Decellularised whole ovine testis as a potential bio-scaffold for tissue engineering

2019 ◽  
Vol 31 (11) ◽  
pp. 1665 ◽  
Author(s):  
Aram Akbarzadeh ◽  
Maral Kianmanesh ◽  
Kiarad Fendereski ◽  
Maryam Ebadi ◽  
Seyedeh Sima Daryabari ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Sodium Dodecyl ◽  
Three Dimensional ◽  
Optimal Time ◽  
Second Phase ◽  
Dapi Staining ◽  
Extracellular Matrix Components ◽  
Time Required ◽  
Triton X

The aim of this study was to determine an efficient whole-organ decellularisation protocol of a human-sized testis by perfusion through the testicular arteries. In the first step of this study, we determined the most efficient detergent agent, whereas the second phase delineated the optimal time required for the decellularisation process. Initially sheep testes were decellularised by one of three different detergent agents: sodium dodecyl sulphate (SDS), Triton X-100 and trypsin-ethylenediamine tetraacetic acid (EDTA) solutions, each perfused for 6h. In the second phase, the selected detergent agent was applied for different time periods. A total number of 20 organs were processed during this investigation. The efficacy of the decellularisation process and the preservation of the extracellular matrix components and structure were evaluated by histopathological examinations, 4′,6′-diamidino-2-phenylindole (DAPI) staining, DNA quantification, hydroxyproline measurement, magnetic resonance imaging and scanning electron microscopy. Organ perfusion with 1% SDS solution for 6 to 8h demonstrated the most desirable outcomes regarding decellularisation and extracellular matrix preservation. The 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay was used to determine the toxicity of the scaffold and its potential for further application in tissue-engineering investigations. This investigation introduces an efficient method to produce a three-dimensional testicular bio-scaffold resembling the properties of the native organ that could be employed in tissue-engineering studies.

scholarly journals Optimization of decellularized human placental macroporous scaffolds for spermatogonial stem cells homing

2021 ◽  
Vol 32 (5) ◽  
Author(s):  
Fatemeh Asgari ◽  
Hamid Reza Asgari ◽  
Mohammad Najafi ◽  
Behnaz Sadat Eftekhari ◽  
Mina Vardiani ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Incubation Time ◽  
Colony Formation ◽  
Sodium Dodecyl ◽  
Three Dimensional ◽  
Spermatogonial Stem Cells ◽  
Negative Effects ◽  
Swelling Properties ◽  
Extracellular Matrix Components

AbstractDecellularized scaffolds have been found to be excellent platforms for tissue engineering applications. The attempts are still being made to optimize a decellularization protocol with successful removal of the cells with minimal damages to extracellular matrix components. We examined twelve decellularization procedures using different concentrations of Sodium dodecyl sulfate and Triton X-100 (alone or in combination), and incubation time points of 15 or 30 min. Then, the potential of the decellularized scaffold as a three-dimensional substrate for colony formation capacity of mouse spermatogonial stem cells was determined. The morphological, degradation, biocompatibility, and swelling properties of the samples were fully characterized. The 0.5%/30 SDS/Triton showed optimal decellularization with minimal negative effects on ECM (P ≤ 0.05). The swelling ratios increased with the increase of SDS and Triton concentration and incubation time. Only 0.5%/15 and 30 SDS showed a significant decrease in the SSCs viability compared with other groups (P < 0.05). The SSCs colony formation was clearly observed under SEM and H&E stained slides. The cells infiltrated into the subcutaneously implanted scaffold at days 7 and 30 post-implantation with no sign of graft rejection. Our data suggest the %0.5/30 SDS/Triton as an excellent platform for tissue engineering and reproductive biology applications.

A novel method for providing scaffold: Decellularization of parathyroid capsule

2021 ◽  
pp. 088532822110543
Author(s):  
Nisa İrem Büyük ◽  
Kardelen Tüfekçi ◽  
Alev Cumbul ◽  
Erhan Ayşan ◽  
Gamze Torun Köse
Keyword(s):  
Extracellular Matrix ◽  
Sodium Dodecyl Sulphate ◽  
Dna Analysis ◽  
Structural Integrity ◽  
Sodium Dodecyl ◽  
Parathyroid Tissue ◽  
The Body ◽  
Extracellular Matrix Components ◽  
Triton X ◽  
Sulphated Glycosaminoglycan

This study aimed to generate a novel biomatrix from the decellularized human parathyroid capsule using different methods and to compare the efficiency of decellularization in the means of cell removal, structural integrity and extracellular matrix preservation. The parathyroid capsules, which were carefully dissected from the parathyroid tissue, were randomly divided into four groups and then decellularized using three different protocols: freeze-thaw only, sodium dodecyl sulphate and Triton X-100 treatments after freeze-thawing. Quantitative DNA analysis, agarose gel electrophoresis, sulphated glycosaminoglycan assay, histological analysis, immunohistochemistry and scanning electron microscopy were used to observe the efficiency of parathyroid capsule decellularization and preservation of extracellular matrix components. Considering all the results, it can be said that only freeze-thawing is not an effective method in parathyroid capsule decellularization. When the tissue was treated with a detergent agent in addition to freeze-thawing, the amount of DNA decreased by 90% while sulphated glycosaminoglycan amount maintained 50% compared to untreated tissue. Comparing the effects of the two detergents on the preservation of extracellular matrix such as collagen and sulphated glycosaminoglycan, it was seen that the integrity of tissues treated with Triton X-100 was preserved more than tissues treated with sodium dodecyl sulphate. It is concluded that Triton X-100 treatment with freeze-thawing is the most suitable and effective method for decellularizing the human parathyroid capsule. The biomatrix obtained with this method can be applied in the transplantation of parathyroid tissue and other endocrine tissue types in the body.

scholarly journals In Vitro Generation of Novel Functionalized Biomaterials for Use in Oral and Dental Regenerative Medicine Applications

Materials ◽  
2020 ◽  
Vol 13 (7) ◽  
pp. 1692 ◽  
Author(s):  
Cristina Blanco-Elices ◽  
Enrique España-Guerrero ◽  
Miguel Mateu-Sanz ◽  
David Sánchez-Porras ◽  
Óscar García-García ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Regenerative Medicine ◽  
Oral Mucosa ◽  
Dental Tissues ◽  
Extracellular Matrix Components ◽  
Matrix Components ◽  
Time Required ◽  
Vitro Development

Recent advances in tissue engineering offer innovative clinical alternatives in dentistry and regenerative medicine. Tissue engineering combines human cells with compatible biomaterials to induce tissue regeneration. Shortening the fabrication time of biomaterials used in tissue engineering will contribute to treatment improvement, and biomaterial functionalization can be exploited to enhance scaffold properties. In this work, we have tested an alternative biofabrication method by directly including human oral mucosa tissue explants within the biomaterial for the generation of human bioengineered mouth and dental tissues for use in tissue engineering. To achieve this, acellular fibrin–agarose scaffolds (AFAS), non-functionalized fibrin-agarose oral mucosa stroma substitutes (n-FAOM), and novel functionalized fibrin-agarose oral mucosa stroma substitutes (F-FAOM) were developed and analyzed after 1, 2, and 3 weeks of in vitro development to determine extracellular matrix components as compared to native oral mucosa controls by using histochemistry and immunohistochemistry. Results demonstrate that functionalization speeds up the biofabrication method and contributes to improve the biomimetic characteristics of the scaffold in terms of extracellular matrix components and reduce the time required for in vitro tissue development.

scholarly journals Harnessing the Topography of 3D Spongy-Like Electrospun Bundled Fibrous Scaffold via a Sharply Inclined Array Collector

Polymers ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 1444 ◽  
Author(s):  
Sun Hee Cho ◽  
Jeong In Kim ◽  
Cheol Sang Kim ◽  
Chan Hee Park ◽  
In Gi Kim
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Three Dimensional ◽  
Living System ◽  
Pivotal Role ◽  
Skin Tissue ◽  
Target Tissue ◽  
Fibrous Scaffold ◽  
Electrospun Scaffolds ◽  
Directional Change

To date, many researchers have studied a considerable number of three-dimensional (3D) cotton-like electrospun scaffolds for tissue engineering, including the generation of bone, cartilage, and skin tissue. Although numerous 3D electrospun fibrous matrixes have been successfully developed, additional research is needed to produce 3D patterned and sophisticated structures. The development of 3D fibrous matrixes with patterned and sophisticated structures (FM-PSS) capable of mimicking the extracellular matrix (ECM) is important for advancing tissue engineering. Because modulating nano to microscale features of the 3D fibrous scaffold to control the ambient microenvironment of target tissue cells can play a pivotal role in inducing tissue morphogenesis after transplantation in a living system. To achieve this objective, the 3D FM-PSSs were successfully generated by the electrospinning using a directional change of the sharply inclined array collector. The 3D FM-PSSs overcome the current limitations of conventional electrospun cotton-type 3D matrixes of random fibers.

scholarly journals A Strategy to Enhance Secretion of Extracellular Matrix Components by Stem Cells: Relevance to Tissue Engineering

2018 ◽  
Vol 24 (1-2) ◽  
pp. 145-156 ◽  
Author(s):  
Navaneethakrishnan Krishnamoorthy ◽  
Yuan‐Tsan Tseng ◽  
Poornima Gajendrarao ◽  
Padmini Sarathchandra ◽  
Ann McCormack ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Extracellular Matrix ◽  
Extracellular Matrix Components ◽  
Matrix Components

scholarly journals Fabrication of 3D Printing Scaffold with Porcine Skin Decellularized Bio-Ink for Soft Tissue Engineering

Materials ◽  
2020 ◽  
Vol 13 (16) ◽  
pp. 3522
Author(s):  
Su Jeong Lee ◽  
Jun Hee Lee ◽  
Jisun Park ◽  
Wan Doo Kim ◽  
Su A Park
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
3D Printing ◽  
Three Dimensional ◽  
3D Bioprinting ◽  
Research Groups ◽  
Porcine Skin ◽  
Alginate Hydrogel ◽  
Chemical Treatments ◽  
Soft Tissue Engineering

Recently, many research groups have investigated three-dimensional (3D) bioprinting techniques for tissue engineering and regenerative medicine. The bio-ink used in 3D bioprinting is typically a combination of synthetic and natural materials. In this study, we prepared bio-ink containing porcine skin powder (PSP) to determine rheological properties, biocompatibility, and extracellular matrix (ECM) formation in cells in PSP-ink after 3D printing. PSP was extracted without cells by mechanical, enzymatic, and chemical treatments of porcine dermis tissue. Our developed PSP-containing bio-ink showed enhanced printability and biocompatibility. To identify whether the bio-ink was printable, the viscosity of bio-ink and alginate hydrogel was analyzed with different concentration of PSP. As the PSP concentration increased, viscosity also increased. To assess the biocompatibility of the PSP-containing bio-ink, cells mixed with bio-ink printed structures were measured using a live/dead assay and WST-1 assay. Nearly no dead cells were observed in the structure containing 10 mg/mL PSP-ink, indicating that the amounts of PSP-ink used were nontoxic. In conclusion, the proposed skin dermis decellularized bio-ink is a candidate for 3D bioprinting.

scholarly journals Electrospun nanofibers for the fabrication of engineered vascular grafts

2019 ◽  
Vol 13 (1) ◽  
Author(s):  
Sonia Fathi Karkan ◽  
Soodabeh Davaran ◽  
Reza Rahbarghazi ◽  
Roya Salehi ◽  
Abolfazl Akbarzadeh
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Physicochemical Properties ◽  
Review Paper ◽  
Vascular Grafts ◽  
Three Dimensional ◽  
Electrospun Nanofibers ◽  
Electrospun Fibers

Abstract Attention has recently increased in the application of electrospun fibers because of their putative capability to create nanoscale platforms toward tissue engineering. To some extent, electrospun fibers are applicable to the extracellular matrix by providing a three-dimensional microenvironment in which cells could easily acquire definite functional shape and maintain the cell-to-cell connection. It is noteworthy to declare that placement in different electrospun substrates with appropriate physicochemical properties enables cells to promote their bioactivities, dynamics growth and differentiation, leading to suitable restorative effects. This review paper aims to highlight the application of biomaterials in engineered vascular grafts by using electrospun nanofibers to promote angiogenesis and neovascularization

scholarly journals Spheroidal aggregate culture of rat liver cells: histotypic reorganization, biomatrix deposition, and maintenance of functional activities.

1985 ◽  
Vol 101 (3) ◽  
pp. 914-923 ◽  
Author(s):  
J Landry ◽  
D Bernier ◽  
C Ouellet ◽  
R Goyette ◽  
N Marceau
Keyword(s):  
Extracellular Matrix ◽  
Rat Liver ◽  
Single Cells ◽  
Three Dimensional ◽  
Cell Types ◽  
Tyrosine Aminotransferase ◽  
Liver Cells ◽  
Specific Pattern ◽  
Extracellular Matrix Components ◽  
Matrix Components

Liver cells isolated from newborn rats and seeded on a non-adherent plastic substratum were found to spontaneously re-aggregate and to form, within a few days, spheroidal aggregates that eventually reached a plateaued diameter of 150-175 micron. Analyses on frozen sections from these spheroids by immunofluorescence microscopy using antibodies to various cytoskeletal elements and extracellular matrix components revealed a sorting out and a histotypic reorganization of three major cell types. A first type consisted of cells that segregated out on the aggregate surface forming a monolayer cell lining; a second type was identified as hepatocytes that regrouped in small islands often defining a central lumen; and a third group of cells reorganized into bile duct-like structures. This intercellular organization in the aggregates was paralleled by the accumulation of extracellular matrix components (laminin, fibronectin, and collagen) and their deposition following a specific pattern around each cell population structure. Determinations of albumin secretion and tyrosine aminotransferase induction by dexamethasone and glucagon at various times after the initiation of the cultures revealed a maintenance of the hepatocyte-differentiated functions for at least up to 2 mo at the levels measured at 3-5 d. It is concluded that cells dispersed as single cells from newborn rat liver conserve in part the necessary information to reconstruct a proper three-dimensional cyto-architecture and that the microenvironment so generated most likely represents a basic requirement for the optimal functioning of these differentiated cells.

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