Comparison of homemade mask designs based on calculated infection risk, using actual COVID-19 infection scenarios.

During pandemics such as COVID-19, shortages of approved respirators necessitate the use of alternative masks, including homemade designs. The effectiveness of the masks is often quantified in terms of the ability to filter particles. However, to formulate public policy the efficacy of the mask in reducing the risk of infection for a given population is considerably more useful than its filtration efficiency (FE). The effect of the mask on the infection profile is complicated to estimate as it depends strongly upon the behavior of the affected population. A recently introduced tool known as the dynamic-spread model is well suited for performing population-specific risk assessment. The dynamic-spread model was used to simulate the performance of a variety of mask designs (all used for source control only) in different COVID-19 scenarios. The efficacy of different masks was found to be highly scenario dependent. Switching from a cotton T-shirt of 8% FE to a 3-layer cotton-gauze-cotton mask of 44% FE resulted in a decrease in number of new infections of about 30% in the New York State scenario and 60% in the Harris County, Texas scenario. The results are valuable to policy makers for quantifying the impact upon the infection rate for different intervention strategies, e.g., investing resources to provide the community with higher-filtration masks.

Vessel Rupture Thresholds for Vessel-Bubble Interactions Using an Earthworm Vasculature Model.

OBJECTIVE: Intravenous microbubble oscillation in the presence of ultrasound has the potential to yield a wide range of therapeutic benefits. However, the likelihood of vessel damage caused by mechanical effects has not been quantified as a function of the numerous important parameters in therapeutic ultrasound procedures. In this study, we examined the effects of microbubbles injected into the vasculature of the earthworm. It was found that the elastic properties of earthworm blood vessels are similar to those of arteries in older humans, and that earthworms are well suited to the large number of experiments necessary to investigate safety of procedures involving microbubble oscillation in sonicated vessels. METHODS: Microbubbles were infused into earthworm vessels, and the rupture time during sonication was recorded as a function of ultrasound frequency, pulse repetition frequency and acoustic pressure. DISCUSSION: A modified mechanical index (MMI) was defined that successfully captured the trends in rupture probability and rupture time for the different parameter values, creating a database of vessel rupture thresholds. In the absence of bubbles, the product of MMI squared and rupture time was approximately constant, indicating a possible radiation-force effect. CONCLUSION: The MMI was an effective correlating parameter in the presence of bubbles, though the mathematical dependence is not yet apparent. The results of the study are expected to be valuable in designing more refined studies in vertebrate models, as well as informing computational models.

Comparison of effectiveness of enhanced infection countermeasures in different scenarios, using a dynamic-spread-function model.

When formulating countermeasures to epidemics such as those generated by COVID-19, estimates of the benefits of a given intervention for a specific population are highly beneficial to policy makers. A recently introduced tool, known as the "dynamic-spread" SIR model, can perform population-specific risk assessment. Behavior is quantified by the dynamic-spread function, which includes the mechanisms of droplet reduction using facemasks and transmission control due to social distancing. The spread function is calibrated using infection data from a previous wave of the infection, or other data felt to accurately represent the population behaviors. The model then computes the rate of spread of the infection for different hypothesized interventions, over the time window for the calibration data. The dynamic-spread model was used to assess the benefit of three enhanced intervention strategies - increased mask filtration efficiency, higher mask compliance, and elevated social distancing - in four COVID-19 scenarios occurring in 2020: the first wave (i.e. until the first peak in numbers of new infections) in New York City; the first wave in New York State; the spread aboard the Diamond Princess Cruise Liner; and the peak occurring after re-opening in Harris County, Texas. Differences in the efficacy of the same intervention in the different scenarios were estimated. As an example, when the average outward filtration efficiency for facemasks worn in New York City was increased from an assumed baseline of 67% to a hypothesized 90%, the calculated peak number of new infections per day decreased by 40%. For the same baseline and hypothesized filtration efficiencies aboard the Diamond Princess Cruise liner, the calculated peak number of new infections per day decreased by about 15%. An important factor contributing to the difference between the two scenarios is the lower mask compliance (derivable from the spread function) aboard the Diamond Princess.

Material Properties, Dissolution and Time Evolution of PEGylated Lipid-Shelled Microbubbles: Effects of the Polyethylene Glycol Hydrophilic Chain Configurations.

Polyethylene glycol (PEG) is often added to the lipid coating of a contrast microbubble to prevent coalescence and improve circulation. At high surface density, PEG chains are known to undergo a transition from a mushroom configuration to an extended brush configuration. We investigated the effects of PEG chain configuration on attenuation and dissolution of microbubbles by varying the molar ratio of the PEGylated lipid in the shell with three (0%, 2% and 5%) in the mushroom configuration and two (10% and 20%) in the brush configuration. We measured attenuation through the bubble suspensions and used it to obtain the characteristic rheological properties of their shells according to two interfacial rheological models. The interfacial elasticity was found to be significantly lower in the brush regime (∼0.6 N/m) than in the mushroom regime (∼1.3 N/m), but similar in value within each regime. The dissolution behavior of microbubbles under acoustic excitation inside an air-saturated medium was studied by measuring the time-dependent attenuation. Total attenuation recorded a transient increase because of growth resulting from air influx and an eventual decrease caused by dissolution. Microbubble shell composition with varying PEG concentrations had significant effects on dissolution dynamics.

Modified Bovine Milk Exosomes for Doxorubicin Delivery to Triple-Negative Breast Cancer Cells.

Biological nanoparticles, such as exosomes, offer an approach to drug delivery because of their innate ability to transport biomolecules. Exosomes are derived from cells and an integral component of cellular communication. However, the cellular cargo of human exosomes could negatively impact their use as a safe drug carrier. Additionally, exosomes have the intrinsic yet enigmatic, targeting characteristics of complex cellular communication. Hence, harnessing the natural transport abilities of exosomes for drug delivery requires predictably targeting these biological nanoparticles. This manuscript describes the use of two chemical modifications, incorporating a neuropilin receptor agonist peptide (iRGD) and a hypoxia-responsive lipid for targeting and release of an encapsulated drug from bovine milk exosomes to triple-negative breast cancer cells. Triple-negative breast cancer is a very aggressive and deadly form of malignancy with limited treatment options. Incorporation of both the iRGD peptide and hypoxia-responsive lipid into the lipid bilayer of bovine milk exosomes and encapsulation of the anticancer drug, doxorubicin, created the peptide targeted, hypoxia-responsive bovine milk exosomes, iDHRX. Initial studies confirmed the presence of iRGD peptide and the exosomes' ability to target the αvß3 integrin, overexpressed on triple-negative breast cancer cells' surface. These modified exosomes were stable under normoxic conditions but fragmented in the reducing microenvironment created by 10 mM glutathione. In vitro cellular internalization studies in monolayer and three-dimensional (3D) spheroids of triple-negative breast cancer cells confirmed the cell-killing ability of iDHRX. Cell viability of 50% was reached at 10 µM iDHRX in the 3D spheroid models using four different triple-negative breast cancer cell lines. Overall, the tumor penetrating, hypoxia-responsive exosomes encapsulating doxorubicin would be effective in reducing triple-negative breast cancer cells' survival.

Retrospective analysis of interventions to epidemics using dynamic simulation of population behavior.

Retrospective analyses of interventions to epidemics, in which the effectiveness of strategies implemented are compared to hypothetical alternatives, are valuable for performing the cost-benefit calculations necessary to optimize infection countermeasures. SIR (susceptible-infected-removed) models are useful in this regard but are limited by the challenge of deciding how and when to update the numerous parameters as the epidemic changes in response to population behaviors. Behaviors of particular interest include facemask adoption (at various levels) and social distancing. We present a method that uses a "dynamic spread function" to systematically capture the continuous variation in the population behavior and the gradual change in infection evolution, resulting from interventions. No parameter updates are made by the user. We use the tool to quantify the reduction in infection rate realizable from the population of New York City adopting different facemask strategies during COVID-19. Assuming a baseline facemask of 67% filtration efficiency, calculations show that increasing the efficiency to 80% could have reduced the roughly 5000 new infections per day occurring at the peak of the epidemic to around 4000. Population behavior that may not be varied as part of the retrospective analysis, such as social distancing in a facemask analysis, are automatically captured as part of the calibration of the dynamic spread function.

Acoustic Droplet Vaporization of Perfluorocarbon Droplets in 3D-Printable Gelatin Methacrylate Scaffolds.

Scientists face a significant challenge in creating effective biomimetic constructs in tissue engineering with sustained and controlled delivery of growth factors. Recently, the addition of phase-shift droplets inside the scaffolds is being explored for temporal and spatial control of biologic delivery through vaporization using external ultrasound stimulation. Here, we explore acoustic droplet vaporization (ADV) in gelatin methacrylate (GelMA), a popular hydrogel used for tissue engineering applications because of its biocompatibility, tunable mechanical properties and rapid reproducibility. We embedded phase-shift perfluorocarbon droplets within the GelMA resin before crosslinking and characterized ADV and inertial cavitation (IC) thresholds of the embedded droplets. We were successful in vaporizing two different perfluorocarbon---perfluoropentane (PFP) and perfluorohexane (PFH)--cores at 2.25- and 5-MHz frequencies and inside hydrogels with varying mechanical properties. The ADV and IC thresholds for PFP droplets in GelMA scaffolds increased with frequency and in stiffer scaffolds. The PFH droplets exhibited ADV and IC activity only at 5 MHz for the range of excitations below 3MPa investigated here and at threshold values higher than those of PFP droplets. The results provide a proof of concept for the possible use of ADV in hydrogel scaffolds for tissue engineering.

Inhibition of Human Breast Cancer Cell Proliferation by Low-Intensity Ultrasound Stimulation.

OBJECTIVES: Cancer is characterized by uncontrolled cell proliferation, which makes novel therapies highly desired. In this study, the effects of near-field low-intensity pulsed ultrasound (LIPUS) stimulation on T47D human breast cancer cell and healthy immortalized MCF-12A breast epithelial cell proliferation were investigated in monolayer cultures. METHODS: A customized ultrasound (US) exposure setup was used for the variation of key US parameters: intensity, excitation duration, and duty cycle. Cell proliferation was quantified by 5-bromo-2'-deoxyuridine and alamarBlue assays after LIPUS excitation. RESULTS: At a 20% duty cycle and 10-minute excitation period, we varied LIPUS intensity from to 100 mW/cm2 (spatial-average temporal-average) to find a gradual decrease in T47D cell proliferation, the decrease being strongest at 100 mW/cm2 . In contrast, healthy MCF-12A breast cells showed an increase in proliferation when exposed to the same conditions. Above a 60% duty cycle, T47D cell proliferation decreased drastically. Effects of continuous wave US stimulation were further explored by varying the intensity and excitation period. CONCLUSIONS: These experiments concluded that, irrespective of the waveform (pulsed or continuous), LIPUS stimulation could inhibit the proliferation of T47D breast cancer cells, whereas the same behavior was not observed in healthy cells. The study demonstrates the beneficial bioeffects of LIPUS on breast cancer cells and offers the possibility of developing novel US-mediated cancer therapy.

Three-Dimensional Printing: A Catalyst for a Changing Orthopaedic Landscape.

Three-dimensional (3D) printing is an emerging tool in provider and patient education, surgical planning, and the design and implementation of medical devices and implants. Recent decreases in the cost of 3D printers along with advances in and cost reduction of printable materials have elevated 3D printing within the medical device industry. The advantages of 3D printing over traditional means of implant manufacturing lie in its ability to use a wide array of materials, its fine control of the macro- and microarchitecture, and its unprecedented customizability. Barriers to the widespread adoption of 3D-printed implants include questions of implant durability, U.S. Food and Drug Administration (FDA) approval for patient-specific implants, and insurance coverage of those implants.

Echogenic Exosomes as ultrasound contrast agents.

Exosomes are naturally secreted extracellular bilayer vesicles (diameter 40-130 nm), which have recently been found to play a critical role in cell-to-cell communication and biomolecule delivery. Their unique characteristics-stability, permeability, biocompatibility and low immunogenicity-have made them a prime candidate for use in delivering cancer therapeutics and other natural products. Here we present the first ever report of echogenic exosomes, which combine the benefits of the acoustic responsiveness of traditional microbubbles with the non-immunogenic and small-size morphology of exosomes. Microbubbles, although effective as ultrasound contrast agents, are restricted to intravascular usage due to their large size. In the current study, we have rendered bovine milk-derived exosomes echogenic by freeze drying them in the presence of mannitol. Ultrasound imaging and direct measurement of linear and nonlinear scattered responses were used to investigate the echogenicity and stability of the prepared exosomes. A commercial scanner registered enhancement (28.9% at 40 MHz) in the brightness of ultrasound images in presence of echogenic exosomes at 5 mg/mL. The exosomes also showed significant linear and nonlinear scattered responses-11 dB enhancement in fundamental, 8.5 dB in subharmonic and 3.5 dB in second harmonic all at 40 µg/mL concentration. Echogenic exosomes injected into the tail vein of mice and the synovial fluid of rats resulted in significantly higher brightness-as much as 300%-of the ultrasound images, showing their promise in a variety of in vivo applications. The echogenic exosomes, with their large-scale extractability from bovine milk, lack of toxicity and minimal immunogenic response, successfully served as ultrasound contrast agents in this study and offer an exciting possibility to act as an effective ultrasound responsive drug delivery system.

Exosomes as Drug Carriers for Cancer Therapy.

Exosomes, biological extracellular vesicles, have recently begun to find use in targeted drug delivery in solid tumor research. Ranging from 30-120 nm in size, exosomes are secreted from cells and isolated from bodily fluids. Exosomes provide a unique material platform due to their characteristics, including physical properties such as stability, biocompatibility, permeability, low toxicity, and low immunogenicity-all critical to the success of any nanoparticle drug delivery system. In addition to traditional chemotherapeutics, natural products and RNA have been encapsulated for the treatment of breast, pancreatic, lung, prostate cancers, and glioblastoma. This review discusses current research on exosomes for drug delivery to solid tumors.

Enhanced Osteogenic Differentiation of Human Mesenchymal Stem Cells Using Microbubbles and Low Intensity Pulsed Ultrasound on 3D Printed Scaffolds.

Lipid-coated microbubbles, clinically approved as contrast enhancing agents for ultrasound imaging, are investigated for the first time for their possible applications in bone tissue engineering. Effects of microbubbles (average diameter 1.1 µm) coated by a mixture of lipids (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000], and 1,2-dipalmitoyl-3-trimethylmmonium-propane) in the presence of low intensity pulsed ultrasound (LIPUS) on human mesenchymal stem cells seeded on 3D printed poly(lactic acid) porous scaffolds are investigated. LIPUS stimulation (30 mW cm-2 , 1.5 MHz, 20% duty cycle) for 3 min a day with 0.5% v/v microbubbles results in a significant increase in proliferation (up to 19.3%) when compared to control after 1, 3, and 5 d. A 3-week osteogenic differentiation study shows a significant increase in total protein content (up to 27.5%), calcium deposition (up to 4.3%), and alkaline phosphatase activity (up to 43.1%) initiated by LIPUS with and without the presence of microbubbles. The microbubbles are found to remain stable during exposure, and their sustained oscillations demonstrably help focus the LIPUS energy toward enhanced cellular response. Integrating LIPUS and microbubbles promises to be a novel and effective strategy for bone tissue engineering and regeneration therapies.

On the nature of the Cu-rich aggregates in brain astrocytes.

Fulfilling a bevy of biological roles, copper is an essential metal for healthy brain function. Cu dyshomeostasis has been demonstrated to be involved in some neurological conditions including Menkes and Alzheimer's diseases. We have previously reported localized Cu-rich aggregates in astrocytes of the subventricular zone (SVZ) in rodent brains with Cu concentrations in the hundreds of millimolar. Metallothionein, a cysteine-rich protein critical to metal homeostasis and known to participate in a variety of neuroprotective and neuroregenerative processes, was proposed as a binding protein. Here, we present an analysis of metallothionein(1,2) knockout (MTKO) mice and age-matched controls using X-ray fluorescence microscopy. In large structures such as the corpus callosum, cortex, and striatum, there is no significant difference in Cu, Fe, or Zn concentrations in MTKO mice compared to age-matched controls. In the astrocyte-rich subventricular zone where Cu-rich aggregates reside, approximately 1/3 as many Cu-rich aggregates persist in MTKO mice resulting in a decrease in periventricular Cu concentration. Aggregates in both wild-type and MTKO mice show XANES spectra characteristic of CuxSy multimetallic clusters and have similar [S]/[Cu] ratios. Consistent with assignment as a CuxSy multimetallic cluster, the astrocyte-rich SVZ of both MTKO and wild-type mice exhibit autofluorescent bodies, though MTKO mice exhibit fewer. Furthermore, XRF imaging of Au-labeled lysosomes and ubiquitin demonstrates a lack of co-localization with Cu-rich aggregates suggesting they are not involved in a degradation pathway. Overall, these data suggest that Cu in aggregates is bound by either metallothionein-3 or a yet unknown protein similar to metallothionein.