Advances in the application of iron oxide nanoparticles (IONs and SPIONs) in three-dimensional cell culture systems.

BACKGROUND: The field of tissue engineering has remarkably progressed through the integration of nanotechnology and the widespread use of magnetic nanoparticles. These nanoparticles have resulted in innovative methods for three-dimensional (3D) cell culture platforms, including the generation of spheroids, organoids, and tissue-mimetic cultures, where they play a pivotal role. Notably, iron oxide nanoparticles and superparamagnetic iron oxide nanoparticles have emerged as indispensable tools for non-contact manipulation of cells within these 3D environments. The variety and modification of the physical and chemical properties of magnetic nanoparticles have profound impacts on cellular mechanisms, metabolic processes, and overall biological function. This review article focuses on the applications of magnetic nanoparticles, elucidating their advantages and potential pitfalls when integrated into 3D cell culture systems. This review aims to shed light on the transformative potential of magnetic nanoparticles in terms of tissue engineering and their capacity to improve the cultivation and manipulation of cells in 3D environments.

Cross-Cultural Validation of the Thai Oral Health Impact Profile for Temporomandibular Disorders.

OBJECTIVES: The Oral Health Impact Profile for Temporomandibular Disorders (OHIP-TMDs) is a validated condition-specific outcome measure to help guide decision-making in the management of the condition. There is no Thai version of OHIP-TMDs, and therefore the original English version needs cross-cultural adaptation translation, and validation with a Thai population to reduce the anomalies due to language and cultural differences. This study aimed to develop the Thai-language version of OHIP-TMDs, perform a cross-cultural adaptation to Thailand, and assess its content validity, internal consistency, reliability, and construct validity. METHODS: The original English version of OHIP-TMDs was forward and backward translated into Thai language using the International Network for Orofacial Pain and Related Disorders methodology (INfORM) protocol for cross-cultural adaptation. The Content Validity Index (CVI) was performed by 5 orofacial pain (OFP) specialists to establish content validity. The OHIP-TMDs-T was then tested in 2 groups of Thai dental patients including 110 TMD patients and 110 control participants. The internal reliability and test-retest reliability (n = 30) were investigated in the TMD group using Cronbach alpha coefficient and intraclass correlation coefficient (2-way mixed effect model), respectively. The difference in OHIP-TMDs-T score between the TMD group and control group was investigated for known group validity. RESULTS: Cronbach alpha and intraclass correlation coefficients were 0.942 and 0.797, respectively. The CVI collected from the OFP specialists was 0.92. There was a statistical difference in the OHIP-TMDs-T overall score between the TMD group (95% CI, 40-46) and control group (95% CI, 2.0-3.4) (Z = 9.060, r = 1, P < .001). CONCLUSIONS: The OHIP-TMDs-T is a valid and reliable tool for evaluating the quality of life and the impact on oral health in Thai patients with TMD.

Efficacy of oropharyngeal exercises as an adjuvant therapy for obstructive sleep apnea: A randomized controlled trial.

PURPOSE: Some patients with mild-to-moderate obstructive sleep apnea (OSA) undergoing mandibular advancement device (MAD) therapy may exhibit residual sleep-breathing abnormalities. These cases require adjunctive treatment to further reduce the remaining airway inadequacy, which may affect the health and quality of life. This clinical trial was conducted since the oropharyngeal exercise, which aims to enhance the muscular function of the upper airway, combined with MAD, is unknown. We assessed the possible improvements in respiratory event parameters and lip and tongue physiological properties in patients with OSA who received oropharyngeal exercises for 2 months as an adjunct to MAD therapy. METHODS: Twenty-three participants with OSA who had been using MAD but still had residual apnea-hypopnea were randomly allocated to either the oropharyngeal exercise (OE group; N=12) or the sham exercise group (N=11). Lip and tongue strength, endurance, daytime sleepiness, respiratory event index (REI), and the lowest oxygen saturation (SpO2 nadir) were determined. RESULTS: Within and between the treatment groups, no significant improvement in the REI or SpO2 nadir was found. In both groups, there was a significant reduction in the Epworth Sleepiness Scale at 2 months (OE, P = 0.02; control, P = 0.02). In the OE group, lip endurance (P = 0.03), anterior tongue strength (P = 0.02), and endurance (P = 0.02) increased after 2-month of exercise, but only anterior tongue endurance increased significantly compared with the sham control (P = 0.01). CONCLUSIONS: This study found that adjunctive OE to MAD showed no additive effect in treating patients with moderate OSA who had a mild residual apnea-hypopnea index following MAD treatment. However, the exercises successfully enhanced lip and anterior tongue endurance, as well as anterior tongue strength.

A Nonsurgical Treatment Strategy for Longstanding, Nonreducible, Bilateral Temporomandibular Joint Dislocation: A Case Report.

Temporomandibular joint (TMJ) dislocation is a condition in which the mandibular condyle is displaced from the glenoid fossa and must be manipulated to return it to its normal position. Among several diagnostic subtypes, long-standing or chronically protracted TMJ dislocation is rare and challenging to manage. Treatment options include various nonsurgical and surgical approaches. This case report describes the successful nonsurgical management of a longstanding bilateral TMJ dislocation using elastic traction with a removable appliance and posterior bite planes. This technique should be attempted before performing additional surgical interventions.

Imaging of muscle activity-induced morphometric changes in fibril network of myofascia by two-photon microscopy.

Myofascia, deep fascia enveloping skeletal muscles, consists of abundant collagen and elastin fibres that play a key role in the transmission of muscular forces. However, understanding of biomechanical dynamics in myofascia remains very limited due to less quantitative and relevant approaches for in vivo examination. The purpose of this study was to evaluate the myofascial fibril structure by means of a quantitative approach using two-photon microscopy (TPM) imaging in combination with intravital staining of Evans blue dye (EBD), a far-red fluorescence dye, which potentially labels elastin. With focus on myofascia of the tibial anterior (TA) muscle, the fibril structure intravitally stained with EBD was observed at the depth level of collagen fibrous membrane above the muscle belly. The EBD-labelled fibril structure and orientation in myofascia indicated biomechanical responses to muscle activity and ageing. The orientation histograms of EBD-labelled fibrils were significantly modified depending upon the intensity of muscle activity and ageing. Moreover, the density of EBD-labelled fibrils in myofascia decreased with habitual exercise but increased with muscle immobilization or ageing. In particular, the diameter of EBD-labelled fibrils in aged mice was significantly higher. The orientation histograms of EBD-labelled fibrils after habitual exercise, muscle immobilization and ageing showed significant differences compared to control. Indeed, the histograms in bilateral TA myofascia of exercise mice made simple waveforms without multiple sharp peaks, whilst muscular immobilization or ageing significantly shifted a histogram with sustaining multiple sharp peaks. Therefore, the dynamics of fibre network with EBD fluorescence in response to the biomechanical environment possibly indicate functional tissue adaptation in myofascia. Furthermore, on the basis of the knowledge that neutrophil recruitment occurs locally in working muscles, we suggested the unique reconstruction mechanism involving neutrophilic elastase in the myofascial fibril structure. In addition to the elastolytic susceptibility of EBD-labelled fibrils, distinct immunoreactivities and activities of neutrophil elastase in the myofascia were observed after electric pulse stimulation-induced muscle contraction for 15 min. Our findings of EBD-labelled fibril dynamics in myofascia through quantitative approach using TPM imaging and intravital fluorescence labelling potentially brings new insights to examine muscle physiology and pathology.

Exercise-evoked intramuscular neutrophil-endothelial interactions support muscle performance and GLUT4 translocation: a mouse gnawing model study.

KEY POINTS: Fractalkine receptor antagonist inhibited neutrophil recruitment to masseter muscles and exacerbated fatigability during masticatory activity. Fractalkine-mediated neutrophil recruitment is required for both upregulation of myokines (CXCL1, interleukin-6) and enhanced GLUT4 translocation in response to masticatory activity. Fractalkine and intercellular adhesion molecule-1 expression in endothelial cells increased in response to masticatory activity. In vitro experiments demonstrated that contracting myotubes lack the ability to upregulate fractalkine but revealed that endothelial fractalkine upregulation is induced using a conditioned medium of contracting myotubes. ABSTRACT: Physical exercise stimulates neutrophil recruitment within working skeletal muscle, although its underlying mechanisms remain ill-defined. By employing a masticatory behaviour (gnawing) model, we demonstrate the importance of intramuscular paracrine and autocrine systems that are triggered by muscle contractile activity and reliant upon fractalkine/CX3CL1-mediated signals. These signals were revealed to be required for achieving proper GLUT4 translocation and glucose uptake to meet the glucose demands for fatigue alleviation. Specifically, fractalkine expression and neutrophil recruitment both increased in the masseter muscle tissues upon masticatory activity. Importantly, a fractalkine antagonist inhibited neutrophil accumulation and exacerbated fatigability during masticatory activity. We found that fractalkine-dependent neutrophil recruitment is required for both upregulation of myokines (i.e. CXCL1 and interleukin-6) and enhanced GLUT4 translocation in response to gnawing activity. Immunofluorescence analysis of masseter muscles demonstrated that fractalkine and intercellular adhesion molecule-1 expression are both upregulated in endothelial cells but not in myofibres. The in vitro exercise model further revealed that contractile activity failed to stimulate fractalkine upregulation in myotubes, implying that fractalkine is not a myokine (myofibre-derived factor). Nevertheless, endothelial fractalkine expression was markedly stimulated by a conditioned medium from the contracting myotubes. Moreover, intercellular adhesion molecule-1, a key adhesion molecule for neutrophils, was upregulated in endothelial cells by fractalkine. Taken together, our findings strongly suggest that endothelial fractalkine serves as a key factor for organizing a physiologically beneficial intramuscular microenvironment by recruiting neutrophils in response to relatively mild exercise (i.e. masticatory muscle activity).

Neutrophils Provide a Favorable IL-1-Mediated Immunometabolic Niche that Primes GLUT4 Translocation and Performance in Skeletal Muscles.

Metabolic immunomodulation involving IL-1 has been investigated for unfavorable metabolic effects, including obesity, but a potentially favorable role for IL-1 remains unclear. Here, we find mechanistic interactions between working skeletal muscles and locally recruited neutrophils expressing IL-1ß, which supports muscle performance through priming exercise-dependent GLUT4 translocation. Thus, during exercise, both IL-1α/ß-deficient and neutrophil-depleted mice similarly exhibit increased fatigability associated with impaired muscle glucose homeostasis due to GLUT4 dysregulation. Deficiency of IL-1-producing neutrophils results in intrinsic abnormalities represented by aberrant Rac1 signaling and irregular GLUT4-storage vesicles, suggesting that these properties are maintained by local IL-1 produced by recruited neutrophils upon exercise, possibly on a daily basis. We propose that neutrophils are highly engaged in skeletal muscle performance via IL-1 regulation, which coordinates favorable inflammatory microenvironments supporting muscle glucose metabolism.

Roles of IL-1α/ß in regeneration of cardiotoxin-injured muscle and satellite cell function.

Skeletal muscle regeneration after injury is a complex process involving interactions between inflammatory microenvironments and satellite cells. Interleukin (IL)-1 is a key mediator of inflammatory responses and exerts pleiotropic impacts on various cell types. Thus, we aimed to investigate the role of IL-1 during skeletal muscle regeneration. We herein show that IL-1α/ß-double knockout (IL-1KO) mice exhibit delayed muscle regeneration after cardiotoxin (CTX) injection, characterized by delayed infiltrations of immune cells accompanied by suppressed local production of proinflammatory factors including IL-6 and delayed increase of paired box 7 (PAX7)-positive satellite cells postinjury compared with those of wild-type (WT) mice. A series of in vitro experiments using satellite cells obtained from the IL-1KO mice unexpectedly revealed that IL-1KO myoblasts have impairments in terms of both proliferation and differentiation, both of which were reversed by exogenous IL-1ß administration in culture. Intriguingly, the delay in myogenesis was not attributable to the myogenic transcriptional program since MyoD and myogenin were highly upregulated in IL-1KO cells, instead appearing, at least in part, to be due to dysregulation of cellular fusion events, possibly resulting from aberrant actin regulatory systems. We conclude that IL-1 plays a positive role in muscle regeneration by coordinating the initial interactions among inflammatory microenvironments and satellite cells. Our findings also provide compelling evidence that IL-1 is intimately engaged in regulating the fundamental function of myocytes.

Involvement of neutrophils and interleukin-18 in nociception in a mouse model of muscle pain.

Muscle pain is a common condition that relates to various pathologies. Muscle overuse induces muscle pain, and neutrophils are key players in pain production. Neutrophils also play a central role in chronic pain by secreting interleukin (IL)-18. The aim of this study was to investigate the involvement of neutrophils and IL-18 in a mouse model of muscle pain. The right hind leg muscles of BALB/c mice were stimulated electrically to induce excessive muscle contraction. The left hind leg muscles were not stimulated. The pressure pain threshold, number of neutrophils, and IL-18 levels were investigated. Furthermore, the effects of the IL-18-binding protein and Brilliant Blue G on pain were investigated. In stimulated muscles, pressure pain thresholds decreased, and neutrophil and IL-18 levels increased compared with that in non-stimulated muscles. The administration of IL-18-binding protein and Brilliant Blue G attenuated hyperalgesia caused by excessive muscle contraction. These results suggest that increased IL-18 secretion from larger numbers of neutrophils elicits mechanical hyperalgesia.

Effective Fluoride Mouthwash Delivery Methods as an Alternative to Rinsing.

BACKGROUND: Special-needs patients with high caries risk cannot benefit from the proven caries-reducing effect of fluoride mouthrinse because of poor rinsing compliance and a lack of home-use fluoride application recommendations. OBJECTIVES: This study aimed to evaluate whether two modified delivery methods-spray or swab application-could raise the salivary fluoride to levels similar to that of rinsing. METHODS: Five healthy men and 16 healthy women, ages 18-22 years, participated in this crossover study. The subjects performed a standardized brushing routine twice a day with 1,000-ppm fluoride dentifrice 1 week before and during the experimental period. The three fluoride mouthwash (0.05% NaF; 226-ppm fluoride) administration methods consisted of rinsing, spray, and cotton swab application. Each application was separated by at least 1 week. Unstimulated whole saliva was sampled for 2 minutes at baseline and at 0, 5, 10, 20, 30, 60, and 120 minutes after each fluoride mouthwash application. Salivary fluoride level (ppm) was measured by a blinded investigator using an ion-specific electrode (ORION EA940). The differences in salivary fluoride concentration for each application method were analyzed by repeated measures ANOVA at a nominal significance level of .05. RESULTS: The newly designed fluoride mouthwash delivery methods yielded a similar fluoride level in whole saliva compared to that of rinsing at all time points (p > .05). The t half-life of fluoride concentration ranged from 1.62 minutes (SD = 0.53) in the spray group to 2.08 minutes (SD = 0.87) in the cotton swab group; the values were not significantly different. DISCUSSION: Our results indicate that the spray and cotton swab fluoride delivery methods were effective alternatives to conventional mouth rinsing. Caregivers of individuals with acute and chronic illness, including developmental disabilities affecting cognitive or physical abilities, may consider using these alternative fluoride delivery methods to help promote dental hygiene.