Mitochondrial Transport from Mesenchymal Stromal Cells to Chondrocytes Increases DNA Content and Proteoglycan Deposition In Vitro in 3D Cultures.

OBJECTIVE: Allogeneic mesenchymal stromal cells (MSCs) are used in the 1-stage treatment of articular cartilage defects. The aim of this study is to investigate whether transport of mitochondria exists between chondrocytes and MSCs and to investigate whether the transfer of mitochondria to chondrocytes contributes to the mechanism of action of MSCs. DESIGN: Chondrocytes and MSCs were stained with MitoTracker, and CellTrace was used to distinguish between cell types. The uptake of fluorescent mitochondria was measured in cocultures using flow cytometry. Transport was visualized using fluorescence microscopy. Microvesicles were isolated and the presence of mitochondria was assessed. Mitochondria were isolated from MSCs and transferred to chondrocytes using MitoCeption. Pellets of chondrocytes, chondrocytes with transferred MSC mitochondria, and cocultures were cultured for 28 days. DNA content and proteoglycan content were measured. Mitochondrial DNA of cultured pellets and of repair cartilage tissue was quantified. RESULTS: Mitochondrial transfer occurred bidirectionally within the first 4 hours until 16 hours of coculture. Transport took place via tunneling nanotubes, direct cell-cell contact, and extracellular vesicles. After 28 days of pellet culture, DNA content and proteoglycan deposition were higher in chondrocyte pellets to which MSC mitochondria were transferred than the control groups. No donor mitochondrial DNA was traceable in the biopsies, whereas an increase in MSC mitochondrial DNA was seen in the pellets. CONCLUSIONS: These results suggest that mitochondrial transport plays a role in the chondroinductive effect of MSCs on chondrocytes in vitro. However, in vivo no transferred mitochondria could be traced back after 1 year.

Platelet-Rich Plasma Does Not Inhibit Inflammation or Promote Regeneration in Human Osteoarthritic Chondrocytes In Vitro Despite Increased Proliferation.

OBJECTIVE: The aims of the study were to assess the anti-inflammatory properties of platelet-rich plasma (PRP) and investigate its regenerative potential in osteoarthritic (OA) human chondrocytes. We hypothesized that PRP can modulate the inflammatory response and stimulate cartilage regeneration. DESIGN: Primary human chondrocytes from OA knees were treated with manually prepared PRP, after which cell migration and proliferation were assessed. Next, tumor necrosis factor-α-stimulated chondrocytes were treated with a range of concentrations of PRP. Expression of genes involved in inflammation and chondrogenesis was determined by real-time polymerase chain reaction. In addition, chondrocytes were cultured in PRP gels and fibrin gels consisting of increasing concentrations of PRP. The production of cartilage extracellular matrix (ECM) was assessed. Deposition and release of glycosaminoglycans (GAG) and collagen was quantitatively determined and visualized by (immuno)histochemistry. Proliferation was assessed by quantitative measurement of DNA. RESULTS: Both migration and the inflammatory response were altered by PRP, while proliferation was stimulated. Expression of chondrogenic markers COL2A1 and ACAN was downregulated by PRP, independent of PRP concentration. Chondrocytes cultured in PRP gel for 28 days proliferated significantly more when compared with chondrocytes cultured in fibrin gels. This effect was dose dependent. Significantly less GAGs and collagen were produced by chondrocytes cultured in PRP gels when compared with fibrin gels. This was qualitatively confirmed by histology. CONCLUSIONS: PRP stimulated chondrocyte proliferation, but not migration. Also, production of cartilage ECM was strongly downregulated by PRP. Furthermore, PRP did not act anti-inflammatory on chondrocytes in an in vitro inflammation model.

Preclinical Feasibility of the Bio-Airbrush for Arthroscopic Cell-Based Cartilage Repair.

IMPACT STATEMENT: This study shows that a bio-airbrush can be a clinically feasible instrument for the use in cell transplantations to repair knee cartilage defects. This technology would enable a shift from cartilage repair procedures, which are mostly performed using an arthrotomy, toward a fully arthoscopic approach. In addition, the technology presented could possibly be translated to other fields of research where controlled in situ deposition of cells is required, such as treatment of burn wounds/ulcers/others. Our research has shown that the use of an airbrush is safe, without harming cell viability and performance.

The feeling of throwing good money after bad: The role of affective reaction in the sunk-cost fallacy.

Continuing investing in a failing plan (i.e., the sunk-cost fallacy) is a common error that people are inclined to make when making decisions. It is impossible to get resources back that already have been invested. Hence, economic theory implies that decision makers' decisions should only be guided by future gains and losses. According to the literature, the sunk-cost fallacy is driven by negative affect. Previous studies focused on negative incidental affect. We investigated, in contrast, whether the sunk-cost fallacy is caused by integral affect elicited by the specific decision context. Study 1 demonstrated a positive relationship between affective reaction and the sunk-cost fallacy. Study 2 replicated the finding in Study 1 in a within-subjects design, and demonstrated a full mediation of type of scenario (invest vs. non-invest) on the sunk-cost effect, mediated by integral affective reaction. A mediation using a within-subjects design additionally demonstrated that the effect is mediated by integral emotional responses experienced in relation to each scenario, and not by incidental emotional states that are unrelated to the scenarios. Study 3 replicated findings in the previous studies, and demonstrated that the relation between the sunk-cost fallacy and affect is moderated by justification. Participants who justified their decision were more resistant to the sunk-cost fallacy, and showed less negative affect elicited by the scenarios, than participants who did not justify their decision. Study 4 provided supporting evidence for our hypothesis by hindering conscious deliberation, and promoting reliance on affect, via cognitive load. The results showed that the relation between affect and the sunk-cost fallacy was stronger for participants under high cognitive load, than under low-load. The paper discussed how this research leads to new ways to protect against the sunk-cost fallacy in the discussion.

A Rate-Reduced Neuron Model for Complex Spiking Behavior.

We present a simple rate-reduced neuron model that captures a wide range of complex, biologically plausible, and physiologically relevant spiking behavior. This includes spike-frequency adaptation, postinhibitory rebound, phasic spiking and accommodation, first-spike latency, and inhibition-induced spiking. Furthermore, the model can mimic different neuronal filter properties. It can be used to extend existing neural field models, adding more biological realism and yielding a richer dynamical structure. The model is based on a slight variation of the Rulkov map.

Cross-scale effects of neural interactions during human neocortical seizure activity.

Small-scale neuronal networks may impose widespread effects on large network dynamics. To unravel this relationship, we analyzed eight multiscale recordings of spontaneous seizures from four patients with epilepsy. During seizures, multiunit spike activity organizes into a submillimeter-sized wavefront, and this activity correlates significantly with low-frequency rhythms from electrocorticographic recordings across a 10-cm-sized neocortical network. Notably, this correlation effect is specific to the ictal wavefront and is absent interictally or from action potential activity outside the wavefront territory. To examine the multiscale interactions, we created a model using a multiscale, nonlinear system and found evidence for a dual role for feedforward inhibition in seizures: while inhibition at the wavefront fails, allowing seizure propagation, feedforward inhibition of the surrounding centimeter-scale networks is activated via long-range excitatory connections. Bifurcation analysis revealed that distinct dynamical pathways for seizure termination depend on the surrounding inhibition strength. Using our model, we found that the mesoscopic, local wavefront acts as the forcing term of the ictal process, while the macroscopic, centimeter-sized network modulates the oscillatory seizure activity.

Arthroscopic Airbrush-Assisted Cell Spraying for Cartilage Repair: Design, Development, and Characterization of Custom-Made Arthroscopic Spray Nozzles.

INTRODUCTION: Airbrush-assisted cell spraying would facilitate fully arthroscopic filling of cartilage defects, thereby providing a minimally invasive procedure for cartilage repair. This study provides the development and characterization of custom-made spray nozzles that could serve as a foundation for the development of a BioAirbrush, a platform technology for the arthroscopic application of (cell laden) hydrogels. MATERIALS AND METHODS: Custom-made spray nozzles were designed and produced with 3D printing technology. A commercially available spraying system was used for comparison. Sprays were characterized based on spray angle, cone width, droplet size, velocity, and density. This was performed with conventional and high-speed imaging. Furthermore, cell survival of chondrocytes and mesenchymal stromal cells, as well as the chondrogenic capacity of chondrocytes after spraying were evaluated. RESULTS: Changing nozzle design from internal to external mixing significantly increased cell survival after spraying. Custom-made spray nozzles provide larger droplets compared to the current commercially available technology, potentially improving cell survival. Sufficient mixing of two gel components was confirmed for the custom-made nozzles. Overall, custom-made nozzles improved cell survival after spraying, without significantly affecting the chondrogenic capacity of the cells. CONCLUSIONS: This study provides a platform for the development of a BioAirbrush for spray-assisted cell implantations in arthroscopic cartilage repair procedures. Evaluation of the fundamental characteristics of a spray as well as a study of cell survival after spraying have further expanded the knowledge regarding cell spraying for cartilage repair. Nozzle design and air pressure characteristics are essential parameters to consider for the clinical implementation of spray-assisted cell implantations.

A Biophysical Model for Cytotoxic Cell Swelling.

We present a dynamic biophysical model to explain neuronal swelling underlying cytotoxic edema in conditions of low energy supply, as observed in cerebral ischemia. Our model contains Hodgkin-Huxley-type ion currents, a recently discovered voltage-gated chloride flux through the ion exchanger SLC26A11, active KCC2-mediated chloride extrusion, and ATP-dependent pumps. The model predicts changes in ion gradients and cell swelling during ischemia of various severity or channel blockage with realistic timescales. We theoretically substantiate experimental observations of chloride influx generating cytotoxic edema, while sodium entry alone does not. We show a tipping point of Na+/K+-ATPase functioning, where below cell volume rapidly increases as a function of the remaining pump activity, and a Gibbs-Donnan-like equilibrium state is reached. This precludes a return to physiological conditions even when pump strength returns to baseline. However, when voltage-gated sodium channels are temporarily blocked, cell volume and membrane potential normalize, yielding a potential therapeutic strategy. SIGNIFICANCE STATEMENT: Cytotoxic edema most commonly results from energy shortage, such as in cerebral ischemia, and refers to the swelling of brain cells due to the entry of water from the extracellular space. We show that the principle of electroneutrality explains why chloride influx is essential for the development of cytotoxic edema. With the help of a biophysical model of a single neuron, we show that a tipping point of the energy supply exists, below which the cell volume rapidly increases. We simulate realistic time courses to and reveal critical components of neuronal swelling in conditions of low energy supply. Furthermore, we show that, after transient blockade of the energy supply, cytotoxic edema may be reversed by temporary blockade of Na+ channels.

Effects of processing style on responsiveness to affective stimuli and processing fluency.

In the present study, we provide direct evidence for effects of global versus local processing on responsiveness to and reliance on affective information in judgement and decision-making. Results of Experiments 1 and 2 showed an increased responsiveness to affective stimuli among participants in a global processing mode. Experiment 3 showed similar effects for processing fluency; participants adopting a global processing style showed an increased reliance on fluency. Experiment 4 replicated our findings in a more mundane judgement task in which participants judged apartments. We discuss our findings in relation to the distinction between intuitive versus deliberative modes of thinking.