scholarly journals Micropatterning Decellularized ECM as a Bioactive Surface to Guide Cell Alignment, Proliferation, and Migration

2020 ◽  
Vol 7 (3) ◽  
pp. 102 ◽  
Author(s):  
Emily Cady ◽  
Jacob A. Orkwis ◽  
Rachel Weaver ◽  
Lia Conlin ◽  
Nicolas N. Madigan ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Soft Lithography ◽  
Cell Function ◽  
Control Cell ◽  
Three Dimensional ◽  
Protein Composition ◽  
Cell Alignment ◽  
Bioactive Component ◽  
And Migration ◽  
High Degree

Bioactive surfaces and materials have displayed great potential in a variety of tissue engineering applications but often struggle to completely emulate complex bodily systems. The extracellular matrix (ECM) is a crucial, bioactive component in all tissues and has recently been identified as a potential solution to be utilized in combination with biomaterials. In tissue engineering, the ECM can be utilized in a variety of applications by employing the biochemical and biomechanical cues that are crucial to regenerative processes. However, viable solutions for maintaining the dimensionality, spatial orientation, and protein composition of a naturally cell-secreted ECM remain challenging in tissue engineering. Therefore, this work used soft lithography to create micropatterned polydimethylsiloxane (PDMS) substrates of a three-dimensional nature to control cell adhesion and alignment. Cells aligned on the micropatterned PDMS, secreted and assembled an ECM, and were decellularized to produce an aligned matrix biomaterial. The cells seeded onto the decellularized, patterned ECM showed a high degree of alignment and migration along the patterns compared to controls. This work begins to lay the groundwork for elucidating the immense potential of a natural, cell-secreted ECM for directing cell function and offers further guidance for the incorporation of natural, bioactive components for emerging tissue engineering technologies.

scholarly journals A Comparative Study of Rat Lung Decellularization by Chemical Detergents for Lung Tissue Engineering

2017 ◽  
Vol 5 (7) ◽  
pp. 859-865 ◽  
Author(s):  
Hamid Tebyanian ◽  
Ali Karami ◽  
Ebrahim Motavallian ◽  
Jafar Aslani ◽  
Ali Samadikuchaksaraei ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Comparative Study ◽  
Lung Tissue ◽  
Three Dimensional ◽  
Protein Composition ◽  
Nuclear Material ◽  
Detergent Concentration ◽  
Lung Tissue Engineering ◽  
And Migration ◽  
Triton X

BACKGROUND: Lung disease is the most common cause of death in the world. The last stage of pulmonary diseases is lung transplantation. Limitation and shortage of donor organs cause to appear tissue engineering field. Decellularization is a hope for producing intact ECM in the development of engineered organs.AIM: The goal of the decellularization process is to remove cellular and nuclear material while retaining lung three-dimensional and molecular proteins. Different concentration of detergents was used for finding the best approach in lung decellularization.MATERIAL AND METHODS: In this study, three-time approaches (24, 48 and 96 h) with four detergents (CHAPS, SDS, SDC and Triton X-100) were used for decellularizing rat lungs for maintaining of three-dimensional lung architecture and ECM protein composition which have significant roles in differentiation and migration of stem cells This comparative study determined that variable decellularization approaches can cause significantly different effects on decellularized lungs.RESULTS: Results showed that destruction was increased with increasing the detergent concentration. Single detergent showed a significant reduction in maintaining of three-dimensional of lung and ECM proteins (Collagen and Elastin). But, the best methods were mixed detergents of SDC and CHAPS in low concentration in 48 and 96 h decellularization.CONCLUSION: Decellularized lung tissue can be used in the laboratory to study various aspects of pulmonary biology and physiology and also, these results can be used in the continued improvement of engineered lung tissue.

Histological study of an experimentally constructed decellularized rat lung as a model of whole organ three-dimensional natural scaffold

QJM ◽  
2021 ◽  
Vol 114 (Supplement_1) ◽  
Author(s):  
Marium Romany Abdelsayed ◽  
Suzi Sobhy Atalla ◽  
Gehan Khalaf Megahed ◽  
Asmaa Abd El-Monem Abo Zeid
Keyword(s):  
Tissue Engineering ◽  
Histological Study ◽  
Three Dimensional ◽  
Nuclear Material ◽  
Biological Properties ◽  
Basement Membranes ◽  
Elastic Fibers ◽  
Scientific Exploration ◽  
And Migration ◽  
End Stage

Abstract Introduction With the increase of end stage lung diseases and the great problems facing lung transplantation tissue engineering become a promising solution. The first step in lung engineering is to obtain a 3D Extracellular matrix lung scaffold via decellularization. Decellularization aims to remove cells from tissue ultrastructure while preserving the mechanical and biological properties of the tissue. Intact ECM provides critical cues for differentiation and migration of cells that are seeded onto the organ scaffold. Objectives This study aimed to obtain an intact and well-preserved ECM lung scaffold by decellularization of rat lungs. Methods Decellularization of lungs of ten Wistar rats was achieved by perfusing detergents through the pulmonary artery. The resultant scaffolds were fixed and analyzed histologically. Results It was found that the decellularization process effectively removed the cellular and nuclear material while retaining native the 3D ECM of lung tissue. The architecture of the collagen and elastic fibers networks were preserved as comparable to the native lungs. Furthermore, the basement membranes of the bronchiolar and interalveolar septa were intact. Conclusions This methodology is expected to allow decellularization of human lung tissues and permits future scientific exploration in tissue engineering.

Micro-Particle Fabrication for Tissue Engineering Applications Using Rapid Prototyping and Soft Lithography Principles

2005 ◽  
Author(s):  
M. Wettergreen ◽  
J. Scheffe ◽  
A. G. Mikos ◽  
M. A. K. Liebschner
Keyword(s):  
Tissue Engineering ◽  
Rapid Prototyping ◽  
Soft Lithography ◽  
Porous Scaffold ◽  
Three Dimensional ◽  
Volume Ratio ◽  
Engineering Applications ◽  
Three Dimensional Printing ◽  
Micro Particles ◽  
Multiple Materials

The goal of this study was to develop an efficient and repeatable process for fabrication of micro-particles from multiple materials using rapid prototyping and soft lithography. Phase change three-dimensional printing was used to create masters for PDMS molds. A photocrosslinkable polymer was then delivered into these molds to fabricate geometrically complex three-dimensional micro-particles. This repeatable process has demonstrated the ability to generate micro-particles with greater than 95% repeatability with complete pattern transfer. This process was illustrated for three shapes based on the extrusion of two-dimensional shapes. These particles will allow for tailoring of the pore shapes within a porous scaffold utilized in tissue engineering applications. In addition, the different shapes may allow control of drug release by varying the surface to volume ratio, which could modulate drug delivery. While soft lithography is currently used with photolithography, its high precision is offset by high cost of production. The employment of rapid prototyping to a specific resolution offers a much less expensive alternative with increased throughput due to the speed of current rapid prototyping systems.

Nanofibrillar Cellulose as an Enzymatically and Flow Driven Degradable Scaffold for Three-Dimensional Tissue Engineering

2019 ◽  
Vol 2 (4) ◽  
Author(s):  
Melanie Krüger ◽  
Bart Spee ◽  
Andreas Walther ◽  
Laura De Laporte ◽  
Linda M. Kock
Keyword(s):  
Tissue Engineering ◽  
Cellulose Nanofibrils ◽  
Three Dimensional ◽  
Shear Thinning ◽  
Scaffold Material ◽  
Nanofibrillar Cellulose ◽  
And Migration ◽  
Fibroblast Spheroids ◽  
Static Conditions ◽  
Tissue Production

Abstract Nanofibrillar cellulose as a naturally biocompatible scaffold material is very promising for tissue engineering. It is shear thinning but has the downside of not being degradable in animals, it can only be degraded by cellulase enzymes. In this study, a newly developed bioreactor was used to culture fibroblast spheroids under flow conditions inside nanocellulose hydrogels with and without the presence of cellulase. The aim was to control the tissue size and ideally find a match between degradation and tissue formation within this promising material. Both the concentration of cellulase and the flow rate were varied and their influence on the activity and growth of fibroblast clusters was assessed. Cluster diameters, degradation, metabolic activity, and tissue production increase with higher cellulase concentration, although concentrations above 1 g/l does not have an additional benefit. Flow leads to more viable cells, more proliferation and migration, leading to overall larger tissue constructs compared to static conditions. This is most likely due to the shear thinning effect of flow on cellulose nanofibrils (CNFs) in addition to the increased nutrient supply through perfusion. At a constant cellulase concentration of 1 g/l, a flow of 2 ml/min proved to be optimal for tissue production. Therefore, degradation in combination with flow leads to more effective tissue production in CNF hydrogels, which is a very potent scaffold material for tissue engineering.

scholarly journals Collective cell traction force analysis on aligned smooth muscle cell sheet between three-dimensional microwalls

2014 ◽  
Vol 4 (2) ◽  
pp. 20130056 ◽  
Author(s):  
Ying Zhang ◽  
Soon Seng Ng ◽  
Yilei Wang ◽  
Huixing Feng ◽  
Wei Ning Chen ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Smooth Muscle ◽  
Blood Vessels ◽  
Cell Sheet ◽  
Adhesion Formation ◽  
Three Dimensional ◽  
Traction Force ◽  
Cytoskeletal Proteins ◽  
Cell Alignment ◽  
Biomaterial Scaffold

During the past two decades, novel biomaterial scaffold for cell attachment and culture has been developed for applications in tissue engineering, biosensing and regeneration medicine. Tissue engineering of blood vessels remains a challenge owing to the complex three-layer histology involved. In order to engineer functional blood vessels, it is essential to recapitulate the characteristics of vascular smooth muscle cells (SMCs) inside the tunica media, which is known to be critical for vasoconstriction and vasodilation of the circulatory system. Until now, there has been a lack of understanding on the mechanotransduction of the SMC layer during the transformation from viable synthetic to quiescent contractile phenotypes. In this study, microfabricated arrays of discontinuous microwalls coated with fluorescence microbeads were developed to probe the mechanotransduction of the SMC layer. First, the system was exploited for stimulating the formation of a highly aligned orientation of SMCs in native tunica medium. Second, atomic force microscopy in combination with regression analysis was applied to measure the elastic modulus of a polyacrylamide gel layer coated on the discontinuous microwall arrays. Third, the conventional traction force assay for single cell measurement was extended for applications in three-dimensional cell aggregates. Then, the biophysical effects of discontinuous microwalls on the mechanotransduction of the SMC layer undergoing cell alignment were probed. Generally, the cooperative multiple cell–cell and cell–microwall interactions were accessed quantitatively by the newly developed assay with the aid of finite-element modelling. The results show that the traction forces of highly aligned cells lying in the middle region between two opposing microwalls were significantly lower than those lying adjacent to the microwalls. Moreover, the spatial distributions of Von Mises stress during the cell alignment process were dependent on the collective cell layer orientation. Immunostaining of the SMC sheet further demonstrated that the collective mechanotransduction induced by three-dimensional topographic cues was correlated with the reduction of actin and vinculin expression. In addition, the online two-dimensional LC–MS/MS analysis verified the modulation of focal adhesion formation under the influence of microwalls through the regulation in the expression of three key cytoskeletal proteins.

scholarly journals Microvascular fragment spheroids: Three-dimensional vascularization units for tissue engineering and regeneration

2021 ◽  
Vol 12 ◽  
pp. 204173142110355
Author(s):  
Lisa Nalbach ◽  
Danièle Müller ◽  
Selina Wrublewsky ◽  
Wolfgang Metzger ◽  
Michael D Menger ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cell Function ◽  
Smooth Muscle Actin ◽  
Three Dimensional ◽  
Stromal Vascular Fraction ◽  
Muscle Actin ◽  
Tissue Constructs ◽  
First Time ◽  
Tissue Defects

Adipose tissue-derived microvascular fragments (MVF) serve as vascularization units in tissue engineering and regenerative medicine. Because a three-dimensional cellular arrangement has been shown to improve cell function, we herein generated for the first time MVF spheroids to investigate whether this further increases their vascularization potential. These spheroids exhibited a morphology, size, and viability comparable to that of previously introduced stromal vascular fraction (SVF) spheroids. However, MVF spheroids contained a significantly higher number of CD31-positive endothelial cells and α-smooth muscle actin (SMA)-positive perivascular cells, resulting in an enhanced angiogenic sprouting activity. Accordingly, they also exhibited an improved in vivo vascularization and engraftment after transplantation into mouse dorsal skinfold chambers. These findings indicate that MVF spheroids are superior to SVF spheroids and, thus, may be highly suitable to improve the vascularization of tissue defects and implanted tissue constructs.

scholarly journals Innovative Strategies in Tendon Tissue Engineering

Pharmaceutics ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 89
Author(s):  
Eleonora Bianchi ◽  
Marco Ruggeri ◽  
Silvia Rossi ◽  
Barbara Vigani ◽  
Dalila Miele ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Soft Lithography ◽  
Sports Injuries ◽  
Therapeutic Potential ◽  
Tendon Repair ◽  
Three Dimensional ◽  
Incidence Rates ◽  
Innovative Strategies ◽  
Tendon Tissue Engineering ◽  
Manufacturing Techniques

The tendon is a highly aligned connective tissue that transmits force from muscle to bone. Each year, more than 32 million tendon injuries have been reported, in fact, tendinopathies represent at least 50% of all sports injuries, and their incidence rates have increased in recent decades due to the aging population. Current clinical grafts used in tendon treatment are subject to several restrictions and there is a significant demand for alternative engineered tissue. For this reason, innovative strategies need to be explored. Tendon replacement and regeneration are complex since scaffolds need to guarantee an adequate hierarchical structured morphology and mechanical properties to stand the load. Moreover, to guide cell proliferation and growth, scaffolds should provide a fibrous network that mimics the collagen arrangement of the extracellular matrix in the tendons. This review focuses on tendon repair and regeneration. Particular attention has been devoted to the innovative approaches in tissue engineering. Advanced manufacturing techniques, such as electrospinning, soft lithography, and three-dimensional (3D) printing, have been described. Furthermore, biological augmentation has been considered, as an emerging strategy with great therapeutic potential.

HYDROXYAPATITE, ALGINATE, AND CHITOSAN FOR BONE SCAFFOLDS: SPECTROSCOPY STUDY

2016 ◽  
Vol 19 (2) ◽  
pp. 93-100
Author(s):  
Lalita El Milla
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Functional Groups ◽  
Bone Growth ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Tissue Engineering Scaffolds ◽  
Hydroxyapatite Powder ◽  
Materials Used

Scaffolds is three dimensional structure that serves as a framework for bone growth. Natural materials are often used in synthesis of bone tissue engineering scaffolds with respect to compliance with the content of the human body. Among the materials used to make scafffold was hydroxyapatite, alginate and chitosan. Hydroxyapatite powder obtained by mixing phosphoric acid and calcium hydroxide, alginate powders extracted from brown algae and chitosan powder acetylated from crab. The purpose of this study was to examine the functional groups of hydroxyapatite, alginate and chitosan. The method used in this study was laboratory experimental using Fourier Transform Infrared (FTIR) spectroscopy for hydroxyapatite, alginate and chitosan powders. The results indicated the presence of functional groups PO43-, O-H and CO32- in hydroxyapatite. In alginate there were O-H, C=O, COOH and C-O-C functional groups, whereas in chitosan there were O-H, N-H, C=O, C-N, and C-O-C. It was concluded that the third material containing functional groups as found in humans that correspond to the scaffolds material in bone tissue engineering.

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