Üç Boyutlu Biyomimetik Kaliksaren Nanofiber İskeleler Üzerinde Prostat Kanseri Kök Hücrelerinin Geliştirilmesi
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Dosyalar
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi, Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Normal kanser dokularından çeĢitli sebepler doğrultusunda kanser kök hücrelerinin meydana
gelmesi, kök hücrelerin tümörlü dokularda hücre popülasyonunda aĢırı artıĢı tedaviyi olumsuz
etkilemektedir. Bu nedenle kanser tipleri için tedavi stratejilerinin geliĢtirilmesinde kök hücreleri önemli
bir çalıĢma alanı haline gelmiĢtir. Buna karĢın, kök hücrelerin, in vitro ortamda fenotipik özelliklerinin
değiĢmesi yani farklılaĢması kök hücreleri hedefleyerek yapılacak çalıĢmaları zorlaĢtırmaktadır.
Son yıllarda yapılan çalıĢmalarda doku mühendisliği çalıĢmalarının ve teknolojinin ilerlemesiyle
oluĢturulan biyomimetik kültür ortamları ile yapılan kök hücre çalıĢmalarında fenotipinin in vitro ortamda
farklılaĢmadan çalıĢılmasına olanak sağlayacak inovatif teknikler ortaya konmaktadır. p-tertbütilkaliks
[4]aren nanofiber (nanofibril-nanolif) yapılar ile oluĢturulan 3B‟li (boyut) kültür ortamı, kök hücrelerin
fenotipinin değiĢmeden çoğaltılmasına olanak sağlayan ve kök hücreler ile uzun vadede çalıĢılması için
iyi bir materyal olarak değerlendirilmektedir. Tez çalıĢmasının amacı; elektro eğirme metodu kullanılarak
sentezlenen p-tertbütilkaliks [4]aren (5,11,17,23-Tetra-tert-bütil-25,27-bis-Furfuryamid-26,28-dihidroksi
kaliks[4]aren) doku iskelelerinin üzerinde, prostat kanser kök hücrelerinin (benzeri) proliferasyonu ve
geliĢtirlmesi olarak belirlendi. Prostat kanser kök hücresi PC3 prostat kanser hücre grubundan izole
edilmesi planlandı.
p-tert-butilkalks [4] aren sentezlendi ve nanofiber üretilmeden önce XTT metodu ile sitotoksite
analizleri prostat kanser hücre hattı PC3, sağlıklı epitel hücre hattı HUVEC ve sağlıklı fibroblastik hücre
hattı olan L929 üzerinde gerçekleĢtirildi. p-tert-butilkalks [4] aren nanofiber üretildi ve FTIR, 1HNMR,13C-NMR, MS gibi spektroskopik teknikler kullanılarak element analizi yapıldı. Karakterizasyon
analizleri için TGA, DSC, SEM ve TEM gibi teknikleri kullanılarak birçok özelliği (hidrofobikliğini,
temas yüzeyini, gözenek boyutunu ve lifler arası mesafeyi) aydınlatıldı. PC3 hücreleri in vitro model
olarak nanofiber yüzey üzerinde kültüre edildi. Ġki boyutlu ve üç boyutlu hücre kültür ortamlarında hücre
proliferasyonunu karĢılaĢtırma yapmak için XTT metodu uygulandı. Üç boyutlu ortamda büyütülen
hücreler metilen boyama yapılarak invert mikroskopta, DAPI/Phalloidin boyama yapılarak konfokal
mikroskopta görüntülendi. Üç boyutlu hücre kültür ortamında hücre büyümesi ve nano malzeme analizi
SEM kullanılarak analiz edildi. AkıĢ sitometrisi metodu ile prostat kanser kök hücre belirteci olan α2β1
hücre yüzey integrin proteini ve CD44 kök hücre belirteci kullanılarak izole edildi. Nanofiber yüzeyde
geliĢtirilen prostat kanser kök hücreleri konfokal mikroskop SEM analizi ile konfirme edildi. Bu doğrultuda in vitro koĢullarda çok daha uzun sürelerde kök hücreler kullanılarak birçok çalışma
yapılabileceği düşünülmektedir. Prostat kanseri kök hücrelerinin 3B kültür ortamlarında prolifere edilip,
üzerinde yeni tedavi stratejileri önerilecektir ve böylece yeni projelere de alt yapı oluşturulacaktır.
The formation of cancer stem cells from normal cancer tissues for various reasons, and the excessive increase in the cell population of stem cells in tumor tissues adversely affect the treatment. Therefore, stem cells have become an important field of study in the development of treatment strategies for cancer types. On the other hand, the change in the phenotypic properties of stem cells in vitro complicates the studies to be conducted by targeting stem cells. In recent years, innovative techniques have been introduced that will allow the phenotype to be studied in vitro without differentiation in stem cell studies with biomimetic culture media created with the advancement of tissue engineering studies and technology. The 3D (dimensional) culture medium formed with p-tertbutylcalyx [4]arene nanofiber structures is considered as a good material for long-term study with stem cells, which allows the phenotype of stem cells to be reproduced unchanged. The aim of the thesis study; The aim of the thesis study; On p-tertbutylcalyx [4]arene (5,11,17,23-Tetra-tert-butyl-25,27- bis-Furfuryamid-26,28-dihydroxy calyx[4]arene) scaffolds synthesized using the electrospinning method, was determined as the proliferation and development of prostate cancer stem cells (similar). Prostate cancer stem cell was planned to be isolated from PC3 prostate cancer cell group. p-tert-butylcalcx [4]arene was synthesized and cytotoxicity analyzes were performed by XTT method on prostate cancer cell line PC3, healthy epithelial cell line HUVEC and healthy fibroblastic cell line L929 before nanofiber was produced. p-tert-butylcalcx [4]arene nanofiber was produced and elemental analysis was performed using spectroscopic techniques such as FTIR, 1H-NMR, 13C-NMR, MS. Many properties (hydrophobicity, pore size, and fiber spacing) were elucidated using techniques such as TGA, DSC, SEM, and TEM for characterization analyses. PC3 cells were cultured on the nanofiber surface as an in vitro model. XTT method was applied to compare cell proliferation in two-dimensional and three-dimensional cell culture media. Cells grown in three-dimensional media were visualized under an invert microscope by methylene staining, and under a confocal microscope by DAPI/Phalloidin staining. Cell growth and nanomaterial analysis in three-dimensional cell culture medium were analyzed using SEM. α2β1 cell surface integrin protein, which is a prostate cancer stem cell marker, and CD44 stem cell marker were isolated by flow cytometry method. Prostate cancer stem cells developed on the nanofiber surface were confirmed by confocal microscope SEM analysis. In this direction, many studies can be carried out using stem cells for much longer periods in vitro conditions. Prostate cancer stem cells will proliferate in 3D culture media, and new treatment strategies will be proposed on them, thus creating an infrastructure for new projects.
The formation of cancer stem cells from normal cancer tissues for various reasons, and the excessive increase in the cell population of stem cells in tumor tissues adversely affect the treatment. Therefore, stem cells have become an important field of study in the development of treatment strategies for cancer types. On the other hand, the change in the phenotypic properties of stem cells in vitro complicates the studies to be conducted by targeting stem cells. In recent years, innovative techniques have been introduced that will allow the phenotype to be studied in vitro without differentiation in stem cell studies with biomimetic culture media created with the advancement of tissue engineering studies and technology. The 3D (dimensional) culture medium formed with p-tertbutylcalyx [4]arene nanofiber structures is considered as a good material for long-term study with stem cells, which allows the phenotype of stem cells to be reproduced unchanged. The aim of the thesis study; The aim of the thesis study; On p-tertbutylcalyx [4]arene (5,11,17,23-Tetra-tert-butyl-25,27- bis-Furfuryamid-26,28-dihydroxy calyx[4]arene) scaffolds synthesized using the electrospinning method, was determined as the proliferation and development of prostate cancer stem cells (similar). Prostate cancer stem cell was planned to be isolated from PC3 prostate cancer cell group. p-tert-butylcalcx [4]arene was synthesized and cytotoxicity analyzes were performed by XTT method on prostate cancer cell line PC3, healthy epithelial cell line HUVEC and healthy fibroblastic cell line L929 before nanofiber was produced. p-tert-butylcalcx [4]arene nanofiber was produced and elemental analysis was performed using spectroscopic techniques such as FTIR, 1H-NMR, 13C-NMR, MS. Many properties (hydrophobicity, pore size, and fiber spacing) were elucidated using techniques such as TGA, DSC, SEM, and TEM for characterization analyses. PC3 cells were cultured on the nanofiber surface as an in vitro model. XTT method was applied to compare cell proliferation in two-dimensional and three-dimensional cell culture media. Cells grown in three-dimensional media were visualized under an invert microscope by methylene staining, and under a confocal microscope by DAPI/Phalloidin staining. Cell growth and nanomaterial analysis in three-dimensional cell culture medium were analyzed using SEM. α2β1 cell surface integrin protein, which is a prostate cancer stem cell marker, and CD44 stem cell marker were isolated by flow cytometry method. Prostate cancer stem cells developed on the nanofiber surface were confirmed by confocal microscope SEM analysis. In this direction, many studies can be carried out using stem cells for much longer periods in vitro conditions. Prostate cancer stem cells will proliferate in 3D culture media, and new treatment strategies will be proposed on them, thus creating an infrastructure for new projects.
Açıklama
Anahtar Kelimeler
Elektro Eğirme, Kaliksaren, Kanser Kök Hücresi, Nanofiber, Prostat Kanseri (PC3), 3B Hücre Kültürü, Cancer Stem Cell, Electro Spinning, Kalixarene, Prostate Cancer (PC3), 3D Cell Culture
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Yıldırım G., (2023). Üç Boyutlu Biyomimetik Kaliksaren Nanofiber İskeleler Üzerinde Prostat Kanseri Kök Hücrelerinin Geliştirilmesi. (Doktora Tezi). Selçuk Üniversitesi, Fen Bilimleri Enstitüsü, Konya.
