Kemik Dokusunun Rejenerasyonunda Hücre Tabakalarının Farklı Uygulamaları

Abstract

This study was supported by Hacettepe University Scientific Research Projects Coordination Unit (Project Number: 17055) and Ülkü Bozoğlu was also supported by TÜBİTAK (TÜBİTAK-2211-A). Within the scope of the presented thesis, studies have been carried out to increase the potential of the use of polyetheretherketone (PEEK), which has the potential of alternative use to titanium (Ti) metallic implants, which are widely used , as a dental implant. However, PEEK is inherently inert material and that characteristic limits cellular adhesion and bone integration of PEEK. There are a number of methods that have been offered to enhance the bioactivity of PEEK including coating PEEK with synthetic osteoconductive hydroxyapatite (HAp) and sulfonation. In this work, a nanostructured porous surface is created on PEEK surfaces by sulfonation (SPEEK). Then, PEEK and SPEEK samples were coated nanostructured B doped HAp (B-nHAp) from 10xSBF by microwave-assisted method. Scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier Transform Infrared spectroscopy (ATR-FTIR) reveal the formation of nanopores and nanostructured HAp on the PEEK surface. In vitro cell viability assay and osteogenic differentiation analyses disclose improved adhesion, proliferation and osteo-differentiation of periodontal ligament cells (PDL). Our results indicate that sulfonation and B-nHAp modifications have dual effects on the osteogenic differentiation of PDL and increase of osteointegration of PEEK dental implant. In the second part of the study, PDL cell sheets and PDL and human umbilical cord vein endothelial cells (human umblical and endothelial cell, HUVEC) co-culture cell sheets using the PEEK-PDL / co-culture cell sheets complex was formed. Human-derived PDL and PDL- HUVEC cell sheets were obtained by vitamin C and high cell seeding methods and combined with SPEEK and SPEEK-B-nHAp samples. These complex, consisting of cell sheets and modified PEEK materials, were kept in the osteogenic environment for 21 weeks. The cell sheets were firmly attached to the surface of the material and maintained their vitality for 3 weeks. Evaluation of bone and vascular markers of cell sheets on modified PEEK samples were evaluated using ALP analysis, RT-PCR, Western blot and immunocytochemistry staining analyzes. The osteogenic differentiation of PDL cell sheets was found to be high. In addition, co-culture with HUVEC has been shown to be particularly effective on late- stage osteogenic differentiation. The combining of cell sheets with SPEEK and SPEEK-B- nHAp materials has been demonstrated as a new approach to increase the bioactivity and complementarity of PEEK as dental implant.