Analyzing the scaffold immune microenvironment using flow cytometry: practices, methods and considerations for immune analysis of biomaterials

Hall, Matthew D.; Elisseeff, Jennifer H.

doi:10.1039/c9bm00349e

Cited by 11 publications

(6 citation statements)

References 53 publications

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“…Although macrophages' response has been studied in three-dimensional (3D) collagen or fibrin matrices in dependence of ligand presentation [18] and stiffness via crosslinkers [18][19][20], inconsistency in the results has been reported due to the difference of differentiation protocols and cell source used. Additionally, while these studies attempted to demonstrate the important role of tissue mechanical properties on macrophage response, the latter is often investigated based on a number of phenotypic specific markers without taking into account their cellular functions [21]. Despite the growing interest in this field, the direct effect of tissue density on macrophage phenotypes and functions has not been well established.…”

Section: Introductionmentioning

confidence: 99%

Collagen Fibril Density Modulates Macrophage Activation and Cellular Functions during Tissue Repair

Sapudom¹,

Mohamed

Garcia-Sabaté³

et al. 2020

Bioengineering

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Monocytes circulate in the bloodstream, extravasate into the tissue and differentiate into specific macrophage phenotypes to fulfill the immunological needs of tissues. During the tissue repair process, tissue density transits from loose to dense tissue. However, little is known on how changes in tissue density affects macrophage activation and their cellular functions. In this work, monocytic cell line THP-1 cells were embedded in three-dimensional (3D) collagen matrices with different fibril density and were then differentiated into uncommitted macrophages (MPMA) using phorbol-12-myristate-13-acetate (PMA). MPMA macrophages were subsequently activated into pro-inflammatory macrophages (MLPS/IFNγ) and anti-inflammatory macrophages (MIL-4/IL-13) using lipopolysaccharide and interferon-gamma (IFNγ), and interleukin 4 (IL-4) and IL-13, respectively. Although analysis of cell surface markers, on both gene and protein levels, was inconclusive, cytokine secretion profiles, however, demonstrated differences in macrophage phenotype. In the presence of differentiation activators, MLPS/IFNγ secreted high amounts of IL-1β and tumor necrosis factor alpha (TNFα), while M0PMA secreted similar cytokines to MIL-4/IL-13, but low IL-8. After removing the activators and further culture for 3 days in fresh cell culture media, the secretion of IL-6 was found in high concentrations by MIL-4/IL-13, followed by MLPS/IFNγ and MPMA. Interestingly, the secretion of cytokines is enhanced with an increase of fibril density. Through the investigation of macrophage-associated functions during tissue repair, we demonstrated that M1LPS/IFNγ has the potential to enhance monocyte infiltration into tissue, while MIL-4/IL-13 supported fibroblast differentiation into myofibroblasts via transforming growth factor beta 1 (TGF-β1) in dependence of fibril density, suggesting a M2a-like phenotype. Overall, our results suggest that collagen fibril density can modulate macrophage response to favor tissue functions. Understanding of immune response in such complex 3D microenvironments will contribute to the novel therapeutic strategies for improving tissue repair, as well as guidance of the design of immune-modulated materials.

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Section: Introductionmentioning

confidence: 99%

Collagen Fibril Density Modulates Macrophage Activation and Cellular Functions during Tissue Repair

Sapudom¹,

Mohamed

Garcia-Sabaté³

et al. 2020

Bioengineering

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“…It allows fast, relatively quantitative and multiparameteric analysis of a single cell in a heterogeneous cell population as well as isolation of those cells with a process called cell sorting (83,84). Such sorted speci c cell types can be sequenced or used for gene expression analysis on a population or a single-cell level, and as well combined with ELISA, proteomics, or Western blots analysis directly (85). Any foreign material implanted in the body will stimulate an immune response, and ow cytometry has been emerged as a critical tool in the study of the immune system.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…Any foreign material implanted in the body will stimulate an immune response, and ow cytometry has been emerged as a critical tool in the study of the immune system. With the growing appreciation of the immune system in tissue engineering and regenerative medicine, ow cytometry is increasingly being implement in the analysis of biomaterials response (85,86). e host's innate and adaptive immune systems are the major mediators in the response to foreign implants, and a surgical implantation of an osteoinductive device that contains carriers such as natural and synthetic polymers and inorganic materials into the human body have the ability to trigger host responses (20,87).…”

Section: Flow Cytometrymentioning

confidence: 99%

A simple rodent subcutaneous assay for identification of new osteoinductive molecules: The key method for screening of novel bone regeneration implants

Štoković¹,

Ivanjko²,

Milešević³

et al. 2022

Rad Hrvatske akademije znanosti i umjetnosti

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Treatment of large bone defects and degenerative diseases of the spine is among the most challenging and still unresolved issues in clinical medicine. erefore, substantial e ort has been devoted to the development of novel bone regenerative therapies. Due to their potent osteoinductive properties, Bone Morphogenetic Proteins (BMPs) have been the basis for the development of novel strategies for bone regeneration. e use of animal models is an indispensable part of the preclinical testing of novel therapeutic solutions. e rat subcutaneous assay became the initial screening procedure for the evaluation of promising BMP-based osteoinductive devices for bone regeneration because only osteogenic BMPs can induce new bone at any ectopic rodent site. Moreover, this model is used for research on the mechanisms of ectopic bone formation as well as for the evaluation of the in ammatory response to di erent materials. In this review, we provided an overview of the assay development and previously conducted studies with di erent methods ( ow cytometry, histological and microCT analyses) for the study outcome evaluation. Moreover, we addressed essential issues in the experimental design such as the follow-up period and the sample size. e rat subcutaneous bone induction assay layed the foundation for isolation and identi cation of BMPs followed by testing of new osteogenic devices in higher animal species and humans.

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“…That is due to the differences in microporous topological structure, degradation cycle and degradation products among different hernia mesh. Scientific research on tissue remodeling and regeneration also includes topics such as collagen structure proportion [ 127 ], biomarkers related to host cell proliferation, differentiation and regulation [ 127 , 128 ], characterization parameters and test methods of tissue-remodeling microenvironment [ 129 ], and advanced manufacturing technology of biomimetic ECM [ 126 , 130 ].…”

Section: Future Perspectivesmentioning

confidence: 99%

Regulatory science for hernia mesh: Current status and future perspectives

Liu

Xie

Zheng

et al. 2021

Bioactive Materials

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Regulatory science for medical devices aims to develop new tools, standards and approaches to assess the safety, effectiveness, quality and performance of medical devices. In the field of biomaterials, hernia mesh is a class of implants that have been successfully translated to clinical applications. With a focus on hernia mesh and its regulatory science system, this paper collected and reviewed information on hernia mesh products and biomaterials in both Chinese and American markets. The current development of regulatory science for hernia mesh, including its regulations, standards, guidance documents and classification, and the scientific evaluation of its safety and effectiveness was first reported. Then the research prospect of regulatory science for hernia mesh was discussed. New methods for the preclinical animal study and new tools for the evaluation of the safety and effectiveness of hernia mesh, such as computational modeling, big data platform and evidence-based research, were assessed. By taking the regulatory science of hernia mesh as a case study, this review provided a research basis for developing a regulatory science system of implantable medical devices, furthering the systematic evaluation of the safety and effectiveness of medical devices for better regulatory decision-making. This was the first article reviewing the regulatory science of hernia mesh and biomaterial-based implants. It also proposed and explained the concepts of evidence-based regulatory science and technical review for the first time.

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Analyzing the scaffold immune microenvironment using flow cytometry: practices, methods and considerations for immune analysis of biomaterials

Abstract: The immune system has evolved as a powerful tool for our body to combat infections, and is being engineered for new treatments in cancer, autoimmune disease, and trauma.

Cited by 11 publications

References 53 publications

Collagen Fibril Density Modulates Macrophage Activation and Cellular Functions during Tissue Repair

Collagen Fibril Density Modulates Macrophage Activation and Cellular Functions during Tissue Repair

A simple rodent subcutaneous assay for identification of new osteoinductive molecules: The key method for screening of novel bone regeneration implants

Regulatory science for hernia mesh: Current status and future perspectives

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