Electrospun fibers/branched-clusters as building units for tissue engineering

2017 ◽  
Vol 1 (1) ◽  
Author(s):  
Benjamin A. Minden-Birkenmaier ◽  
Gretchen S. Selders ◽  
Kasyap Cherukuri ◽  
Gary L. Bowlin
Keyword(s):  
Tissue Engineering ◽  
Tissue Regeneration ◽  
Fiber Diameter ◽  
Drug Loading ◽  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
Polymer Composition ◽  
Electrospun Fibers ◽  
Branch Lengths ◽  
Normal Human

AbstractAlthough electrospun templates are effective at mimicking the extracellular matrix (ECM) of native tissue due to the tailorability of parameters such as fiber diameter, polymer composition, and drug loading, these templates are often limited with regards to cell infiltration and the tailorability of the microenvironments within the structures. Thus, there remains a need for a flexible threedimensional template system which could be combined with cell suspensions to promote three-dimensional tissue regeneration, and ultimately allow cells to freely reorganize and modify their microenvironment. In this study, a mincing process was designed and optimized to create mixtures of electrospun fibers/branched-clusters for use as fundamental tissue engineering building units. These fiber/branched-cluster elements were characterized with regards to fiber and branch lengths, and a method was optimized to combine them with normal human dermal fibroblasts (nHDFs) in culture to create interconnected template constructs. Sectioning and imaging of these constructs revealed cell/fiber integration as well as even cell distribution throughout the construct interior. These fiber/branched-cluster elements represent an innovative flexible tissue regeneration template system.

scholarly journals Additive Manufacturing of Biopolymers for Tissue Engineering and Regenerative Medicine: An Overview, Potential Applications, Advancements, and Trends

2021 ◽  
Vol 2021 ◽  
pp. 1-20 ◽  
Author(s):  
Dhinakaran Veeman ◽  
M. Swapna Sai ◽  
P. Sureshkumar ◽  
T. Jagadeesha ◽  
L. Natrayan ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Additive Manufacturing ◽  
3D Printing ◽  
Regenerative Medicine ◽  
Tissue Regeneration ◽  
Three Dimensional ◽  
Porous Scaffolds ◽  
Wide Range ◽  
Potential Applications ◽  
Pharmaceutical Industries

As a technique of producing fabric engineering scaffolds, three-dimensional (3D) printing has tremendous possibilities. 3D printing applications are restricted to a wide range of biomaterials in the field of regenerative medicine and tissue engineering. Due to their biocompatibility, bioactiveness, and biodegradability, biopolymers such as collagen, alginate, silk fibroin, chitosan, alginate, cellulose, and starch are used in a variety of fields, including the food, biomedical, regeneration, agriculture, packaging, and pharmaceutical industries. The benefits of producing 3D-printed scaffolds are many, including the capacity to produce complicated geometries, porosity, and multicell coculture and to take growth factors into account. In particular, the additional production of biopolymers offers new options to produce 3D structures and materials with specialised patterns and properties. In the realm of tissue engineering and regenerative medicine (TERM), important progress has been accomplished; now, several state-of-the-art techniques are used to produce porous scaffolds for organ or tissue regeneration to be suited for tissue technology. Natural biopolymeric materials are often better suited for designing and manufacturing healing equipment than temporary implants and tissue regeneration materials owing to its appropriate properties and biocompatibility. The review focuses on the additive manufacturing of biopolymers with significant changes, advancements, trends, and developments in regenerative medicine and tissue engineering with potential applications.

scholarly journals Validation of in vitro assays in three-dimensional human dermal constructs

2018 ◽  
Vol 41 (11) ◽  
pp. 779-788 ◽  
Author(s):  
Ayesha Idrees ◽  
Valeria Chiono ◽  
Gianluca Ciardelli ◽  
Siegfried Shah ◽  
Richard Viebahn ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
In Vitro Assays ◽  
Dimensional System ◽  
Type I ◽  
Human Dermal Fibroblasts ◽  
Cell Viability Assay ◽  
Normal Human ◽  
Normal Human Dermal Fibroblasts

Three-dimensional cell culture systems are urgently needed for cytocompatibility testing of biomaterials. This work aimed at the development of three-dimensional in vitro dermal skin models and their optimization for cytocompatibility evaluation. Initially “murine in vitro dermal construct” based on L929 cells was generated, leading to the development of “human in vitro dermal construct” consisting of normal human dermal fibroblasts in rat tail tendon collagen type I. To assess the viability of the cells, different assays CellTiter-Blue®, RealTime-Glo™ MT, and CellTiter-Glo® (Promega) were evaluated to optimize the best-suited assay to the respective cell type and three-dimensional system. Z-stack imaging (Live/Dead and Phalloidin/DAPI-Promokine) was performed to visualize normal human dermal fibroblasts inside matrix revealing filopodia-like morphology and a uniform distribution of normal human dermal fibroblasts in matrix. CellTiter-Glo was found to be the optimal cell viability assay among those analyzed. CellTiter-Blue reagent affected the cell morphology of normal human dermal fibroblasts (unlike L929), suggesting an interference with cell biological activity, resulting in less reliable viability data. On the other hand, RealTime-Glo provided a linear signal only with a very low cell density, which made this assay unsuitable for this system. CellTiter-Glo adapted to three-dimensional dermal construct by optimizing the “shaking time” to enhance the reagent penetration and maximum adenosine triphosphate release, indicating 2.4 times higher viability value by shaking for 60 min than for 5 min. In addition, viability results showed that cells were viable inside the matrix. This model would be further advanced with more layers of skin to make a full thickness model.

scholarly journals Alginate/Gelatin Hydrogels Reinforced with TiO2 and β-TCP Fabricated by Microextrusion-based Printing for Tissue Regeneration

Polymers ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 457 ◽  
Author(s):  
Rodrigo Urruela-Barrios ◽  
Erick Ramírez-Cedillo ◽  
A. Díaz de León ◽  
Alejandro Alvarez ◽  
Wendy Ortega-Lara
Keyword(s):  
Mechanical Properties ◽  
Tissue Engineering ◽  
Tissue Regeneration ◽  
Average Pore Size ◽  
Three Dimensional ◽  
Engineering Applications ◽  
Average Pore ◽  
Viscous Deformation ◽  
Freeze Dried ◽  
3D Printed

Three-dimensional (3D) printing technologies have become an attractive manufacturing process to fabricate scaffolds in tissue engineering. Recent research has focused on the fabrication of alginate complex shaped structures that closely mimic biological organs or tissues. Alginates can be effectively manufactured into porous three-dimensional networks for tissue engineering applications. However, the structure, mechanical properties, and shape fidelity of 3D-printed alginate hydrogels used for preparing tissue-engineered scaffolds is difficult to control. In this work, the use of alginate/gelatin hydrogels reinforced with TiO2 and β-tricalcium phosphate was studied to tailor the mechanical properties of 3D-printed hydrogels. The hydrogels reinforced with TiO2 and β-TCP showed enhanced mechanical properties up to 20 MPa of elastic modulus. Furthermore, the pores of the crosslinked printed structures were measured with an average pore size of 200 μm. Additionally, it was found that as more layers of the design were printed, there was an increase of the line width of the bottom layers due to its viscous deformation. Shrinkage of the design when the hydrogel is crosslinked and freeze dried was also measured and found to be up to 27% from the printed design. Overall, the proposed approach enabled fabrication of 3D-printed alginate scaffolds with adequate physical properties for tissue engineering applications.

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 3D Nanoprinting Technologies for Tissue Engineering Applications

2015 ◽  
Vol 2015 ◽  
pp. 1-14 ◽  
Author(s):  
Jin Woo Lee
Keyword(s):  
Tissue Engineering ◽  
Tissue Regeneration ◽  
Three Dimensional ◽  
Photon Absorption ◽  
Tissue Formation ◽  
Original Function ◽  
Cell Dynamics ◽  
Scaffold Fabrication ◽  
Two Photon Absorption ◽  
Two Photon

Tissue engineering recovers an original function of tissue by replacing the damaged part with a new tissue or organ regenerated using various engineering technologies. This technology uses a scaffold to support three-dimensional (3D) tissue formation. Conventional scaffold fabrication methods do not control the architecture, pore shape, porosity, or interconnectivity of the scaffold, so it has limited ability to stimulate cell growth and to generate new tissue. 3D printing technologies may overcome these disadvantages of traditional fabrication methods. These technologies use computers to assist in design and fabrication, so the 3D scaffolds can be fabricated as designed and standardized. Particularly, because nanofabrication technology based on two-photon absorption (2PA) and on controlled electrospinning can generate structures with submicron resolution, these methods have been evaluated in various areas of tissue engineering. Recent combinations of 3D nanoprinting technologies with methods from molecular biology and cell dynamics have suggested new possibilities for improved tissue regeneration. If the interaction between cells and scaffold system with biomolecules can be understood and controlled and if an optimal 3D environment for tissue regeneration can be realized, 3D nanoprinting will become an important tool in tissue engineering.

Proliferation and Fiber Formation of Human Dermal Fibroblasts on Patterned Differentially Adherent Substrates

2008 ◽  
Author(s):  
Nathan R. Schiele ◽  
Douglas B. Chrisey ◽  
David T. Corr
Keyword(s):  
Tissue Engineering ◽  
Soft Tissue ◽  
Cellular Proliferation ◽  
Critical Role ◽  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
Fiber Formation ◽  
Specific Cell ◽  
Fibroblast Cells ◽  
Myoblast Cells

Fibroblast cells are crucial in the human body for maintenance of the extracellular matrix, including synthesizing macromolecules like collagen, and they play a critical role in wound healing of soft tissues such as skin [1]. Directing fibroblast growth is an important step in tissue engineering where the focus has gone from a top-down approach of homogeneously introducing cells into a pre-formed scaffold to a bottom-up approach in which the tissue construct is built on a cell-by-cell basis with ability to manipulate specific cell environments through location, proximity, and geometry. The ability to direct cell proliferation to encourage organized tissue formation can provide tissue engineers a means of controlling the architectural and mechanical properties of soft tissue scaffolds. This approach to functional tissue engineering represents a novel direction for the development of replacement tissues. Previous attempts of directed growth have proven successful with C2C12 mouse myoblast cells. Using laser micromachined channels in agarose hydrogel lined with a basement membrane matrix, myoblast cells were guided to align and produce myotubes [2]. The objective of the current study was to apply similar principals to direct fibroblast cell growth and proliferation, ultimately leading to their growth into three-dimensional fibers, on differentially adherent substrates. Channels (widths ranging from 60 μm to 200 μm) were laser micromachined in agarose gel to explore an optimal geometry for cellular proliferation and fiber formation. The fibroblast cells used range in size from roughly 20–30 μm. Thus, the width of each channel was chosen to explore which multiple of cell width would allow for directional alignment parallel to the channel and subsequent fiber growth. The ability to direct fibroblast cells to align and produce fibers through manipulation of their environment is critical to our laboratory’s ongoing efforts to develop three-dimensional customized tissue replacement constructs to be used in many soft tissue applications such as ligament and skin grafts.

scholarly journals Piezo- and Magnetoelectric Polymers as Biomaterials for Novel Tissue Engineering Strategies

MRS Advances ◽  
2018 ◽  
Vol 3 (30) ◽  
pp. 1671-1676 ◽  
Author(s):  
C. Ribeiro ◽  
D.M. Correia ◽  
S. Ribeiro ◽  
M. M. Fernandes ◽  
S. Lanceros-Mendez
Keyword(s):  
Tissue Engineering ◽  
Muscle Tissue ◽  
Tissue Regeneration ◽  
Three Dimensional ◽  
Electrical Signal ◽  
Active Materials ◽  
Living Tissues ◽  
Electrical Stimuli ◽  
Technological Opportunities

ABSTRACTTissue engineering and regenerative medicine are increasingly taking advantage of active materials, allowing to provide specific clues to the cells. In particular, the use of electroactive polymers that deliver an electrical signal to the cells upon mechanical solicitation, open new scientific and technological opportunities, as they in fact mimic signals and effects that occur in living tissues, allowing the development of suitable microenvironments for tissue regeneration. Thus, a novel overall strategy for bone and muscle tissue engineering was developed based on the fact that these cells type are subjected to mechano-electrical stimuli in their in vivo microenvironment and that piezo- and magnetoelectric polymers, used as scaffolds, are suitable for delivering those cues. The processing and functional characterizations of piezoelectric and magnetoelectric polymers based on poly(vinylindene fluoride) and poly-L-lactic acid in a variety of shapes, from microspheres to electrospun mats and three dimensional scaffolds, are shown as well as their performance in the development of novel bone and muscle tissue engineering.

Abstract 9487: Implantation of Scaffold-Free Tubular Tissue Using a Bio-3D Printer Augments Vascular Remodeling and Endothelialization

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Manabu Itoh ◽  
Koichi Nakayama ◽  
Ryo Noguchi ◽  
Keiji Kamohara ◽  
Kojirou Furukawa ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Vascular Grafts ◽  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
3D Printer ◽  
Multicellular Spheroids ◽  
Biological Features ◽  
Normal Human ◽  
Umbilical Vein Endothelial Cells

Introduction: Small caliber synthetic vascular grafts are not clinically available. We developed a novel method to create scaffold-free tubular tissue from multicellular spheroids (MCS) using a “Bio-3D printer”-based system, which enables the creation of various three-dimensional structures pre-designed using a computer system. With this system, we created a tubular structure (Fig. 1), and studied its biological features. Methods: We made 1.5 mm in diameter scaffold-free tubular tissues from MCS (1.25 x 10[[Unable to Display Character: ⁷]] cells) composed of human umbilical vein endothelial cells (40%), human aortic smooth muscle cells (10%) and normal human dermal fibroblasts (50%) using a Bio-3D printer. The vessels were cultured in a perfusion system. We implanted grafts into the abdominal aortas of F344 nude rats, and assessed the flow by ultrasonography and performed histological examinations on the second (N=5) and fifth (N=5) days after implantation. Results: All grafts were patent. Remodeling of the vessel (enlargement of the lumen area and thinning of the wall) was observed (Fig. 2). A layer of endothelial cells was developed after implantation of the graft (Fig. 3). Conclusions: The scaffold-free vascular grafts made of MCS using a Bio-3D printer showed biological features comparable to native vessels. Further studies are warranted toward the clinical application of this novel technology.

scholarly journals Enzyme-Crosslinked Electrospun Fibrous Gelatin Hydrogel for Potential Soft Tissue Engineering

Polymers ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 1977
Author(s):  
Kexin Nie ◽  
Shanshan Han ◽  
Jianmin Yang ◽  
Qingqing Sun ◽  
Xiaofeng Wang ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Soft Tissue ◽  
Tissue Regeneration ◽  
Water Solution ◽  
Cellular Adhesion ◽  
Fibrous Structure ◽  
Electrospun Fibers ◽  
Hydrogel Scaffolds ◽  
Cell Functions ◽  
Soft Tissue Engineering

Soft tissue engineering has been seeking ways to mimic the natural extracellular microenvironment that allows cells to migrate and proliferate to regenerate new tissue. Therefore, the reconstruction of soft tissue requires a scaffold possessing the extracellular matrix (ECM)-mimicking fibrous structure and elastic property, which affect the cell functions and tissue regeneration. Herein, an effective method for fabricating nanofibrous hydrogel for soft tissue engineering is demonstrated using gelatin–hydroxyphenylpropionic acid (Gel–HPA) by electrospinning and enzymatic crosslinking. Gel–HPA fibrous hydrogel was prepared by crosslinking the electrospun fibers in ethanol-water solution with an optimized concentration of horseradish peroxidase (HRP) and H2O2. The prepared fibrous hydrogel held the soft and elastic mechanical property of hydrogels and the three-dimensional (3D) fibrous structure of electrospun fibers. It was proven that the hydrogel scaffolds were biocompatible, improving the cellular adhesion, spreading, and proliferation. Moreover, the fibrous hydrogel showed rapid biodegradability and promoted angiogenesis in vivo. Overall, this study represents a novel biomimetic approach to generate Gel–HPA fibrous hydrogel scaffolds which have excellent potential in soft tissue regeneration applications.

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