Real-time monitoring of human Schwann cells on heparin-collagen coatings reveals enhanced adhesion and growth factor response

Human mesenchymal stromal cells (hMSCs) are multipotent cells that have been proposed for cell therapies due to their immunosuppressive capacity that can be enhanced in the presence of interferon-gamma (IFN-γ). In this study, multilayers of heparin (HEP) and collagen (COL) (HEP/COL) were used as a bioactive surface to enhance the immunomodulatory activity of hMSCs using soluble IFN-γ. Multilayers were formed, via layer-by-layer assembly, varying the final layer between COL and HEP and supplemented with IFN-γ in the culture medium. We evaluated the viability, adhesion, real-time growth, differentiation, and immunomodulatory activity of hMSCs on (HEP/COL) multilayers. HMSCs viability, adhesion, and growth were superior when cultured on (HEP/COL) multilayers compared to tissue culture plastic. We also confirmed that hMSCs osteogenic and adipogenic differentiation remained unaffected when cultured in (HEP/COL) multilayers in the presence of IFN-γ. We measured the immunomodulatory activity of hMSCs by measuring the level of indoleamine 2,3-dioxygenase (IDO) expression. IDO expression was higher on (HEP/COL) multilayers treated with IFN-γ. Lastly, we evaluated the suppression of peripheral blood mononuclear cell (PBMC) proliferation when co-cultured with hMSCs on (HEP/COL) multilayers with IFN-γ. hMSCs cultured in (HEP/COL) multilayers in the presence of soluble IFN-γ have a greater capacity to suppress PBMC proliferation. Altogether, (HEP/COL) multilayers with IFN-γ in culture medium provides a potent means of enhancing and sustaining immunomodulatory activity to control hMSCs immunomodulation.

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“…Therefore, the higher the CI, the greater the number of cells adhered to the bottom of the well [ 39 ]. Fig.…”

Section: Resultsmentioning

confidence: 99%

“…(HEP/COL) multilayers were constructed as described in our previous works [ 12 , 29 , 39 , 40 ]. Heparin sodium (HEP) purchased from Celsus Laboratories, Inc. (Cat.…”

Section: Methodsmentioning

confidence: 99%

Immunomodulatory functions of human mesenchymal stromal cells are enhanced when cultured on HEP/COL multilayers supplemented with interferon-gamma

Haseli

Castilla‐Casadiego

Pinzón-Herrera

et al. 2022

Materials Today Bio

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Section: Anticipated Resultsmentioning

confidence: 99%

“…At a molecular level, the mechanism is identical to the layer-by-layer assembly on flat substrates (see Figure ). − ,,, …”

Section: Methods 1 Layer-by-layer Film Construction and Hmscs Seeded ...mentioning

confidence: 99%

“…Our group has recently demonstrated infrared spectroscopic ellipsometry (IRVASE) as a viable method for the simultaneous characterization of physical, chemical, and thermal properties characterization of heparin/collagen multilayers . Furthermore, our group has demonstrated that heparin/collagen multilayers may be used to modulate hMSC response to soluble factors. − In addition, our group has demonstrated the heparin/collagen multilayers improve Schwann cell adhesion and response to growth factors . The work presented here describes the various experimental and characterization techniques to construct and characterize thin LbL films and the interaction of hMSCs with these films.…”

Section: Introductionmentioning

confidence: 99%

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Methods for the Assembly and Characterization of Polyelectrolyte Multilayers as Microenvironments to Modulate Human Mesenchymal Stromal Cell Response

Castilla‐Casadiego

Timsina

Haseli

et al. 2020

ACS Biomater. Sci. Eng.

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Thin films are of interest in materials design because they allow for the modification of surface properties of materials while the bulk properties of the material are largely unaffected. In this work, we outline methods for the assembly of thin films using a technique known as layer-by-layer (LbL). Furthermore, their interactions with human mesenchymal stromal cells (hMSCs) are discussed. hMSCs are a subject of growing interest because of their potential to treat or cure diseases, given their immunosuppressive properties, multipotent differentiation capabilities, and tissue regeneration capabilities. Numerous improvements and modifications have been suggested for the harvesting, treatment, and culture of hMSCs prior to their administration in human subjects. Here, we discuss methods to assess the interactions of hMSCs with thin LbL-assembled films of heparin and collagen. Three different methods are discussed. The first details the preparation of heparin/collagen multilayers on different surfaces and the seeding of cells on these multilayers. The second method details the characterization of multilayers, including techniques to assess the thickness, roughness, and surface charge of the multilayers, as well as in situ deposition of multilayers. The third method details the analysis of cell interactions with the multilayers, including techniques to assess proliferation, viability, real-time monitoring of hMSC behavior, analysis of hMSC-adhesive proteins on the multilayers, immunomodulatory factor expression of hMSCs, and cytokine expression on heparin/collagen multilayers. We propose that the methods described in this work will assist in the design and characterization of LbL-assembled thin films and the analysis of hMSCs cultured on these thin films.

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Surface Modification of Nerve Guide Conduits with ECM Coatings and Investigating Their Impact on Schwann Cell Response

Pinzon‐Herrera,

Magness,

Apodaca‐Reyes

et al. 2024

Adv Healthcare Materials

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In this study, we investigated surface modification through the layer‐by‐layer method as a straightforward and highly stable approach for depositing layers of natural polymers onto a commercial nerve guide conduit (NGC), thereby mimicking the extracellular matrix. The application of NGCs for nerve regeneration is constrained by the size of the lesion, limiting their use to small gaps (1‐2 cm). The principal aim of this modification is to augment the cellular response specific to peripheral nerve injury (PNI) repair processes and enhance the overall performance of NGCs. To achieve this objective, we conducted in vitro studies examining the stability, degradation over time, and bioactivity of six bilayers of heparin/collagen layer‐by‐layer coatings, denoted as (HEP/COL)6, after incubating the coatings with cell culture medium prior to cell seeding. The stability study revealed that (HEP/COL)6 is stable after incubating the coatings in cell media for up to 21 days. We evaluated the impact of (HEP/COL)6 on human Schwann cells (hSCs), focusing on viability, protein expression, and cell migration. The presence or absence of the nerve growth factor (NGF) was considered during all the experiments. These assays showed that hSCs cultured in (HEP/COL)6 had enhanced protein expression and migration. This condition increased the expression of neurotrophic and immunomodulatory factors up to 1.5‐fold compared to controls, and hSCs migrated 1.34 times faster than in the uncoated surfaces. Finally, (HEP/COL)6 was also applied to a commercial collagen‐based NGC, NeuraGen®, and hSC viability and adhesion were studied after six days of culture. The morphology of NeuraGen® is not altered by the presence of (HEP/COL)6 and a nearly 170% increase of the cell viability was observed in the condition where NeuraGen® was used with (HEP/COL)6. Additionally, we successfully demonstrated cell adhesion on the coated samples.This article is protected by copyright. All rights reserved

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Real-time monitoring of human Schwann cells on heparin-collagen coatings reveals enhanced adhesion and growth factor response

Abstract: In this work, we evaluate the enhancing effect of six bilayers of heparin/collagen (HEP/COL)6 layer-by-layer coatings on human Schwann cell (hSCs) adhesion and proliferation in the presence or absence of...

Cited by 13 publications

References 41 publications

Immunomodulatory functions of human mesenchymal stromal cells are enhanced when cultured on HEP/COL multilayers supplemented with interferon-gamma

Immunomodulatory functions of human mesenchymal stromal cells are enhanced when cultured on HEP/COL multilayers supplemented with interferon-gamma

Methods for the Assembly and Characterization of Polyelectrolyte Multilayers as Microenvironments to Modulate Human Mesenchymal Stromal Cell Response

Surface Modification of Nerve Guide Conduits with ECM Coatings and Investigating Their Impact on Schwann Cell Response

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