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Öğe 3D Printing of Extracellular Matrix-Based Multicomponent, All-Natural, Highly Elastic, and Functional Materials toward Vascular Tissue Engineering(John Wiley and Sons Inc, 2023) Işık, Melis; Karakaya, Ece; Arslan, Tugba Sezgin; Atila, Deniz; Erdoğan, Yaşar Kemal; Arslan, Yavuz Emre3D printing offers an exciting opportunity to fabricate biological constructs with specific geometries, clinically relevant sizes, and functions for biomedical applications. However, successful application of 3D printing is limited by the narrow range of printable and bio-instructive materials. Multicomponent hydrogel bioinks present unique opportunities to create bio-instructive materials able to display high structural fidelity and fulfill the mechanical and functional requirements for in situ tissue engineering. Herein, 3D printable and perfusable multicomponent hydrogel constructs with high elasticity, self-recovery properties, excellent hydrodynamic performance, and improved bioactivity are reported. The materials' design strategy integrates fast gelation kinetics of sodium alginate (Alg), in situ crosslinking of tyramine-modified hyaluronic acid (HAT), and temperature-dependent self-assembly and biological functions of decellularized aorta (dAECM). Using extrusion-based printing approach, the capability to print the multicomponent hydrogel bioinks with high precision into a well-defined vascular constructs able to withstand flow and repetitive cyclic compressive loading, is demonstrated. Both in vitro and pre-clinical models are used to show the pro-angiogenic and anti-inflammatory properties of the multicomponent vascular constructs. This study presents a strategy to create new bioink whose functional properties are greater than the sum of their components and with potential applications in vascular tissue engineering and regenerative medicineÖğe A novel method for constructing an acellular 3D biomatrix from bovine spinal cord for neural tissue engineering applications(Wiley, 2019) Arslan, Yavuz Emre; Efe, Burcu; Arslan, Tugba SezginIn this study, we aimed at generating 3-dimensional (3D) decellularized bovine spinal cord extracellular matrix-based scaffolds (3D-dCBS) for neural tissue engineering applications. Within this scope, bovine spinal cord tissue pieces were homogenized in 0.1 M NaOH and this viscous mixture was molded to attain 3D bioscaffolds. After resultant bioscaffolds were chemically crosslinked, the decellularization process was conducted with detergent, buffer, and enzyme solutions. Nuclear remnants in the native tissue and 3D-dCBS were determined with DNA content analysis and agarose gel electrophoresis. Afterward, 3D-dCBS were biochemically characterized in depth via glycosaminoglycan (GAG) content, hydroxyproline (HYP) assay, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Cellular survival of human adipose-derived mesenchymal stem cells (hAMSCs) on the 3D-dCBS for 3rd, 7th, and 10th days was assessed via MTT assay. Scaffold and cell/scaffold constructs were also evaluated with scanning electron microscopy and histochemical studies. DNA contents for native and 3D-dCBS were respectively found to be 520.76 +/- 18.11 and 28.80 +/- 0.20 ng/mg dry weight (n = 3), indicating a successful decellularization process. GAG content, HYP assay, and SDS-PAGE results proved that the extracellular matrix was substantially preserved during the decellularization process. In conclusion, it is believed that the novel decellularization method may allow fabricating 3D bioscaffolds with desired geometry from soft nervous system tissues.Öğe A Novel Protocol to Generate Decellularized Bovine Spinal Cord Extracellular Matrix-based Scaffolds (3D-dCBS)(Bio-Protocol, 2019) Arslan, Yavuz E.; Efe, Burcu; Arslan, Tugba SezginExtracellular matrix (ECM)-based tissue engineering scaffolds have an essential role in promoting tissue regeneration. Nerve tissue engineering aims at facilitating the repair of permanent damage to the peripheral and central nervous systems, which are difficult to heal. For this purpose, a variety of biomaterials are being developed consisting of numerous synthetic and/or natural polymers to provide axonal reinnervation and to direct the growth of axons. Here, we present a novel protocol that enables to fabricate a 3-dimensional (3D) decellularized scaffold derived from the bovine spinal cord (BSC) ECM (3D-dCBS) for neural tissue engineering applications. In this protocol, a viscous ECMderived gel from BSC is prepared, molded, and chemically crosslinked with EDC/NHS (3D-CBS) before decellularization process. Decellularization of 3D-CBS is performed with 1% SDS to attain 3D-dCBS. As compared with other available methods, our protocol is a novel decellularization method that preserves a more significant part of the ECM. We believe that the mentioned protocol has the potential to produce a bioengineered scaffold from spinal cord tissue with desired geometry for regenerative medicine applications related to neural tissue engineering.Öğe Decellularized Bone Extracellular Matrix-Coated Electrospun PBAT Microfibrous Membranes with Cell Instructive Ability and Improved Bone Tissue Forming Capacity(Wiley-V C H Verlag Gmbh, 2022) Karakaya, Ece; Erdogan, Yasar Kemal; Arslan, Tugba Sezgin; Arslan, Yavuz Emre; Odabas, Sedat; Ercan, Batur; Emregul, EmelCurrent approaches to develop bone tissue engineering scaffolds have some limitations and shortcomings. They mainly suffer from combining mechanical stability and bioactivity on the same platform. Synthetic polymers are able to produce mechanically stable sturctures with fibrous morphology when they are electrospun, however, they cannot exhibit bioactivity, which is crucial for tissue engineering and regenerative medicine. One current strategy to bring bioactivity in synthetic materials is to combine extracellular matrix (ECM)-sourced materials with biologically inert synthetic materials. ECM-sourced materials without any modifications are mechanically unstable; therefore, reinforcing them with mechanically stable platforms is indispensable. In order to overcome this bifacial problem, we have demonstrated that poly(butylene adipate-co-terephthalate) (PBAT) electrospun microfibrous membranes can be successfully modified with decellularized bone ECM to endow fibers with bioactive hydrogel and mimic natural micro-features of the native bone tissue. The developed structures have been shown to support osteogenesis, confirmed by histochemical staining and gene expression studies. Furthermore, ECM-coated PBAT fibers, when they were aligned, supplied an improved level of osteogenesis. The strategy demonstrated can be adapted to any other tissues, and the emerging microfibrous, mechanically stable, and bioactive materials can find implications in the specific fields of tissue engineering and regenerative medicine.Öğe Detergent-free decellularization of bovine costal cartilage for chondrogenic differentiation of human adipose mesenchymal stem cells in vitro(Royal Soc Chemistry, 2016) Erten, Evren; Arslan, Tugba Sezgin; Derkus, Burak; Arslan, Yavuz EmreIn this study, we report a novel, detergent-free decellularization protocol for the preparation of intact cartilage ECM-based scaffolds (CEbS) during an effective decalcification process. On treatment with 10 mM Na(2)EDTA, the amount of calcium lost was around 55% +/- 5% (percent +/- S.D.%) (n = 3) and nearly 84% of the nuclear material was removed; however, the most effective removal was observed on treatment with 10 mM Na2EDTA combined with 0.5% Triton X-100 for 48 hours. Notably, our proposed method decreased the GAG content by only 5% compared to untreated CEbS (380.37 +/- 16.02 mu g mg(-1) dry weight). There was no significant difference in hydroxyproline content between the untreated (13.04 +/- 1.51 mg mg(-1) dry weight) sample and our proposed method (12.95 +/- 1.55 mg mg(-1) dry weight). The scaffold morphology and cell attachment were evaluated using SEM micrographs, and the cells that were inoculated with detergent-free decellularized CEbS for 14, 21 and 28 days covered the scaffold area, including the porous cavities. Microscopic observations showed that the cell density increased day by day and there was no cytotoxic evidence for the scaffolds, which is a desirable environment for cells. The histochemical and immunohistochemical assessments are supported by glycosaminoglycan and hydroxyproline assays. The proposed detergent-free decellularization technique could be a promising method for cartilage tissue regeneration.Öğe Development of Ductile-Sticky Bone Fillers from Biodegradable Hydrolyzed Wool-Keratin and Silk Fibroin(Wiley-V C H Verlag Gmbh, 2024) Bekar, Serife; Arslan, Tugba Sezgin; Arslan, Yavuz EmreIn the present study, a method is proposed for preparing novel ductile-sticky materials that can be used as bone void fillers using hydrolyzed wool-keratin (WK) and silk fibroin (SF). This methodology uses citric acid as a cross-linking agent in preparing keratin paste (KP) owing to its non-toxicity and plasticizing properties. The Keratin paste-silk fibroin structure (KPSF) is obtained by adding SF, which possesses biocompatible and superior mechanical properties. Methanol treatment is employed on the KPSF mixture to convert the Silk I structure in the SF to Silk II, resulting in a water-insoluble and tightly packed proteinaceous structure. The physicochemical properties of both bioscaffolds are investigated and discussed in detail by comparison. Based on the findings, the presence of SF in the KPSF structure contributed to properties such as flexibility and porosity. In ovo CAM analysis reveals that both materials exhibit proangiogenic properties and are biocompatible. KP and KPSF bioscaffolds can be converted into ductile-sticky forms by adding water. It believes that these forms can easily apply to bone defect areas, particularly cavitary bone defects. Furthermore, KPSF bioscaffolds, with better mechanical properties, can be considered candidates for use in non-load-bearing bone tissue engineering applications. This study introduces a strategy using citric acid as a crosslinker and plasticizer to create ductile-sticky Keratin Paste (KP) and Keratin paste-silk fibroin (KPSF) bone fillers. This study proposes that moldable ductile-sticky KP and KPSF materials are suitable for cavitary bone defects, while lyophilized forms of KPSF bioscaffolds, offering flexibility and robustness, are ideal for non-load-bearing bone tissue engineering applications. imageÖğe Fabrication of human hair keratin/jellyfish collagen/eggshell-derived hydroxyapatite osteoinductive biocomposite scaffolds for bone tissue engineering: From waste to regenerative medicine products(Elsevier, 2017) Arslan, Yavuz Emre; Arslan, Tugba Sezgin; Derkus, Burak; Emregul, Emel; Emregul, Kaan C.In the present study, we aimed at fabricating an osteoinductive biocomposite scaffold using keratin obtained from human hair, jellyfish collagen and eggshell-derived nano-sized spherical hydroxyapatite (nHA) for bone tissue engineering applications. Keratin, collagen and nHA were characterized with the modified Lowry method, free-sulfhydryl groups and hydroxyproline content analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR) and thermal gravimetric analysis (TGA) which confirmed the success of the extraction and/or isolation processes. Human adipose mesenchymal stem cells (hAMSCs) were isolated and the cell surface markers were characterized via flow cytometry analysis in addition to multi lineage differentiation capacity. The undifferentiated hAMSCs were highly positive for CD29, CD44, CD73, CD90 and CD105, but were not seen to express hematopoietic cell surface markers such as CD14, CD34 and CD45. The cells were successfully directed towards osteogenic, chondrogenic and adipogenic lineages in vitro. The microarchitecture of the scaffolds and cell attachment were evaluated using scanning electron microscopy (SEM). The cell viability on the scaffolds was assessed by the MTT assay which revealed no evidence of cytotoxicity. The osteogenic differentiation of hAMSCs on the scaffolds was determined histologically using alizarin red S, osteopontin and osteonectin stainings. Early osteogenic differentiation markers of hAMSCs were significantly expressed on the collagen-keratin-nHA scaffolds. In conclusion, it is believed that collagen-keratin-nHA osteoinductive biocomposite scaffolds have the potential of being used in bone tissue engineering. (C) 2017 Elsevier B.V. All rights reserved.Öğe Sophisticated Biocomposite Scaffolds from Renewable Biomaterials for Bone Tissue Engineering(Springer International Publishing Ag, 2019) Arslan, Yavuz Emre; Ozudogru, Eren; Arslan, Tugba Sezgin; Derkus, Burak; Emregul, Emel; Emregul, Kaan C.[Anstract Not Available]Öğe Trans-differentiation of human adipose-derived mesenchymal stem cells into cardiomyocyte-like cells on decellularized bovine myocardial extracellular matrix-based films(Springer, 2018) Arslan, Yavuz Emre; Galata, Yusuf Furkan; Arslan, Tugba Sezgin; Derkus, BurakIn this study, we aimed at fabricating decellularized bovine myocardial extracellular matrix-based films (dMEbF) for cardiac tissue engineering (CTE). The decellularization process was carried out utilizing four consecutive stages including hypotonic treatment, detergent treatment, enzymatic digestion and decontamination, respectively. In order to fabricate the dMEbF, dBM were digested with pepsin and gelation process was conducted. dMEbF were then crosslinked with N-hydroxysuccinimide/1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (NHS/EDC) to increase their durability. Nuclear contents of native BM and decellularized BM (dBM) tissues were determined with DNA content analysis and agarose-gel electrophoresis. Cell viability on dMEbF for 3rd, 7th, and 14th days was assessed by MTT assay. Cell attachment on dMEbF was also studied by scanning electron microscopy. Trans-differentiation capacity of human adipose-derived mesenchymal stem cells (hAMSCs) into cardiomyocyte-like cells on dMEbF were also evaluated by histochemical and immunohistochemical analyses. DNA contents for native and dBM were, respectively, found as 886.11 +/- 164.85 and 47.66 +/- 0.09 ng/mg dry weight, indicating a successful decellularization process. The results of glycosaminoglycan and hydroxyproline assay, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), performed in order to characterize the extracellular matrix (ECM) composition of native and dBM tissue, showed that the BM matrix was not damaged during the proposed method. Lastly, regarding the histological study, dMEbF not only mimics native ECM, but also induces the stem cells into cardiomyocyte-like cells phenotype which brings it the potential of use in CTE.
