Atopik dermatit tedavisi için yeni bir katyonik nano boyutlu taşıyıcı geliştirilmesi
Küçük Resim Yok
Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/embargoedAccess
Özet
Atopik Dermatit (AD), çocukları %20’ye, erişkinleri %3’e kadar etkileyen kronik ekzematöz bir deri hastalığıdır. AD görülen hastalarda, aynı zamanda TNF-α sitokin seviyesinin de arttığı bulunmuştur. Bu nedenle TNF-α, enflamasyonu düzenleyen anahtar regülatör olarak düşünülmüştür. TNF-α, makrofajlar, T hücreleri, B hücreleri, NK hücreleri, nötrofiller, mast hücreleri, endotelyal hücreler, adiposit hücreleri ve epidermal kerotinositler tarafından üretilen pro-enflamatuvar bir sitokindir. Deksametazon gibi topikal kortikosteroidler (TC’ler) AD fare modelinde güçlü anti-enflamatuvar etki göstermelerine karşın, ciddi yan etkiler sergiler. Bu sebeple, TNF-α’nın hücredeki üretimini baskılayıcı ajanlar ile kombinasyonu yeni bir tedavi yaklaşımı olarak düşünülebilir.
Antikor bazlı terapötikler, kullanımlarında yüksek maliyet ve sınırlı hedeflendirme gibi kısıtlayıcı faktörler olsa da, kronik enflamatuvar ile karakterize deri hastalıklarının tedavisinde önemli rol oynarlar. Kronik enflamatuvar deri hastalıklarından biri olan AD tedavisinde, antikor bazlı terapötiklere alternatif olarak siRNA kullanılabilir. siRNA aracılı gen susturması, mRNA seviyesinde pro-enflamatuvar sitokinlerinin kronik enflamatuvar koşullarını önlemeye alternatif bir tedavi stratejisi önerir. siRNA, antikor bazlı terapötiklerle kıyaslandığında düşük maliyet, kolay üretim ve spesifik hedeflendirme gibi avantajlar gösterir. siRNA, TNF-α’nın sitokin gen ekspresyonunu susturarak psoriyazis tedavisinde deriye uygulanabilmiştir. siRNA’ların bilinen dezavantajları, biyolojik sıvılarda düşük stabilite göstermesi ve düşük hücre penetrasyonuna sahip olmasıdır.
Bu tez çalışmasında, TC ve TNF-α hedeflendirilmiş siRNA ile tedavi kombinasyonu oluşturularak olası istenmeyen yan etkilerin önlenmesi ve daha güvenilir bir sistem oluşturulması hedeflenmiştir. Bu amaçla siRNA, deksametazon palmitat nanopartiküllerini (DXP-NP) içeren katyonik lipitle iyonik etkileşim yoluyla ilişkilendirilmiştir.
DXP-NP’leri, emülsiyon evaporasyon metodu ile hazırlanmıştır. Hazırlanan DXP-NP, TNF-α hedefli siRNA ile kombine edilerek farklı Nitrojen/Fosfat (N/P) oranlarında lipopleksler oluşturulmuştur. DXP-NP ve lipoplekslerin fizikokimyasal karakterizasyon çalışmaları gerçekleştirilmiştir. Sitotoksisite, anti-enflamavutar etkinlik ve hücre içe alım çalışmaları içeren hücre kültürü çalışmaları ise RAW264.7 makrofaj hücre hattında araştırılmıştır.
Atopic dermatitis (AD) is a common eczematous skin disorder affecting up to 20% of children and up to 3% of adults. Patients with atopic dermatitis also have elevated TNF-α (tumor necrosis factor alpha) levels in the skin. TNF-α is thought to be a key regulator of inflammation in AD. TNF-α is a representative pro-inflammatory cytokine produced by macrophages, T-cells, B-cells, NK-cells, neutrophils, mast cells, endothelial cells, adipocytes and epidermal keratinocytes. In an AD mouse model, Topical corticosteroid (TCs) such as dexamethasone have shown very good anti-inflammatory properties, however, they have strong side effects. Therefore, combination therapy of TCs and anti- TNF-α is now being approached. Antibody-based therapeutics have improved the treatment of chronic inflammatory skin diseases such as psoriasis, atopic dermatitis, and eczema, however, there are limiting factors with this therapy such as higher cost and limited targeting ability. As an alternative to antibodies, small interfering RNA (siRNA) might also considerably improve the treatment of AD. SiRNA-mediated knockdown of pro-inflammatory cytokines at mRNA level offers an alternative therapeutic strategy to overcome chronic inflammatory conditions. SiRNA present several advantages over antibodies as well as a very good specificity including low cost and ease of production. SiRNA could nicely be applied to skin knocking down cytokine expression of TNF-α in psoriasis. The only drawbacks of siRNAs are their poor stability in biological fluids and their low cellular penetration which can both be circumvent by nanotechnologies. In this project we would like to combine treatment of TCs with small interfering RNA targeting TNF-α to reduce the undesired side effects. For this purpose, siRNA will be associated through ionic interaction to Dexamethasone palmitate nanoparticles (DXP-NPs) covered by cationic lipids. DXP-NPs have prepared by emulsion evaporation method and Lipoplexes have obtained at different Nitrogen-to-Phosphate (N/P) ratios. Physicochemical characterizations were performed for both DXP-NPs and lipoplexes. Cell culture studies those are cytotoxicity, anti-inflammatory activity and cellular uptake have investigated on the macrophage cell line RAW264.7.
Atopic dermatitis (AD) is a common eczematous skin disorder affecting up to 20% of children and up to 3% of adults. Patients with atopic dermatitis also have elevated TNF-α (tumor necrosis factor alpha) levels in the skin. TNF-α is thought to be a key regulator of inflammation in AD. TNF-α is a representative pro-inflammatory cytokine produced by macrophages, T-cells, B-cells, NK-cells, neutrophils, mast cells, endothelial cells, adipocytes and epidermal keratinocytes. In an AD mouse model, Topical corticosteroid (TCs) such as dexamethasone have shown very good anti-inflammatory properties, however, they have strong side effects. Therefore, combination therapy of TCs and anti- TNF-α is now being approached. Antibody-based therapeutics have improved the treatment of chronic inflammatory skin diseases such as psoriasis, atopic dermatitis, and eczema, however, there are limiting factors with this therapy such as higher cost and limited targeting ability. As an alternative to antibodies, small interfering RNA (siRNA) might also considerably improve the treatment of AD. SiRNA-mediated knockdown of pro-inflammatory cytokines at mRNA level offers an alternative therapeutic strategy to overcome chronic inflammatory conditions. SiRNA present several advantages over antibodies as well as a very good specificity including low cost and ease of production. SiRNA could nicely be applied to skin knocking down cytokine expression of TNF-α in psoriasis. The only drawbacks of siRNAs are their poor stability in biological fluids and their low cellular penetration which can both be circumvent by nanotechnologies. In this project we would like to combine treatment of TCs with small interfering RNA targeting TNF-α to reduce the undesired side effects. For this purpose, siRNA will be associated through ionic interaction to Dexamethasone palmitate nanoparticles (DXP-NPs) covered by cationic lipids. DXP-NPs have prepared by emulsion evaporation method and Lipoplexes have obtained at different Nitrogen-to-Phosphate (N/P) ratios. Physicochemical characterizations were performed for both DXP-NPs and lipoplexes. Cell culture studies those are cytotoxicity, anti-inflammatory activity and cellular uptake have investigated on the macrophage cell line RAW264.7.
Açıklama
Anahtar Kelimeler
siRNA, TNF-α, Atopik Dermatit, Glukokortikoidler, Katı Lipit Nanopartiküller, Deksametazon, siRNA Delivery, TNF-α, Atopic Dermatitis, Glucocorticoids, Solid Lipid Nanoparticles, Dexamethasone
