Publication: Doku mühendisliği için multiferroik nanopartikül yüklü doku iskelelerin 3B yazıcı ile üretimi ve karakterizasyonu
Abstract
Doku ve rejenerasyon mühendisliği, yapı iskeleleri, hücreler ve biyofiziksel ve biyokimyasal uyaranlar dahil olmak üzere, mühendislik yaklaşımlarını kullanarak hasarlı, dejenere olmuş veya kusurlu dokuları değiştirmeyi, onarmayı veya yenilemeyi amaçlar. Bu çalışmada Biyokompozit olarak kullanılabilen biyolojik olarak parçalanabilen metakrilatlanmış İpek fibroin (SilkMA) ve keratin metakrilat (KerMA) iskele sentezini ve fiziko-kimyasal karakterizasyonu yapılmış, doku mühendisliğinin amaçladığı ve hedeflediği özellikler analiz edilmiştir. SilkMA, belirlenen bir doku için iyi biyouyumluluk, reolojik özellikleri ve mekanik özellikler sergiler. Bu proje kapsamında metakrilatlanmış olarak, özgünlük ve biyouyumluluk açısından KerMA malzemesi seçilmiştir ama bu malzeme tek başına reolojik özellikleri ve mekanik dayanımı bakımından zayıftır. KerMA malzemesinin reolojik özelliklerini ve fibroblast hücre canlılığını geliştirmek için farklı konsantrasyonlarda SilkMA ile kompozit oluşturulmuştur. Biyolojik aktif maddelerin vücut içinde taşınma kabiliyetini arttırmak için manyetik demir oksit nanoparçacık protein jellerine (akışkan formda) ilave edildi. Bu manyetik nanoparçacıklara dışardan müdahale olarak 3 eksenli bir Helmholtz bobini tasarlanarak üretilmiş olup, kullanımı içinde 3B cihaza entegre edildi. Bobinin entegrasyonu ve SilkMA/ KerMA iskelesi üretimi ise, 3B SLA yazıcı ile yapıldı. Polimer zincirlerinin iç yapıların düzenli ve anizotropik olacak şekilde yönlendirilme işlemi, 3B baskı sırasında 3 eksenli Helmholtz bobinin oluşturduğu 15mT manyetik alan ile gerçekleştirildi. SilkMA/ KerMA yapıların, reolojik ve elektrik iletkenliği gibi değerlerin testleri yapılarak, manyetik olarak yönlendirilip, iskelelerin 1,3 ve 7 gün boyunca üstünde fibroblast hücreler iskele üzerinde toksik etki göstermediği ve yönlendirilmiş yapılarda hücre uyumluluğu özelliklerini geliştirildiği, yapılan testler ve analizler ile değerlendirildi. Ayrıca hücrelerin çoğalmasını ve yapışması desteklediğini görmek için SEM ve DAPI testleri yapıldı. Anizotropik oryantasyonun etkisini deneysel olarak görmek için, SEM ve atomik kuvvet mikroskobunda (AFM) görüntüleme çalışmaları ve incelemeleri yapıldı. İskelelere basma testi uygulanarak, mekanik gerilmelere karşı dayanımı ve elastisitesi hesaplanıp ayrıca şişme ve bozunma testleri yapılarak, iskelelerin yüksek şişme özelliği sergilediği ve 14 gün boyunca çözünmediği tespit edildi.
Tissue and regeneration engineering aims to replace, repair or regenerate damaged, degenerated or defective tissues using engineering approaches, including scaffolds, cells, and biophysical and biochemical stimuli. In this study, the synthesis and physico-chemical characterization of biodegradable methacrylate Silk fibroin (SilkMA) and keratin methacrylate (KerMA) scaffolds, which can be used as biocomposites, were performed, and tissue engineering aimed and targeted properties were analyzed. SilkMA exhibits good biocompatibility, rheological properties and mechanical properties for a given tissue. Within the scope of this project, KerMA material was chosen as methacrylate in terms of originality and biocompatibility, but this material alone is weak in terms of rheological properties and mechanical strength. In order to improve the rheological properties of KerMA material and fibroblast cell viability, a composite was formed with SilkMA at different concentrations. Magnetic iron oxide nanoparticle was added to protein gels (in fluid form) to increase the ability of biological active substances to be transported within the body. As an external intervention to these magnetic nanoparticles, a 3-axis Helmholtz coil was designed and produced and integrated into the 3D device in use. Integration of the coil and SilkMA/ KerMA scaffold production were done with a 3D SLA printer. Orientation of the polymer chains in such a way that the internal structures are ordered and anisotropic was carried out with a 15mT magnetic field created by the 3-axis Helmholtz coil during 3D printing. It was evaluated by tests and analyzes that SilkMA/ KerMA structures were magnetically oriented, did not show any toxic effects on the scaffolds for 1,3 and 7 days, and cell compatibility properties were improved in oriented structures. In addition, SEM and DAPI tests were performed to confirm that scaffolds support cell proliferation and adhesion. To experimentally see the effect of anisotropic orientation, imaging studies and examinations in SEM and atomic force microscopy (AFM) were performed. It was determined that the scaffolds exhibited high swelling properties and did not dissolve for 14 days by applying compression test to the scaffolds, calculating its resistance and elasticity against mechanical stresses, and also performing swelling and degradation tests.
Tissue and regeneration engineering aims to replace, repair or regenerate damaged, degenerated or defective tissues using engineering approaches, including scaffolds, cells, and biophysical and biochemical stimuli. In this study, the synthesis and physico-chemical characterization of biodegradable methacrylate Silk fibroin (SilkMA) and keratin methacrylate (KerMA) scaffolds, which can be used as biocomposites, were performed, and tissue engineering aimed and targeted properties were analyzed. SilkMA exhibits good biocompatibility, rheological properties and mechanical properties for a given tissue. Within the scope of this project, KerMA material was chosen as methacrylate in terms of originality and biocompatibility, but this material alone is weak in terms of rheological properties and mechanical strength. In order to improve the rheological properties of KerMA material and fibroblast cell viability, a composite was formed with SilkMA at different concentrations. Magnetic iron oxide nanoparticle was added to protein gels (in fluid form) to increase the ability of biological active substances to be transported within the body. As an external intervention to these magnetic nanoparticles, a 3-axis Helmholtz coil was designed and produced and integrated into the 3D device in use. Integration of the coil and SilkMA/ KerMA scaffold production were done with a 3D SLA printer. Orientation of the polymer chains in such a way that the internal structures are ordered and anisotropic was carried out with a 15mT magnetic field created by the 3-axis Helmholtz coil during 3D printing. It was evaluated by tests and analyzes that SilkMA/ KerMA structures were magnetically oriented, did not show any toxic effects on the scaffolds for 1,3 and 7 days, and cell compatibility properties were improved in oriented structures. In addition, SEM and DAPI tests were performed to confirm that scaffolds support cell proliferation and adhesion. To experimentally see the effect of anisotropic orientation, imaging studies and examinations in SEM and atomic force microscopy (AFM) were performed. It was determined that the scaffolds exhibited high swelling properties and did not dissolve for 14 days by applying compression test to the scaffolds, calculating its resistance and elasticity against mechanical stresses, and also performing swelling and degradation tests.