scholarly journals Accounting for Material Changes in Decellularized Tissue with Underutilized Methodologies

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Ryan A. Behmer Hansen ◽  
Xinming Wang ◽  
Gitanjali Kaw ◽  
Valinteshley Pierre ◽  
Samuel E. Senyo
Keyword(s):  
Tissue Engineering ◽  
Function Analysis ◽  
Mechanical Analysis ◽  
Physical Characterization ◽  
Molecular Profile ◽  
Tissue Properties ◽  
Engineering Research ◽  
Functional Features ◽  
The Impact

Tissue decellularization has rapidly developed to be a practical approach in tissue engineering research; biological tissue is cleared of cells resulting in a protein-rich husk as a natural scaffold for growing transplanted cells as a donor organ therapy. Minimally processed, acellular extracellular matrix reproduces natural interactions with cells in vitro and for tissue engineering applications in animal models. There are many decellularization techniques that achieve preservation of molecular profile (proteins and sugars), microstructure features such as organization of ECM layers (interstitial matrix and basement membrane) and organ level macrofeatures (vasculature and tissue compartments). While structural and molecular cues receive attention, mechanical and material properties of decellularized tissues are not often discussed. The effects of decellularization on an organ depend on the tissue properties, clearing mechanism, chemical interactions, solubility, temperature, and treatment duration. Physical characterization by a few labs including work from the authors provides evidence that decellularization protocols should be tailored to specific research questions. Physical characterization beyond histology and immunohistochemistry of the decellularized matrix (dECM) extends evaluation of retained functional features of the original tissue. We direct our attention to current technologies that can be employed for structure function analysis of dECM using underutilized tools such as atomic force microscopy (AFM), cryogenic electron microscopy (cryo-EM), dynamic mechanical analysis (DMA), Fourier-transform infrared spectroscopy (FTIR), mass spectrometry, and rheometry. Structural imaging and mechanical functional testing combined with high-throughput molecular analyses opens a new approach for a deeper appreciation of how cellular behavior is influenced by the isolated microenvironment (specifically dECM). Additionally, the impact of these features with different decellularization techniques and generation of synthetic material scaffolds with desired attributes are informed. Ultimately, this mechanical profiling provides a new dimension to our understanding of decellularized matrix and its role in new applications.

scholarly journals Cell-derived Extracellular Matrix as maintaining Biomaterial for adipogenic differentiation

2020 ◽  
Vol 6 (3) ◽  
pp. 410-413
Author(s):  
Petra J. Kluger ◽  
Svenja Nellinger ◽  
Simon Heine ◽  
Ann-Cathrin Volz
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Adipogenic Differentiation ◽  
Cellular Activity ◽  
Adipose Tissue Engineering ◽  
Physical Support ◽  
Supporting Effect ◽  
Brdu Assay ◽  
The Impact

AbstractThe extracellular matrix (ECM) naturally surrounds cells in humans, and therefore represents the ideal biomaterial for tissue engineering. ECM from different tissues exhibit different composition and physical characteristics. Thus, ECM provides not only physical support but also contains crucial biochemical signals that influence cell adhesion, morphology, proliferation and differentiation. Next to native ECM from mature tissue, ECM can also be obtained from the in vitro culture of cells. In this study, we aimed to highlight the supporting effect of cell-derived- ECM (cdECM) on adipogenic differentiation. ASCs were seeded on top of cdECM from ASCs (scdECM) or pre-adipocytes (acdECM). The impact of ECM on cellular activity was determined by LDH assay, WST I assay and BrdU assay. A supporting effect of cdECM substrates on adipogenic differentiation was determined by oil red O staining and subsequent quantification. Results revealed no effect of cdECM substrates on cellular activity. Regarding adipogenic differentiation a supporting effect of cdECM substrates was obtained compared to control. With these results, we confirm cdECM as a promising biomaterial for adipose tissue engineering.

Bioprinted Nanoparticles for Tissue Engineering Applications

2009 ◽  
Author(s):  
Kivilcim Buyukhatipoglu ◽  
Robert Chang ◽  
Wei Sun ◽  
Alisa Morss Clyne
Keyword(s):  
Tissue Engineering ◽  
Tissue Growth ◽  
Bioactive Components ◽  
Engineering Applications ◽  
Tissue Properties ◽  
Native Tissue ◽  
3D Architecture

Tissue engineering may require precise patterning of cells and bioactive components to recreate the complex, 3D architecture of native tissue. However, it is difficult to image and track cells and bioactive factors once they are incorporated into the tissue engineered construct. These bioactive factors and cells may also need to be moved during tissue growth in vitro or after implantation in vivo to achieve the desired tissue properties, or they may need to be removed entirely prior to implantation for biosafety concerns.

scholarly journals Cerebrospinal Fluid Pulsation Stress Promotes the Angiogenesis of Tissue-Engineered Laminae

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Linli Li ◽  
Yiqun He ◽  
Han Tang ◽  
Wei Mao ◽  
Haofei Ni ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cerebrospinal Fluid ◽  
Engineering Technology ◽  
Expression Levels ◽  
Fluid Pulsation ◽  
Endothelial Growth Factor ◽  
The Impact

Background. Angiogenesis is a prerequisite step to achieve the success of bone regeneration by tissue engineering technology. Previous studies have shown the role of cerebrospinal fluid pulsation (CSFP) stress in the reconstruction of tissue-engineered laminae. In this study, we investigated the role of CSFP stress in the angiogenesis of tissue-engineered laminae. Methods. For the in vitro study, a CSFP bioreactor was used to investigate the impact of CSFP stress on the osteogenic mesenchymal stem cells (MSCs). For the in vivo study, forty-eight New Zealand rabbits were randomly divided into the CSFP group and the Non-CSFP group. Tissue-engineered laminae (TEL) was made by hydroxyapatite-collagen I scaffold and osteogenic MSCs and then implanted into the lamina defect in the two groups. The angiogenic and osteogenic abilities of newborn laminae were examined with histological staining, qRT-PCR, and radiological analysis. Results. The in vitro study showed that CSFP stress could promote the vascular endothelial growth factor A (VEGF-A) expression levels of osteogenic MSCs. In the animal study, the expression levels of angiogenic markers in the CSFP group were higher than those in the Non-CSFP group; moreover, in the CSFP group, their expression levels on the dura mater surface, which are closer to the CSFP stress stimulation, were also higher than those on the paraspinal muscle surface. The expression levels of osteogenic markers in the CSFP group were also higher than those in the Non-CSFP group. Conclusion. CSFP stress could promote the angiogenic ability of osteogenic MSCs and thus promote the angiogenesis of tissue-engineered laminae. The pretreatment of osteogenic MSC with a CSFP bioreactor may have important implications for vertebral lamina reconstruction with a tissue engineering technique.

scholarly journals Synthesis and Properties of Flexible Polyurethane Using Ferric Catalyst for Hypopharyngeal Tissue Engineering

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Zhisen Shen ◽  
Jian Wang ◽  
Dakai Lu ◽  
Qun Li ◽  
Chongchang Zhou ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Matrix Material ◽  
Biological Properties ◽  
Hexamethylene Diisocyanate ◽  
Engineering Research ◽  
Biodegradable Polyurethane ◽  
Flexible Polyurethane ◽  
Hydrophilic Polyurethane

Biodegradable polyurethane is an ideal candidate material to fabricate tissue engineered hypopharynx from its good mechanical properties and biodegradability. We thus synthesized a hydrophilic polyurethane via reactions among polyethylene glycol (PEG), e-caprolactone (e-CL) and hexamethylene diisocyanate (HDI), and thrihydroxymethyl propane (TMP). The product possessed a fast degradability due to its good wettability and good mechanical parameters with the elongations at break (137 ± 10%) and tensile strength (4.73 ± 0.46 MPa), which will make it a good matrix material for soft tissue like hypopharynx. Its biological properties were evaluated viain vitroandin vivotests. The results showed that this hydrophilic polyurethane material can support hypopharyngeal fibroblast growth and owned good degradability and low inflammatory reaction in subcutaneous implantation. It will be proposed as the scaffold for hypopharyngeal tissue engineering research in our future study.

scholarly journals Three-dimensional printed bone scaffolds: The role of nano/micro-hydroxyapatite particles on the adhesion and differentiation of human mesenchymal stem cells

2017 ◽  
Vol 231 (6) ◽  
pp. 555-564 ◽  
Author(s):  
Marco Domingos ◽  
Antonio Gloria ◽  
Jorge Coelho ◽  
Paulo Bartolo ◽  
Joaquim Ciurana
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Calcium Phosphates ◽  
Three Dimensional ◽  
Human Mesenchymal Stem Cells ◽  
Mechanical Tests ◽  
Engineering Research

Bone tissue engineering is strongly dependent on the use of three-dimensional scaffolds that can act as templates to accommodate cells and support tissue ingrowth. Despite its wide application in tissue engineering research, polycaprolactone presents a very limited ability to induce adhesion, proliferation and osteogenic cell differentiation. To overcome some of these limitations, different calcium phosphates, such as hydroxyapatite and tricalcium phosphate, have been employed with relative success. This work investigates the influence of nano-hydroxyapatite and micro-hydroxyapatite (nHA and mHA, respectively) particles on the in vitro biomechanical performance of polycaprolactone/hydroxyapatite scaffolds. Morphological analysis performed with scanning electron microscopy allowed us to confirm the production of polycaprolactone/hydroxyapatite constructs with square interconnected pores of approximately 350 µm and to assess the distribution of hydroxyapatite particles within the polymer matrix. Compression mechanical tests showed an increase in polycaprolactone compressive modulus ( E) from 105.5 ± 11.2 to 138.8 ± 12.9 MPa (PCL_nHA) and 217.2 ± 21.8 MPa (PCL_mHA). In comparison to PCL_mHA scaffolds, the addition of nano-hydroxyapatite enhanced the adhesion and viability of human mesenchymal stem cells as confirmed by Alamar Blue assay. In addition, after 14 days of incubation, PCL_nHA scaffolds showed higher levels of alkaline phosphatase activity compared to polycaprolactone or PCL_mHA structures.

scholarly journals Engineered Hypopharynx from Coculture of Epithelial Cells and Fibroblasts Using Poly(ester urethane) as Substratum

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Zhisen Shen ◽  
Jingjing Chen ◽  
Cheng Kang ◽  
Changfeng Gong ◽  
Yabin Zhu
Keyword(s):  
Tissue Engineering ◽  
Epithelial Cells ◽  
Silk Fibroin ◽  
Porous Scaffold ◽  
Engineering Research ◽  
Polymeric Scaffolds ◽  
Cell Tissue ◽  
Silkworm Cocoon

Porous polymeric scaffolds have been much investigated and applied in the field of tissue engineering research. Poly(ester urethane) (PEU) scaffolds, comprising pores of 1–20 μm in diameter on one surface and ≥200 μm on the opposite surface and in bulk, were fabricated using phase separation method for hypopharyngeal tissue engineering. The scaffolds were grafted with silk fibroin (SF) generated from natural silkworm cocoon to enhance the scaffold’s hydrophilicity and further improve cytocompatibility to both primary epithelial cells (ECs) and fibroblasts of human hypopharynx tissue. Coculture of ECs and fibroblasts was conducted on the SF-grafted PEU scaffold (PEU-SF) to evaluate itsin vitrocytocompatibility. After co-culture for 14 days, ECs were lined on the scaffold surface while fibroblasts were distributed in scaffold bulk. The results ofin vivoinvestigation showed that PEU porous scaffold possessed good biocompatibility after it was grafted by silk fibroin. SF grafting improved the cell/tissue infiltration into scaffold bulk. Thus, PEU-SF porous scaffold is expected to be a good candidate to support the hypopharynx regeneration.

Cardiovascular Tissue Engineering: A New Laminar Flow Chamber for In Vitro Improvement of Mechanical Tissue Properties

ASAIO Journal ◽  
2002 ◽  
Vol 48 (1) ◽  
pp. 8-11 ◽  
Author(s):  
Stefan Jockenhoevel ◽  
Gregor Zund ◽  
Simon P. Hoerstrup ◽  
Andrea Schnell ◽  
Marco Turina
Keyword(s):  
Tissue Engineering ◽  
Laminar Flow ◽  
Flow Chamber ◽  
Tissue Properties ◽  
Cardiovascular Tissue ◽  
Cardiovascular Tissue Engineering ◽  
Mechanical Tissue

scholarly journals Alternating Air-Medium Exposure in Rotating Bioreactors Optimizes Cell Metabolism in 3D Novel Tubular Scaffold Polyurethane Foams

2017 ◽  
Vol 15 (2) ◽  
pp. 122-132 ◽  
Author(s):  
Claudia Tresoldi ◽  
Ilaria Stefani ◽  
Gaia Ferracci ◽  
Serena Bertoldi ◽  
Alessandro F. Pellegata ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Single Phase ◽  
Polyurethane Foams ◽  
Dynamic Conditions ◽  
Matrix Deposition ◽  
Dynamic Culture ◽  
Double Phase ◽  
Tubular Scaffold ◽  
The Impact

Background In vitro dynamic culture conditions play a pivotal role in developing engineered tissue grafts, where the supply of oxygen and nutrients, and waste removal must be permitted within construct thickness. For tubular scaffolds, mass transfer is enhanced by introducing a convective flow through rotating bioreactors with positive effects on cell proliferation, scaffold colonization and extracellular matrix deposition. We characterized a novel polyurethane-based tubular scaffold and investigated the impact of 3 different culture configurations over cell behavior: dynamic (i) single-phase (medium) rotation and (ii) double-phase exposure (medium-air) rotation; static (iii) single-phase static culture as control. Methods A new mixture of polyol was tested to create polyurethane foams (PUFs) as 3D scaffold for tissue engineering. The structure obtained was morphologically and mechanically analyzed tested. Murine fibroblasts were externally seeded on the novel porous PUF scaffold, and cultured under different dynamic conditions. Viability assay, DNA quantification, SEM and histological analyses were performed at different time points. Results The PUF scaffold presented interesting mechanical properties and morphology adequate to promote cell adhesion, highlighting its potential for tissue engineering purposes. Results showed that constructs under dynamic conditions contain enhanced viability and cell number, exponentially increased for double-phase rotation; under this last configuration, cells uniformly covered both the external surface and the lumen. Conclusions The developed 3D structure combined with the alternated exposure to air and medium provided the optimal in vitro biochemical conditioning with adequate nutrient supply for cells. The results highlight a valuable combination of material and dynamic culture for tissue engineering applications.

scholarly journals Function Analysis of MBF1, a Factor Involved in the Response to Amino Acid Starvation and Virulence in Candida albicans

2021 ◽  
Vol 2 ◽  
Author(s):  
Sara Amorim-Vaz ◽  
Alix T. Coste ◽  
Van Du T. Tran ◽  
Marco Pagni ◽  
Dominique Sanglard
Keyword(s):  
Gene Expression ◽  
Candida Albicans ◽  
Amino Acid ◽  
Fungal Pathogens ◽  
Galleria Mellonella ◽  
Function Analysis ◽  
Amino Acid Starvation ◽  
Infection Model ◽  
The Impact

Candida albicans is a commensal of human mucosae, but also one of the most common fungal pathogens of humans. Systemic infections caused by this fungus, mostly affecting immunocompromised patients, are associated to fatality rates as high as 50% despite the available treatments. In order to improve this situation, it is necessary to fully understand how C. albicans is able to cause disease and how it copes with the host defenses. Our previous studies have revealed the importance of the C. albicans gene MBF1 in virulence and ability to colonize internal organs of mammalian and insect hosts. MBF1 encodes a putative transcriptional regulator, and as such it likely has an impact in the regulation of C. albicans gene expression during host infection. Here, recent advances in RNA-seq technologies were used to obtain a detailed analysis of the impact of MBF1 on C. albicans gene expression both in vitro and during infection. MBF1 was involved in the regulation of several genes with a role in glycolysis and response to stress, particularly to nutritional stress. We also investigated whether an interaction existed between MBF1 and GCN4, a master regulator of response to starvation, and found that both genes were needed for resistance to amino acid starvation, suggesting some level of interaction between the two. Reinforcing this idea, we showed that the proteins encoded by both genes could interact. Consistent with the role of MBF1 in virulence, we also established that GCN4 was necessary for virulence in the mouse model of systemic infection as well as in the Galleria mellonella infection model.

scholarly journals Review of additive manufactured tissue engineering scaffolds: relationship between geometry and performance

2018 ◽  
Vol 6 ◽  
Author(s):  
Andrew Gleadall ◽  
Dafydd Visscher ◽  
Jing Yang ◽  
Daniel Thomas ◽  
Joel Segal
Keyword(s):  
Tissue Engineering ◽  
Additive Manufacturing ◽  
Free Form ◽  
Additive Manufacture ◽  
Scaffold Design ◽  
Structural Geometry ◽  
Engineering Research ◽  
Wide Range ◽  
Biological Performance ◽  
The Impact

Abstract Material extrusion additive manufacturing has rapidly grown in use for tissue engineering research since its adoption in the year 2000. It has enabled researchers to produce scaffolds with intricate porous geometries that were not feasible with traditional manufacturing processes. Researchers can control the structural geometry through a wide range of customisable printing parameters and design choices including material, print path, temperature, and many other process parameters. Currently, the impact of these choices is not fully understood. This review focuses on how the position and orientation of extruded filaments, which sometimes referred to as the print path, lay-down pattern, or simply “scaffold design”, affect scaffold properties and biological performance. By analysing trends across multiple studies, new understanding was developed on how filament position affects mechanical properties. Biological performance was also found to be affected by filament position, but a lack of consensus between studies indicates a need for further research and understanding. In most research studies, scaffold design was dictated by capabilities of additive manufacturing software rather than free-form design of structural geometry optimised for biological requirements. There is scope for much greater application of engineering innovation to additive manufacture novel geometries. To achieve this, better understanding of biological requirements is needed to enable the effective specification of ideal scaffold geometries.

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