Publication: Alfa-13-karboksikromanolün nanopartikül aracılı makrofaja iletiminin in vitro eğerlendirilmesi
Abstract
Amaç : Bu çalışma, uzun zincirli Vitamin E metaboliti alfa-13-Karboksikromanolün (α-13′COOH) inflamasyon süreçlerindeki etkilerini, makrofajlara spesifik hedeflenmiş nanopartikül (NP) sistemleri aracılığıyla incelemeyi amaçladı. Çalışmada, α-13′COOH’ün makrofaj hücreleri ile etkileşiminin artırılması, metabolitin biyolojik aktivitesinin daha iyi anlaşılması ve inflamasyona modülatör etkilerinin değerlendirilmesi hedeflendi. Geliştirilen NP tabanlı taşıma sistemleri, α-13′COOH’ün düşük stabilite ve hızlı metabolize olma gibi sınırlamalarını aşarak, metabolitin makrofajlar üzerindeki potansiyel terapötik faydalarını analiz etmek ve inflamasyonun düzenlenmesinde yeni stratejiler sunmak amacıyla tasarlandı. Gereç ve Yöntem : Makrofaj hedefli S2P kaplı α-13′COOH metaboliti yüklü ve yüksüz NP’ler önceden temin edildi. Ardından, primer hücre kültürü çalışmalarıyla makrofaj hücreleri farklılaştırıldı ve makrofaj-NP etkileşimi floresan mikroskobi yöntemi ile incelendi. Hücre canlılığı MTT testi ve α-13′COOH seviyeleri LC-MS/ MS yöntemi ile ölçüldü ayrıca TNF-α, TGF-β, IL-10, IL-6 ve IL-1β mRNA düzeyleri qRT-PCR yöntemiyle analiz edildi. Bulgular : Makrofajlarla etkileşim sonrası NP birikimi incelendi ve MTT testi sonuçlarında gruplar arasında anlamlı bir fark bulunmadı. LC-MS/ MS analizinde, hedeflendirilmiş, NP’lerin hedeflendirilmemiş NP’lere kıyasla α-13′COOH düzeylerini anlamlı derecede artırdığı ve konsantrasyon artışıyla bu etkinin güçlendiği tespit edildi. qRT-PCR sonuçlarına göre, TNF-α ekspresyonunda hedeflendirilmiş 0,5 µM NP grubunda istatistiksel olarak anlamlı bir azalma belirlendi. Sonuç : Hedeflendirilmiş NP’lerin α-13′COOH düzeylerini artırdığı belirlendi ayrıca TNF-α ekspresyonunun baskılandığı saptandı. Bu bulgular, α-13′COOH'ün inflamasyonun düzenlenmesindeki iyileştirici potansiyelini ve NP sistemlerinin biyoteknolojik tedavi yaklaşımlarında kullanımını desteklemektedir.
Objective : This study aims to investigate the effects of the long-chain Vitamin E metabolite alpha-13’carboxychromanol (α-13′COOH) on inflammation processes through macrophage-specific targeted nanoparticle (NP) systems. The study focuses on enhancing the interaction between α-13′COOH and macrophage cells, gaining a deeper understanding of the metabolite's biological activity, and evaluating its modulatory effects on inflammation. The developed NP-based delivery systems are designed to overcome the limitations of α-13′COOH, such as low stability and rapid metabolism, to analyze its potential therapeutic benefits on macrophages and propose novel strategies for regulating inflammation. Materials and Methods : Macrophage-targeted S2P-coated α-13′COOH-loaded and unloaded NPs were pre-prepared, followed by the differentiation of macrophage cells through primary cell culture studies. The interaction between macrophages and NPs was examined using fluorescence microscopy. Cell viability was assessed via the MTT assay, and α-13′COOH levels were measured using LC-MS/ MS. Additionally, mRNA levels of TNF-α, TGF-β, IL-10, IL-6, and IL-1β were analyzed using the qRT-PCR method. Results : Following interaction with macrophages, NP accumulation was examined, and MTT assay results showed no significant differences between the groups. LC-MS/ MS analysis revealed that targeted NPs significantly increased α-13′COOH levels compared to non-targeted NPs, with this effect becoming more pronounced as the concentration increased. According to qRT-PCR results, a statistically significant decrease in TNF-α expression was observed in the targeted 0.5 µM NP group. Conclusion : Targeted NPs were found to enhance α-13′COOH levels and suppress TNF-α expression. These findings support the therapeutic potential of α-13′COOH in inflammation modulation and highlight the utility of NP systems in biotechnological therapeutic approaches.
Objective : This study aims to investigate the effects of the long-chain Vitamin E metabolite alpha-13’carboxychromanol (α-13′COOH) on inflammation processes through macrophage-specific targeted nanoparticle (NP) systems. The study focuses on enhancing the interaction between α-13′COOH and macrophage cells, gaining a deeper understanding of the metabolite's biological activity, and evaluating its modulatory effects on inflammation. The developed NP-based delivery systems are designed to overcome the limitations of α-13′COOH, such as low stability and rapid metabolism, to analyze its potential therapeutic benefits on macrophages and propose novel strategies for regulating inflammation. Materials and Methods : Macrophage-targeted S2P-coated α-13′COOH-loaded and unloaded NPs were pre-prepared, followed by the differentiation of macrophage cells through primary cell culture studies. The interaction between macrophages and NPs was examined using fluorescence microscopy. Cell viability was assessed via the MTT assay, and α-13′COOH levels were measured using LC-MS/ MS. Additionally, mRNA levels of TNF-α, TGF-β, IL-10, IL-6, and IL-1β were analyzed using the qRT-PCR method. Results : Following interaction with macrophages, NP accumulation was examined, and MTT assay results showed no significant differences between the groups. LC-MS/ MS analysis revealed that targeted NPs significantly increased α-13′COOH levels compared to non-targeted NPs, with this effect becoming more pronounced as the concentration increased. According to qRT-PCR results, a statistically significant decrease in TNF-α expression was observed in the targeted 0.5 µM NP group. Conclusion : Targeted NPs were found to enhance α-13′COOH levels and suppress TNF-α expression. These findings support the therapeutic potential of α-13′COOH in inflammation modulation and highlight the utility of NP systems in biotechnological therapeutic approaches.