Piezo1 is a mechanosensitive cation channel with widespread physiological importance; however, its role in the heart poorly understood. Cardiac fibroblasts help preserve myocardial integrity and play key regulating repair remodeling following stress or injury. Here we investigated expression function cultured human mouse cardiac fibroblasts. RT-PCR experiments confirmed that mRNA expressed at levels similar to those endothelial cells. The results of Fura-2 intracellular Ca2+ assay validated...

The aetiology of intervertebral disc (IVD) degeneration remains poorly understood. Painful IVD is associated with an acidic intradiscal pH but the response NP cells to this aberrant microenvironmental factor be fully characterised. aim here was address hypothesis that pH, similar found in degenerate IVDs, leads altered cell/functional phenotype observed during degeneration, and investigate involvement acid-sensing ion channel (ASIC) -3 response. Human were treated a range from non-degenerate...

Abstract Background Intervertebral disc (IVD) degeneration is a common cause of low back pain, and the most symptomatic patients with neural compression need surgical intervention to relieve symptoms. Current techniques used diagnose IVD degeneration, such as magnetic resonance imaging (MRI), do not detect changes in tissue extracellular matrix (ECM) progresses. Improved techniques, combination blood biomarkers, are needed monitor progression for more effective treatment plans. Methods To...

A water-soluble azide-functionalised chitosan was crosslinked with propiolic acid ester-functional poly(ethylene glycol) using copper-free click chemistry. The resultant hydrogel materials were formed within 5-60 min at 37 °C and resulted in mechanically robust tuneable properties such as swelling, mechanical strength degradation. Importantly, the hydrogels supported mesenchymal stem cell attachment proliferation also non-toxic to encapsulated cells. As these studies indicate that have...

Studies of cellular mechano-signaling have often utilized static models that do not fully replicate the dynamics living tissues. Here, we examine time-dependent response primary human mesenchymal stem cells (hMSCs) to cyclic tensile strain (CTS). At low-intensity (1 h, 4% CTS at 1 Hz), cell characteristics mimic responses increased substrate stiffness. As regime is intensified (frequency 5 characterize rapid establishment a broad, structured and reversible protein-level response, even as...

Mechanical loads are important for homeostasis of the intervertebral disc (IVD) cell matrix, with physiologic and nonphysiologic leading to matrix anabolism catabolism, respectively. Previous investigations into effects load on cells have predominantly used animal models, limited number human studies focusing primarily nucleus pulposus cells. The aim this study was examine effect cyclic tensile strain (CTS) anulus fibrosus (AF) ascertain whether response frequency-dependent compare AF...

The mechanical environment can influence cell behaviour, including changes to transcriptional and proteomic regulation, morphology and, in the case of stem cells, commitment lineage. However, current tools for characterizing substrates' properties, such as atomic force microscopy (AFM), often do not fully recapitulate length time scales over which cells 'feel' substrates. Here, we show that an immortalised, clonal line human mesenchymal (MSCs) maintains responsiveness substrate mechanics...

Recent evidence suggests that intervertebral disc (IVD) cells derived from degenerative tissue are unable to respond physiologically relevant mechanical stimuli in the 'normal' anabolic manner, but instead by increasing matrix catabolism. Understanding nature of biological processes which allow sense and (a process termed 'mechanotransduction') is important ascertain whether these signalling pathways differ with disease. The aim here was investigate involvement interleukin (IL)-1 IL-4...

A new class of degradable aliphatic poly(carbonate) resins for use in microstereolithographic process is described. Using a biologically inert photo-inhibiting dye, exemplar 3-dimensional structures were produced using thiol-ene chemistry via microstereolithography. Fabricated constructs demonstrated good biological compatibility with cells and had tensile properties that render them suitable as tissue engineering scaffolds.

Cells respond to stress by synthesizing chaperone proteins that seek correct protein misfolding and maintain function. However, abrogation of homeostasis is a hallmark aging, leading loss function the formation proteotoxic aggregates characteristic pathology. Consequently, discovering underlying molecular causes this deterioration in proteostasis key designing effective interventions disease or maintaining cell health regenerative medicine strategies. Here, we examined primary human...

A novel method for the production of inhibitor- and solvent-free resins suitable three-dimensional (3D) microstereolithography is reported. Using an exemplar poly(ethylene glycol)-based resin, control features in X, Y, Z planes demonstrated such that complex structures can be manufactured. Human mesenchymal stem cells cultured on manufactured scaffolds remained viable during 7 day assessment period, with proliferation rates comparable to those observed tissue culture polystyrene. These data...

Intervertebral disc (IVD) cells derived from degenerate tissue respond aberrantly to mechanical stimuli, potentially due altered mechanotransduction pathways. Elucidation of the altered, or alternative, pathways operating with degeneration could yield novel targets for treatment IVD disease. Our aim here was investigate involvement RGD-recognising integrins and associated signalling molecules in response cyclic tensile strain (CTS) human annulus fibrosus (AF) non-degenerate IVDs. AF IVDs...

Mechanical stimulation is known to influence intervertebral disc (IVD) cell behavior and function, but the effect on cells routinely considered in isolation from other microenvironmental factors. Acidic pH has been shown be a prominent detrimental factor present degenerate IVDs, its human mechanoresponse never studied. We investigated response of agarose‐encapsulated nucleus pulposus (NP) 0.004 MPa, 1.0 Hz 1 hour compression (Flexcell FX4000 Compression System) under conditions...

Growth differentiation factor (GDF) family members have been implicated in the development and maintenance of healthy nucleus pulposus (NP) tissue, making them promising therapeutic candidates for treatment intervertebral disc (IVD) degeneration associated back pain. GDF6 has shown to promote discogenic mesenchymal stem cells, but its effect on NP cells remains largely unknown. Our aim was investigate signalling adult human derived from degenerate tissue determine signal transduction...

Summary Our current understanding of cellular mechano-signalling is based on static models, which do not replicate the dynamics living tissues. Here, we examined time-dependent response primary human mesenchymal stem cells (hMSCs) to cyclic tensile strain (CTS). At low-intensity (1 hour, 4% CTS at 1 Hz) morphological changes mimicked responses increased substrate stiffness. As regime was intensified (frequency 5 Hz), characterised rapid establishment a broad, structured and reversible...

In this issue of Biotechnology Journal, Harkness et al. [1] demonstrate a technology that combines patterned substrates, for the purpose controlling cell geometry, with multi-well plate layouts to allow high-throughput analysis inputs affect behavior.

Abstract The mechanical environment can influence cell behaviour, including changes to transcriptional and proteomic regulation, morphology and, in the case of stem cells, commitment lineage. However, current tools for characterizing substrates’ properties, such as atomic force microscopy (AFM), often do not fully recapitulate length time scales over which cells ‘feel’ substrates. Here, we show that an immortalised, clonal line human mesenchymal (MSCs) maintained responsiveness substrate...

Abstract Background Stem cell‐based therapies show promise as a means of repairing the degenerate intervertebral disc, with growth factors often used alongside cells to help direct differentiation toward nucleus pulposus (NP)‐like phenotype. We previously demonstrated adipose‐derived stem cell (ASC) GDF6 optimal for generating NP‐like through evaluating end‐stage parameters. Here we conducted time‐resolved transcriptomic characterization ASCs response stimulation understand early drivers...

ABSTRACT Cells respond to stress by synthesising chaperone proteins that correct protein misfolding maintain function. However, homeostasis is lost in ageing, leading aggregates characteristic of protein-folding diseases. Whilst much known about how these diseases progress, discovering what causes deteriorate could be key their prevention. Here, we examined primary human mesenchymal stem cells (hMSCs), cultured a point replicative senescence and subjected heat shock, as an vitro model the...

Abstract Piezo1 is a mechanosensitive cation channel with widespread physiological importance; however its role in the heart poorly understood. Cardiac fibroblasts are responsible for preserving structural integrity of myocardium and play key regulating repair remodeling following stress or injury. We investigated expression function cultured human mouse cardiac fibroblasts. RT-PCR studies confirmed mRNA at similar levels to endothelial cells. Fura-2 intracellular Ca 2+ measurements...

ABSTRACT Tissues are maintained by homeostatic feedback mechanisms where cells respond to, but also modify, the chemical and mechanical properties of surrounding extracellular matrix. Mesenchymal stem (MSCs) resident in marrow niche experience a diverse environment, ageing can affect composition quality bone tissues. Here we quantified effect replication-induced senescence on MSC morphology their ability to correctly different substrate stiffnesses. The matrix proteome was found be sensitive...