Hücresizleştirilmiş Bitki İçeren 3B Baskılı Doku İskelelerinin Hazırlanması ve Temel Hücre Etkileşimlerinin İncelenmesi

Abstract

Hücresizleştirme işlemi, dokularda biyomekanik destek sağlayan ve hücre dışı matristen (ECM) oluşan 3B biyomalzemelerin elde edilmesinde kullanılmak üzere, dokudaki hücresel içeriğin dokunun yapısal bütünlüğünü bozmadan uzaklaştırılması işlemidir. Literatürde yapılan çalışmalar, hücresizleştirilmiş hayvan kaynaklı dokuların, çeşitli biyopolimerlerle farklı teknikler kullanılarak birleştirilerek kompozit doku iskeleleri hazırlandığını ortaya koymuştur. Son yıllarda yapısal özellikleri nedeniyle hayvan dokularının yanı sıra bitki kaynaklı dokuların da hücresizleştirme çalışmaları ön plana çıkmıştır. Bu çalışmada, hücresizleştirilmiş sukulent dokuları aljinat ile birleştirilerek hibrit doku iskeleleri hazırlanması amaçlanmıştır. Çalışmanın amacı mezenkimal kök hücreleri kullanılarak temel hücre-malzeme etkileşimlerini ve kıkırdak farklılaşma parametrelerini araştırmaktır. Kullanılan metoda göre, etli bitki yaprakları hücresizleştirilip su tutan kısımlar diğer dokulardan ayrıldı, liyofilize edildi ve toz haline getirildi. Bu yaklaşım, su tutma kapasitesine sahip biyomolekülleri toz halinde korumak için kullanıldı. Hücresizleştirme sürecinin verimliliğini belirlemek için DNA ve protein miktarları değerlendirilip karşılaştırıldı. Yüksek su tutma kapasiteleri nedeniyle etli bitkilerin su tutan yapıları, 3B baskıya uygun bir mürekkep hazırlamak için çeşitli konsantrasyonlarda aljinat biyopolimeri ile birleştirildi. Baskıdan sonra, elde edilen iskelelerin bozunma ve şişme davranışı, kimyasal bileşimi, yapısal karakterizasyonu ve termal özellikleri incelendi. Son aşamada, 3B yazdırılmış iskeleler üzerinde L929 fare fibroblast hücreleri ve insan mezenkimal kök hücreleri kullanılarak hücre-malzeme etkileşimi üzerine temel bir çalışma yürütüldü. Hazırlanan hibrit iskelelerinde hücre-malzeme etkileşimi temel sitotoksisite testleri kullanılarak incelenmiştir.

Keywords

• Sukulent bitkisi
• bitki hücresizleştirme işlemi
• biyomalzemeler

Development of 3D Printed Scaffolds Containing Decellularized Plants and Investigation of Their Basic Cell Interactions

Abstract

The decellularization process fundamentally removes the cellular content of the tissue (nuclear material and other nucleic acid components) without disrupting the structural integrity of the tissue. It is an effective approach, especially for obtaining three-dimensional (3D) biomaterials composed of the extracellular matrix (ECM), which provides tissue biomechanical support. In the literature, studies have shown that after the decellularization process, animal-derived decellularized tissues have been combined with various biopolymers to prepare composite scaffolds using different techniques. In recent years, due to their structural features, decellularization studies of plant-derived tissues have also gained prominence alongside animal tissues. In this study, succulent plants were chosen as the plant tissue, and the purpose was to prepare hybrid scaffolds by combining decellularized succulent tissues with alginate structures. The study aimed to investigate the fundamental cell-material interactions and cartilage-specific differentiation parameters using mesenchymal stem cells. Succulent plant leaves were decellularized using a solution containing Triton X-100 and SDS. The water-retaining parts were separated from other tissues, lyophilized, and turned into a powder. This approach was employed to preserve biomolecules with water-retaining capacity in powdered form. To determine the efficiency of the decellularization process, the quantities of DNA and proteins were assessed and compared. Due to their high water-absorbing capacity, the succulent plants' water-retaining structures were combined with alginate biopolymer at various viscosity levels to prepare an ink suitable for 3D printing. After printing, the resulting scaffolds' degradation and swelling behavior, chemical composition, structural characterization, and thermal properties were examined. In the final phase, a fundamental investigation was carried out on cell-material interactions using L929 mouse fibroblast cells and human mesenchymal stem cells on 3D printed scaffolds. The interactions within the prepared hybrid scaffolds were analyzed through basic cytotoxicity tests.

Keywords

• Succulent plant
• plant decellularization
• biomaterials

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