A Novel Single-Step Self-Assembly Approach for the Fabrication of Tissue-Engineered Vascular Constructs

Gauvin, Robert; Ahsan, Taby; Larouche, Danielle; Lévesque, Philippe; Dubé, Jean; Auger, François A.; Nerem, Robert M.; Germain, Lucie

doi:10.1089/ten.tea.2009.0313

Cited by 104 publications

(100 citation statements)

References 43 publications

(60 reference statements)

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“…13 Compared with other ECM derived from smooth muscle cells and hMSCs, fibroblast-derived ECM is stronger and denser and contains more elastin that makes the ECM more elastic. 31 Therefore, the ECM scaffolds derived from fibroblast cell sheets could serve as a stronger and more effective scaffold for tissue regeneration. The purpose of decellularization is to reduce cellular components that potentially hinder tissue remodeling and induce immune responses.…”

Section: Discussionmentioning

confidence: 99%

Decellularization of Fibroblast Cell Sheets for Natural Extracellular Matrix Scaffold Preparation

Xing

Yates

Tahtinen

et al. 2015

Tissue Engineering Part C: Methods

167

131

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The application of cell-derived extracellular matrix (ECM) in tissue engineering has gained increasing interest because it can provide a naturally occurring, complex set of physiologically functional signals for cell growth. The ECM scaffolds produced from decellularized fibroblast cell sheets contain high amounts of ECM substances, such as collagen, elastin, and glycosaminoglycans. They can serve as cell adhesion sites and mechanically strong supports for tissue-engineered constructs. An efficient method that can largely remove cellular materials while maintaining minimal disruption of ECM ultrastructure and content during the decellularization process is critical. In this study, three decellularization methods were investigated: high concentration (0.5 wt%) of sodium dodecyl sulfate (SDS), low concentration (0.05 wt%) of SDS, and freeze-thaw cycling method. They were compared by characterization of ECM preservation, mechanical properties, in vitro immune response, and cell repopulation ability of the resulted ECM scaffolds. The results demonstrated that the high SDS treatment could efficiently remove around 90% of DNA from the cell sheet, but significantly compromised their ECM content and mechanical strength. The elastic and viscous modulus of the ECM decreased around 80% and 62%, respectively, after the high SDS treatment. The freeze-thaw cycling method maintained the ECM structure as well as the mechanical strength, but also preserved a large amount of cellular components in the ECM scaffold. Around 88% of DNA was left in the ECM after the freeze-thaw treatment. In vitro inflammatory tests suggested that the amount of DNA fragments in ECM scaffolds does not cause a significantly different immune response. All three ECM scaffolds showed comparable ability to support in vitro cell repopulation. The ECM scaffolds possess great potential to be selectively used in different tissue engineering applications according to the practical requirement.

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Section: Discussionmentioning

confidence: 99%

Decellularization of Fibroblast Cell Sheets for Natural Extracellular Matrix Scaffold Preparation

Xing

Yates

Tahtinen

et al. 2015

Tissue Engineering Part C: Methods

167

131

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“…[10][11][12][13] With our current standard protocol, the production of the dermal substitute of the SASS requires 3-4 weeks and is the most time consuming step. 12,14 Our aim was to modify the SASS production process to obtain a significant reduction in the dermal production time from the point where the patient's cells have been harvested and expanded in culture, allowing quicker grafting of the skin substitute on the patient.…”

Section: Introductionmentioning

confidence: 99%

Production of a Bilayered Self-Assembled Skin Substitute Using a Tissue-Engineered Acellular Dermal Matrix

Cloutier

Guignard

Bernard

et al. 2015

Tissue Engineering Part C: Methods

Self Cite

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Our bilayered self-assembled skin substitutes (SASS) are skin substitutes showing a structure and functionality very similar to native human skin. These constructs are used, in life threatening burn wounds, as permanent autologous grafts for the treatment of such affected patients even though their production is exacting. We thus intended to shorten their current production time to improve their clinical applicability. A self-assembled decellularized dermal matrix was used. It allowed the production of an autologous skin substitute from patient's cells. The characterization of SASS reconstructed using a decellularized dermal matrix (SASS-DM) was performed by histology, immunofluorescence, transmission electron microscopy and uniaxial tensile analysis.Using the SASS-DM, it was possible to reduce the standard production time from about eight to four weeks and a half. The structure, cell differentiation and mechanical properties of the new skin substitutes were shown to be similar to the SASS. The decellularization process had no influence on the final microstructure and mechanical properties of the dermal matrix. This model, by enabling the production of a skin substitute in a shorter time frame without compromising its intrinsic tissue properties, represents a promising addition to the currently available burn and wound treatments.

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Section: Introductionmentioning

confidence: 99%

“…Several promising approaches to engineering blood vessels have been explored, with many recent studies focusing on development and analysis of cell-based methods. [2][3][4][5] Herein, we present a method to rapidly self-assemble cells into 3Dtissue rings that can be used in vitro to model vascular tissues.To do this, suspensions of smooth muscle cells are seeded into round-bottomed annular agarose wells. The non-adhesive properties of the agarose allow the cells to settle, aggregate and contract around a post at the center of the well to form a cohesive tissue ring.…”

mentioning

confidence: 99%

Directed Cellular Self-Assembly to Fabricate Cell-Derived Tissue Rings for Biomechanical Analysis and Tissue Engineering

Gwyther

Billiar

et al. 2011

JoVE

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Each year, hundreds of thousands of patients undergo coronary artery bypass surgery in the United States.1 Approximately one third of these patients do not have suitable autologous donor vessels due to disease progression or previous harvest. The aim of vascular tissue engineering is to develop a suitable alternative source for these bypass grafts. In addition, engineered vascular tissue may prove valuable as living vascular models to study cardiovascular diseases. Several promising approaches to engineering blood vessels have been explored, with many recent studies focusing on development and analysis of cell-based methods. [2][3][4][5] Herein, we present a method to rapidly self-assemble cells into 3Dtissue rings that can be used in vitro to model vascular tissues.To do this, suspensions of smooth muscle cells are seeded into round-bottomed annular agarose wells. The non-adhesive properties of the agarose allow the cells to settle, aggregate and contract around a post at the center of the well to form a cohesive tissue ring. 6,7 These rings can be cultured for several days prior to harvesting for mechanical, physiological, biochemical, or histological analysis. We have shown that these cell-derived tissue rings yield at 100-500 kPa ultimate tensile strength 8 which exceeds the value reported for other tissue engineered vascular constructs cultured for similar durations (<30 kPa). 9,10 Our results demonstrate that robust cell-derived vascular tissue ring generation can be achieved within a short time period, and offers the opportunity for direct and quantitative assessment of the contributions of cells and cell-derived matrix (CDM) to vascular tissue structure and function. Video LinkThe video component of this article can be found at https://www.jove.com/video/3366/ Protocol Cell seeding mold fabricationBegin by milling a 1/2" thick piece of polycarbonate to create 15, round-bottomed, annular wells with a center post diameter of 2 mm. The milled channels are 6 mm deep and 3.75 mm wide. Clean and dry the polycarbonate mold to remove any plastic debris from the milling process.Mix polydimethylsiloxane (PDMS) at a 10:1 ratio (w/w) of base to curing agent, degas to remove all air bubbles, and pour onto the polycarbonate mold. Degas again to remove any remaining bubbles, and cure in the oven at 60°C for 4 hours.Once cured, carefully remove the PDMS template by slowly peeling it away from the polycarbonate, wash with soap and water, and autoclave. Also autoclave a solution of two percent agarose (w/v) dissolved in Dulbecco's modified Eagle medium (DMEM).Place the PDMS template on a level surface and fill with molten agarose by first pipetting agarose into each of the center post molds, and then pipetting into the space around it. Allow the agarose to solidify (approximately 15 minutes), then invert the mold to release the agarose from the PDMS. Cut excess agarose from around each of the wells, and place the agarose wells into 12-well plates. Cell culture and ring seedingAdd cell culture media (DMEM with 10% ...

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A Novel Single-Step Self-Assembly Approach for the Fabrication of Tissue-Engineered Vascular Constructs

Cited by 104 publications

References 43 publications

Decellularization of Fibroblast Cell Sheets for Natural Extracellular Matrix Scaffold Preparation

Decellularization of Fibroblast Cell Sheets for Natural Extracellular Matrix Scaffold Preparation

Production of a Bilayered Self-Assembled Skin Substitute Using a Tissue-Engineered Acellular Dermal Matrix

Directed Cellular Self-Assembly to Fabricate Cell-Derived Tissue Rings for Biomechanical Analysis and Tissue Engineering

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