Tümör hedefli silika nanoparçacıkların bifonksiyonel ajan potansiyelinin incelenmesi
Küçük Resim Yok
Tarih
2018
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Kanser, DNA hasarı veya mutasyon sonucu vücut hücrelerinin kontrolsüz ve anormal bir şekilde çoğalmasıdır. Pankreas kanseri, kanser türleri arasında teşhis ve tedavisi en zor olanlarındandır. Bununla birlikte teşhisinden sonra sağ kalım oranı oldukça düşüktür. Bu nedenle pankreas kanserinin tedavisinde, erken teşhis oldukça önemlidir. Son yıllarda, klasik tedavi yöntemlerinin tek başına etkili olmadığı görülmüştür. Bunun yerine kombine tedavi yöntemlerinin uygulanması artmıştır. Aynı zamanda, nanoteknolojinin medikal alanda kullanılmasıyla, hem teşhis hem de tedaviye olanak sağlayan teranostik ajanların geliştirilmesi hız kazanmıştır. Bu tez çalışmasında, Zn ftalosiyanin (ZnPc) yüklü ve cetuximab antikoru ile hedeflenmiş gözenekli silika nanoparçacıklar hazırlanmış (MSNP5) ve 131I radyonüklidi ile işaretlenmiştir. Nanoparçacıklar içerisinde bulunan ZnPc, fotodinamik terapinin (FDT) gerçekleştirilmesine olanak sağlamaktadır. Bununla birlikte, nanoparçacıkların 131I ile işatlenmesiyle, fotodinamik terapi, nükleer görüntüleme ve/veya nükleer terapide kullanılabilicek bir teranostik ajan oluşturulmuştur. Sentezlenen nanoparçacıkların boyut, zeta potansiyel ve yüzey modifikasyonu analizleri Fourier Dönüşümlü Kızılötesi Spektroskopisi (FTIR), Taramalı Elektron Mikroskobu (SEM), Geçirimli Elektron Mikroskobu (TEM) ve Dinamik Işık Saçılımı (DLS) cihazları ile gerçekleştirilmiştir. Gerçekleştirilen karakterizasyon çalışmalarıyla, Zn ftalosiyaninin enkapsülasyonunun ve yüzey modifikasyonlarının başarılı bir şekilde gerçekleştirildiği belirlenmiştir. Aynı zamanda, MSNP5 nanoparçacıklarının hidrodinamik çapı 185,6±22,6 nm olarak tespit edilmiştir. Fotodinamik tedavi, fotodinamik etkiye dayanan non-invaziv ve alternatif bir tedavidir. Bu tez çalışmasıyla, ZnPc yüklü nanoparçacıkların [MSNP4 (cetuximab içermeyen) ve MSNP5 (cetuximab içeren)], pankreatik kanser hücrelerine karşı singlet oksijen (1O2) üretimi, radyoişaretleme verimi, hücre tutulum ve in vitro fotodinamik tedavi potansiyelinin belirlenmesi amaçlanmıştır. ZnPc'nin tolüendeki 1O2 kuantum verim (??) değeri 0,60 olarak bulunmuştur. In vitro FDT çalışmaları, ZnPc ve nanoparçacıkların karanlık toksisite ve fototoksisitesini belirlemek için pankreatik kanser hücreleri (MIA PaCa-2, AsPC-1 ve PANC-1) kullanılarak gerçekleştirilmiştir. Sonuçlara göre, tüm hücre hatlarında, ZnPc ve nanoparçacıklar için karanlık toksisite gözlenmemiştir. Bununla birlikte, MSNP5 nanoparçacıkları, MSNP4 nanoparçacıklarından daha yüksek fototoksik etki göstermiştir. MIA PaCa-2, AsPC-1 ve PANC-1 hücrelerindeki en yüksek fototoksik etkiler, 5 µM konsantrasyon ve 40 J/cm2 ışık dozunda sırasıyla, % 16± 5, % 13±3 ve % 5±1 hücre canlılığı olarak gözlenmiştir. Florometrik yöntemler ile belirlenen hücre tutulum çalışmalarıyla, MSNP5 nanoparçacıklarının tutulum potansiyelinin MSNP4 nanoparçacıklarından daha yüksek olduğu belirlenmiştir. Nanoparçacıkların 131I ile radyoişaretlenmesi iyodojen metoduna göre gerçekleştirilmiştir. MSNP5 nanoparçacıklarının radyoişaretlenme verimi, optimum koşullarda (pH 9 ve 60 dakika reaksiyon süresi) oldukça yüksek (% 95,5±1,2) bulunmuştur. Buna ek olarak, MIA PaCa-2, AsPC-1 ve PANC-1 hücrelerinde 131I-MSNP5'in en yüksek hücre içi tutulum verimleri 24 saat inkübasyon süresinde sırasıyla, % 43,9±3,8, % 41,8±0,2 ve % 37,9±1,3 olarak tespit edilmiştir. In vivo FDT çalışmaları, AsPC-1 pankreas kanseri hücreleri ile tümör oluşturulmuş Nude fareler ile gerçkeleştirilmiştir. MSNP5 nanoparçacıkları, tümörün altından subkutan olarak verilmiştir. Fotodinamik terapi için 100 J/cm2 ışık dozu, 685 nm kırmızı lazer ışık kullanılarak uygulanmıştır. Gerçekleştirilen histopatalojik incelemelerle, MSNP5 nanoparçacıkları uygulanan tedavi grubunda tümör dokusundaki nekroz oranının kontrol grubundan yüksek olduğu tespit edilmiştir. Sonuç olarak, hedefleyici cetuximab antikoru içeren MSNP5 nanoparçacıkları, pankreatik tümörlerin nükleer görüntüleme/terapi ve fotodinamik tedavisi için uygun bir teranostik ajan potansiyeli göstermiştir. Anahtar Kelimeler: Zn (II) ftalosiyanin, gözenekli silika nanoparçacıklar, cetuximab, singlet oksijen, fotodinamik tedavi
Cancer, is an abnormal and uncontrolled proliferation of body cells result of DNA damage or mutation. Pancreatic cancer is the most difficult to diagnose and treatment among cancer types. In addition, survival rate after diagnosis is very low. For this reason, early diagnosis is very important in the treatment of pancreatic cancer. In recent years, it has been seen that alone conventional treatment methods have not been effective. Instead of that, the application of combined treatment methods has increased. Also, using of nanotechnology in the medical field, the development of teranostic agents that enable both diagnosis and treatment has gained pace. In this thesis, mesoporous silica nanoparticles loaded with Zn phthalocyanine (ZnPc) and targeted with cetuximab antibody (MSNP5) were prepared and labeled with 131I radionuclide. ZnPc in nanoparticles allows to carry out the photodynamic therapy (PDT). However, by labeling nanoparticles with 131I, a teranuteric agent was formed which can be used in photodynamic therapy, nuclear imaging and/or nuclear therapy. The size, zeta potential and surface modification analyzes of the synthesized nanoparticles were performed with Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS) devices. It was determined with the characterization that the encapsulation of Zn phthalocyanine and the surface modification were successfully carried out. At the same time, the hydrodynamic diameter of MSNP5 nanoparticles was determined to be 185.6±22.6 nm. Photodynamic therapy (PDT) is a non-invasive and an alternative therapy based on the photodynamic effect. In this thesis, we sought to determine the singlet oxygen (1O2) production, radiolabeling efficiency, intracellular uptake and in vitro photodynamic therapy potential of Zn phthalocyanine-loaded mesoporous silica nanoparticles [MSNP4 (without cetuximab) and MSNP5 (with cetuximab)] against pancreatic cancer cells. The 1O2 quantum yield (??) value of ZnPc was found to be 0.60 in toluene. In vitro PDT studies were performed to determine the dark and phototoxicity of ZnPc and nanoparticles by using pancreatic cancer cells (MIA PaCa-2, AsPC-1 and PANC-1). According to the results, no dark toxicity was observed for ZnPc and nanoparticles in any cell line. However, MSNP5 nanoparticles showed quite high phototoxic effect, more pronounced than MSNP4 nanoparticles. The highest phototoxic effect was observed at an exposure of 40 J/cm2, namely 16±5%,13±3% and 5±1% cell viability at 5 µM concentrations in MIA PaCa-2, AsPC-1 and PANC-1 cells, respectively. It was observed from the intracellular uptake studies determined by Fluorometric methods that MSNP5 nanoparticles' uptake potential was higher than MSNP4 nanoparticles. Radiolabelling of the nanoparticles with 131I was performed according to the iodogen method. The radiolabeling efficiency of MSNP5 nanoparticles was found to be quite high (95.5±1.2%) in optimum conditions (pH 9 and 60 minutes reaction time). In addition to this, the highest intracellular uptake yields of 131I-MSNP5 in MIA PaCa-2, AsPC-1 and PANC-1cells were determined as 43.9±3.8%, %41.8±0.2 and %37.9±1.3, respectively at 24 hours incubation time. In vivo PDT studies were performed with tumor-bearing Nude mice (AsPC-1 pancreatic cancer cells). MSNP5 nanoparticles were subcutaneously administered under the tumor. For photodynamic therapy, 100 J/cm2 light dose was applied using 685 nm red laser light. Histopathological studies revealed that the ratio of necrosis in tumor tissue was higher in the treatment group treated with MSNP5 nanoparticles than in the control group. In conclusion, MSNP5 nanoparticles with targerting cetuximab antibody appear as suitable a theranostic agent for nuclear imaging/therapy and photodynamic therapy of pancreatic tumors. Keywords: Zn(II) phthalocyanine, mesoporous silica nanoparticles, cetuximab, singlet oxygen, photodynamic therapy.
Cancer, is an abnormal and uncontrolled proliferation of body cells result of DNA damage or mutation. Pancreatic cancer is the most difficult to diagnose and treatment among cancer types. In addition, survival rate after diagnosis is very low. For this reason, early diagnosis is very important in the treatment of pancreatic cancer. In recent years, it has been seen that alone conventional treatment methods have not been effective. Instead of that, the application of combined treatment methods has increased. Also, using of nanotechnology in the medical field, the development of teranostic agents that enable both diagnosis and treatment has gained pace. In this thesis, mesoporous silica nanoparticles loaded with Zn phthalocyanine (ZnPc) and targeted with cetuximab antibody (MSNP5) were prepared and labeled with 131I radionuclide. ZnPc in nanoparticles allows to carry out the photodynamic therapy (PDT). However, by labeling nanoparticles with 131I, a teranuteric agent was formed which can be used in photodynamic therapy, nuclear imaging and/or nuclear therapy. The size, zeta potential and surface modification analyzes of the synthesized nanoparticles were performed with Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS) devices. It was determined with the characterization that the encapsulation of Zn phthalocyanine and the surface modification were successfully carried out. At the same time, the hydrodynamic diameter of MSNP5 nanoparticles was determined to be 185.6±22.6 nm. Photodynamic therapy (PDT) is a non-invasive and an alternative therapy based on the photodynamic effect. In this thesis, we sought to determine the singlet oxygen (1O2) production, radiolabeling efficiency, intracellular uptake and in vitro photodynamic therapy potential of Zn phthalocyanine-loaded mesoporous silica nanoparticles [MSNP4 (without cetuximab) and MSNP5 (with cetuximab)] against pancreatic cancer cells. The 1O2 quantum yield (??) value of ZnPc was found to be 0.60 in toluene. In vitro PDT studies were performed to determine the dark and phototoxicity of ZnPc and nanoparticles by using pancreatic cancer cells (MIA PaCa-2, AsPC-1 and PANC-1). According to the results, no dark toxicity was observed for ZnPc and nanoparticles in any cell line. However, MSNP5 nanoparticles showed quite high phototoxic effect, more pronounced than MSNP4 nanoparticles. The highest phototoxic effect was observed at an exposure of 40 J/cm2, namely 16±5%,13±3% and 5±1% cell viability at 5 µM concentrations in MIA PaCa-2, AsPC-1 and PANC-1 cells, respectively. It was observed from the intracellular uptake studies determined by Fluorometric methods that MSNP5 nanoparticles' uptake potential was higher than MSNP4 nanoparticles. Radiolabelling of the nanoparticles with 131I was performed according to the iodogen method. The radiolabeling efficiency of MSNP5 nanoparticles was found to be quite high (95.5±1.2%) in optimum conditions (pH 9 and 60 minutes reaction time). In addition to this, the highest intracellular uptake yields of 131I-MSNP5 in MIA PaCa-2, AsPC-1 and PANC-1cells were determined as 43.9±3.8%, %41.8±0.2 and %37.9±1.3, respectively at 24 hours incubation time. In vivo PDT studies were performed with tumor-bearing Nude mice (AsPC-1 pancreatic cancer cells). MSNP5 nanoparticles were subcutaneously administered under the tumor. For photodynamic therapy, 100 J/cm2 light dose was applied using 685 nm red laser light. Histopathological studies revealed that the ratio of necrosis in tumor tissue was higher in the treatment group treated with MSNP5 nanoparticles than in the control group. In conclusion, MSNP5 nanoparticles with targerting cetuximab antibody appear as suitable a theranostic agent for nuclear imaging/therapy and photodynamic therapy of pancreatic tumors. Keywords: Zn(II) phthalocyanine, mesoporous silica nanoparticles, cetuximab, singlet oxygen, photodynamic therapy.
Açıklama
Anahtar Kelimeler
Radyoloji ve Nükleer Tıp, Radiology and Nuclear Medicine
