L-glutamic acid-g-p(HEMA) polimerik nanopartiküllerinin sentezi, karakterizasyonu, in vitro ve in vivo toksisite potensiyeli
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Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Nanomateryaller fiziksel ve kimyasal olarak modifiye edilebilen malzemeler olup, tıp, biyomedikal, endüstri gibi birçok alanda yaygın olarak kullanımlarıyla, son yıllarda büyük ölçüde araştırmalara konu olmuştur. Sürfaktansız emülsiyon polimerizasyon yöntemiyle elde edilen p(HEMA) ile L-glutamik asit graftlama tekniği ile sentezlenmiştir. Polimerik nanopartiküllün karakterizasyon analizleri, boyut ve morfolojik özelliklerini incelemek amacıyla SEM (Scanning Electron Microscopy) görüntülemesi, polar sıvılar içerisindeki davranışlarını belirlemek amacıyla ZETA boyut- potansiyel analizi, yüzey topografyasını görüntülemek amacıyla AFM (Atomik Kuvvet Mikroskobu) analizi ve yapıdaki bileşenlerin yüzey kimyası ile etkileşimlerini belirlemek amacıyla FTIR Absorbsiyon Spektroskopisi gerçekleştirilmiştir. Karakterizasyon analiz sonuçlarına göre 194.6 nm boyutunda olan L-glutamik asit-g-p(HEMA), -18 Mv yüzey yüküne sahiptir. Elde edilen polimerik nanopartikülün in vitro ve in vivo deneysel yöntemlerle toksisitesi belirlenmiştir. L-glutamik asit-g-p(HEMA)'nın, 16HBE (Human bronchial epithelial cell line) ve THP-1 (human monocytic cell line) hücre hatları üzerinde üzerinde WST-1 ve LDH testleri ile sitotoksik etki göstermediği, mutajenik potansiyelin belirlenmesi amacıyla Salmonella typhimurium suşları üzerinde gerçekleştirilen Salmonella/AMES testi sonucunda mutajenik olmadığı saptanmıştır. Genotoksik potansiyelini belirlemek amacıyla 16HBE ve THP-1 hücre hatları ile yapılan COMET testi, ve in vivo kemik iliği ile mikronukleus testi sonucunda genotoksik etki göstermediği belirlenmiştir. 16HBE hücre hattında hücre migrasyon testi gerçekleştirilmiş kapanma yüzdesi %21 olarak belirlenmiştir. HET-CAM testi ile uygulanan konsantrasyonlarda irritasyon potansiyeli ve MIC yöntemi ile antimikrobiyal etkinliği ortaya konulmuştur. In vivo tek doz akut toksisite testi ile LD50 değeri ≥ 2000 mg/kg olarak belirlenmiş, 28 gün tekrarlı doz toksisite (subakut) çalışması sonucunda hayvan ve organ ağırlık verileriyle birlikte biyokimyasal veriler sağlanmış, histopatolojik incelemeler gerçekleştirilmiş ve herhangi bir toksik bulguya rastlanmamıştır. Polimerik nanopartikülün vücut içerisinde biyodağılımı in vivo görüntüleme sistemi olan IVIS spektrum ile ortaya konmuştur. Sonuç olarak sentezlediğimiz ve karakterize ettiğimiz polimerik nanopartikül olan L-glutamik asit-g-p(HEMA)'nın biyouyumlu olduğu ve ilaç taşıma sistemi olarak kullanılabilme potansiyeli bulunduğu ortaya konmuştur.
Nanomaterials can be physically and chemically modified and have been used commonly in recent years with their widespread use in many areas as medicine, biomedical and industry. p(HEMA) was obtained with surfactant free emulsion polymerization method and it was synthesized with L-glutamic acid by grafting technique. Characterization analysis of polymeric nanoparticles were investigated by SEM (Scanning Electron Microscopy) imaging to determine their size and morphological characteristics, ZETA size-potential analysis to determine their behavior in polar liquids, AFM (Atomic Force Microscopy) analysis to display surface topography and their interactions with surface chemistry by FTIR Absorption Spectroscopy. According to the results of characterization analysis, L-glutamic acid-g-p (HEMA) has 194.6 nm size and -18 Mv negative surface value. The toxicity of polymeric nanoparticle was determined with in vitro and in vivo assays. L-glutamic acid-g-p (HEMA) has no cytotoxic effect on 16HBE (Human bronchial epithelial cell line) and THP-1 (human monocytic cell line) cell lines with WST-1 and LDH tests and in order to determine the mutagenic potential on Salmonella typhimurium strains which was not showed mutagenic effect with Salmonella/AMES test. In order to determine genotoxic potential, it has been identified that there is no genotoxic effect with COMET test on 16HBE and THP-1 cell lines and in vivo micronucleus test on bone marrow. Cell migration test was performed on 16HBE cell line and the percent closure rate was determined as 21%. The irritation potential was obtained with applied concentration by HET-CAM test and antimicrobial activity were determined with MIC method. The LD50 value was determined as ≥ 2000 mg / kg by the in vivo single dose acute toxicity test, also biochemical data and histopathological investigation were obtained with the animal and organ weight data as a nontoxic result of the 28 days repeated dose toxicity (subacute) study. The biodistribution of the polymeric nanoparticle in the body was demonstrated by IVIS spectrum which is in vivo imaging system. As a result, L-glutamic acid-g-p (HEMA), a polymeric nanoparticle which was synthesized and characterized by us, has been found to be biocompatible and has the potential to be used as a drug delivery system.
Nanomaterials can be physically and chemically modified and have been used commonly in recent years with their widespread use in many areas as medicine, biomedical and industry. p(HEMA) was obtained with surfactant free emulsion polymerization method and it was synthesized with L-glutamic acid by grafting technique. Characterization analysis of polymeric nanoparticles were investigated by SEM (Scanning Electron Microscopy) imaging to determine their size and morphological characteristics, ZETA size-potential analysis to determine their behavior in polar liquids, AFM (Atomic Force Microscopy) analysis to display surface topography and their interactions with surface chemistry by FTIR Absorption Spectroscopy. According to the results of characterization analysis, L-glutamic acid-g-p (HEMA) has 194.6 nm size and -18 Mv negative surface value. The toxicity of polymeric nanoparticle was determined with in vitro and in vivo assays. L-glutamic acid-g-p (HEMA) has no cytotoxic effect on 16HBE (Human bronchial epithelial cell line) and THP-1 (human monocytic cell line) cell lines with WST-1 and LDH tests and in order to determine the mutagenic potential on Salmonella typhimurium strains which was not showed mutagenic effect with Salmonella/AMES test. In order to determine genotoxic potential, it has been identified that there is no genotoxic effect with COMET test on 16HBE and THP-1 cell lines and in vivo micronucleus test on bone marrow. Cell migration test was performed on 16HBE cell line and the percent closure rate was determined as 21%. The irritation potential was obtained with applied concentration by HET-CAM test and antimicrobial activity were determined with MIC method. The LD50 value was determined as ≥ 2000 mg / kg by the in vivo single dose acute toxicity test, also biochemical data and histopathological investigation were obtained with the animal and organ weight data as a nontoxic result of the 28 days repeated dose toxicity (subacute) study. The biodistribution of the polymeric nanoparticle in the body was demonstrated by IVIS spectrum which is in vivo imaging system. As a result, L-glutamic acid-g-p (HEMA), a polymeric nanoparticle which was synthesized and characterized by us, has been found to be biocompatible and has the potential to be used as a drug delivery system.
Açıklama
Anahtar Kelimeler
p(HEMA), Polimerik Nanopartikül, İn Vitro Toksisite, Biyodağılım, Polymeric Nanoparticle, İn Vitro Toxicity, Biodistribution
