Chemical decellularization: a promising approach for preparation of extracellular matrix

Hrebíková, Hana; Díaz, Daniel; Mokrý, Jaroslav

doi:10.5507/bp.2013.076

Cited by 41 publications

(43 citation statements)

References 59 publications

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“…A removal of cells or nuclei was here confirmed by various microscope techniques, in accordance to other studies producing biological scaffolds (Crapo et al, ; Hrebikova, Diaz, & Mokry, ). However, our quantitative survey likewise resulted in a dramatic decrease of dsDNA in decellularized bovine cotyledons that was well within the established parameters of less than 50 ng dsDNA per mg of biological scaffold (Crapo et al, ; Hrebikova et al, ).…”

Section: Discussionsupporting

confidence: 90%

Decellularized bovine cotyledons may serve as biological scaffolds with preserved vascular arrangement

Barreto

Romagnolli

Mess

et al. 2017

J Tissue Eng Regen Med

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Technically produced scaffolds are common to establish transplantable tissues for regenerative medicine, but also biological ones that are closer to the natural condition become of interest. Placentas are promising, because they represented available, complete organs with rich extracellular matrix (ECM) and well-developed vasculature that easily could build anastomoses to a host's organ. Only placentas from larger animal models such as the bovine meet the dimensions large enough for most organs but are not adequately described yet. We here studied the nature of the ECM in 27 natural and decellularized bovine cotyledons, that is, the fetal part of the placentomes, by means of histology, immunohistochemistry, and electron microscopy. Successful decellularization was done by perfusion with 0.01%, 0.1%, and 0.5% sodium dodecyl sulfate each and subsequent immersion in 1% Triton X-100, resulting in a removal of cells and DNA, whereas the structure of the allantochorionic surface and villi was preserved. Although some fibres disappeared, also the arrangement of the main ECM proteins was largely similar before and after decellularization: Along the larger vessels, a densely packed network of thick fibres occurred, organized in layers without cells or spaces in between. Collagen IV, fibronectin, and laminin contributed to those areas. In contrast, collagen I and III characterized the meshwork of medium-sized and thin fibres in the mesenchyme, respectively. In conclusion, decellularized bovine cotyledons indeed had characteristics of a biological scaffold and provide an interesting alternative to develop large-scale scaffolds with complex vascular architecture for tissue engineering purposes.

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Section: Discussionsupporting

confidence: 90%

Decellularized bovine cotyledons may serve as biological scaffolds with preserved vascular arrangement

Barreto

Romagnolli

Mess

et al. 2017

J Tissue Eng Regen Med

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“…Further loss of water-soluble GAGs occurred due to the process conducted in an aqueous solution which can easily wash away the separated GAGs from the tissues 44. According to Hrebíková, the chemical detergent SDS has a deleterious effect on the GAG content within the tissues 49. From the previous study, it revealed that GAG reduction after the decellularization process is a crucial step for future in vitro study.…”

Section: Discussionmentioning

confidence: 99%

<p>Development of decellularized meniscus using closed sonication treatment system: potential scaffolds for orthopedics tissue engineering applications</p>

Yusof¹,

Sha'ban²,

Azhim³

2019

IJN

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Purpose Meniscus is a fibrocartilagenous tissue that cannot effectively heal due to its complex structure and presence of avascular zone. Thus, tissue engineering and regenerative medicine offer an alternative for the regeneration of meniscus tissues using bioscaffolds as a replacement for the damaged one. The aim of this study was to prepare an ideal meniscus bioscaffold with minimal adverse effect on extracellular matrix components (ECMs) using a sonication treatment system. Methods The decellularization was achieved using a developed closed sonication treatment system for 10 hrs, and continued with a washing process for 5 days. For the control, a simple immersion treatment was set as a benchmark to compare the decellularization efficiency. Histological and biochemical assays were conducted to investigate the cell removal and retention of the vital extracellular matrix. Surface ultrastructure of the prepared scaffolds was evaluated using scanning electron microscope at 5,000× magnification viewed from cross and longitudinal sections. In addition, the biomechanical properties were investigated through ball indentation testing to study the stiffness, residual forces and compression characteristics. Statistical significance between the samples was determined with p -value =0.05. Results Histological and biochemical assays confirmed the elimination of antigenic cellular components with the retention of the vital extracellular matrix within the sonicated scaffolds. However, there was a significant removal of sulfated glycosaminoglycans. The surface histoarchitecture portrayed the preserved collagen fibril orientation and arrangement. However, there were minor disruptions on the structure, with few empty micropores formed which represented cell lacunae. The biomechanical properties of bioscaffolds showed the retention of viscoelastic behavior of the scaffolds which mimic native tissues. After immersion treatment, those scaffolds had poor results compared to the sonicated scaffolds due to the inefficiency of the treatment. Conclusion In conclusion, this study reported that the closed sonication treatment system had high capabilities to prepare ideal bioscaffolds with excellent removal of cellular components, and retained extracellular matrix and biomechanical properties.

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“…Decellularized organ-derived extracellular matrix (dECM) has been used for many years in tissue engineering and regenerative medicine (Conklin et al, 2002;Dahl et al, 2003;Schenke-Layland et al, 2003). Decellularization of the extracellular matrix can be achieved through different physical, chemical or biological processes, including freeze/thaw cycles, use of organic detergents, and mild treatment with proteolytic enzymes (Crapo et al, 2011;Hrebikova et al, 2015;Keane et al, 2015;Leonel et al, 2017;Kawecki et al, 2018). Commonly, different processes are combined or used sequentially, so as to decrease the damage caused by each of them to the extracellular matrix but, at the same time, to potentiate the decellularization protocol (Crapo et al, 2011;Kawecki et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Molecular and Biomechanical Clues From Cardiac Tissue Decellularized Extracellular Matrix Drive Stromal Cell Plasticity

Liguori

Moraes

et al. 2020

Front. Bioeng. Biotechnol.

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Decellularized-organ-derived extracellular matrix (dECM) has been used for many years in tissue engineering and regenerative medicine. The manufacturing of hydrogels from dECM allows to make use of the pro-regenerative properties of the ECM and, simultaneously, to shape the material in any necessary way. The objective of the present project was to investigate differences between cardiovascular tissues (left ventricle, mitral valve, and aorta) with respect to generating dECM hydrogels and their interaction with cells in 2D and 3D. The left ventricle, mitral valve, and aorta of porcine hearts were decellularized using a series of detergent treatments (SDS, Triton-X 100 and deoxycholate). Mass spectrometry-based proteomics yielded the ECM proteins composition of the dECM. The dECM was digested with pepsin and resuspended in PBS (pH 7.4). Upon warming to 37 • C, the suspension turns into a gel. Hydrogel stiffness was determined for samples with a dECM concentration of 20 mg/mL. Adipose tissue-derived stromal cells (ASC) and a combination of ASC with human pulmonary microvascular endothelial cells (HPMVEC) were cultured, respectively, on and in hydrogels to analyze cellular plasticity in 2D and vascular network formation in 3D. Differentiation of ASC was induced with 10 ng/mL of TGF-β1 and SM22α used as differentiation marker. 3D vascular network formation was evaluated with confocal microscopy after immunofluorescent staining of PECAM-1. In dECM, the most abundant protein was collagen VI for the left ventricle and mitral valve and elastin for the aorta. The stiffness of the hydrogel derived from the aorta (6,998 ± 895 Pa) was significantly higher than those derived from the left ventricle (3,384 ± 698 Pa) and the mitral valve (3,233 ± 323 Pa) (One-way ANOVA, p = 0.0008). Aorta-derived dECM hydrogel drove non-induced (without TGF-β1) differentiation, while hydrogels derived from the left ventricle and mitral valve inhibited TGF-β1-induced differentiation. All hydrogels supported vascular network formation within 7 days of culture, but ventricular dECM hydrogel demonstrated more Liguori et al. Tissue-Specific Hydrogels Drive Cell Plasticity robust vascular networks, with thicker and longer vascular structures. All the three main cardiovascular tissues, myocardium, valves, and large arteries, could be used to fabricate hydrogels from dECM, and these showed an origin-dependent influence on ASC differentiation and vascular network formation.

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Chemical decellularization: a promising approach for preparation of extracellular matrix

Cited by 41 publications

References 59 publications

Decellularized bovine cotyledons may serve as biological scaffolds with preserved vascular arrangement

Decellularized bovine cotyledons may serve as biological scaffolds with preserved vascular arrangement

<p>Development of decellularized meniscus using closed sonication treatment system: potential scaffolds for orthopedics tissue engineering applications</p>

Molecular and Biomechanical Clues From Cardiac Tissue Decellularized Extracellular Matrix Drive Stromal Cell Plasticity

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