Fabrication of Single and Multi-Layer Fibrous Biomaterial Scaffolds for Tissue Engineering

2008 ◽  
Author(s):  
Amrinder S. Nain ◽  
Eric Miller ◽  
Metin Sitti ◽  
Phil Campbell ◽  
Cristina Amon
Keyword(s):  
Tissue Engineering ◽  
Fiber Diameter ◽  
Living Cells ◽  
Cellular Adhesion ◽  
C2c12 Cells ◽  
Fiber Axis ◽  
Cellular Interactions ◽  
Fiber Spacing ◽  
Biomaterial Scaffolds ◽  
And Migration

For regenerative medicine applications, we need to expand our understanding of the mechanisms by which nature assembles and functionalizes specialized complex tissues to form a complete organism. The first step towards this goal involves understanding the underlying complex mechanisms of highly organized behavior spanning not only diverse scientific fields, but also nano, micro and macro length-scales. For example, an engineered fibrous biomaterial scaffold possessing the hierarchal spatial properties of a native extracellular matrix (ECM) can serve as a building block upon which living cells are seeded for repair or regeneration. The hierarchical nature of ECM along with the inherent topological constraints of fiber diameter, fiber spacing, multi-layer configurations provide different pathways for living cells to adapt and conform to the surrounding environment. Our previously developed Spinneret based Tunable Engineered Parameters (STEP) technique to deposit biomaterial scaffolds in aligned configurations has been used for the first time to deposit single and multi-layer biological scaffolds of fibrinogen. Fibrinogen is a very well established tissue engineering scaffold material, as it improves cellular interactions and allows scaffold remodeling compared to synthetic polymers. Current state-of-the-art fiber deposition techniques lack the ability to fabricate scaffolds of desired fiber dimensions and orientations and in this study we present fabrication and aligned deposition of fibrinogen fiber arrays with diameters ranging from sub-200 nm to sub-microns and several millimeters in length. The fabricated scaffolds are then cultured with pluripotent mouse C2C12 cells for seven days and cells on the scaffolds are observed to elongate resembling myotube morphology along the fiber axis, spread along intersecting layers and fuse into bundles at the macroscale. Additionally, we demonstrate the ability to deposit poly (lactic-co-glycolic acid) (PLGA), Polystyrene (PS) biomaterial scaffolds of different diameters to investigate the effects of topological variations on cellular adhesion, proliferation and migration. Previous studies have indicated cells making right angle transitions upon encountering perpendicular double layer fibers and cellular motion is thwarted in the vicinity of diverging fibers. Current ongoing studies are aimed at determining the effects of fiber diameter and fiber spacing on mouse C2C12 cellular adhesion and migration, which are envisioned to aid in the design of future scaffolds for tissue engineering possessing appropriate material and geometrical properties.

scholarly journals Trends in Articular Cartilage Tissue Engineering: 3D Mesenchymal Stem Cell Sheets as Candidates for Engineered Hyaline-Like Cartilage

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 643
Author(s):  
Hallie Thorp ◽  
Kyungsook Kim ◽  
Makoto Kondo ◽  
Travis Maak ◽  
David W. Grainger ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Hyaline Cartilage ◽  
Cell Sheet ◽  
Cellular Adhesion ◽  
Joint Disease ◽  
Cartilage Tissue ◽  
Cellular Interactions ◽  
Tissue Integration ◽  
Defect Sites

Articular cartilage defects represent an inciting factor for future osteoarthritis (OA) and degenerative joint disease progression. Despite multiple clinically available therapies that succeed in providing short term pain reduction and restoration of limited mobility, current treatments do not reliably regenerate native hyaline cartilage or halt cartilage degeneration at these defect sites. Novel therapeutics aimed at addressing limitations of current clinical cartilage regeneration therapies increasingly focus on allogeneic cells, specifically mesenchymal stem cells (MSCs), as potent, banked, and available cell sources that express chondrogenic lineage commitment capabilities. Innovative tissue engineering approaches employing allogeneic MSCs aim to develop three-dimensional (3D), chondrogenically differentiated constructs for direct and immediate replacement of hyaline cartilage, improve local site tissue integration, and optimize treatment outcomes. Among emerging tissue engineering technologies, advancements in cell sheet tissue engineering offer promising capabilities for achieving both in vitro hyaline-like differentiation and effective transplantation, based on controlled 3D cellular interactions and retained cellular adhesion molecules. This review focuses on 3D MSC-based tissue engineering approaches for fabricating “ready-to-use” hyaline-like cartilage constructs for future rapid in vivo regenerative cartilage therapies. We highlight current approaches and future directions regarding development of MSC-derived cartilage therapies, emphasizing cell sheet tissue engineering, with specific focus on regulating 3D cellular interactions for controlled chondrogenic differentiation and post-differentiation transplantation capabilities.

Quantitative Diameter Measurements of Chromatin and TMV Fibers in the Freeze-Dried State

1988 ◽  
Vol 46 ◽  
pp. 170-171
Author(s):  
B. D. Athey ◽  
A. L. Stout ◽  
M. F. Smith ◽  
J. P. Langmore
Keyword(s):  
Reliable Method ◽  
Fiber Diameter ◽  
Quantitative Agreement ◽  
One Dimension ◽  
Fiber Axis ◽  
Two Dimensional ◽  
Fiber Density ◽  
Variable Diameter ◽  
Freeze Dried ◽  
Expected Values

Although there is general agreement that Inactive chromosome fibers consist of helically packed nucleosomes, the pattern of packing is still undetermined. Only one of the proposed models, the crossed-linker model, predicts a variable diameter dependent on the length of DNA between nucleosomes. Measurements of the fiber diameter of negatively-stained and frozen- hydrated- chromatin from Thyone sperm (87bp linker) and Necturus erythrocytes (48bp linker) have been previously reported from this laboratory. We now introduce a more reliable method of measuring the diameters of electron images of fibrous objects. The procedure uses a modified version of the computer program TOTAL, which takes a two-dimensional projection of the fiber density (represented by the micrograph itself) and projects it down the fiber axis onto one dimension. We illustrate this method using high contrast, in-focus STEM images of TMV and chromatin from Thyone and Necturus. The measured diameters are in quantitative agreement with the expected values for the crossed-linker model for chromatin structure

Biomaterial scaffolds for tissue engineering

10.2741/e620 ◽  
2013 ◽  
Vol 5 (1) ◽  
pp. 341-360 ◽  
Author(s):  
Kajal K Mallick
Keyword(s):  
Tissue Engineering ◽  
Biomaterial Scaffolds

scholarly journals Light Cross-Linkable Marine Collagen for Coaxial Printing of a 3D Model of Neuromuscular Junction Formation

Biomedicines ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 16
Author(s):  
Borja Sanz ◽  
Ane Albillos Sanchez ◽  
Bonnie Tangey ◽  
Kerry Gilmore ◽  
Zhilian Yue ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Neuromuscular Junction ◽  
Disease Transmission ◽  
Low Cost ◽  
Attractive Alternative ◽  
Waste Products ◽  
Environmentally Sustainable ◽  
Zoonotic Disease Transmission ◽  
And Migration

Collagen is a major component of the extracellular matrix (ECM) that modulates cell adhesion, growth, and migration, and has been utilised in tissue engineering applications. However, the common terrestrial sources of collagen carry the risk of zoonotic disease transmission and there are religious barriers to the use of bovine and porcine products in many cultures. Marine based collagens offer an attractive alternative and have so far been under-utilized for use as biomaterials for tissue engineering. Marine collagen can be extracted from fish waste products, therefore industry by-products offer an economical and environmentally sustainable source of collagen. In a handful of studies, marine collagen has successfully been methacrylated to form collagen methacrylate (ColMA). Our work included the extraction, characterization and methacrylation of Red Snapper collagen, optimisation of conditions for neural cell seeding and encapsulation using the unmodified collagen, thermally cross-linked, and the methacrylated collagen with UV-induced cross-linking. Finally, the 3D co-axial printing of neural and skeletal muscle cell cultures as a model for neuromuscular junction (NMJ) formation was investigated. Overall, the results of this study show great potential for a novel NMJ in vitro 3D bioprinted model that, with further development, could provide a low-cost, customizable, scalable and quick-to-print platform for drug screening and to study neuromuscular junction physiology and pathogenesis.

scholarly journals XPS Modeling of Immobilized Recombinant Angiogenin and Apoliprotein A1 on Biodegradable Nanofibers

Nanomaterials ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 879
Author(s):  
Anton Manakhov ◽  
Elizaveta Permyakova ◽  
Sergey Ershov ◽  
Svetlana Miroshnichenko ◽  
Mariya Pykhtina ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Growth Factor ◽  
Photoelectron Spectroscopy ◽  
Antioxidant Activities ◽  
Fluorescent Microscopy ◽  
X Ray ◽  
Vascular Growth ◽  
Vascular Growth Factor ◽  
And Migration ◽  
Protein Density

The immobilization of viable proteins is an important step in engineering efficient scaffolds for regenerative medicine. For example, angiogenin, a vascular growth factor, can be considered a neurotrophic factor, influencing the neurogenesis, viability, and migration of neurons. Angiogenin shows an exceptional combination of angiogenic, neurotrophic, neuroprotective, antibacterial, and antioxidant activities. Therefore, this protein is a promising molecule that can be immobilized on carriers used for tissue engineering, particularly for diseases that are complicated by neurotrophic and vascular disorders. Another highly important and viable protein is apoliprotein A1. Nevertheless, the immobilization of these proteins onto promising biodegradable nanofibers has not been tested before. In this work, we carefully studied the immobilization of human recombinant angiogenin and apoliprotein A1 onto plasma-coated nanofibers. We developed a new methodology for the quantification of the protein density of these proteins using X-ray photoelectron spectroscopy (XPS) and modeled the XPS data for angiogenin and apoliprotein A1 (Apo-A1). These findings were also confirmed by the analysis of immobilized Apo-A1 using fluorescent microscopy. The presented methodology was validated by the analysis of fibronectin on the surface of plasma-coated poly(ε-caprolactone) (PCL) nanofibers. This methodology can be expanded for other proteins and it should help to quantify the density of proteins on surfaces using routine XPS data treatment.

Preparation of Fibrous Scaffolds Containing Calcium and Silicon Species

2011 ◽  
Vol 493-494 ◽  
pp. 840-843
Author(s):  
Akiko Obata ◽  
Hiroki Ozasa ◽  
Julian R. Jones ◽  
Toshihiro Kasuga
Keyword(s):  
Tissue Engineering ◽  
Hybrid Materials ◽  
Fiber Diameter ◽  
Fiber Spinning ◽  
High Porosity ◽  
Cotton Wool ◽  
Tissue Engineering Scaffolds ◽  
Large Defect ◽  
Nanofibrous Structure ◽  
Versatile Technique

Materials for bone defect filling should have 3D macroporous structure and be flexible to be packed into complex defects with limited entrance space. Tissue engineering scaffolds should also mimic the structure and morphology of the host tissue. Electrospinning is a versatile technique to produce materials with micro/nanofibrous structure, large surface area and high porosity. Electrospun materials are very promising for tissue engineering due to the possibility of mimicking the fibrous structure of natural extra cellular matrix (ECM). Siloxane-containing vaterite (SiV)/poly (L-lactic acid) (PLLA) hybrids (SiPVH) with controlled silicate and calcium ions releasing ability has been produced in our group. They have also demonstrated good cell infiltration into the electrospun hybrid materials that had fiber diameters greater than 10 μm. However, these electrospun hybrid materials were planar (2D) and are not suitable for large defect regeneration. In this work, the development of a fabrication technique for the production of 3D cotton wool-like structures with fiber diameter in the range of 10 μm was performed. SiPVH cotton wool-like structure containing 0, 30 and 60 wt % SiV were prepared by blowing air in the direction perpendicular to fiber spinning. Si-vaterite particles and small pores were found on the surface of the fibers. The fiber diameter of the samples were found to be in the range of 10 ~ 20 μm. Stretch tests showed more than 50 % extension for the SiPVH cotton wool-like material containing 30 wt % SiV (SiPVH30). This extension was similar to that observed for the PLLA cotton wool-like material. The results suggest that the SiPVH30 cotton wool-like material are good candidates for bone tissue engineering scaffolds.

scholarly journals Binary Biocompatible CNC–Gelatine Hydrogel as 3D Scaffolds Suitable for Cell Culture Adhesion and Growth

Applied Nano ◽  
2021 ◽  
Vol 2 (2) ◽  
pp. 118-127
Author(s):  
Luca Zoia ◽  
Anna Binda ◽  
Laura Cipolla ◽  
Ilaria Rivolta ◽  
Barbara La Ferla
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
Cell Cultures ◽  
Cellulose Nanocrystals ◽  
Potential Application ◽  
Chemical Properties ◽  
Cellular Adhesion ◽  
Chemical Crosslinking ◽  
3D Scaffolds ◽  
Physical Chemical

Binary nano-biocomposite 3D scaffolds of cellulose nanocrystals (CNCs)—gelatine were fabricated without using chemical crosslinking additives. Controlled oxidative treatment allowed introducing carboxyl or carbonyl functionalities on the surface of CNCs responsible for the crosslinking of gelatine polymers. The obtained composites were characterized for their physical-chemical properties. Their biocompatibility towards different cell cultures was evaluated through MTT and LDH assays, cellular adhesion and proliferation experiments. Gelatine composites reinforced with carbonyl-modified CNCs showed the most performing swelling/degradation profile and the most promising adhesion and proliferation properties towards cell lines, suggesting their potential application in the field of tissue engineering.

scholarly journals Effects of a New Bioceramic Material on Human Apical Papilla Cells

2018 ◽  
Vol 9 (4) ◽  
pp. 74 ◽  
Author(s):  
Diana Sequeira ◽  
Catarina Seabra ◽  
Paulo Palma ◽  
Ana Cardoso ◽  
João Peça ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mineral Trioxide Aggregate ◽  
Cellular Adhesion ◽  
Cellular Migration ◽  
Cellular Viability ◽  
Migration Rates ◽  
Apical Papilla ◽  
And Migration ◽  
Proliferation And Migration

Background: The development of materials with bioregenerative properties is critically important for vital pulp therapies and regenerative endodontic procedures. The aim of this study was to evaluate the cytocompatibility and cytotoxicity of a new endodontic biomaterial, PulpGuard, in comparison with two other biomaterials widely used in endodontic procedures, ProRoot Mineral Trioxide Aggregate (MTA) and Biodentine. Methods: Apical papilla cells (APCs) were isolated from third molars with incomplete rhizogenesis from patients with orthodontic indication for dental extraction. Cultured APCs were incubated for 24, 48, or 72 h with different dilutions of eluates prepared from the three materials. Cellular viability, mobility, and proliferation were assessed in vitro using the Alamar Blue assay and a wound-healing test. The cells were also cultured in direct contact with the surface of each material. These were then analyzed via Scanning Electron Microscopy (SEM), and the surface chemical composition was determined by Energy-Dispersive Spectroscopy (EDS). Results: Cells incubated in the presence of eluates extracted from ProRoot MTA and PulpGuard presented rates of viability comparable to those of control cells; in contrast, undiluted Biodentine eluates induced a significant reduction of cellular viability. The wound-healing assay revealed that eluates from ProRoot MTA and PulpGuard allowed for unhindered cellular migration and proliferation. Cellular adhesion was observed on the surface of all materials tested. Consistent with their disclosed composition, EDS analysis found high relative abundance of calcium in Biodentine and ProRoot MTA and high abundance of silicon in PulpGuard. Significant amounts of zinc and calcium were also present in PulpGuard discs. Concerning solubility, Biodentine and ProRoot MTA presented mild weight loss after eluate extraction, while PulpGuard discs showed significant water uptake. Conclusions: PulpGuard displayed a good in vitro cytocompatibility profile and did not significantly affect the proliferation and migration rates of APCs. Cells cultured in the presence of PulpGuard eluates displayed a similar profile to those cultured with eluates from the widely used endodontic cement ProRoot MTA.

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