Factors influencing hospital charges for tonsillectomy to treat obstructive sleep apnea in children.

PURPOSE: This study investigates the impact of patient characteristics and demographics on hospital charges for tonsillectomy as a treatment for pediatric obstructive sleep apnea (OSA). The aim is to identify potential disparities in hospital charges and contribute to efforts for equitable access to care. METHODS: Data from the 2016 Healthcare Cost and Utilization Project (HCUP) Kid Inpatient Database (KID) was analyzed. The sample included 3,304 pediatric patients undergoing tonsillectomy ± adenoidectomy for OSA. Variables such as age, race, length of stay, hospital region, residential location, payer information, and median household income were collected. The primary outcome variable was hospital charge. Statistical analyses, including t-tests, ANOVA, and multiple linear regression, were conducted. RESULTS: Among 3,304 pediatric patients undergoing tonsillectomy for OSA. The average total charges for tonsillectomy were $26,400, with a mean length of stay of 1.70 days. Significant differences in charges were observed based on patient race, hospital region, and payer information. No significant differences were found based on gender, discharge quarter, residential location, or median household income. Multiple linear regression showed race, hospital region, and residential location were significant predictors of total hospital charges. CONCLUSION: This study highlights the influence of patient demographics and regional factors on hospital charges for pediatric tonsillectomy in OSA cases. These findings underscore the importance of addressing potential disparities in healthcare access and resource allocation to ensure equitable care for children with OSA. Efforts should be made to promote fair and affordable treatment for all pediatric OSA patients, regardless of their demographic backgrounds.

AI-generated text in otolaryngology publications: a comparative analysis before and after the release of ChatGPT.

PURPOSE: This study delves into the broader implications of artificial intelligence (AI) text generation technologies, including large language models (LLMs) and chatbots, on the scientific literature of otolaryngology. By observing trends in AI-generated text within published otolaryngology studies, this investigation aims to contextualize the impact of AI-driven tools that are reshaping scientific writing and communication. METHODS: Text from 143 original articles published in JAMA Otolaryngology - Head and Neck Surgery was collected, representing periods before and after ChatGPT's release in November 2022. The text from each article's abstract, introduction, methods, results, and discussion were entered into ZeroGPT.com to estimate the percentage of AI-generated content. Statistical analyses, including T-Tests and Fligner-Killeen's tests, were conducted using R. RESULTS: A significant increase was observed in the mean percentage of AI-generated text post-ChatGPT release, especially in the abstract (from 34.36 to 46.53%, p = 0.004), introduction (from 32.43 to 45.08%, p = 0.010), and discussion sections (from 15.73 to 25.03%, p = 0.015). Publications of authors from non-English speaking countries demonstrated a higher percentage of AI-generated text. CONCLUSION: This study found that the advent of ChatGPT has significantly impacted writing practices among researchers publishing in JAMA Otolaryngology - Head and Neck Surgery, raising concerns over the accuracy of AI-created content and potential misinformation risks. This manuscript highlights the evolving dynamics between AI technologies, scientific communication, and publication integrity, emphasizing the urgent need for continued research in this dynamic field. The findings also suggest an increasing reliance on AI tools like ChatGPT, raising questions about their broader implications for scientific publishing.

Low concentration of polyethylene glycol facilitates separation of extracellular vesicles from bronchoalveolar lavage fluid.

Extracellular vesicles (EVs) in bodily fluids play an essential role in cell-cell cross talk and potentially serve as novel biomarkers in "liquid biopsy." It is crucial to have a consistent, efficient, and reliable method to separate EVs from bodily fluids. Currently, there is no universally accepted, "best" method to separate EVs. Besides differential ultracentrifugation (UC), polyethylene glycol (PEG) is among the commonly used methods for EV separation from bodily fluids. However, the optimal concentration of PEG to be used remains inadequately addressed. We initially observed that the concentration of PEG has a significant impact on the amount of separated EVs and EV-cargos, which are recovered from bronchoalveolar lavage fluid (BALF). To determine the optimal PEG concentration to be used in EV separation from BALF, we first separated the BALF and serum from wild-type C57BL/6 mice. Next, various concentrations of PEG (5%, 10%, and 15% PEG), a commercial kit, and UC were used to obtain EVs from BALF and serum. EVs were characterized, and EV-cargo protein, RNA, and miRNA levels were determined. We found that high concentration of PEG (10% and 15%) altered various EV parameters that are frequently used in EV studies, including EV yield, purity, and morphology. Using miR-15a, miR-142, and miR-223 as examples, we found that 10% and 15% PEG robustly reduced the detected levels of EV-cargo miRNAs compared with those in the EVs separated using UC or 5% PEG. Collectively, low concentration of PEG facilitates the optimal BALF EV separation.

Single-step RT-qPCR for detection of extracellular vesicle microRNAs in vivo: a time- and cost-effective method.

Emerging evidence suggests that extracellular vesicle (EV)-associated microRNAs (miRNAs) are a potential diagnostic tool for liquid biopsy in various human diseases. However, the experimental procedure for the detection of EV-associated miRNAs (EV-miRNAs) from body fluids is relatively complex and not cost-effective. Due to the limited amount of EVs and EV-RNAs, a column-based RNA purification, which is an expensive approach, is often used to detect EV-miRNAs via reverse transcription-quantitative real-time PCR (RT-qPCR). Here, we developed and validated a simple and cost-effective method (single-step RT-qPCR) in which we directly detect EV-miRNAs without RNA purification from the EVs. We validated this protocol using the EVs isolated from mouse broncho-alveolar lavage fluid (BALF) and serum. The obtained EVs were first lysed in the EV-lysis buffer, followed by RT-qPCR without isolation and purification of RNAs. We successfully detected the designated miRNAs from lysed EVs; 106 to 107 EVs were optimal to detect the EV-miRNAs using the single-step RT-qPCR. In our previously published work, using the conventional RT-qPCR method, we have reported that miR-142 and -223 are dramatically upregulated in both BALF and serum EVs after lung infection. Hence, we reassessed and confirmed the level of EV-miR-142/223 using the newly developed single-step RT-qPCR. Notably, inhibition of RNase activity in the lysed EVs remains crucial for the detection of EV-miRNAs. Moreover, repeated freeze-thaw cycling significantly interferes the EV-miRNA quantification. Collectively, the single-step RT-qPCR for the detection of EV-miRNAs in vivo will potentially provide a fast, accurate, and convenient way to quantify circulating and/or body fluid-derived EV-miRNAs. This method may potentially be applied to the diagnostic blood testing used in the medical centers or research laboratories.

Isolation and characterization of extracellular vesicles from Broncho-alveolar lavage fluid: a review and comparison of different methods.

Extracellular vesicles (EVs) are cell-derived membranous vesicles secreted by cells into the extracellular space, which play a role in cell to cell communication. EVs are categorized into 3 groups depending on their size, surface marker, and method of release from the host cell. Recently, EVs have become of interest in the study of multiple disease etiologies and are believed to be potential biomarkers for many diseases. Multiple different methods have been developed to isolate EVs from different samples such as cell culture medium, serum, blood, and urine. Once isolated, EVs can be characterized by technology such as nanotracking analysis, dynamic light scattering, and nanoscale flow cytometry. In this review, we summarize the current methods of EV isolation, provide details into the three methods of EV characterization, and provide insight into which isolation approaches are most suitable for EV isolation from bronchoalveolar lavage fluid (BALF).

Advances in the Use of Hypoglossal Nerve Stimulator in Adolescents With Down Syndrome and Persistent Obstructive Sleep Apnea-A Systematic Review.

INTRODUCTION: The Hypoglossal Nerve Stimulator (HNS) is a novel therapy that has been extensively studied in adults and more recently, it has been incorporated in children with Down Syndrome (DS) with persistent obstructive sleep apnea after adenotonsillectomy and trial of continuous positive airway pressure treatment. This systematic review article aims to examine the existing literature on HNS use in children to explore the benefits, efficacy, and parental experiences. METHODS: MEDLINE, Web of Science and EMBASE were searched to include all studies published up to March 2nd, 2023, on the topic of HNS use in pediatric population under 21 years old. RESULTS: A total of 179 studies were initially identified from which 10 articles were consistent with the inclusion criteria. Nine articles addressed outcomes after implantation of the HNS device in children with DS and 1 article explored the parental experiences. Findings were similar across studies where after implantation of HNS, there was marked improvement in polysomnographic outcomes and quality of life scores with high level of compliance. CONCLUSIONS: HNS holds promise as an effective treatment option for pediatric patients with DS and persistent OSA after AT and CPAP trials. It significantly improves sleep-disordered breathing, quality of life, and neurocognitive measures, leading to substantial and sustained benefits for these children. While the findings are encouraging, further research is needed to explore the potential of HNS in other pediatric populations without DS and to raise awareness among healthcare providers about this treatment option. Overall, HNS may offer significant long-term benefits for the overall well-being and health of pediatric patients with DS and persistent OSA.

Speech Outcomes of Frenectomy for Tongue-Tie Release: A Systematic Review and Meta-Analysis.

OBJECTIVE: Tongue-tie, which is also known as ankyloglossia, is a common condition where the lingual frenulum is unusually tight or short. While most literature investigates the impact of tongue-tie on breastfeeding, recent articles have examined its role in speech production in children. However, these have not previously been reviewed systematically. This study aims to determine the impact of tongue-tie on speech outcomes and assess whether frenectomy can improve speech function. METHODS: In this systematic review, we conducted a comprehensive search of PubMed/MEDLINE, Cochrane Library, Embase, and speechBITE to analyze primary studies investigating the impact of frenectomy for tongue-tie on speech outcomes. We extracted data regarding patient age, male to female ratio, procedure type, follow-up time, and speech outcomes and ran statistical analyses to determine if frenectomy for tongue-tie leads to improvement in speech issues in pediatric patients. Speech outcomes extracted were subjectively measured based on the interpretation of a speech and language pathologist or parent. RESULTS: Our analysis included 10 studies with an average patient age of 4.10 years, and average cohort size of 22.17 patients. Overall, frenectomy for tongue-tie was associated with an improvement in speech articulation (0.78; 95% CI: 0.64-0.87; P < .01). Increasing patient age was found to be negatively correlated with post-frenectomy speech outcomes (P = .01). However, this relationship disappeared in the adjusted model. CONCLUSION: Overall, we conclude that frenectomy is a suitable treatment to correct speech issues in select patients with tongue-tie if caught early in childhood. Despite the limited investigations around speech outcomes post-frenectomy, these results are informative to providers treating tongue-tie.

Comparison of ChatGPT knowledge against 2020 consensus statement on ankyloglossia in children.

OBJECTIVE: This paper evaluates ChatGPT's accuracy and consistency in providing information on ankyloglossia, a congenital oral condition. Assessing alignment with expert consensus, the study explores potential implications for patients relying on AI for medical information. METHODS: Statements from the 2020 clinical consensus statement on ankyloglossia were presented to ChatGPT, and its responses were scored using a 9-point Likert scale. The study analyzed the mean and standard deviation of ChatGPT scores for each statement. Statistical analysis was conducted using Excel. RESULTS: Among the 63 statements assessed, 67 % of ChatGPT responses closely aligned with expert consensus mean scores. However, 17 % (11/63) were statements in which the ChatGPT mean response was different from the CCS mean by 2.0 or greater, raising concerns about ChatGPT's potential influence in disseminating uncertain or debated medical information. Variations in mean scores highlighted discrepancies, with some statements showing significant deviations from expert opinions. CONCLUSION: While ChatGPT mirrored medical viewpoints on ankyloglossia, alignment with non-consensus statements raises caution in relying on it for medical advice. Future research should refine AI models, address inaccuracies, and explore diverse user queries for safe integration into medical decision-making. Despite potential benefits, ongoing examination of ChatGPT's power and limitations is crucial, considering its impact on health equity and information access.

Assessing ChatGPT's Responses to Otolaryngology Patient Questions.

OBJECTIVE: This study aims to evaluate ChatGPT's performance in addressing real-world otolaryngology patient questions, focusing on accuracy, comprehensiveness, and patient safety, to assess its suitability for integration into healthcare. METHODS: A cross-sectional study was conducted using patient questions from the public online forum Reddit's r/AskDocs, where medical advice is sought from healthcare professionals. Patient questions were input into ChatGPT (GPT-3.5), and responses were reviewed by 5 board-certified otolaryngologists. The evaluation criteria included difficulty, accuracy, comprehensiveness, and bedside manner/empathy. Statistical analysis explored the relationship between patient question characteristics and ChatGPT response scores. Potentially dangerous responses were also identified. RESULTS: Patient questions averaged 224.93 words, while ChatGPT responses were longer at 414.93 words. The accuracy scores for ChatGPT responses were 3.76/5, comprehensiveness scores were 3.59/5, and bedside manner/empathy scores were 4.28/5. Longer patient questions did not correlate with higher response ratings. However, longer ChatGPT responses scored higher in bedside manner/empathy. Higher question difficulty correlated with lower comprehensiveness. Five responses were flagged as potentially dangerous. CONCLUSION: While ChatGPT exhibits promise in addressing otolaryngology patient questions, this study demonstrates its limitations, particularly in accuracy and comprehensiveness. The identification of potentially dangerous responses underscores the need for a cautious approach to AI in medical advice. Responsible integration of AI into healthcare necessitates thorough assessments of model performance and ethical considerations for patient safety.

Appointment Factors Contributing to Children with Speech Disorders Missing Speech and Language Pathology Appointments.

This study explores missed pediatric speech and language pathology (SLP) appointments to identify barriers for patients with speech disorders. Data from 839 referrals at Boston Medical Center, including demographics, appointment details, COVID-19 lockdown, and number of items on patient problem lists, were analyzed using chi-square tests and logistic regression. The findings revealed that lockdown status, appointment timing, appointment type (in-person vs telemedicine), referral department (ear, nose, and throat [ENT] vs non-ENT), sex, race, primary language, birthplace, and primary care provider presence had no significant impact on attendance. However, the number of patient-listed problems, prior cancelations, and missed appointments were significant predictors of patients who did not keep appointments. In conclusion, this research emphasizes the patient's problem list and past appointment behavior as critical factors in predicting missed SLP appointments for pediatric speech disorder patients. These insights can guide targeted interventions to improve attendance and enhance SLP engagement.

Bacterial Outer Membrane Vesicles Promote Lung Inflammatory Responses and Macrophage Activation via Multi-Signaling Pathways.

Emerging evidence suggests that Gram-negative bacteria release bacterial outer membrane vesicles (OMVs) and that these play an important role in the pathogenesis of bacterial infection-mediated inflammatory responses and organ damage. Despite the fact that scattered reports have shown that OMVs released from Gram-negative bacteria may function via the TLR2/4-signaling pathway or induce pyroptosis in macrophages, our study reveals a more complex role of OMVs in the development of inflammatory lung responses and macrophage pro-inflammatory activation. We first confirmed that various types of Gram-negative bacteria release similar OMVs which prompt pro-inflammatory activation in both bone marrow-derived macrophages and lung alveolar macrophages. We further demonstrated that mice treated with OMVs via intratracheal instillation developed significant inflammatory lung responses. Using mouse inflammation and autoimmune arrays, we identified multiple altered cytokine/chemokines in both bone marrow-derived macrophages and alveolar macrophages, suggesting that OMVs have a broader spectrum of function compared to LPS. Using TLR4 knock-out cells, we found that OMVs exert more robust effects on activating macrophages compared to LPS. We next examined multiple signaling pathways, including not only cell surface antigens, but also intracellular receptors. Our results confirmed that bacterial OMVs trigger both surface protein-mediated signaling and intracellular signaling pathways, such as the S100-A8 protein-mediated pathway. In summary, our studies confirm that bacterial OMVs strongly induced macrophage pro-inflammatory activation and inflammatory lung responses via multi-signaling pathways. Bacterial OMVs should be viewed as a repertoire of pathogen-associated molecular patterns (PAMPs), exerting more robust effects than Gram-negative bacteria-derived LPS.

The effect of frenectomy for tongue-tie, lip-tie, or cheek-tie on breastfeeding outcomes: A systematic review of articles over time and suggestions for management.

OBJECTIVE: Tongue-tie, or ankyloglossia, is a common condition characterized by an abnormally short or tight lingual frenulum and is known to cause breastfeeding difficulties, leading to damage to the nipple, early discontinuation of breastfeeding, and delayed infant growth. In addition to tongue-tie, abnormal frenulums such as the labial frenulum and buccal frenulum can cause lip-tie and cheek-tie, respectively. While both of these conditions have been reported to potentially cause similar issues related to breastfeeding as tongue-tie, limited research has been conducted to understand their effects and how we should treat these conditions. METHODS: In this systematic review, we conducted a comprehensive search of MEDLINE to analyze the trend in publications of all three of these conditions and their impact on breastfeeding for the past 36 years. Keywords included, "tongue-tie", "lip-tie", "cheek-tie", and "breastfeeding outcomes". RESULTS: We found that publications describing the effect of only tongue-ties on breastfeeding have increased exponentially over time while less focus has been on other oral ties. It was also discovered that the majority of studies describing only lip-tie or tongue-tie were editorials, commentary, perspectives, or consensus statements. Finally, we found that articles describing more than one abnormal frenulum were more likely to be cited and articles describing tongue-tie only were published in the highest impact factor journals. CONCLUSION: This study revealed a significant increase in publications discussing tongue-tie and a lack of research on lip-tie and cheek-tie in relation to breastfeeding. The findings highlight the need for more comprehensive research and attention to lip-tie and cheek-tie, as well as standardized diagnostic criteria. Ongoing debate surrounding management of these conditions stem from the lack of investigations on the impact of these abnormal frenulums and outcomes post-frenectomy. Future high-quality studies, specifically prospective cohort studies and randomized controlled trials, are necessary to provide more robust evidence and guide clinical practice.

Extracellular Vesicles: An Emerging Clinical Opportunity in Musculoskeletal Disease.

Extracellular vesicles (EVs) are important mediators of cell-to-cell communication in the extracellular space. These membranous nanoparticles carry various molecules, often referred to as "cargo," which are delivered to nearby target cells. In the past decade, developments in nanotechnology have allowed for various new laboratory techniques for the increased utilization of EVs in cellular and animal studies. Such techniques have evolved for the isolation, characterization, and delivery of EVs to biological tissues. This emerging technology has immense clinical potential for both diagnostic and therapeutic applications. Various EV cargo molecules, including DNA, RNA, and proteins, can act as pathological biomarkers. Furthermore, EVs derived from certain cell sources have shown therapeutic benefit in certain pathologies. In addition to their native therapeutic benefit, EVs can be engineered to carry and selectively deliver therapeutic agents. While EVs have gained increasing interest in various pathologies, few studies have compiled their clinical potential in musculoskeletal pathologies. To bridge this gap, we present an overview of EVs, introduce current laboratory preparation techniques, and outline the most recent literature regarding the potential therapeutic applications of EVs in musculoskeletal pathologies.

An Analysis of Laryngomalacia and Its Interplay With Obesity and Obstructive Sleep Apnea in Pediatric Inpatients.

Objective  This study aimed to investigate the potential relationship between laryngomalacia and obesity as well as explore the interplay between laryngomalacia and obstructive sleep apnea using the Kids' Inpatient Database (KID) for the year 2016. Methods The Healthcare Cost and Utilization Project (HCUP) KID for 2016 provided a dataset for analysis. Patient demographics, diagnoses, and hospital characteristics were considered. Patients less than three years old were included due to the high prevalence of laryngomalacia in this age group. Results Among 4,512,196 neonatal admissions, 1,341 obesity cases and 11,642 laryngomalacia cases were analyzed. The frequency of laryngomalacia in patients with obesity was 578.1% higher than in the general population. Patients with obstructive sleep apnea (OSA) exhibited a 5,243.2% increase in laryngomalacia frequency compared to the overall population. Combining obesity and laryngomalacia resulted in a 6,738.5% increase in OSA frequency. Conclusions This study identified a significant correlation between obesity and increased laryngomalacia risk. The findings have important clinical implications for pediatric care, emphasizing the need to prevent childhood obesity to reduce laryngomalacia risk. Additionally, understanding these risk factors enables better risk stratification for laryngomalacia and potential OSA development.

Artificial Intelligence in Orthopaedic Surgery: Can a Large Language Model "Write" a Believable Orthopaedic Journal Article?

ABSTRACT: ➢ Natural language processing with large language models is a subdivision of artificial intelligence (AI) that extracts meaning from text with use of linguistic rules, statistics, and machine learning to generate appropriate text responses. Its utilization in medicine and in the field of orthopaedic surgery is rapidly growing.➢ Large language models can be utilized in generating scientific manuscript texts of a publishable quality; however, they suffer from AI hallucinations, in which untruths or half-truths are stated with misleading confidence. Their use raises considerable concerns regarding the potential for research misconduct and for hallucinations to insert misinformation into the clinical literature.➢ Current editorial processes are insufficient for identifying the involvement of large language models in manuscripts. Academic publishing must adapt to encourage safe use of these tools by establishing clear guidelines for their use, which should be adopted across the orthopaedic literature, and by implementing additional steps in the editorial screening process to identify the use of these tools in submitted manuscripts.

Exploring the appointment factors affecting pediatric patients with swallow disorders: Implications for speech and language pathology attendance.

INTRODUCTION: Feeding and swallowing disorders have become increasingly prevalent among children, necessitating effective management to prevent long-term complications. Speech and language pathology (SLP) services play a crucial role in diagnosing and treating these disorders. The objective of this study was to explore the factors that influence patient attendance to SLP appointments for swallow disorders. METHODS: This study was conducted at Boston Medical Center, involving 359 pediatric patients referred to SLP for swallow-related concerns. De-identified patient and appointment information was obtained from the electronic medical record. Various factors such as age, gender, race/ethnicity, primary language, appointment date/time, and COVID-19 lockdown status were analyzed to determine their impact on patient no-shows. Statistical analyses, including Chi-Square tests and binary logistic regression, were conducted using appropriate methodologies. RESULTS: 355 individual patient records were included in the analysis. Lockdown status and appointment time of day did not significantly affect patient no-shows. However, appointments conducted through telemedicine showed a significant difference in attendance. Patient referral department, gender, race, language, and being born at the medical center did not significantly influence patient attendance. Notably, having a primary care provider (PCP) at the medical center significantly affected patient attendance. Furthermore, previous appointment cancellations made a patient more likely to no-show. CONCLUSION: This study provides valuable insights into the factors influencing patient attendance at SLP appointments for pediatric swallowing disorders. Having a PCP at the medical center and utilizing telemedicine appointments were associated with higher attendance rates. Addressing appointment cancellations and investigating underlying reasons behind missed appointments should be prioritized in future research. Understanding these factors will facilitate the development of interventions to optimize patient attendance and improve the delivery of SLP services in pediatric populations.

Extracellular vesicles in respiratory disease.

Extracellular vesicles (EVs) are membranous nanoparticles secreted by nearly all cell types and play a critical role in cell-to-cell crosstalk. EVs can be categorized based on their size, surface markers, or the cell type from which they originate. EVs carry "cargo," including but not limited to, RNA, DNA, proteins, and small signaling molecules. To date, many methods have been developed to isolate EVs from biological fluids, such as blood plasma, urine, bronchoalveolar lavage fluid, and urine. Once isolated, EVs can be characterized by dynamic light scattering, nanotracking analysis, nanoscale flow cytometry, and transmission electron microscopy. Given the ability of EVs to transport cargo between cells, research has recently focused on understanding their role in various human diseases. As understanding of their significance to disease processes grows, insight into the mechanisms behind the physiological role of their cargo in target cells can facilitate the development of a new type of biomarker and therapeutic target for diseases in future. In addition, their ability to deliver their cargo selectively to target cells within the human body means that they could serve as therapeutic agents or methods of drug delivery. In this review, we will first introduce EVs and the cargo they carry, outline current methods for EV isolation and characterization, and discuss their potential use as biomarkers and therapeutic agents in the near future.

The Role of Extracellular Vesicles in Diseases of the Ear, Nose, and Throat.

Extracellular vesicles (EVs) are membranous nanoparticles produced by most cell types into the extracellular space and play an important role in cell-to-cell communication. Historically, EVs were categorized based on their methods of biogenesis and size into three groups: exosomes, microvesicles, and apoptotic bodies. Most recently, EV nomenclature has evolved to categorize these nanoparticles based on their size, surface markers, and/or the cell type which secreted them. Many techniques have been adopted in recent years which leverage these characteristics to isolate them from cell culture media and biological fluids. EVs carry various "cargo", including DNA, RNA, proteins, and small signaling molecules. After isolation, EVs can be characterized by various methods to analyze their unique cargo profiles which define their role in cell-to-cell communication, normal physiology, and disease progression. The study of EV cargo has become more common recently as we continue to delineate their role in various human diseases. Further understanding these mechanisms may allow for the future use of EVs as novel biomarkers and therapeutic targets in diseases. Furthermore, their unique cargo delivery mechanisms may one day be exploited to selectively deliver therapeutic agents and drugs. Despite the growing research interest in EVs, limited studies have focused on the role of EVs in the diseases of the ear, nose, and throat. In this review, we will introduce EVs and their cargo, discuss methods of isolation and characterization, and summarize the most up-to-date literature thus far into the role of EVs in diseases of the ear, nose, and throat.

Microvesicle-Derived miRNAs Regulate Proinflammatory Macrophage Activation in the Lung Following Ozone Exposure.

Ozone is a ubiquitous air pollutant that causes lung damage and altered functioning. Evidence suggests that proinflammatory macrophages contribute to ozone toxicity. Herein, we analyzed the role of extracellular vesicles (EVs) and microRNA (miRNA) cargo in ozone-induced macrophage activation. Exposure of mice to ozone (0.8 ppm, 3 h) resulted in increases in bronchoalveolar lavage fluid EVs, which were comprised predominantly of microvesicles (MVs). NanoFACS analysis revealed that MVs generated following both air and ozone exposure was largely from CD45+ myeloid cells; these MVs were readily taken up by macrophages. Functionally, MVs from ozone, but not air treated mice, upregulated mRNA expression of inflammatory proteins in macrophages including inducible nitric oxide synthase (iNOS), CXCL-1, CXCL-2, and interleukin (IL)-1ß. The miRNA profile of MVs in bronchoalveolar lavage fluid (BALF) was altered after ozone exposure; thus, increases in miR-21, miR-145, miR320a, miR-155, let-7b, miR744, miR181, miR-17, miR-92a, and miR-199a-3p were observed, whereas miR-24-3p and miR-20 were reduced. Ingenuity pathway analysis revealed that these miRNAs regulate pathways that promote inflammatory macrophage activation, and predicted that let-7a-5p/let-7b, miR-24-3p, miR-21-5p, miR-17, and miR-181a-5p are key upstream regulators of inflammatory proteins. After ozone exposure, miR-199a-3p, but not precursor miR-199a-3p, was increased in lung macrophages, indicating that it is derived from MV-mediated delivery. Furthermore, lung macrophage mRNA expression of IL-1ß was upregulated after administration of MVs containing miR-199a-3p mimic but downregulated by miR-199a-3p inhibitor. Collectively, these data suggest that MVs generated following ozone exposure contribute to proinflammatory macrophage activation via MV-derived miRNAs including miR-199a-3p. These findings identify a novel pathway regulating macrophage inflammatory responses to inhaled ozone.