

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