Challenges in the Development and Application of Organ-on-Chips for Intranasal Drug Delivery Studies.

With the growing demand for the development of intranasal (IN) products, such as nasal vaccines, which has been especially highlighted during the COVID-19 pandemic, the lack of novel technologies to accurately test the safety and effectiveness of IN products in vitro so that they can be delivered promptly to the market is critically acknowledged. There have been attempts to manufacture anatomically relevant 3D replicas of the human nasal cavity for in vitro IN drug tests, and a couple of organ-on-chip (OoC) models, which mimic some key features of the nasal mucosa, have been proposed. However, these models are still in their infancy, and have not completely recapitulated the critical characteristics of the human nasal mucosa, including its biological interactions with other organs, to provide a reliable platform for preclinical IN drug tests. While the promising potential of OoCs for drug testing and development is being extensively investigated in recent research, the applicability of this technology for IN drug tests has barely been explored. This review aims to highlight the importance of using OoC models for in vitro IN drug tests and their potential applications in IN drug development by covering the background information on the wide usage of IN drugs and their common side effects where some classical examples of each area are pointed out. Specifically, this review focuses on the major challenges of developing advanced OoC technology and discusses the need to mimic the physiological and anatomical features of the nasal cavity and nasal mucosa, the performance of relevant drug safety assays, as well as the fabrication and operational aspects, with the ultimate goal to highlight the much-needed consensus, to converge the effort of the research community in this area of work.

Approach to Sinonasal Masses: A Comprehensive Review

The complex anatomy and exposure to various potential carcinogens lead to the development of tumors and tumor-like pathologies of the nose and paranasal sinuses. Delays in the diagnosis of sinonasal masses are common, leading to significant morbidity and mortality. Often the radiologist is the first person to recognize a neoplasm in a suspected inflammatory condition. Computed tomography and magnetic resonance imaging play a synergistic role in the assessment of pathologies in the sinonasal region, and their importance has become increasingly prevalent in the current scenario of coronavirus disease 2019 associated mucormycosis. Besides, imaging is essential for mapping the exact extent of the pathology and delineating vascular supply of hypervascular masses, thus facilitating the operating surgeon and interventional radiologist in management. The cases presented in this article have been accrued over the past three decades and analyzed as a retrospective observational study, with clinical, radiological, and pathological data having been extracted from the existing database in the institution. We present the imaging spectrum of sinonasal masses in the pediatric and adult population, highlight the key radiological features of the common pathologies and discuss an imaging template for reporting these masses, with special focus on the surgically relevant points to be included in the report. The educational goal of this review is to explore a meticulous and systematic imaging approach toward soft tissue lesions in the sinonasal region, which would enable the radiologist to reach a diagnosis or point toward the possible etiology and nature of the lesions.

Anatomical and Vision-Guided Path Generation Method for Nasopharyngeal Swabs Sampling

Due to the global COVID-19 pandemic, there is a strong demand for pharyngeal swab sampling and nucleic acid testing. Research has shown that the positive rate of nasopharyngeal swabs is higher than that of oropharyngeal swabs. However, because of the high complexity and visual obscuring of the interior nasal cavity, it is impossible to obtain the sampling path information directly from the conventional imaging principle. Through the combination of anatomical geometry and spatial visual features, in this paper, we present a new approach to generate nasopharyngeal swabs sampling path. Firstly, this paper adopts an RGB-D camera to identify and locate the subject's facial landmarks. Secondly, the mid-sagittal plane of the subject's head is fitted according to these landmarks. At last, the path of the nasopharyngeal swab movement in the nasal cavity is determined by anatomical geometry features of the nose. In order to verify the validity of the method, the location accuracy of the facial landmarks and the fitting accuracy of mid-sagittal plane of the head are verified. Experiments demonstrate that this method provides a feasible solution with high efficiency, safety and accuracy. Besides, it can solve the problem that the nasopharyngeal robot cannot generate path based on traditional imaging principles. It also provides a key method for automatic and intelligent sampling of nasopharyngeal swabs, and it is of great clinical value to reduce the risk of cross-infection. © 2023, The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd.

Simulating and Optimizing Nasopharyngeal Swab Insertion Paths for use in Robotics

The nasopharyngeal swab is the standardized method of collecting specimens for diagnosing COVID-19, among numerous other respiratory illnesses. While there has been interest from the robotics community in the design of robots and manipulators for performing swab collections, detailed simulation and planning for swab insertion trajectories through the nasal cavity is less studied. In this work, we propose a simulation environment with the swab modelled as an Euler-Bernoulli beam, subject to linear elastic collisions coming from the nasal cavity. We evaluate the impact of inserting the swab with different amounts of force. We also leverage the simulation environment to pose an optimization problem that finds trajectories that minimize strain on the swab during the insertion. We find that the optimized trajectories adhere to qualitative clinical advice. © 2022 IEEE.

Simulation of Virus-Laden Droplets Transmitted from Lung to Lung

In this study, we conducted a computational fluid dynamics analysis to estimate the trajectory of the virus-laden droplets. As numerical models, two human body models with airways were prepared. These models are represented by unstructured grids. Having calculated the unsteady airflow in the room, we simulated the trajectory of droplets emitted by the human speaking. In addition, inhaling the droplets into the lung of the conversation partner was simulated. The number of the droplets adhered to the respiratory lining of the partner was counted separately on the nasal cavity, oral cavity, trachea, bronchi, and bronchial inlet surface. The diameters of the droplets were also investigated in the same manner. It was noticeable that more than 80% of the droplets inhaled by the conversation partner adhered to the bronchial inlet surface. Also, the conversation partner did not inhale droplets larger than 35 μm in diameter. It was found that when the distance between two people was 0.75 m, more droplets adhered to the partner’s torso. © 2022, The Author(s), under exclusive license to Springer Nature Switzerland AG.

Nasal vaccine as a booster shot: a viable solution to restrict pandemic?

The coronavirus disease 2019 (COVID-19) pandemic revolutionized the vaccine market and initiated the momentum for alternative routes of administration for vaccines. The intranasal route of immunization is one such possibility that appears to be the most promising since it has some significant advantages, particularly in the prevention of respiratory infection. To analyze and summarize the role of nasal vaccines over conventional vaccines during COVID-19 and the need for the nasal vaccine as a booster shot. In this narrative review, the required data was retrieved using keywords "COVID-19," "Intranasal," "Immunity," "Nasal spray," and "Mucosal" in databases including PubMed, Scopus, Embase, Science Direct, and Web of Sciences. The results of the study showed that the nasal vaccines were both effective and protective according to the current researches approaching during the COVID-19 period and the preclinical and clinical phase trials prove the intranasal vaccination elicits more robust and cross-protective immunity than conventional vaccines. In this narrative review article, mechanisms across the nasal mucosa will be briefly presented and the current status of nasal vaccines during the COVID-19 pandemic is summarized, and advantages over traditional vaccines are provided. Furthermore, after exploring the primary benefits and kinetics of nasal vaccine, the potential for consideration of nasal vaccine as a booster dose is also discussed.

Lipid nanoparticle formulations for optimal RNA-based topical delivery to murine airways.

INTRODUCTION: Lipid nanoparticles (LNP) have been successfully used as a platform technology for delivering nucleic acids to the liver. To broaden the application of LNPs in targeting non-hepatic tissues, we developed LNP-based RNA therapies (siRNA or mRNA) for the respiratory tract. Such optimized LNP systems could offer an early treatment strategy for viral respiratory tract infections such as COVID-19. METHODS: We generated a small library of six LNP formulations with varying helper lipid compositions and characterized their hydrodynamic diameter, size distribution and cargo entrapment properties. Next, we screened these LNP formulations for particle uptake and evaluated their potential for transfecting mRNA encoding green fluorescence protein (GFP) or SARS-CoV2 nucleocapsid-GFP fusion reporter gene in a human airway epithelial cell line in vitro. Following LNP-siGFP delivery, GFP protein knockdown efficiency was assessed by flow cytometry to determine %GFP+ cells and median fluorescence intensity (MFI) for GFP. Finally, lead LNP candidates were validated in Friend leukemia virus B (FVB) male mice via intranasal delivery of an mRNA encoding luciferase, using in vivo bioluminescence imaging. RESULTS: Dynamic light scattering revealed that all LNP formulations contained particles with an average diameter of <100 nm and a polydispersity index of <0.2. Human airway epithelial cell lines in culture internalized LNPs with differential GFP transfection efficiencies (73-97%). The lead formulation LNP6 entrapping GFP or Nuc-GFP mRNA demonstrated the highest transfection efficiency (97%). Administration of LNP-GFP siRNA resulted in a significant reduction of GFP protein expression. For in vivo studies, intranasal delivery of LNPs containing helper lipids (DSPC, DOPC, ESM or DOPS) with luciferase mRNA showed significant increase in luminescence expression in nasal cavity and lungs by at least 10 times above baseline control. CONCLUSION: LNP formulations enable the delivery of RNA payloads into human airway epithelial cells, and in the murine respiratory system; they can be delivered to nasal mucosa and lower respiratory tract via intranasal delivery. The composition of helper lipids in LNPs crucially modulates transfection efficiencies in airway epithelia, highlighting their importance in effective delivery of therapeutic products for airways diseases.

Development of a Robotic Arm and End-Effector for COVID-19 Testing

Health-care professionals play an essential role in fighting the Covid-19 pandemic. Currently, most of the collected Covid-19 specimens are collected manually by health care workers and by using sterile cotton swab. The close contact between the health workers and the patients put them at high risk of being infected. One of approaches toward protecting the health workers, is to reduce the direct contact between the health care workers and the Covid-19 patients through deploying robots. In this study, a robotic system for Covid-19 swab sampling is designed and built. The robotic system consists of a 3 DOF arm, 2 DOF end effector with force sensor, and head holder. The robot is controlled remotely and from a safe distance. The preliminary experimental tests show that the robot is able to position the swab tip at the opening of the nasal cavity with the orientation and location that makes the swab insertion parallel to the palate. A force sensor is integrated in the end effector design to measure the normal contact force between the swab tip and the interior of the nasal cavity. The experimental results show that the normal contact force was successfully controlled and maintained within the acceptable force range, between 0.35N and 0.68N, as reported in literature. Maintaining the contact force within such range is essential in protecting the interior tissue of the nasal cavity from any injury © 2021 IEEE.

Developing a Novel Method for Disinfection and Sterilization Using a Charged Ozonated mist

The COVID-19 virus has caused a large-scale global outbreak and has become a major public health issue 1. Although there are several vaccines, herd immunity will likely take a long time to establish, and it is not clear whether the existing vaccines are completely effective against evolved versions of the virus. The COVID-19 virus as well as other respiratory viruses can be spread through coughing, sneezing, skin contact, etc., and can enter the human body from the eyes, nasal cavity, and oral cavity. © 2021 IEEE.

Exhaled Jet and Viral-Laden Aerosol Transport from Nasal Sneezing

The recognition that the spread of COVID-19 is primarily through airborne transmission has brought renewed urgency to understand the spread of aerosols generated from patients. Viral-laden aerosols generated from oral coughs have been well studied;however, aerosols generated from nasal sneezing has been overlooked. This scenario arises from patients who suffer allergenic rhinosinusitis, or the nasal cavity is irritated, particularly during naso-endoscopy. Nasal sneezing is characterised by an explosive blast of air exiting the nostrils, which can be considered as dual jets, resulting in the spread of viral-laden aerosols remaining suspended in the air. This study used computational fluid dynamics consisting of a hybrid RANS-LES turbulence method to model the airflow and the discrete phase model to track aerosol dispersion during nasal sneezing. The results demonstrated that the exhaled airflow jets during nasal sneezing resemble the flow characteristics of two parallel jets in co-flow. These two jets interfere with each other in the merging zone, and after they merge, the sneeze plume expands radially. The nasal sneeze forms a V-shaped plume with smaller particles in the core region. At the end of the sneeze, when the exhaled jets have lost their initial momentum, the large particle dispersion is dominated by gravity. We detected the presence of a 'sneeze puff' that transport droplets away from the body, similar to the buoyant puff observed in recent COVID-19 studies of oral coughs.

Evaluation of the Diagnostic Accuracy of Nasal Cavity and Nasopharyngeal Swab Specimens for SARS-CoV-2 Detection via Rapid Antigen Test According to Specimen Collection Timing and Viral Load.

The rapid diagnosis of SARS-CoV-2 is an essential aspect in the detection and control of the spread of COVID-19. We evaluated the accuracy of the rapid antigen test (RAT) using samples from the nasal cavity and nasopharynx based on sample collection timing and viral load. We enrolled 175 patients, of which 71 patients and 104 patients had tested positive and negative, respectively, based on real time-PCR. Nasal cavity and nasopharyngeal swab samples were tested using STANDARD Q COVID-19 Ag tests (Q Ag, SD Biosensor, Korea). The sensitivity of the Q Ag test was 77.5% (95% confidence interval [CI], 67.8-87.2%) for the nasal cavity and 81.7% (95% [CI, 72.7-90.7%) for the nasopharyngeal specimens. The RAT results showed a substantial agreement between the nasal cavity and nasopharyngeal specimens (Cohen's kappa index = 0.78). The sensitivity of the RAT for nasal cavity specimens exceeded 89% for <5 days after symptom onset (DSO) and 86% for Ct of E and RdRp < 25. The Q Ag test performed fairly well, especially in the early DSO when a high viral load was present, and the nasal cavity swab can be considered an alternative site for the rapid diagnosis of COVID-19.

Hamsters Expressing Human Angiotensin-Converting Enzyme 2 Develop Severe Disease following Exposure to SARS-CoV-2.

The rapid emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has created a global health emergency. While most human disease is mild to moderate, some infections lead to a severe disease characterized by acute respiratory distress, hypoxia, anosmia, ageusia, and, in some instances, neurological involvement. Small-animal models reproducing severe disease, including neurological sequela, are needed to characterize the pathophysiological mechanism(s) of disease and to identify medical countermeasures. Transgenic mice expressing the human angiotensin-converting enzyme 2 (hACE2) viral receptor under the control of the K18 promoter develop severe and lethal respiratory disease subsequent to SARS-CoV-2 intranasal challenge when high viral doses are used. Here, we report on SARS-CoV-2 infection of hamsters engineered to express the hACE2 receptor under the control of the K18 promoter. K18-hACE2 hamsters infected with a relatively low dose of 100 or 1,000 PFU of SARS-CoV-2 developed a severe and lethal disease, with most animals succumbing by day 5 postinfection. Hamsters developed severe lesions and inflammation within the upper and lower respiratory system, including infection of the nasal cavities causing marked destruction of the olfactory epithelium as well as severe bronchopneumonia that extended deep into the alveoli. Additionally, SARS-CoV-2 infection spread to the central nervous system (CNS), including the brain stem and spinal cord. Wild-type (WT) hamsters naturally support SARS-CoV-2 infection, with the primary lesions present in the respiratory tract and nasal cavity. Overall, infection in the K18-hACE2 hamsters is more extensive than that in WT hamsters, with more CNS involvement and a lethal outcome. These findings demonstrate the K18-hACE2 hamster model will be valuable for studying SARS-CoV-2. IMPORTANCE The rapid emergence of SARS-CoV-2 has created a global health emergency. While most human SARS-CoV-2 disease is mild, some people develop severe, life-threatening disease. Small-animal models mimicking the severe aspects of human disease are needed to more clearly understand the pathophysiological processes driving this progression. Here, we studied SARS-CoV-2 infection in hamsters engineered to express the human angiotensin-converting enzyme 2 viral receptor under the control of the K18 promoter. SARS-CoV-2 produces a severe and lethal infection in transgenic hamsters that mirrors the most severe aspects of COVID-19 in humans, including respiratory and neurological injury. In contrast to other animal systems, hamsters manifest disease with levels of input virus more consistent with natural human infection. This system will be useful for the study of SARS-CoV-2 disease and the development of drugs targeting this virus.

Can the Nasal Cavity Help Tackle COVID-19?

Despite the progress made in the fight against the COVID-19 pandemic, it still poses dramatic challenges for scientists around the world. Various approaches are applied, including repurposed medications and alternative routes for administration. Several vaccines have been approved, and many more are under clinical and preclinical investigation. This review aims to systemize the available information and to outline the key therapeutic strategies for COVID-19, based on the nasal route of administration.

Determinants of SARS-CoV-2 entry and replication in airway mucosal tissue and susceptibility in smokers.

Understanding viral tropism is an essential step toward reducing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission, decreasing mortality from coronavirus disease 2019 (COVID-19) and limiting opportunities for mutant strains to arise. Currently, little is known about the extent to which distinct tissue sites in the human head and neck region and proximal respiratory tract selectively permit SARS-CoV-2 infection and replication. In this translational study, we discover key variabilities in expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2), essential SARS-CoV-2 entry factors, among the mucosal tissues of the human proximal airways. We show that SARS-CoV-2 infection is present in all examined head and neck tissues, with a notable tropism for the nasal cavity and tracheal mucosa. Finally, we uncover an association between smoking and higher SARS-CoV-2 viral infection in the human proximal airway, which may explain the increased susceptibility of smokers to developing severe COVID-19. This is at least partially explained by differences in interferon (IFN)-ß1 levels between smokers and non-smokers.

Immunohistochemical and qPCR Detection of SARS-CoV-2 in the Human Middle Ear Versus the Nasal Cavity: Case Series.

Viral infections have already been implicated with otitis media and sudden sensorineural hearing loss. However, the pathophysiology of COVID-19 as it relates to otologic disorders is not well-defined. With the spread of SARS-CoV-2, it is important to evaluate its colonization of middle ear mucosa. Middle ear and nasal tissue samples for quantitative RT-PCR and histologic evaluations were obtained from post-mortem COVID-19 patients and non-diseased control patients. Here we present evidence that SARS-CoV-2 colonizes the middle ear epithelium and co-localizes with the primary viral receptor, angiotensin-converting enzyme 2 (ACE2). Both middle ear and nasal epithelial cells show relatively high expression of ACE2, required for SARS-CoV-2 entry. The epithelial cell adhesion molecule (EpCAM) was use as a biomarker of epithelia. Furthermore, we found that the viral load in the middle ear is lower than that present in the nasal cavity.

Complications of nasal SARS-CoV-2 testing: a review.

Transnasal swab testing for the detection of SARS-CoV-2 is well established. The Centers for Disease Control and Prevention advocates swabbing either of the anterior nares, middle turbinate, or nasopharynx for specimen collection depending on available local resources. The purpose of this review is to investigate complications related to transnasal SARS-CoV-2 testing with specific attention to specimen collection site and swab approach. The literature demonstrates that while nasopharyngeal swabbing is associated with an increased risk of complications, it should remain the gold-standard test due to greater diagnostic accuracy relative to anterior nasal and middle turbinate swabs.

Precision silver nitrate cautery of the posterior nasal cavity.

BACKGROUND: This technical note describes a novel method of cauterising the posterior nasal cavity through the use of a plastic straw and silver nitrate. OBJECTIVE: This technique aims to prevent unwanted damage to surrounding nasal mucosa. METHODS: Once the nasal cavity has been prepared for cauterisation, the silver nitrate stick is navigated to the bleeding point covered by the plastic straw. The silver nitrate stick is then advanced onto the bleeding point allowing precise cauterisation of the nasal mucosa, without effecting surrounding healthy mucosa.

A Dry Powder Platform for Nose-to-Brain Delivery of Dexamethasone: Formulation Development and Nasal Deposition Studies.

Nasal route of administration offers a unique opportunity of brain targeted drug delivery via olfactory and trigeminal pathway, providing effective CNS concentrations at lower doses and lower risk for adverse reactions compared to systemic drug administration. Therefore, it has been recently proposed as a route of choice for glucocorticoids to control neuroinflammation processes in patients with severe Covid-19. However, appropriate delivery systems tailored to enhance their efficacy yet need to emerge. In this work we present the development of sprayable brain targeting powder delivery platform of dexamethasone sodium phosphate (DSP). DSP-loaded microspheres, optimised employing Quality-by-Design approach, were blended with soluble inert carriers (mannitol or lactose monohydrate). Powder blends were characterized in terms of homogeneity, flow properties, sprayability, in vitro biocompatibility, permeability and mucoadhesion. Nasal deposition studies were performed using 3D printed nasal cavity model. Mannitol provided better powder blend flow properties compared to lactose. Microspheres blended with mannitol retained or enlarged their mucoadhesive properties and enhanced DSP permeability across epithelial model barrier. DSP dose fraction deposited in the olfactory region reached 17.0% revealing the potential of developed powder platform for targeted olfactory delivery. The observed impact of nasal cavity asymmetry highlighted the importance of individual approach when aiming olfactory region.

Community Use of Face Masks against the Spread of COVID-19.

The role of face masks to prevent and control COVID-19 is critical, especially since asymptomatic or pre-symptomatic infected individuals can shed high loads of SARS-CoV-2 in the surrounding environment. In addition to being a two-way barrier to protect against virions droplets both in terms of "source control" (for the benefits of the community) and "physical protection" (for wearer), face masks also allow maintaining physiological temperatures and humidity of the nasal cavity and mouth, independently from the external environmental conditions. Beyond compromising the viral transmission speed, exposure to cold environments could have a detrimental effect on the host's susceptibility to SARS-CoV-2. The innate human immune system becomes in fact weaker with cooler nose temperatures and thus more vulnerable to viral replication. Furthermore, there is evidence that warm, humid climates are associated with reduced spread of SARS-CoV-2, while cold dry conditions favor its stability and transmissibility. In the early stage of a viral infection, a physiological body temperature in the upper airways supports the innate immune system, endorsing the muco-ciliary clearance, inhibiting, or deactivating any first settlement of viruses. Face masks are therefore strongly recommended also outdoors, especially under cold weather conditions, not only as a physical barrier against the transmission of SARS-CoV-2, but also to prevent the rapid cooling of the nasal mucosa and the inhibition of the human innate defense of the upper airways.

A comparison of swab types on sample adequacy, suspects comfort and provider preference in COVID-19.

AIM: This study was aimed to compare the virological, suspect reported outcomes and provider preferences during COVID-19 swab taking procedure used for sampling. METHODS: The COVID-19 suspects are subjected to nasopharyngeal (NP) and oropharyngeal (OP) swabs for testing. Two types of swabs (Nylon and Dacron) are used for sample collection. Prospectively each suspect's response is collected and assessed for self-reported comfort level. The provider's experience with each suspect and virological outcomes recorded separately. The sample adequacy was compared based on swab types and demographic characteristics. RESULTS: A total of 1008 COVID-19 suspects were considered for comparison of various outcomes. Dacron and flocked Nylon swab sticks are used for taking 530 and 478 samples, respectively. Suspects who underwent the procedure using Nylon swabs were six times more likely to have pain/discomfort compared to when Dacron swab was used (Adj RR (95% CI: 6.76 (3.53 to 13, p=0.0001))). The providers perceived six times more resistance with the Nylon swabs compared to Dacron Swabs (Adj RR (95% CI: 5.96 (3.88 to 9.14, p=0.0001))). The pediatric population had a higher rate of blood staining in Dacron swab [Dacron 66 (80.5%); Nylon 51 (54.8%) p=0.0001]. The sample adequacy rate and laboratory positivity rate were not significantly different from each other. CONCLUSIONS: Given the comparable virological outcomes, the difference in suspect and providers comfort should drive swab selection based on characteristics of the suspects. The bulbous Nylon swab caused more pain/discomfort in adults compared to Dacron.