Publication: Kitosan nano-parçacıkların pseudomonas aerugınosa üzerine antibakteriyel etkisinin ın sılıco ve ın vıtro ortamlarda incelenmesi
Abstract
KİTOSAN NANO-PARÇACIKLARIN PSEUDOMONAS AERUGINOSA ÜZERİNE ANTİBAKTERİYEL ETKİSİNİN IN SILICO VE IN VITRO ORTAMLARDA İNCELENMESİ Bakteriler, biyofilm formuna geçerek birçok tıbbi vakanın oluşmasına sebep olmaktadırlar. Fırsatçı patojenlerden Pseudomonas aeruginosa da biyofilm oluşturarak ciddi enfeksiyonlara sebep olabilmektedir. P. aeruginosa’nın biyofilm oluşturmasında rol oynayan Quorum Sensing (QS) mekanizmasının aydınlatılması yeni ilaçların geliştirilmesine öncülük edebilecektir. İlaç geliştirmede uygun hedefin belirlenmesi önem taşımaktadır. Son yıllarda uygun hedef bölgenin belirlenmesinde in silico çalışmalar ön plana çıkmaktadır. In silico yöntemler ile ilaç moleküllerinin in vivo ya da in vitro olarak test edilmesi için harcanan zamandan ve maliyetten tasarruf sağlanması beklenmektedir. Bu bilgiler ışığında P. aeruginosa PAO1 suşunda antimikrobiyal etkinliği bilinen kitosan ve kitosan nanoparçacıklarının antibakteriyel ve anti-QS etkisinin in silico ve in vitro ortamlarda incelenmesi amaçlanmıştır. Bu amaçla P. aeruginosa PAO1 suşunun QS ve ikili bileşen sistemine ait proteinlerinin üç boyutlu yapıları ile kitosan ve kitosan nanoparçacık moleküllerinin üç boyutlu yapıları çeşitli veri bankalarından elde edilerek moleküler kenetlenme işlemine tabi tutulmuştur. Bununla birlikte kitosanın ve kitosan nanoparçacıkların in vitro olarak P. aeruginosa PAO1 suşu üzerinde antibakteriyel ve anti-QS etkileri incelenmiştir. QS mekanizmasına ait proteinlerle gerçekleştirilen in silico işlemler sonucunda, kitosan molekülleri kitosan nanoparçacık molekülüne göre daha iyi skor vermiştir. Antibakteriyel aktivitede etkili proteinlerde ise kitosan nanoparçacık molekülü, kitosan moleküllerine göre daha iyi skor vermiştir. In vitro deneylerde uygulanan düşük dozda kitosan ve kitosan nanoparçacık çözeltilerinin P. aeruginosa PAO1 suşu üzerinde antibakteriyel veya anti-QS etkisine rastlanılmamışken, yüksek dozda uygulanan çözeltilerde sırasıyla HMW > MMW > Kitosan > LMW olarak antimikrobiyal aktivite gözlenmiştir. Sonuç olarak, in silico uygulamalarda kullanılan moleküllerin stabilitesi, boyutları, programların algoritmaları arasındaki farklılıklar gibi etmenlerin in vitro şartlarla uyum sağladığı durumlarda, in silico verilerle in vitro verilerin birbirini destekleyebileceği görülmüştür.
IN SILICO AND IN VITRO INVESTIGATION OF CHITOSAN NANOPARTICLES FOR ANTIBACTERIAL EFFECTS ON PSEUDOMONAS AERUGINOSA Bacteria may cause many different medical circumstances through their biofilm forms. Pseudomonas aeruginosa, one of the opportunistic pathogens, causes serious infections via biofilm formation as well. Elucidation of the Quorum Sensing (QS) mechanism, which plays a role in biofilm formation of P. aeruginosa, will be able to lead the development of new drugs. Determination of the appropriate target is carry weight in the developement of new drugs. Recently, in silico methods and studies have come into prominence for determination of the appropriate targets. In silico methods are expected to provide saving the cost and the time spent on testing the drug molecule candidates in vitro and in vivo. In the light of this information, the aim of our study is to investigate the antibacterial and anti-QS effects of the chitosan and chitosan nanoparticles, which are known for their antimicrobial effects, both in silico and in vitro on Pseudomonas aeruginosa PAO1 strain. The 3D structures of chitosan, chitosan nanoparticles and the proteins of P. aeruginosa PAO1 Quorum Sensing and Two Component System were downloaded from several databases, and several molecular docking studies were performed to this end. In addition, the antibacterial and anti-QS activities of chitosan and chitosan nanoparticles on P. aeruginosa PAO1 strain were investigated in vitro. The in silico results showed that chitosan molecules had better scores than chitosan nanoparticles on the proteins that play a role in QS mechanism. On the other hand, the chitosan nanoparticles had a better score than chitosan molecules on the proteins that are relevant to the antibacterial activity. As a result of in vitro experiments, neither antibacterial nor anti-QS effects on P.aeruginosa PAO1 strain were observed on the lower doses of chitosan and chitosan nanoparticles solutions, but we observed antimicrobial activities on the higher doses of HMW, MMW, Chitosan, LMW solutions, respectively. In conclusion, it seems possible that in silico and in vitro data could support each other under the appropriate conditions of adapted factors such as stability, size and dimension of the molecules in addition to the differences between the algorithms of programs used for in silico methods.
IN SILICO AND IN VITRO INVESTIGATION OF CHITOSAN NANOPARTICLES FOR ANTIBACTERIAL EFFECTS ON PSEUDOMONAS AERUGINOSA Bacteria may cause many different medical circumstances through their biofilm forms. Pseudomonas aeruginosa, one of the opportunistic pathogens, causes serious infections via biofilm formation as well. Elucidation of the Quorum Sensing (QS) mechanism, which plays a role in biofilm formation of P. aeruginosa, will be able to lead the development of new drugs. Determination of the appropriate target is carry weight in the developement of new drugs. Recently, in silico methods and studies have come into prominence for determination of the appropriate targets. In silico methods are expected to provide saving the cost and the time spent on testing the drug molecule candidates in vitro and in vivo. In the light of this information, the aim of our study is to investigate the antibacterial and anti-QS effects of the chitosan and chitosan nanoparticles, which are known for their antimicrobial effects, both in silico and in vitro on Pseudomonas aeruginosa PAO1 strain. The 3D structures of chitosan, chitosan nanoparticles and the proteins of P. aeruginosa PAO1 Quorum Sensing and Two Component System were downloaded from several databases, and several molecular docking studies were performed to this end. In addition, the antibacterial and anti-QS activities of chitosan and chitosan nanoparticles on P. aeruginosa PAO1 strain were investigated in vitro. The in silico results showed that chitosan molecules had better scores than chitosan nanoparticles on the proteins that play a role in QS mechanism. On the other hand, the chitosan nanoparticles had a better score than chitosan molecules on the proteins that are relevant to the antibacterial activity. As a result of in vitro experiments, neither antibacterial nor anti-QS effects on P.aeruginosa PAO1 strain were observed on the lower doses of chitosan and chitosan nanoparticles solutions, but we observed antimicrobial activities on the higher doses of HMW, MMW, Chitosan, LMW solutions, respectively. In conclusion, it seems possible that in silico and in vitro data could support each other under the appropriate conditions of adapted factors such as stability, size and dimension of the molecules in addition to the differences between the algorithms of programs used for in silico methods.