A silk fibroin/decellularized extract of Wharton’s jelly hydrogel intended for cartilage tissue engineering

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Abstract

A hybrid hydrogel was obtained from decellularized extract from Wharton’s jelly (DEWJ) and silk fibroin (SF) and characterized for cartilage tissue engineering. Wharton’s jelly was used due to its similarity with articular cartilage in extracellular matrix composition. Also, silk fibroin has good mechanical properties which make this construct appropriate for cartilage repair. Decellularization of Wharton’s jelly was verified by DAPI staining, DNA quantification, and PCR analysis. Then, the biochemical composition of DEWJ was determined by ELISA kits for total proteins, collagens, sulfated glycosaminoglycans (sGAG), and transforming growth factor β1 (TGF-β1). After fabricating pure SF and SF/DEWJ hybrid hydrogels, their physical and mechanical properties were characterized by FESEM, Fourier-transform infrared spectroscopy (FTIR) and rheological assays (amplitude and frequency sweeps). Furthermore, cell viability and proliferation were assessed by MTT assay. The results have shown that DEWJ in hybrid hydrogels enhances mechanical properties of the construct relative to pure SF hydrogels. Also, this extract at its 40% concentration in culture media and 20% or 40% concentrations in SF/DEWJ hybrid hydrogels significantly increases population of the cells compared to control and pure SF hydrogel after 7 days. In conclusion, this study proposes the potential of SF/DEWJ hybrid hydrogels for cartilage tissue engineering applications.

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Effect of the Microenvironment on Mesenchymal Stem Cell Paracrine Signaling: Opportunities to Engineer the Therapeutic Effect.

Gina D. Kusuma, James Carthew, Rebecca Lim … (2017)

Cues from the extracellular environment, including physical stimuli, are well known to affect mesenchymal stem cell (MSC) properties in terms of proliferation and differentiation. Many therapeutic strategies are now targeting this knowledge to increase the efficacy of cell therapies, typically employed to repair tissue functions in the event of injury, either by direct engraftment into the target tissue or differentiation into mature tissues. However, it is now envisioned that harnessing the repertoire of factors secreted by MSCs (termed the secretome) may provide an alternate to these cell therapies. Of current interest are both direct protein secretions and two major subpopulations of bioactive extracellular vesicles (EVs), namely exosomes and microvesicles. EVs released by MSCs are reflective of their cells of origin, able to impact upon the activities of other cells in the local microenvironment, making the rational design of MSC paracrine activities an encouraging strategy to reproducibly modulate cell therapies. The precise mechanisms by which the secretome is modulated by the microenvironment, however, remain elusive. Controlling MSC growth conditions with oxygen tension, growth factor composition, and mechanical properties may serve to directly influence paracrine activity. Our growing understanding implicates components of the mechanotransduction machinery in translating both mechanical and chemical cues from the environment into alterations in gene regulation and varied paracrine activity. As technologies are developed to manufacture MSCs, advances in bioengineering and novel insight of how the extracellular environment affects MSC paracrine activity will play a pivotal role in the generation of widespread, successful, clinical MSC therapies.

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Sonication-induced gelation of silk fibroin for cell encapsulation.

Xiaoqin Wang, Jonathan Kluge, Gary Leisk … (2008)

Purified native silk fibroin forms beta-sheet-rich, physically cross-linked, hydrogels from aqueous solution, in a process influenced by environmental parameters. Previously we reported gelation times of days to weeks for aqueous native silk protein solutions, with high ionic strength and temperature and low pH responsible for increasing gelation kinetics. Here we report a novel method to accelerate the process and control silk fibroin gelation through ultrasonication. Depending on the sonication parameters, including power output and time, along with silk fibroin concentration, gelation could be controlled from minutes to hours, allowing the post-sonication addition of cells prior to final gel setting. Mechanistically, ultrasonication initiated the formation of beta-sheets by alteration in hydrophobic hydration, thus accelerating the formation of physical cross-links responsible for gel stabilization. K(+) at physiological concentrations and low pH promoted gelation, which was not observed in the presence of Ca(2+). The hydrogels were assessed for mechanical properties and proteolytic degradation; reported values matched or exceeded other cell-encapsulating gel material systems. Human bone marrow derived mesenchymal stem cells (hMSCs) were successfully incorporated into these silk fibroin hydrogels after sonication, followed by rapid gelation and sustained cell function. Sonicated silk fibroin solutions at 4%, 8%, and 12% (w/v), followed by mixing in hMSCs, gelled within 0.5-2 h. The cells grew and proliferated in the 4% gels over 21 days, while survival was lower in the gels with higher protein content. Thus, sonication provides a useful new tool with which to initiate rapid sol-gel transitions, such as for cell encapsulation.

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Silk protein-based hydrogels: Promising advanced materials for biomedical applications.

Subhas C. Kundu, Sonia Kapoor (2016)

Hydrogels are a class of advanced material forms that closely mimic properties of the soft biological tissues. Several polymers have been explored for preparing hydrogels with structural and functional features resembling that of the extracellular matrix. Favourable material properties, biocompatibility and easy processing of silk protein fibers into several forms make it a suitable material for biomedical applications. Hydrogels made from silk proteins have shown a potential in overcoming limitations of hydrogels prepared from conventional polymers. A great deal of effort has been made to control the properties and to integrate novel topographical and functional characteristics in the hydrogel composed from silk proteins. This review provides overview of the advances in silk protein-based hydrogels with a primary emphasis on hydrogels of fibroin. It describes the approaches used to fabricate fibroin hydrogels. Attempts to improve the existing properties or to incorporate new features in the hydrogels by making composites and by improving fibroin properties by genetic engineering approaches are also described. Applications of the fibroin hydrogels in the realms of tissue engineering and controlled release are reviewed and their future potentials are discussed.

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Author and article information

Contributors

Mohamadreza Baghaban Eslaminejad: +98 2123562524 , eslami@royaninstitute.org

Jafar Ai:

ORCID: http://orcid.org/0000-0003-3892-325X

+98 2143052000-110 , jafar_ai@tums.ac.ir

Journal

Journal ID (nlm-ta): Prog Biomater

Journal ID (iso-abbrev): Prog Biomater

Title: Progress in Biomaterials

Publisher: Springer Berlin Heidelberg (Berlin/Heidelberg )

ISSN (Print): 2194-0509

ISSN (Electronic): 2194-0517

Publication date (Electronic): 31 January 2019

Publication date PMC-release: 31 January 2019

Publication date Collection: March 2019

Volume: 8

Pages: 31-42

Affiliations

[1 ]ISNI 0000 0001 0166 0922, GRID grid.411705.6, Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, , Tehran University of Medical Sciences, ; Tehran, Iran

[2 ]ISNI 0000 0000 9562 2611, GRID grid.420169.8, National Cell Bank of Iran, Pasteur Institute of Iran, ; Tehran, Iran

[3 ]GRID grid.411600.2, Department of Biotechnology, School of Advanced Technologies in Medicine, , Shahid Beheshti University of Medical Sciences, ; Tehran, Iran

[4 ]ISNI 0000 0001 0166 0922, GRID grid.411705.6, Sina Trauma and Surgery Reasearch Center, , Tehran University of Medical Sciences, ; Tehran, Iran

[5 ]ISNI 0000 0004 0612 4397, GRID grid.419336.a, Department of Stem Cells and Developmental Biology, Cell Science Research Center, , Royan Institute for Stem Cell Biology and Technology, ACECR, ; Tehran, Iran

Author information

Jafar Ai http://orcid.org/0000-0003-3892-325X

Article

Publisher ID: 108

DOI: 10.1007/s40204-019-0108-7

PMC ID: 6424998

PubMed ID: 30706299

SO-VID: 6ac9e09a-2310-44f8-b188-f9dcbdf8fe7c

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Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

History

Date received : 16 December 2018

Date accepted : 23 January 2019

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Keywords: decellularization,wharton’s jelly,silk fibroin,hydrogel,cartilage tissue engineering

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Keywords: decellularization, wharton’s jelly, silk fibroin, hydrogel, cartilage tissue engineering

A silk fibroin/decellularized extract of Wharton’s jelly hydrogel intended for cartilage tissue engineering

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Most cited references 42

Effect of the Microenvironment on Mesenchymal Stem Cell Paracrine Signaling: Opportunities to Engineer the Therapeutic Effect.

Sonication-induced gelation of silk fibroin for cell encapsulation.

Silk protein-based hydrogels: Promising advanced materials for biomedical applications.

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