Pregnancy-associated hemodynamic overload and hormonal changes induce hypertrophy and metabolic remodeling of the maternal heart. Mitochondrial motility, mediated by ras homolog family member T (RHOT) 1 and RHOT2, is essential for cardiac adaptation to increased workload, cardiomyocyte hypertrophy, and sarcomere maturation. To test the hypothesis that Rhot1/2 expression is required for pregnancy- and postpartum-associated adaptations of the maternal heart, female mice with tamoxifen-inducible, cardiomyocyte-selective deletion of Rhot1 and Rhot2 (iRhot1/2-KO) were mated. Following gene deletion in adult mice, cardiac tissue and function were analyzed after three to five successive pregnancies and postpartum nursing periods. Age-matched nulliparous iRhot1/2-KO mice and age-matched mice expressing Rhot1 and Rhot2 served as controls. Motility of mitochondria isolated from iRhot1/2-KO hearts was impaired, as determined by the number of mobile mitochondria in an in vitro motor protein-driven single mitochondrion motility assay performed on surface-immobilized microtubules. Despite loss of Rhot1/2 expression, contractile function assessed by transthoracic echocardiography, mRNA expression of peripartum-associated heart failure markers, cardiac structure, mitochondrial morphology, mitochondrial enzymatic activity, and mitochondrial DNA content were all comparable to controls expressing Rhot1/2 at the investigated time points. RNA sequencing-based gene profiling identified a transcriptional program through which RHOT proteins preserve cardiac energetic and contraction gene expression during pregnancy and postpartum. Together, cardiomyocyte-selective loss of Rhot1/2 expression in the adult heart does not cause peripartum-associated heart failure, despite reduced cardiac energetic and contraction gene expression.
High-content screening (HCS) is a powerful approach for rapidly and efficiently assessing the harmfulness of numerous compounds across a wide range of cultured cell types. We recently developed a fully automated, miniaturized HCS wet-plus-dry pipeline (MITOMATICS) which leverages mitochondrial morphology as a sensitive and dynamic biomarker of cellular health or damage. Mitochondria are indeed not only vital for energy production and homeostasis, but also serve as critical gatekeepers of apoptotic cell death. MITOMATICS incorporates a proprietary software tool (MitoRadar) designed in-house to perform fast, comprehensive and cost-effective analysis of mitochondrial morphology in live cells. Together, the pipeline and its associated big data analytics software provide a valuable framework for early detection of acute mitotoxic effects of chemicals agents or physical stressors. To illustrate this, we present here a complete protocol for quantifying the impact of the pesticide chlorpyrifos-methyl on mitochondrial morphology of human lung epithelial BEAS-2B cells. Our results show that chlorpyrifos-methyl, even as a single compound, induces profound disruptions in mitochondrial subcellular structure. Beyond this case study, MitoRadar opens up promising avenues for investigating mitotoxicity across diverse cell types and environmental exposures, paving the way for a new generation of cellular diagnostics that could be of interest to the cell death community.
The choroid plexus epithelial cells (CPECs) at the blood-cerebrospinal fluid (CSF) interface possess an exceptionally high mitochondrial content to support CNS homeostasis. Oncocytic CPECs (O-CPECs), characterized by enlarged and granular eosinophilic cytoplasm composed of excessive abnormal mitochondria, likely contribute to an energetic failure of this energy-demanding tissue. The relationship between O-CPECs and other CPEC pathologies in humans, such as Biondi body (BB) amyloid inclusions, remains poorly defined. In the present study, using H&E-stained sections from 68 postmortem cases, we classified O-CPECs by quantitative size criteria and cytological features, and found an increase in the prevalence of O-CPECs with age after adjusting for sex and tissue source. After excluding two influential control cases, there was evidence for a further increase associated with Alzheimer's disease. Using antibodies to ATP synthase beta chain to classify O-CPECs, and thioflavin-S to identify BBs, we revealed an increased prevalence of BBs in O-CPECs compared to neighboring non-oncocytic cells. Small multiple BB inclusions were responsible for the increase in O-CPECs, while the prevalence of larger inclusions was decreased in O-CPECs. Together, our data support a clear age-associated oncocytic transformation of CPECs and implicate mitochondrial dysfunction-amyloid interactions.
Mitochondrial dysfunction is not merely a byproduct of transformation but a driver of tumorigenesis, metastasis, and therapeutic resistance. Recent advancements in intercellular communication have identified Extracellular Vesicles (EVs) or exosomes as critical mediators that bridge the gap between the tumor and its microenvironment (TME). These EVs contain a complex repertoire of bioactive cargo, including proteins, lipids, and RNAs. Among the class of RNAs, small non-coding RNAs, microRNAs (miRNAs), are the most abundantly expressed bioactive compounds that are selectively packaged and delivered to recipient cells. EV-delivered miRNAs can target nuclear-encoded mitochondrial genes and have also been reported to localize to mitochondria (mitomiRs), where they function as post-transcriptional regulators of bioenergetic and mitochondrial dynamic adaptations that support tumor progression. This review explores the "EV-miRNA-Mitochondria Axis", delineating the molecular mechanisms by which EV-carried miRNAs reprogram the "Mitochondrial Information Processing System" (MIPS) - a signaling network where mitochondria integrate metabolic cues (e.g., ROS, calcium flux) to dictate critical biological outcomes, such as immune regulation and cell survival. We summarized specific sorting machineries (e.g., hnRNPA2B1, Lupus La) that package oncogenic miRNAs into EVs and how these cargoes hijack mitochondrial function upon delivery. Specifically, we discussed how EV-miRNAs induce metabolic shifts, manipulate mitochondrial dynamics (fission/fusion), and inhibit the intrinsic apoptosis to drive cancer progression. Finally, we highlighted the dual utility of these EV-miRNAs as drivers of pathogenesis and promising non-invasive biomarkers for early diagnosis, prognostic and therapeutic monitoring.
Neuronal aging is a key but often overlooked part of Alzheimer's disease. It links age-related loss of cellular energy to long-lasting problems in neuronal function. Even though neurons don't usually divide, ongoing stressors like oxidative damage, mitochondrial dysfunction, and problems with protein handling can make them appear old. This is seen as lasting DNA damage, calcium imbalance, and the release of substances that cause inflammation. These changes are closely tied to loss of balance in cell energy and redox function. Mitochondria, which make most neurons' energy, are both a main source and a target for reactive oxygen species (ROS). Long-term redox imbalance damages energy production, lowers NAD+, and disrupts SIRT1 regulation. Eventually, this leads to energy failure and loss of synaptic function. In AD, excessive ROS production and redox imbalances interact with amyloid-β toxicity, tau hyperphosphorylation, and metal ion disturbances. This convergence increases mitochondrial damage and fragmentation. When mitophagy is impaired, dysfunctional mitochondria are not removed, leading to ROS accumulation that further damages cellular structures and reinforces oxidative stress. This forms a self-perpetuating cycle that accelerates neuronal aging and neurodegeneration. Notably, research shows that energy failure and redox imbalance often precede the formation of amyloid plaques and tangles, suggesting that these mechanisms may initiate disease onset. This chapter reviews core mechanisms underlying redox signaling and neuronal senescence. It details how altered mitochondrial function disrupts neuronal energy homeostasis and triggers a cascade of molecular events that underlie Alzheimer's disease. The proposed framework connects redox failure, cellular senescence, and neurodegeneration as interdependent drivers of disease progression.
Perrault syndrome is a genetically and clinically diverse autosomal recessive disorder characterized by sensorineural hearing loss in both sexes and primary ovarian insufficiency in females. This comprehensive review synthesizes data from various studies to map the genetic architecture of Perrault syndrome, highlighting mutations in fifteen principal genes: HSD17B4, HARS2, CLPP, LARS2, TWNK, ERAL1, RMND1, DAP3, PRORP, MRPL50, MRPL49, MRPS7, PEX6, GGPS1, and TFAM. Each of these genes plays a critical role either in mitochondrial function or peroxisomal processes, central to cellular energy metabolism and biosynthesis pathways. The review not only documents the spectrum of mutations found within these genes but also correlates specific genetic alterations with the range of phenotypes observed in patients, emphasizing the syndrome's allelic, locus, and clinical heterogeneity. The cohort demonstrates a distribution of 56.1% homozygous and 43.9% compound heterozygous variants, reflecting diverse ancestral backgrounds and potential selective pressures against deleterious alleles. The findings underscore the necessity for advanced genetic screening techniques in accurate diagnosis and the potential for gene-specific therapies that may mitigate some of the clinical manifestations of this complex condition.
Background: Hashimoto's thyroiditis (HT) is a common autoimmune disorder characterized by chronic inflammation and metabolic alterations. Mitochondria-derived peptides (MDPs), particularly mitochondrial open-reading frame of the 12S rRNA-c (MOTS-c), have emerged as key regulators of cellular metabolism, insulin sensitivity, oxidative stress, and inflammatory responses. This study aimed to investigate the association between circulating MOTS-c levels and HT and to explore its potential role in thyroid autoimmunity and metabolic regulation. Methods: In this cross-sectional study, patients diagnosed with HT (n: 90) were compared with age- and sex-matched healthy controls (n: 90). Results: A total of 180 participants were included, comprising 90 patients with HT and 90 age- and sex-matched healthy controls. Circulating MOTS-c levels were significantly lower in patients with HT compared to controls (p < 0.001). MOTS-c levels demonstrated significant inverse correlations with body mass index, fasting glucose, HbA1c, HOMA-IR, thyroid-stimulating hormone, C-reactive protein, and thyroid autoantibody levels (all p < 0.05). In subgroup analyses, these associations remained significant within the HT cohort, particularly for HOMA-IR and thyroid autoantibodies. Multivariable regression analysis identified HT (β = -30.04, p < 0.001) and HOMA-IR (β = -0.85, p < 0.001) as independent determinants of reduced circulating MOTS-c levels. Levothyroxine (LT4) use was not associated with significant differences in MOTS-c concentrations. Conclusions: Circulating MOTS-c levels are markedly reduced in patients with HT and are independently associated with insulin resistance and autoimmune burden. These findings suggest that impaired mitochondrial signaling may play a role in the pathophysiology of thyroid autoimmunity and highlight MOTS-c as a promising biomarker linking metabolic dysfunction and immune dysregulation.
Sepsis is a dynamic syndrome of infection-driven metabolic and immune dysregulation in which oxidative stress can escalate into an "oxidative storm," promoting organ dysfunction and maladaptive host responses. Within this context, ferroptosis represents a metabolically constrained form of regulated necrotic cell death driven by iron-dependent lipid peroxidation, linking redox collapse to tissue injury in sepsis. Emerging evidence suggests that autophagy critically shapes ferroptosis susceptibility by regulating intracellular iron mobilization, membrane lipid substrate availability, mitochondrial quality control, and energy-stress signaling. This review therefore frames autophagy-ferroptosis crosstalk in sepsis as a host metabolic vulnerability and discusses how mechanism-guided, host-directed antioxidant nanomedicine may help preserve tissue integrity while limiting interference with antimicrobial defense. We explored how autophagy modulates ferroptosis susceptibility by regulating iron metabolism, lipid substrate availability, and mitochondrial quality control. Building on this framework, we evaluated emerging antioxidant nanomedicines targeting key intervention points, including iron chelation, catalytic ROS/RNS scavenging, membrane-localised radical trapping, mitochondria-targeted source control, and enhancement of endogenous defences. Organ- and immune-specific effects are highlighted, emphasizing the need for aligned biochemical readouts, flux-aware autophagy evaluation, and stage-specific therapeutic targeting. Finally, we outline translational priorities for precision redox modulation in sepsis, focusing on biomarker-guided patient stratification, compartment-specific delivery, and biosafety considerations.
Adipocyte lipid metabolism is coordinated by circadian rhythms, diet, and environmental temperature. Yet how these diverse signals are molecularly integrated remains unknown. Here we show that clock, diet, and temperature cues converge on the orphan mitochondrial transporter, SLC25A34, to orchestrate thermogenic cycling of lipid synthesis and oxidation. During sleep, the clock suppresses Slc25a34 transcription through REV-ERBα. Waking, lipid-rich diets, or cold exposure abolish this repression, allowing lipolytic signals to stimulate Slc25a34 expression via PPARα. SLC25A34 then imports oxaloacetate into mitochondria to accelerate the export of substrates used for acetyl-CoA production in the cytosol. This feeds into cytosolic lipid synthesis and transcriptional induction of mitochondrial biogenesis, which collectively promote mitochondrial lipid oxidation. Thus, SLC25A34 confers circadian, dietary, and environmental control of thermogenic metabolism through interorganellar lipid cycling.
The rotenone adjuvant kindling paradigm replicates key clinical features of drug-resistant epilepsy (DRE), including broad-spectrum pharmacoresistance, neuroinflammation, and oxidative stress, by combining mitochondrial complex I inhibition with pentylenetetrazol (PTZ) or corneal kindling. PTZ, a GABAA receptor antagonist, induces seizures by reducing inhibitory neurotransmission and is widely used to model kindling and seizure susceptibility; in this paradigm, its repeated subconvulsive dosing facilitates progressive epileptogenesis and enhances network hyperexcitability. Rotenone-induced microglial activation and mitochondrial dysfunction further potentiate PTZ sensitivity, limiting the penetration and effectiveness of antiseizure drugs by promoting cytokine release, disrupting the blood-brain barrier, and overexpressing efflux transporters. In comparison to traditional DRE models, this paradigm's construct, face, and predictive validity are strengthened by its recapitulation of neuropsychiatric comorbidities and spontaneous recurrent seizures. Comparative findings suggest greater clinical relevance and improved suitability for evaluating mechanism-based therapies targeting mitochondrial dysfunction, GABAergic imbalance, and inflammatory signaling; however, concerns regarding systemic toxicity, mortality, and inter-animal variability remain important limitations. Overall, the rotenone adjuvant PTZ-kindling model represents a promising, though imperfect, translational platform for the development of novel therapies for DRE.
O-GlcNAcylation is a ubiquitous post-translational modification regulated by O-GlcNAcase (OGA) and O-GlcNAc transferase (OGT) in response to environmental and genetic alterations. It occurs in the nucleus, mitochondrion, and cytoplasm and is implicated in cardiovascular disease (CVD) development. O-GlcNAcylation modulates diverse cellular processes, including metabolic pathways, signaling networks, and transcriptional programs. Acute increase in O-GlcNAcylation serves as an adaptive response that preserves cardiac function, whereas chronic elevation leads to persistent metabolic dysregulation and promotes pathological cardiac remodeling. In this review, we provide a comprehensive overview of the role of O-GlcNAcylation across diverse disease contexts. We also summarize the current understanding of its complex interplay with CVD, including the underlying mechanisms. Finally, we highlight existing knowledge gaps and discuss the therapeutic potential of targeting O-GlcNAcylation in various cardiovascular events, emphasizing key priorities for future research.
White adipose browning is a promising route to restore energy balance; however, how inorganic anion signals engage intracellular organelle networks to drive this process remains unclear. Here, we identify Sialin2 as a nitrate sensor that converts dietary nitrate into a spatially confined thermogenic program by coupling ER-mitochondria Ca2+ transfer with lipid routing into mitochondrial oxidation. Sialin2 localizes to mitochondria and the endoplasmic reticulum (ER), where it strengthens ER-mitochondria contacts and engages the inositol 1,4,5-trisphosphate receptor type 1 (IP3R1)-voltage-dependent anion channel 1 (VDAC1)-mitochondrial calcium uniporter 1 (MCU1) conduit to enhance inducible mitochondrial Ca2+ uptake. In parallel, Sialin2 associates with lysosomal acid lipase (LIPA), acyl-CoA synthetase long-chain family member 3 (ACSL3), and carnitine palmitoyltransferase 1 A (CPT1A) to channel lipid-droplet-derived fatty acids into β-oxidation, thereby fueling the tricarboxylic acid cycle and uncoupling protein 1 (UCP1)-dependent respiration. Loss of Slc17a5 abolishes nitrate-evoked browning and metabolic benefits, whereas nitrate supplementation improves adipose thermogenesis and systemic metabolic indices in male mice with diet-induced obesity without adrenergic stimulation. Together, these findings identify an organelle-specific nitrate-sensing mechanism that couples inorganic anion signalling to substrate routing in adipocytes and establish a non-hormonal pathway for restoring metabolic homeostasis.
Ribosome stalling caused by polyproline (PPs) motifs is common. Their translation is enhanced by accessory proteins such as YebC in bacteria, whose homolog, TRANSLATIONAL ACTIVATOR OF CYTOCHROME C OXIDASE 1 (TACO1), aids the translation of mitochondria-encoded proteins. The prevalence of PP motifs across plastid-encoded genes and their impact on the translation of photosynthesis-relevant proteins remains unexplored. Equally, a translation-enhancer of PP motifs equivalent to TACO1 for plastid ribosomes has not been reported. Here, we show that plastid genomes encode 24 proteins with a minimum of one PP motif on average, half of which are conserved in their cyanobacterial homologs, and that the vast majority of eukaryotes, including plants, encode a single TACO1 that we demonstrate to be dually targeted to mitochondria and plastids of Marchantia polymorpha . We resolved the MpTACO1 structure at 2.34 Å by X-ray crystallography and the flexibility by small-angle X-ray scattering. Through modelling, we demonstrate that MpTACO1 can fit into the peptidyl transfer centre of plant chlororibosomes in a similar manner as human TACO1 in the mitoribosome. The identification and structure determination of the first plastid-targeted YebC/TACO1 allows us to sketch a unified model for the function and evolution of this ancient family of ribosomal accessory proteins, underscoring their indispensable role in the translation of bioenergetic membrane proteins reaching back almost 4 billion years. Dozens of GC-rich polyproline (PP) encoding regions are retained by AT-rich genomesPP motif conservation hints at regulatory mechanisms and required translation pausesChloroplast targeting of a (mitochondrial) translation enhancer of PP motifsMpTACO1 structure at 2.34 Å resolution demonstrates its high level of conservation.
In this study, bis-1,10-phenanthroline (Biphen) was synthesized via a hydrogen transfer-mediated coupling reaction in a single step. The resulting compound was demonstrated, for the first time, to function as a selective fluorescent probe for Ni2+ ions. The presence of Ni2+ at a 2:1 molar ratio of Biphen to Ni2+ results in complete fluorescence quenching, with a detection limit of 4.34 × 10-9 M in aqueous medium. Fluorescence is restored upon the introduction of a suitable chelating agent, producing an "on-off-on" fluorescence switching response. Furthermore, fluorescence co-localization studies demonstrate that Biphen functions as a mitochondria-targeted fluorescent probe with excellent cell membrane permeability, enabling rapid, reversible imaging and ultratrace detection of Ni2+ in mitochondria of live cells. Overall, this work demonstrates highly selective ultratrace detection of Ni2+ in both aqueous and biological environments, providing a promising platform for mitochondrial imaging and potential diagnostic applications.
UFMylation is a post-translational modification that conjugates ubiquitin-fold modifier 1 (UFM1) to substrate proteins, regulating fundamental processes including ribosomal homeostasis, the endoplasmic reticulum (ER) stress response and DNA damage repair. While loss-of-function mutations in the UFMylation cascade cause lethality in mammals, they are viable in Caenorhabditis elegans, offering a unique opportunity to investigate its physiological role at the organismal level. We demonstrate that UFM-1 expression progressively increases from larval stages to adulthood, with predominant localization in intestinal cells. Its expression is upregulated during ER stress and autophagy induction, linking it to these pathways. We used CRISPR/Cas9 to create a targeted ufm-1 loss-of-function mutant, which revealed that UFMylation is crucial for lifespan, development and reproduction, with mutants exhibiting increased gonadal dysfunction and sterility. Deletion of ufm-1 enhanced tolerance to various stressors, a resilience potentially arising from a hormetic response to persistent ER stress. Loss of ufm-1 selectively activated the unfolded protein response in the ER but not in mitochondria. Notably, ufm-1 loss exacerbated proteotoxicity in C. elegans muscle-expressed models of protein aggregation, accelerating paralysis and increasing the number and size of amyloid-β, α-synuclein and polyQ aggregates. Furthermore, mutant worms displayed impaired locomotion, including altered swimming patterns resembling those of aging worms, stemming from accelerated, age-dependent sensory neuron dysfunction and structural neurodegeneration.
Organelle contact sites are increasingly recognized as regulatory interfaces that coordinate lipid transfer, ion signaling, and metabolic adaptation. In neurons, communication among the endoplasmic reticulum (ER), lysosomes, and mitochondria is essential for cellular homeostasis. Recent studies have identified vacuolar protein sorting 13 homolog C (VPS13C), a lipid transport protein, as a key mediator of ER-lysosome tethering and as an important component of the response to lysosomal stress. Structural analyses show that VPS13 family proteins form elongated lipid transport channels that are proposed to facilitate phospholipid transfer between adjacent membranes. Following lysosomal damage, VPS13C is recruited to ER-lysosome contact interfaces, where it forms tethering bridges that may support membrane repair by enabling high-capacity lipid transfer from the ER to lysosomal membranes. Beyond membrane repair, these contact interfaces may also participate in broader organelle communication networks. ER-lysosome contacts can occur in proximity to ER-mitochondria junctions, potentially forming multi organelle signaling hubs that coordinate lipid redistribution, calcium signaling, and mitochondrial adaptation. These signals may influence downstream responses, including activation of TFEB and TFE3, which regulate lysosomal biogenesis and autophagy. Disruption of this contact site network has emerged as a potential contributor to Parkinson's disease. Loss of VPS13C function is associated with altered lysosomal homeostasis and intersects with pathogenic pathways involving α-synuclein aggregation, PINK1/Parkin-mediated mitophagy, and LRRK2 signaling. This review presents a framework in which ER-lysosome tethering is considered part of a staged cellular damage response linking membrane repair, metabolic coordination, and transcriptional adaptation.
Mitochondria are major sources of intracellular reactive oxygen species (ROS), and act as central signaling hubs in maintaining homeostasis of cellular oxidative states. Mitochondrial permeability transition (MPT) is coordinately mediated by mitochondrial outer membrane permeabilization (MOMP) and opening of the permeability transition pore (PTP). MPT is highly sensitive to ROS, and serves as a critical checkpoint in redox balances and cell death. This review will summarize the regulatory systems of mitochondrial and intracellular redox homeostasis, as well as the recent advances in understanding of MPT regulatory mechanisms. Furthermore, this review highlights the functional roles of MPT in redox homeostasis and ferroptosis, a form of iron-dependent, lipid peroxidation-driven cell death. The PTP is a critical molecular switch, which can convert from a defender against mitochondrial redox stress and cell death processes, including specifically iron-dependent, lipid peroxidation-driven cell death, known as ferroptosis, into a ROS amplifier and cell death promoter depending on its open states. MOMP causes the uncoupling of the mitochondrial respiratory chain, and increases ROS production, leading to oxidative stress. The most recent work suggests that the interplay between MTCH2 and F-ATP synthase coordinates MOMP and the PTP opening to mediate the occurrence of MPT. This review provides insight on molecular switches that regulate MPT, determining redox state and cell death.
The global rise in metabolic disorders demands novel interventions targeting starch digestion. This study investigated two dietary phenolic acids (caffeic acid (CA) and p-hydroxycinnamic acid (p-HA)) as inhibitors of α-amylase and α-glucosidase through integrated experimental and computational approaches. Molecular docking showed distinct binding modes, and CA formed stable hydrogen bonds with catalytic residues of α-glucosidase, while p-HA interacted mainly with α-amylase via hydrophobic contacts. Enzyme kinetics revealed concentration-dependent mixed-type inhibition, with CA being more potent against α-glucosidase and p-HA against α-amylase. Spectroscopic analysis indicated both acids induced structural changes in the enzymes, with CA causing greater α-helix reduction (Δ7.03% vs. Δ2.10%) by altering the tryptophan microenvironment. Moreover, both compounds significantly suppress glucose absorption in the proximal small intestine in an ex vivo everted gut sac model, with p-HA exhibiting exceptional efficacy in the duodenum. These findings clarify structure-activity relationships and support the potential use of CA and p-HA as local intestinal agents for modulating carbohydrate absorption.
Stress-induced premature senescence (SIPS) of endothelial cells can cause endothelial dysfunction. As a first-line antidiabetic agent, the specific role of metformin in SIPS has not yet been clarified. In this study, an in vitro SIPS model was induced by exposing human umbilical vein endothelial cells (HUVECs) to hydrogen peroxide (H2O2), and the effects of metformin on cell senescence, proliferation, migration, tube formation, and mitochondrial function were evaluated. Gene expressions altered by metformin were profiled via transcriptome sequencing. Specifically, the potential involvement of migrasome-mediated mitocytosis in metformin-driven effects was examined using confocal microscopy and siRNA-mediated silencing. The results showed that metformin significantly reduced SA-β-gal activity and restored the migration and tube-forming capacities of H2O2-induced senescent HUVECs. Moreover, metformin regulated mitochondrial dynamics, restored mitochondrial membrane potential, and attenuated intracellular oxidative stress. Notably, transcriptomic and functional analyses suggested that metformin enhanced migrasome formation and migrasome-mediated mitocytosis. Inhibition of migrasome formation by siTSPAN4 abolished the protective effect of metformin against SIPS. Collectively, these findings demonstrate that metformin alleviates early SIPS-associated changes in HUVECs and suggest that migrasome-mediated mitocytosis contributes to this protection by ameliorating mitochondrial dysfunction. This provides novel mechanistic insight into the vascular protective effects of metformin.
Photobiomodulation (PBM) is a noninvasive light therapy that penetrates deeper skin layers, using wavelengths such as near-infrared. Different wavelengths target specific cellular components and pathways, influencing wound healing. Light absorption by both mitochondria and light-sensitive ion channels increases intracellular reactive oxygen species (ROS) levels, thereby affecting inflammation, platelet activation, angiogenesis, tissue remodeling, and cell viability. Concurrent use of multiple wavelengths has an advantage, activating distinct pathways involved in the wound-healing process. This in vitro study aimed to investigate the effects of concurrent PBM on fibrablasts using a dual-wavelength system (655 nm and 808 nm). The effects of concurrent PBM application at three energy densities on cell viability, intracellular ROS levels, and cell migration were assessed using two modalities: single- and triple-treatment protocols. Three different energy densities (0.5, 1, and 2 J/cm2) of a dual wavelength system were used, and applications were performed using single or triple applications. Cellular responses were evaluated by assessing viability, ROS generation, and migration. Concurrent PBM application at an energy density of 1 J/cm2 showed a synergistic effect, increasing cell viability by 3%, intracellular ROS levels by 20%, and accelerating wound closure by 89% compared with the untreated group. Concurrent dual-wavelength PBM application may reduce treatment duration and enhance wound healing, offering a promising approach for targeting multiple layers and substructures of complex biological tissue in in vivo models and clinical applications.