Fotobiyomodülasyon ve Düşük Doz Fotodinamik Terapinin HUVEC Hücrelerindeki Anjiyogenez'e Yönelik Olası Terapötik Etkileri: Karşılaştırmalı İn Vitro Analiz

Abstract

Fotobiyomodülasyon (FBM), yara iyileşmesi, hücre proliferasyonu/farklılaşması mekanizmalarını hızlandıran iyonlaştırıcı olmayan bir ışık terapisidir. Son zamanlarda, fotodinamik terapinin (FDT) düşük dozlarda uygulandığında FBM gibi davranabileceği anlaşılmıştır. Bu çalışmada karşılaştırmalı bir analizle anjiyogenezi artırmak için HUVEC'ler üzerinde FBM ve düşük doz FDT’nin etkisi araştırılmıştır. HUVEC’ler 808 nm dalgaboyu ile uyarılmıştır. FDT uygulamalarında fotosensitizan olarak indosiyanin yeşil kullanılmıştır. Her iki yöntem tekli ve üçlü olarak uygulanmıştır. Uygulamaların etkileri hücre canlılığı, hücre içi ROS, MMP değişikliği, NO salınımı ve morfolojik analizler ile incelenmiştir. Vaskülarizasyonla ilgili proteinlerin (VEGF, PECAM-1 ve vWf) ifadeleri, immünofloresan boyama ve qRT-PCR yöntemleri ile belirlenmiştir. Olası termal hasarları belirlemek için ışık uygulamaları sırasındaki sıcaklık değişimleri izlenmiştir. Sonucunda, üçlü düşük doz FDT uygulamasının hücre proliferasyonu ve damar benzeri yapı oluşumunda %20 oranında daha başarılı olduğu gözlenmiştir. Bütün uygulamalarda ROS seviyesi önemli ölçüde değişmezken, üçlü FDT uygulamalarında NO salınımının miktarı kontrol grubunun yaklaşık 5 katı olmuş, bu da NO salınımının bu mekanizmada anahtar bir molekül olarak hareket ettiğini göstermiştir. Vaskülarizasyonla ilgili proteinler de FDT uygulamalarında daha güçlü bir şekilde ifade edilmiştir. Sonuç olarak, düşük doz FDT'nin NO salınımı yoluyla vaskülarizasyonu hızlandırmak için fotobiyomodülatif etki gösterdiği anlaşılmıştır.

Keywords

• Fotobiyomodülasyon
• Fotodinamik Terapi
• 808-nm
• İndosiyanin Yeşil
• HUVEC hücreleri

THE POSSIBLE THERAPEUTIC IMPACTS OF PHOTOBIOMODULATION AND LOW-DOSE PHOTODYNAMIC THERAPY ON HUVECS TOWARDS ANGIOGENESIS: A COMPARATIVE IN VITRO ANALYSIS

Abstract

Photobiomodulation (PBM) is a non-ionizing therapy that promotes faster wound healing and cell proliferation/differentiation. It is recently understood that photodynamic therapy (PDT) may act as PBM when applied at low-level. In this study, a comparative analysis between PBM and low-dose PDT was performed on HUVECs to increase angiogenesis. HUVECs were irradiated at 808-nm of wavelength. Indocyanine green was used as a photosensitizer in PDT applications. Single and triple treatments were employed for both modalities. Their effects were analyzed with cell viability, intracellular ROS, MMP change, NO release, and morphological analysis. The expressions of vascularization-related proteins (VEGF, PECAM-1, and vWf) were determined through immunofluorescence staining and qRT-PCR. Temperature changes during applications were monitored to determine any thermal damages. It was observed that triple PDT application was more successful at increasing cell proliferation and tube-like structure formation with a 20% rate. The level of ROS did not significantly change in all applications. However, the amount of NO release in triple PDT application was nearly 5 times that of the control group, which showed it acted as a key molecule. The vascularization-related proteins were more strongly expressed in PDT applications. It was understood that low-dose PDT can exert a photobiomodulation effect to accelerate vascularization through NO release.

Keywords

• Photobiomodulation
• Photodynamic Therapy
• 808-nm
• Indocyanine Green
• HUVECs

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