Gut inflammation can be triggered by multiple factors, including the loss of intestinal homeostasis and dysregulation of the innate immune system, which compromise the epithelial barrier and lead to tissue damage. Intestinal innate immunity protects the host from invading pathogens and limits microbial translocation while maintaining tolerance toward the commensal microbiota. Disruption of this balance is considered an important contributor to intestinal inflammation in inflammatory bowel diseases (IBD), encompassing ulcerative colitis (UC) and Crohn's disease (CD). While IBD pathogenesis encompasses genetic susceptibility, epigenomic dysregulation, and environmental factors, this mini-review focuses on innate immune and macrophage-driven mechanisms that integrate these upstream signals into chronic mucosal inflammation. In this context, macrophages are key innate immune cells that provide a rapid first line of defense against conserved microbial and danger signals and play a central role in initiating and sustaining inflammatory responses. In this mini-review, we describe how disruption of intestinal homeostasis triggers activation of the innate immune system, including the recruitment and activation of macrophages. Specifically, we examine the functional polarization of macrophages during inflammation and its impact on disease progression in UC and CD. We highlight the role of inflammasomes, central components of innate immune signaling, which mediate the release of pro-inflammatory cytokines and pyroptotic cell death, thereby exacerbating tissue damage and disrupting host-microbiota interactions. We also discuss trained immunity, a process through which macrophages undergo long-lasting changes following repeated inflammatory signals, which may enhance their responses to future stimuli and contribute to persistent inflammation and disease recurrence in IBD. Finally, we review therapeutic strategies targeting macrophages and innate immune pathways. Despite clinical advances, current therapies remain limited and fail to address the complex inflammatory networks underlying IBD. A deeper understanding of innate immune and inflammasome-related pathways will be relevant for the development of multitargeted therapeutic strategies in IBD.
Anterior knee pain (AKP) is a common musculoskeletal condition with heterogeneous presentations, including unilateral and bilateral symptoms. Although altered gait biomechanics have been reported in AKP, it remains unclear whether unilateral and bilateral AKP exhibit distinct biomechanical phenotypes, particularly in terms of interlimb asymmetry. To compare lower-limb kinematics and joint moments during level walking between individuals with unilateral and bilateral AKP, and to quantify interlimb asymmetry using symmetry indices. Seventy-one individuals with AKP (unilateral: n = 36 (16 male and 20 female); bilateral: n = 35 (16 male and 19 female); age range: 11-35) underwent three-dimensional gait analysis at a self-selected speed. Hip, knee, and ankle joint angles (three planes) and normalized joint moments were extracted. The primary kinetic outcome was peak knee extension moment within 0%-60% of the gait cycle. Predefined kinematic and kinetic peak measures for the hip, knee, and ankle joints during defined phases of the stance period were also analyzed. We collected surface electromyographic (sEMG) data from a subset of participants to analyze muscle activation patterns during gait. Interlimb asymmetry was quantified using symmetry indices between more symptomatic (MS) limb for unilateral AKP; and less symptomatic (LS) limb. Between-group comparisons (unilateral vs. bilateral AKP) for baseline characteristics and symmetry indices (SI) were conducted using independent samples t-tests or Mann-Whitney U tests. Within-group differences (more symptomatic vs. less symptomatic limb) for kinematic and kinetic parameters were analyzed using paired t-tests or Wilcoxon signed-rank tests. Both groups demonstrated significantly reduced peak knee flexion angle and knee extension moment in the more symptomatic limb during stance. Distinct distal adaptations were observed: In the unilateral AKP group, the more symptomatic limb showed lower peak ankle plantarflexion moment (0.95 ± 0.21 N·m/kg vs. 1.06 ± 0.22 N·m/kg, p < 0.05) and a greater foot progression angle (10.03° ± 3.93° vs. 7.62° ± 3.34°, p < 0.05) compared to the less symptomatic limb. In the bilateral AKP group, peak ankle dorsiflexion angle was lower on the more symptomatic side (10.89° ± 3.36°) than on the less symptomatic side (12.60° ± 3.86°) (p < 0.05). Symmetry index analysis revealed no significant between-group differences across most variables, indicating comparable overall asymmetry. sEMG findings showed reduced rectus femoris activation in the symptomatic limb. Unilateral and bilateral AKP share a protective gait strategy characterized by reduced sagittal-plane knee loading. However, overall gait asymmetry does not differ significantly between groups, suggesting that bilateral AKP may also involve side-specific compensations or globally conservative movement patterns. These findings highlight that the magnitude of interlimb asymmetry alone may not distinguish between unilateral and bilateral presentations. Instead, the observed differences in distal joint adaptations (e.g., ankle dorsiflexion, plantarflexion moment, foot progression angle) provide more informative biomechanical targets for phenotype-specific rehabilitation strategies.
Age-related gait decline is associated with reduced heel loading, forefoot overloading, and impaired gait stability. Soft wearable robotic suits that assist hip extension may improve gait biomechanics, but evidence for training-induced carry-over effects on plantar pressure distribution remains limited in community-dwelling older adults. This study investigated whether a short-term gait training program using a hip extensor-assistive soft wearable robotic suit could modify plantar pressure and spatiotemporal gait parameters under device-free assessment conditions. In this prospective, single-center, randomized controlled pilot trial, 42 community-dwelling older adults aged 65-79 years were randomly assigned to a robot-assisted training group (n = 21) or a control group (n = 21). Both groups completed a 3-week intervention consisting of six sessions, each including 20 min of treadmill-based gait training and 20 min of strength training. The robot-assisted group wore a cable-driven soft wearable robotic suit providing bilateral hip extension assistance during treadmill training, whereas the control group performed identical training without the device. All post-intervention assessments were conducted without the device. Plantar pressure and spatiotemporal gait parameters were measured using the GAITRite® system at baseline and post-intervention. Compared with the control group, the robot-assisted group showed a greater increase in heel contact area (p = 0.014), greater reductions in forefoot contact area (p = 0.016) and forefoot peak plantar pressure (p = 0.026), and significant time × group interactions for heel contact area (F = 7.273, p = 0.011), forefoot contact area (F = 5.042, p = 0.031), and forefoot peak plantar pressure (F = 5.535, p = 0.024). The robot-assisted group also showed reduced step length (p = 0.005), stride length (p = 0.005), single support time (p = 0.032), and swing phase duration (p = 0.037), along with increased stance phase duration (p = 0.041). Short-term training with a hip extensor-assistive soft wearable robotic suit was associated with changes in plantar loading and gait cycle patterns under device-free assessment conditions, suggesting that gait pattern modifications may occur after repeated training sessions.
Quantitative gait analysis supports clinical decision-making and outcome monitoring, yet its use remains largely restricted to specialized laboratories due to cost and operational complexity. Smartphone-based two-dimensional video approaches using open-source software have shown promising validity, but inter-rater reliability-particularly the influence of examiner expertise across spatiotemporal and kinematic outcomes-remains insufficiently characterized in powered samples. In this observational study, 84 healthy adults were included and recorded during level barefoot walking using a standardized smartphone setup. Four examiners (two experienced, two non-experienced) independently extracted spatiotemporal parameters and hip, knee, and ankle joint angles at different gait events using Kinovea. Inter-rater reliability was assessed with intraclass correlation coefficients (two-way mixed-effects, absolute agreement; ICC [3,1]) with 95% confidence intervals, and measurement error was quantified using the standard error of measurement (SEM) and minimal detectable change (MDC95). Agreement between expert and non-expert examiners was explored using Bland-Altman plots for representative variables. Mean values were descriptively compared with established three-dimensional reference values. Global spatiotemporal descriptors demonstrated excellent reliability with minimal error and limited dependence on examiner expertise (walking speed ICC = 0.96; cadence ICC = 0.98; stride length ICC = 0.96). In contrast, temporal gait phase variables showed poor reliability overall (ICC ≈ 0.31-0.43), with higher-but still limited-agreement among experienced raters. Joint kinematics exhibited joint- and event-dependent reliability following a proximal-to-distal gradient: hip angles showed the highest consistency (ICC up to 0.88), knee angles moderate-to-good agreement (ICC ≈ 0.65-0.85), and ankle angles the greatest variability, particularly for event-dependent measures (e.g., opposite toe-off ICC = 0.45). Bland-Altman analyses revealed negligible bias for spatiotemporal outcomes but wider limits of agreement for kinematic variables. Mean values were biomechanically coherent, although systematic differences in joint angles were observed when compared with 3D reference data. In healthy young adults, a smartphone- and Kinovea-based workflow can provide reliable spatiotemporal gait metrics and moderate-to-good reliability for selected proximal kinematics, whereas temporal gait phase variables and distal joint angles remain less reliable and should be interpreted cautiously in longitudinal or multi-examiner contexts.
Cysteine-rich miniproteins from venomous animals are promising therapeutic scaffolds due to their compact structure, high stability, and low immunogenicity. Chlorotoxin (CTX), a miniprotein derived from scorpion venom, selectively targets glioblastoma by binding matrix metalloproteinase-2 (MMP-2), an invasion-associated enzyme overexpressed in ∼80% of glioblastomas. CTX has been utilized in tumor imaging, drug delivery, and immunotherapy, including its incorporation into chimeric antigen receptor (CAR) constructs. CTX-based CAR show potent MMP-2-dependent cytotoxicity in glioblastoma models. We hypothesized that affinity maturation of CTX via phage display could enhance MMP-2 binding and improve CAR T-cell performance. A CTX-based phage display library (∼1.3 million variants) was screened against immobilized MMP-2. A lead variant, CTXA8, was recombinantly expressed and tested for binding specificity against a panel of off-target proteins. Its cellular uptake and localization were evaluated, followed by incorporation into CAR constructs in both single-unit and tandem formats (eCTXA8-CAR). The cytotoxicity of CTX-, CTXA8-, and eCTXA8-CAR T cells was assessed against glioblastoma cell lines and primary patient-derived tumor cells. Screening identified CTXA8, which demonstrated a 4.4-fold increase in MMP-2 affinity and reduced off-target binding relative to CTX. Fluorescent CTXA8 showed 2.4-3.5-fold greater uptake in glioblastoma cells than CTX. Among the CAR constructs tested, eCTXA8 induced the highest IFN-γ release. eCTXA8-CAR T-cells consistently outperformed CTX-CAR and non-transduced T cells in cytotoxicity assays, especially at low effector-to-target ratios. This study demonstrates that directed evolution of CTX can produce high-affinity, selective MMP-2 ligands suitable for next-generation CAR T-cell therapies. CTXA8-based CARs offer enhanced anti-tumor efficacy, supporting their potential in overcoming challenges in solid tumor immunotherapy.
Minimally invasive surgical treatment is increasingly favored for intra-articular displaced calcaneal fractures with a high risk of soft tissue complications. The calcaneus is the largest tarsal bone in the foot and plays a critical role in weight-bearing and gait mechanics; displaced fractures often result in significant functional impairment. Sanders Type II fractures, characterized by a single major fracture line that divides the posterior articular surface into two major bone fragments, are one of the most common subtypes and present a therapeutic challenge in balancing mechanical stability with soft tissue preservation. A finite element model of Sanders Type II calcaneal fractures was established to compare five fixation methods under physiological tendon loading: (1) a periosteal locking plate, (2) four hollow screws, (3) four K-wires placed along the screw trajectory, (4) two intra-articular K-wires combined with two standard K-wires, and (5) six K-wires placed at dispersed locations. Outcome measures included maximum fragment displacement, implant-induced bone stress, and implant stress. Rigid internal fixation (hollow screws) provided the best initial stability and the least displacement of the bone fragments. A key finding was that under the most severe static loading condition (dorsiflexion with maximum Achilles tendon force), the maximum fragment displacement for all K-wire configurations (≤0.103 mm) was below both the clinically reported acceptable threshold (≤0.5 mm) and the safe healing range (<1 mm), indicating sufficient initial healing stability. Furthermore, K-wires generated significantly lower stress on the bone than the plate, while cannulated screw fixation carried the lowest risk of implant failure. Based on extended observations from clinical literature, K-wire fixation offers potential advantages including a minimally invasive approach, flexible handling, low implant burden, and the possibility of outpatient removal to avoid secondary surgery. Although rigid implants are biomechanically superior, K-wire fixation provides clinically acceptable initial stability and, combined with its minimally invasive characteristics, represents an attractive alternative. The choice of fixation method requires a comprehensive consideration of individual patient factors (such as bone quality and functional demands) and the overall treatment strategy.
Three-dimensional ground reaction force (GRF) is an important biomechanical indicator of weight-bearing, propulsion, and bilateral asymmetry in hemiplegic gait. However, conventional GRF measurement relies on laboratory-based force plates, limiting its use in continuous rehabilitation assessment. A specific methodological challenge is to estimate continuous three-dimensional GRF in post-stroke hemiplegic gait without using force-plate signals as model input, while still preserving whole-body kinematic coordination and affected-unaffected side asymmetry. This study proposed a force-plate-independent, marker-based method for estimating continuous stance-phase three-dimensional GRF in patients with hemiplegia by combining a spatio-temporal graph convolutional network (ST-GCN) with two-stage transfer learning. Data were collected from 30 chronic stroke patients with hemiplegia and 60 healthy controls. The model used 39 raw Plug-in Gait markers as graph nodes, with 10-dimensional node features consisting of three-dimensional position, velocity, acceleration, and laterality encoding. The model was pretrained using healthy participant data and then fine-tuned and evaluated on hemiplegic gait data using leave-one-subject-out cross-validation. The main contribution of this work is the integration of marker-level body topology, explicit kinematic derivatives, pathological laterality encoding, and healthy-to-hemiplegic transfer learning within a unified ST-GCN framework. The proposed ST-GCN achieved Pearson's r values of 0.984, 0.956, and 0.912, and rRMSE values of 5.24%, 8.15%, and 11.05% for vertical, anterior-posterior, and medio-lateral GRF prediction, respectively, outperforming MLP, 2D-CNN, BiLSTM, and lightweight Transformer baselines. Bland-Altman analysis further showed small mean biases for the first vertical peak force, anterior-posterior peak propulsive force, weight-bearing asymmetry index, and propulsion asymmetry index, with all participants falling within the 95% limits of agreement. These findings suggest that the proposed framework can reconstruct overall GRF waveform morphology and preserve group-level kinetic asymmetry features in hemiplegic gait. The method may provide a force-plate-independent, marker-based laboratory framework for kinetic gait assessment, but it should not yet be interpreted as a fully wearable or home-based clinical monitoring system.
Intervertebral disc degeneration is a major structural correlate of low back pain and is increasingly viewed as a disorder of mechanobiological dysregulation rather than simple mechanical overload. Over the past decade, disc cells have been shown to engage a diverse mechanosensory repertoire, including Piezo1, transient receptor potential channels, integrin-focal adhesion complexes, acid-sensing ion channels, primary cilia, and cytoskeletal-nuclear signaling axes. These upstream sensors converge on a more limited set of downstream hubs, most notably Yes-associated protein and transcriptional co-activator with PDZ-binding motif and mitogen-activated protein kinase/nuclear factor kappa B, which then shape cell-fate programs such as senescence, autophagy, pyroptosis, and ferroptosis. The extracellular matrix functions both as the substrate and as the output of mechanotransduction, creating a self-reinforcing loop in which degenerative matrix changes amplify pathological mechanosensing. Despite substantial molecular progress, mechanistic understanding remains concentrated in nucleus pulposus models, whereas matched cross-compartment analyses involving the annulus fibrosus, cartilage endplate, and vertebral interface remain limited. As a result, the relative importance of mechanosensors across compartments and loading contexts is still unresolved. Multiscale platforms, including finite element modeling, tunable hydrogels, organ culture, and quantitative magnetic resonance imaging, have strengthened causal interrogation, but translational progress remains largely preclinical. The clearest advance to date has come from biomaterial-based strategies, supported by early human feasibility data for injectable hydrogel implants. In this Review, we synthesize mechanotransduction in intervertebral disc degeneration, summarize the relative maturity of current mechanistic understanding across compartments and signaling branches, and discuss the importance of systematic cross-compartment comparison alongside single-sensor, single-compartment studies.
Bioimpedance spectroscopy (BIS) has emerged as a versatile, non-invasive technique for real-time electrical characterization of biological systems. By applying alternating current and analyzing the resulting complex impedance, BIS provides insights into cellular structure, tissue properties, and physiological processes, and can also detect pathogenic bacteria. This article presents a scoping review of recent advances in BIS, conducted under the Arksey and O'Malley framework and PRISMA-ScR guidelines, and analyzes 46 studies published between 2015 and 2025 that address BIS applications across biological systems. The strongest evidence was identified in fluid management and hydration monitoring (39.1%), body composition assessment (17.4%), and lymphedema monitoring (15.2%), whereas applications in tissue characterization, cellular systems, agriculture, machine learning-assisted diagnostics, and biosensing technologies remain at lower levels of translational maturity. Recent advances in instrumentation, sensor design, microfluidics, wearable systems, and computational analysis are critically examined. The review identifies major technological barriers, including a lack of standardized acquisition protocols, device-dependent variability, limited interoperability, heterogeneous modeling approaches, and insufficient multicenter validation. Despite challenges related to standardization, modeling of heterogeneous systems, and measurement reproducibility, BIS continues to demonstrate potential across biomedical, biotechnological, and industrial applications. Its ability to provide continuous, real-time, and non-destructive measurements supports its growing use in diagnostic platforms, therapeutic monitoring, and industrial biosensing applications. This review provides an integrated perspective on current developments, limitations, and future directions of BIS-based technologies.
The study's objective is to propose a novel non-invasive method for rapid screening and regular assessment of adolescent idiopathic scoliosis (AIS) through development of a wearable system integrated with multiple inertial measurement units (IMUs) and deep learning models. The system is designed to automatically distinguish between healthy individuals and AIS patients, and subsequently predict the Cobb angle based on continuous temporal kinematic angular sequences acquired during gait. Gait kinematic data were acquired from 124 participants (104 patients with average Cobb angle of 21.62 ± 7.93° and 20 healthy subjects) using a 9-IMU wearable device. Extracted angular features were analyzed to quantify bilateral asymmetry, compare differences across severity subgroups, and evaluate their linear correlation with Cobb angles. A two-stage deep learning framework was implemented with a convolutional neural network (CNN) classification model developed for the rapid screening of scoliosis, and a CNN-Transformer model was designed and compared with other five model architectures to predict Cobb angles from the acquired temporal angular sequences. Scapular kinematics emerged as the most prominent marker of asymmetry, and knee joint kinematics served as the strongest indicator of severity. Meanwhile, angular features of knee, hip and ankle joints demonstrated weak negative linear correlations with Cobb angle. In addition, the scoliosis screening model achieved high predictive performance, with an accuracy of 96.59% and a precision-recall AUC of 0.94 for scoliosis detection. For Cobb angle prediction, the CNN-Transformer model regularized with Gaussian noise during training proved most effective, yielding a mean absolute error of 2.14 ± 0.28° and R 2 value of 0.85 ± 0.03, outperforming other architectural alternatives evaluated in this study. Kinematic analysis of angular data validated the efficacy of the wearable system and effectively captured gait characteristics specific to AIS. The deep learning models accurately distinguished scoliosis patients from healthy cases and predicted Cobb angles using temporal kinematic angular sequences, providing a safe, non-invasive, operator-friendly approach suitable for rapid screening and regular assessment.
Dental hypersensitivity (DH) is a common clinical condition characterized by brief, intense pain. This condition is treated with fluoride-containing specialized dental gels. The effect of calcium ions present in saliva and pH of the oral cavity are indicated as factors affecting their clinical performance. In the presented study, two dental desensitizing gels were evaluated. Fourier Transform Infrared Spectroscopy (FTIR) spectra of the gels were registered and analyzed. Measurement of concentration of measurable free fluoride ions and pH in vitro assessment was performed using selective electrodes. Measurement with addition of TISAB witch CDTA was performed to evaluate the amount of fluoride bound within the gel matrix. Univariate and multifactorial ANOVA tests were used to analyze acquired results of measurable free fluoride in twelve distinct incubation solutions. The highest values of free, measurable fluoride were obtained after addition of TISAB with CDTA. However, in physiological study without ionic strength adjuster and chelator, the highest free, measurable fluoride observed in the artificial saliva solutions was 1,043 ppm for the SensiKIN desensitizing gel at pH 4.5 in the solution without calcium ion addition. The lowest value (658 ppm) was denoted for the Teeth Desensitiser TD gel at pH 7.0 with calcium ion addition. The presented amounts of fluoride pertain to measurable free fluoride in solution. It has been shown that the presence of calcium in saliva reduces the concentration of measurable free fluoride in solution (by as much as 137 ppm). Although the effect of presence of Ca2+ is pronounced, the initial pH remains the most significant variable. Observed levels of measurable free fluoride in solution might reflect initial and preliminary bioavailability under static in vitro conditions. Measurable free fluoride may serve as an indirect analytical proxy for potentially available fluoride under the tested in vitro conditions. However, it does not represent direct biological or clinical bioavailability. The evaluated desensitizing gels showed maximal free, measurable fluoride concentration under acidic conditions. For studies incorporating TISAB/CDTA, the most profound effect on F- concentration was gel composition. Future formulation studies could investigate whether calcium- complexing or buffering components influence measurable solution-phase free fluoride under standardized in vitro conditions.
Even though the cornea is one of the most widely transplanted solid tissues, it is estimated that approximately 98% of patients requiring a transplant lack access to this sight-saving procedure. This critical gap is a direct consequence of a tremendous global donor shortage, highlighting the urgent need for bioengineered alternatives. Despite significant research efforts, creating a functional bioengineered cornea remains a major challenge. The present study investigates a novel biomimetic approach leveraging Plasma Rich in Growth Factors (PRGF) technology in combination with dental pulp stem cells. Human dental pulp stem cells (hDPSCs) were isolated, characterized, and differentiated into epithelial, stromal, and endothelial corneal phenotypes. Two distinct types of corneal grafts were developed, categorized by the differentiation stage of the incorporated hDPSCs. The bioengineered constructs consisted of a central bioactive PRGF-fibrin core embedding cells and sandwiched between a trilaminar anterior layer and a posterior cell sheet. The PRGF-derived supernatant was utilized as a medium supplement. The resulting corneal substitutes were evaluated for their optical and rheological properties, as well as cellular viability. Finally, the structural stability and remodeling kinetics of the bioengineered grafts were histologically examined and quantified. Corneal grafts incorporating undifferentiated hDPSCs exhibited superior elasticity and transparency, while maintaining a higher cell density following the fabrication process. The more pronounced mass reduction observed in these constructs was concomitant with accelerated fibrin degradation and enhanced de novo collagen synthesis. Bioengineered grafts comprising a central PRGF membrane fibrin core integrated with hDPSCs to mimetically replicate the trilaminar cellular architecture of the cornea, represent a dynamic biomimetic candidate for future clinical applications in corneal transplantation surgery.
Sulfonamides remain important to medicinal and fine-chemical production but conventional aniline precursor routes like Pd-catalysed hydrogenation and Béchamp reductions carry sustainability, safety, and chemoselectivity challenges, especially for N-S bond integrity. This review assesses biocatalytic nitroreduction as a selective alternative for nitro-sulfonamides, focusing on p-aminobenzenesulfonamide (p-ABS). We summarise mechanistic and engineering advances in Type I (oxygen-insensitive) flavin-dependent nitroreductases (NTRs), highlight emerging roles and limits of Old Yellow Enzymes, and discuss auxiliary reductive platforms (H2-driven hydrogenases, photo-/electro-biocatalysis) for improved cofactor economy and endpoint selectivity. Process-intensification strategies, whole-cell vs. cell-free operation, immobilisation, packed-bed flow, on-line LC/IR PAT, and NAD(P)H regeneration via GDH/FDH or electroenzymatic modules are mapped to chemoselectivity risks (hydroxylamine accumulation, azo/azoxy formation) and mass-transfer constraints. We highlight development choices with green metrics (PMI, E-factor) and emphasise early LCA integration to avoid burden shifting from buffer salts or mediator residues. Evidence from continuous NTR reactors and immobilised formats supports scalable, aqueous, low-pressure operation; however, direct data on sulfonamide-linked nitroarenes is limited. This motivates a feasibility screen using an NTR panel (including engineered NfsB lineages), water-rich media with low co-solvent, and O2-tolerant settings. We conclude with a proposed flowsheet for p-ABS coupling immobilised NTR with FDH or electro-NAD(P)H supply, real-time analytics, and membrane-based product extraction. As direct data on sulfonamide-linked nitroarenes remain limited, this roadmap provides a practical and critical starting point for substrate-specific feasibility screening and future development.
Adipose tissue derived stromal vascular fraction (SVF) has emerged as cell therapeutic applicable by point-of-care one-step procedures in autologous settings. Even a mechanical isolation, completed within minutes, is typically followed by multiple steps such as cell washing, filtration, erythrocyte lysis and cryopreservation, which may impact the cell isolate in terms of cell amounts and quality. Using the BioMicroMill, a straightforward device providing mechanically isolated SVF suited for multiple therapeutic doses, we aimed to evaluate the impact of relevant processing steps during isolation and cryopreservation on cell quantity and quality in terms of viability, the presence of regenerative cells, pro-regenerative secretome, vascular network formation. The mechanical isolation yielded in the mean 3.4 × 105 ± 1.42 × 105 SVF cells per ml of lipoaspirate with a mean viability of 38% ± 7.2%. It comprised a heterogeneous mixture of single cells, cell aggregates, extracellular matrix, and microvascular fragments enriched from adipose tissue, providing functionally relevant cell populations (CD31, CD34, CD90, CD105). Adipose-derived stromal cells (ASC) exhibited robust outgrowth, proliferation, and differentiation capacity in vitro. The SVF showed pronounced paracrine activity (IL-10, VEGF-A, HGF, IL-6, IL-8, MCP-1), including proangiogenic factors, and supported 3D vascular network formation, demonstrating strong proangiogenic potential. However, all additional SVF processing steps substantially influenced total cell yield, with cumulative cell losses (erylysis 68% ± 16%, filtration 67% ± 16%, washing 54% ± 9%) but no loss of viability or ASC attachment and proliferation in vitro was observed. In contrast to other processing steps, filtration substantially altered SVF composition towards single cells, which was also reflected in an altered paracrine activity (significant increase in IL-10 and reduction to minimal levels of HGF). Furthermore, despite further cell loss (mean 47% ± 16%), cryopreservation maintained functional cell populations including ASC, paracrine activity and support of 3D network formation in vitro. Taken together, our data demonstrate that processing steps can influence both cell yield and quality. Accordingly, protocol selection should be guided by the intended application and the required functional properties and therefore warrants careful consideration.
The eukaryotic cell-based biomedicine landscape is undergoing a paradigm shift, driven by the transition from static culture to highly controlled microbioreactor environments. This mini-review assembles high-value applications, including autologous cell therapies, recombinant protein production, induced pluripotent stem cell expansion, and organoid engineering. Technologically, we highlight how the integration of microfluidics into Chimeric antigen receptor T-cell manufacturing achieves numerical optimization of cell count and phenotypic distribution through automated, closed-loop workflows. Regarding bioproduction, perfusion-based microbioreactors demonstrate a six-fold increase in productivity over traditional T-flasks, utilizing at-line monitoring and advanced cell-retention configurations to minimize shear-induced damage. Beyond production, these systems are active regulators of oxygenation and tissue morphogenesis in the context of human induced pluripotent stem cells while enabling controlled 3D tissue organization. In this connection, microfluidic platforms influence extracellular matrix remodeling and developmental signaling by modulating convective flow and mechanotransduction pathways. Furthermore, automation enhances reproducibility in feeding schedules and differentiation timing. Critical to this endeavor, the ability to generate precise spatiotemporal morphogen gradients enables reproducible lineage commitment and the maturation of complex tissues through synchronized mechanical and electrical stimuli for hosting organoid growth. The microbioreactor's superior mass transfer and low shear stress produced organoids 140% larger than static controls. While automation significantly reduces reagent consumption and inter-batch variability, challenges in scalability and material selection remain. Finally, parameters that push the microscale control boundaries are presented. Collectively, these advancements streamline therapeutic pipelines, support the replacement of animal-based testing, and align with Sustainable Development Goal 3 by accelerating the move toward personalized medicine and ethical drug screening.
In this study, we developed and validated a standardized computational framework for pediatric balance assessment by integrating anthropometric normalization and a dual-track analytical architecture into a unified center-of-pressure (COP) signal-processing pipeline. This study included 90 typically developing children. They were stratified into age groups of 4, 6, and 10 years (n = 30 per group), balanced by sex. The children performed bipedal stance tasks under eyes-open and eyes-closed conditions. Static balance was assessed using a Zebris force platform (120 Hz). COP sway area, mediolateral displacement, and anteroposterior displacement were measured. Two parallel datasets were processed that included raw COP signals and leg length-normalized COP signals. Both datasets were subjected to 2 (sex) × 3 (age) analysis of variance to evaluate scaling sensitivity. Coefficients of variation were computed to assess system-level dispersion. Anthropometric normalization markedly increased parameter sensitivity. Under the eyes-open condition, normalized COP sway area and anteroposterior displacement revealed age effects not detectable in the raw dataset, indicating scaling bias in uncorrected COP signals. The dual-track comparison confirmed that normalization functions as a computational validation component rather than a statistical add-on. For most COP parameters, the coefficients of variation exceeded 50%, indicating substantial interindividual variability consistent with developmental heterogeneity rather than measurement error. The proposed framework advances pediatric balance assessment from a descriptive developmental comparison to a standardized, bias-aware signal-processing architecture. By integrating anthropometric scaling correction and dual-track validation into a unified COP signal-processing pipeline, the framework enhances scaling sensitivity, inferential validity, and reproducibility. It is readily deployable within intelligent clinical screening systems and supports the development of age-stratified reference databases and early detection algorithms for pediatric neuromotor assessment.
Rotator cuff injury (RCI) represents a leading cause of shoulder pain and functional impairment, imposing a considerable burden on patients' quality of life and healthcare systems worldwide. The subacromial bursa (SAB), a critical anatomical structure located between the acromion and rotator cuff tendons, has traditionally been regarded as a primary pathological contributor to pain generation, inflammation, and tendon degeneration. Aberrant inflammatory signaling, fibrotic remodeling, and nociceptive sensitization within the diseased SAB can disrupt the homeostatic tendon microenvironment and impair intrinsic healing capacity, thereby supporting the historical rationale for routine bursectomy during surgical repair. Paradoxically, accumulating evidence challenges this conventional view, suggesting that the SAB also serves as a biologically active, endogenous regenerative reservoir. It contains mesenchymal stem cells (MSCs) with promising proliferative and tenogenic potential, secretes bioactive growth factors, and modulates the local immune microenvironment to promote reparative and anti-inflammatory responses conducive to tendon healing. This dual biological role presents an important clinical dilemma in arthroscopic rotator cuff repair: should surgeons resect pathological bursal tissue to alleviate the inflammatory burden and improve visualization, or selectively preserved to maintain its potential regenerative capacity? In this comprehensive narrative review, we critically synthesize current evidence regarding the pathological and regenerative functions of the SAB in RCI, critically evaluate the ongoing controversy between bursal resection versus preservation, and summarize emerging SAB-targeted therapeutic strategies. Furthermore, we integrate multi-omics insights into bursa biology aimed at moderating the local microenvironment. Finally, we highlight current translational challenges and future directions to inform biologically guided and evidence-based management of the SAB for rotator cuff repair.
Posterior cervical decompression is widely used to relieve spinal cord compression; however, postoperative spinal stability and the risk of adjacent segment degeneration (ASD) remain important clinical concerns. Laminoplasty and laminectomy are the two principal posterior decompression procedures and differ substantially in their biomechanical consequences, yet systematic quantitative comparisons are still limited. In this study, an individualized three-dimensional finite element (FE) model integrating high-resolution magnetic resonance imaging (MRI) and computed tomography (CT) data was developed to compare these two procedures with respect to postoperative stability, load redistribution, adjacent-segment biomechanical risk, and fixation-related mechanical response. Quasi-static simulations of cervical motion were combined with postoperative MRI evaluation and clinical follow-up data for validation. The results showed that laminoplasty was more effective than laminectomy in limiting postoperative increases in cervical mobility, reducing intradiscal pressure (IDP), and alleviating facet joint stress, thereby lowering the biomechanical risk of ASD. In addition, an imaging-mechanics coupled degeneration scoring system showed good predictive performance (area under the curve [AUC] = 0.87). These findings clarify the biomechanical mechanisms by which different surgical strategies shape the postoperative mechanical environment and degeneration risk, and provide a quantitative framework to support individualized surgical planning and precision medicine in cervical spine surgery.
This study aimed to investigate the hemodynamic mechanisms associated with arterial stenosis at the anastomosis floor of arteriovenous fistula (AVF), with a particular focus on characterizing how varying degrees of stenosis affect hemodynamic parameters in downstream venous hyperplastic regions. Computational fluid dynamics (CFD) simulations were conducted to numerically analyze hemodynamic characteristics in AVF using a standardized idealized AVF model. Key hemodynamic parameters, such as wall shear stress (WSS) and oscillatory shear index (OSI), were quantitatively evaluated. Particular attention was given to the variations and spatial distribution of these parameters in the presence of stenotic lesions within the arterial wall at the anastomosis floor, as well as across varying severities of stenosis. The regions of high wall shear stress in AVF are predominantly located at the anastomosis and along the outer wall of the juxta-anastomotic vein, whereas the low wall shear stress regions are primarily observed on the arterial wall at the floor of the anastomosis and along the inner wall of the swinging segment of the cephalic vein. These regions show partial consistency with the areas of high oscillatory shear index. In this idealized model, when the hyperplastic size of the artery at the anastomosis floor reaches approximately 1.0 mm, the wall shear stress level within the hyperplastic region returns to physiological levels, at which point neointimal hyperplasia ceases. At hyperplastic sizes of 0.5 mm and 1.0 mm in the artery at the anastomosis floor, the area of the high wall shear stress region on the outer wall of the juxta-anastomotic vein was reduced by 3.6% and 9.5%, respectively, compared to the non-hyperplastic state, with corresponding decreases in maximum wall shear stress of 7.0% and 10.2%. Similarly, under these hyperplastic conditions, the area of the low wall shear stress region on the inner wall of the swinging segment of the cephalic vein was reduced by 8.6% and 17.4%, respectively, relative to the non-hyperplastic state. In this idealized AVF model, when neointimal hyperplasia develops in the arterial wall at the anastomosis floor and reaches moderate severity, key hemodynamic parameters are restored toward physiological ranges, which is associated with attenuation of neointimal hyperplasia progression. This hyperplasia-induced moderate stenosis of the radial artery helps reduce the hyperplastic region in the cephalic vein near the anastomosis, thereby lowering the risk of stenosis and occlusion in the cephalic vein's swing segment.
Sacroiliac joint screw (SIJS) fixation is widely regarded as the gold standard for treating unstable posterior pelvic ring injuries. However, the technique presents notable challenges, including a high risk of neurovascular injury and a steep learning curve. This study aimed to introduce a novel Posterior Internal Fixation (P-INFIX) system and evaluate its biomechanical performance in the treatment of Denis zone I sacral fractures using finite element analysis. A three-dimensional pelvic model-cortical bone, cancellous bone, and cartilage-was reconstructed from computed tomography data of a healthy volunteer. A Denis zone I sacral fracture was simulated and stabilized using either the P-INFIX system (bilateral pedicle screws connected by a transverse Kirschner wire) or the standard SIJS. Finite element analysis was conducted under simulated sitting, supine, and lateral decubitus conditions to assess fracture fragment displacement and implant stress distribution. The P-INFIX system provided fracture stability comparable to SIJS under most testing conditions. Although SIJS demonstrated a slight statistical advantage in limiting lateral fragment displacement (For example, Z2 in the sitting position: 0.13 mm vs. 0.18 mm, P = 0.001), all measured displacements were far below the clinically significant threshold (<1.3 mm). In terms of implant performance, the peak stress in P-INFIX (sitting: 141 MPa; supine: 320 MPa) was higher than that in SIJS; however, it remained well within the safe range below the yield strength of titanium alloy (1,050 MPa). The novel P-INFIX system demonstrated biomechanical stability comparable to the gold standard SIJS in managing Denis zone I sacral fractures. Given its potential to reduce surgical risks and simplify the procedure through a "safe channel" lateral to the posterior superior iliac spine, P-INFIX represents a promising and more accessible alternative fixation technique.