Extracellular matrix protein microarray-based biosensor with single cell resolution: Integrin profiling and characterization of cell-biomaterial interactions

Gonzalez‐Pujana, Ainhoa; Santos‐Vizcaíno, Edorta; García-Hernando, Maite; Hernaez-Estrada, Beatriz; Pancorbo, Marian M. de; Benito‐Lopez, Fernando; Igartua, Manoli; Basabe‐Desmonts, Lourdes; Hernández, Rosa María

doi:10.1016/j.snb.2019.126954

Cited by 16 publications

(16 citation statements)

References 43 publications

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“… 239 Bioactivity of the biomaterial can be assessed through microarray based biosensor methods. 240 These microarray based methods give insight about cell adhesion kinetics, integrin profiling, and non-specific interactions. This platform enables to perform adhesion profiling of various ECM components with multiple cell types in parallel.…”

Section: Final Characterisation To Investigate Retained Activitymentioning

confidence: 99%

Bioactive potential of natural biomaterials: identification, retention and assessment of biological properties

Joyce

Fabra

Bozkurt

et al. 2021

Sig Transduct Target Ther

126

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Biomaterials have had an increasingly important role in recent decades, in biomedical device design and the development of tissue engineering solutions for cell delivery, drug delivery, device integration, tissue replacement, and more. There is an increasing trend in tissue engineering to use natural substrates, such as macromolecules native to plants and animals to improve the biocompatibility and biodegradability of delivered materials. At the same time, these materials have favourable mechanical properties and often considered to be biologically inert. More importantly, these macromolecules possess innate functions and properties due to their unique chemical composition and structure, which increase their bioactivity and therapeutic potential in a wide range of applications. While much focus has been on integrating these materials into these devices via a spectrum of cross-linking mechanisms, little attention is drawn to residual bioactivity that is often hampered during isolation, purification, and production processes. Herein, we discuss methods of initial material characterisation to determine innate bioactivity, means of material processing including cross-linking, decellularisation, and purification techniques and finally, a biological assessment of retained bioactivity of a final product. This review aims to address considerations for biomaterials design from natural polymers, through the optimisation and preservation of bioactive components that maximise the inherent bioactive potency of the substrate to promote tissue regeneration.

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Section: Final Characterisation To Investigate Retained Activitymentioning

confidence: 99%

Bioactive potential of natural biomaterials: identification, retention and assessment of biological properties

Joyce

Fabra

Bozkurt

et al. 2021

Sig Transduct Target Ther

126

View full text Add to dashboard Cite

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“…We hypothesize that adsorbed proteins may interact with cellular receptors, especially with integrins; thus, triggering the activation of particular signal transduction pathways involved in cellular processes such as proliferation or migration [ 31 , 32 ]. Therefore, the nature and extent of this biologic response will largely depend on the integrin profile expressed by each particular cell type [ 33 ] and the layer of adsorbed proteins presented by the biomaterial surface [ 34 ]. In addition, further still-unraveled mechanisms may be responsible for cell attachment and subsequent biologic responses [ 35 ].…”

Section: Discussionmentioning

confidence: 99%

Overcoming the Inflammatory Stage of Non-Healing Wounds: In Vitro Mechanism of Action of Negatively Charged Microspheres (NCMs)

et al. 2020

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Negatively charged microspheres (NCMs) represent a new therapeutic approach for wound healing since recent clinical trials have shown NCM efficacy in the recovery of hard-to-heal wounds that tend to stay in the inflammatory phase, unlocking the healing process. The aim of this study was to elucidate the NCM mechanism of action. NCMs were extracted from a commercial microsphere formulation (PolyHeal® Micro) and cytotoxicity, attachment, proliferation and viability assays were performed in keratinocytes and dermal fibroblasts, while macrophages were used for the phagocytosis and polarization assays. We demonstrated that cells tend to attach to the microsphere surface, and that NCMs are biocompatible and promote cell proliferation at specific concentrations (50 and 10 NCM/cell) by a minimum of 3 fold compared to the control group. Furthermore, NCM internalization by macrophages seemed to drive these cells to a noninflammatory condition, as demonstrated by the over-expression of CD206 and the under-expression of CD64, M2 and M1 markers, respectively. NCMs are an effective approach for reverting the chronic inflammatory state of stagnant wounds (such as diabetic wounds) and thus for improving wound healing.

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“…There are two main techniques for the isolation of cells as single cells. On one hand, individual cells can be confined in cavities (Hosokawa et al, 2011), and on the other hand, they can be chemically guided to settle at specific locations by protein patterning techniques (Gonzalez-Pujana et al, 2019), enabling high density of single-cell arrays. Single-cell toxicity testing can provide with quantitative analysis.…”

Section: Single Cell Resolutionmentioning

confidence: 99%

“…In the last decades, remarkable advancements have been made in the field of microtechnology to improve analytical processes in biology, through miniaturization, for biosensing DNA (Bulyk et al, 1999;Zhang et al, 2010) and protein arrays (He et al, 2008;Ramachandran et al, 2008;Lopez-Alonso et al, 2013;Gonzalez-Pujana et al, 2019), on-chip electrophoresis (Fritzsche et al, 2010;Ou, et al, 2019), microimmunoassays (Riahi et al, 2016;Hu et al, 2017), microfluidic cell sorting (Shields et al, 2015;Vaidyanathan et al, 2018) and for cellular membrane modelling (Hirano-Iwata et al, 2010;Strulson and Maurer, 2011;Galvez et al, 2020), among others (Beebe, et al, 2002;Sackmann, et al, 2014). In fact, microtechnology enables the precise control of the topography and the surface chemistry, leading to engineered platforms for the study of cellular processes or biosensing and, at the same time, bringing advantages such as time saving, reduced costs and working space, automation of the processes, increased sensitivity and reduced volumes of the required reagents (Wurm et al, 2010;Azuaje-Hualde et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Advances in Microtechnology for Improved Cytotoxicity Assessment

2020

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In vitro cytotoxicity testing is essential in the pharmaceutical and environmental industry to study the effects of potential harmful compounds for human health. Classical assays present several disadvantages: they are commonly based on live-death labelling, are highly time consuming and/or require skilled personnel to be performed. The current trend is to reduce the number of required cells and the time during the analysis, while increasing the screening capability and the accuracy and sensitivity of the assays, aiming single cell resolution. Microfabrication and surface engineering are enabling novel approaches for cytotoxicity assessment, offering high sensitivity and the possibility of automation in order to minimize user intervention. This review aims to overview the different microtechnology approaches available in this field, focusing on the novel developments for high-throughput, dynamic and real time screening of cytotoxic compounds.

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Extracellular matrix protein microarray-based biosensor with single cell resolution: Integrin profiling and characterization of cell-biomaterial interactions

Cited by 16 publications

References 43 publications

Bioactive potential of natural biomaterials: identification, retention and assessment of biological properties

Bioactive potential of natural biomaterials: identification, retention and assessment of biological properties

Overcoming the Inflammatory Stage of Non-Healing Wounds: In Vitro Mechanism of Action of Negatively Charged Microspheres (NCMs)

Advances in Microtechnology for Improved Cytotoxicity Assessment

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