Biting Innovations of Mosquito-Based Biomaterials and Medical Devices.

Mosquitoes are commonly viewed as pests and deadly predators by humans. Despite this perception, investigations of their survival-based behaviors, select anatomical features, and biological composition have led to the creation of several beneficial technologies for medical applications. In this review, we briefly explore these mosquito-based innovations by discussing how unique characteristics and behaviors of mosquitoes drive the development of select biomaterials and medical devices. Mosquito-inspired microneedles have been fabricated from a variety of materials, including biocompatible metals and polymers, to mimic of the mouthparts that some mosquitoes use to bite a host with minimal injury during blood collection. The salivary components that these mosquitoes use to reduce the clotting of blood extracted during the biting process provide a rich source of anticoagulants that could potentially be integrated into blood-contacting biomaterials or administered in therapeutics to reduce the risk of thrombosis. Mosquito movement, vision, and olfaction are other behaviors that also have the potential for inspiring the development of medically relevant technologies. For instance, viscoelastic proteins that facilitate mosquito movement are being investigated for use in tissue engineering and drug delivery applications. Even the non-wetting nanostructure of a mosquito eye has inspired the creation of a robust superhydrophobic surface coating that shows promise for biomaterial and drug delivery applications. Additionally, biosensors incorporating mosquito olfactory receptors have been built to detect disease-specific volatile organic compounds. Advanced technologies derived from mosquitoes, and insects in general, form a research area that is ripe for exploration and can uncover potential in further dissecting mosquito features for the continued development of novel medical innovations.

Perceived healthcare discrimination and well-being among older adults in the United States and Brazil.

Despite well-documented evidence illustrating the relationship between discrimination and health, less is known about the influence of unfair treatment when receiving medical care. Moreover, our current knowledge of cross-national and racial variations in healthcare discrimination is limited in aging populations. This article addresses these gaps using two harmonized data sets of aging populations to clarify the relationship between healthcare discrimination and health in the United States and Brazil. We use nationally representative, harmonized data from the Health and Retirement Study in the United States and the Brazilian Longitudinal Study of Aging to examine and compare perceived discrimination in the healthcare setting and its relationship to self-rated health, depression diagnosis, and depressive symptoms across national contexts. Using Poisson regression models and population attributable risk percent estimates, we found that aging adults reporting healthcare discrimination were at higher risk of poor self-rated health, diagnosed depression, and depressive symptoms. Our results also suggest that reducing perceived healthcare discrimination may contribute to improved self-rated health and mental well-being in later life across racialized societies. In two comparative settings, we highlight the differential impact of healthcare discrimination on self-rated health and depression. We describe the implications of our study's findings for national public health strategies focused on eliminating discrimination in the healthcare setting, particularly among aging countries.

3D printed transwell-integrated nose-on-chip model to evaluate effects of air flow-induced mechanical stresses on mucous secretion.

While there are many chip models that simulate the air-tissue interface of the respiratory system, only a few represent the upper respiratory system. These chips are restricted to unidirectional flow patterns that are not comparable to the highly dynamic and variable flow patterns found in the native nasal cavity. Here we describe the development of a tunable nose-on-chip device that mimics the air-mucosa interface and is coupled to an air delivery system that simulates natural breathing patterns through the generation of bi-directional air flow. Additionally, we employ computational modeling to demonstrate how the device design can be tuned to replicate desired mechanical characteristics within specific regions of the human nasal cavity. We also demonstrate how to culture human nasal epithelial cell line RPMI 2650 within the lab-on-chip (LOC) device. Lastly, Alcian Blue histological staining was performed to label mucin proteins, which play important roles in mucous secretion. Our results revealed that dynamic flow conditions can increase mucous secretion for RPMI 2650 cells, when compared to no flow, or stationary, conditions.

Colorism and classism confounded: Perceptions of discrimination in Latin America.

Despite competing narratives of mestizaje (race-mixing) emphasizing class discrimination and social movements highlighting the existence of racial discrimination in Latin America, little work has examined the overlap of class and color in people's understandings of discrimination. This study moves beyond the color/class binary by examining perceptions of only class, only color, and both class and color discrimination (dual discrimination). I also examine whether individuals have difficulty attributing the causes of discrimination by expanding upon the social psychological concept of attributional ambiguity. Using nationally representative data from the 2010 LAPOP's Americas Barometer survey, I find that color-based explanations have not replaced class-based explanations. Instead, both class and color appear to be part of schemas drawn upon by individuals to understand the unfavorable treatment they perceive-in line with scholarship showing both class disadvantage and color conjointly influence the stratification systems of Latin America. There is also suggestive evidence that individuals may have trouble disentangling the causes of the discrimination they perceive.

Morphometric and computational assessments to evaluate neuron survival and maturation within compartmentalized microfluidic devices: The influence of design variation on diffusion-driven nutrient transport.

Burgeoning use of segregated microfluidic platforms that parse somas and neurites into discrete compartments is fueling unique examinations of neuronal structure and physiology in a manner impossible to achieve with non-compartmentalized systems. However, even though this line of axon-soma polarizing microfluidic devices stems from the same general design of a Campenot chamber set-up, slight deviations in device geometry appear to induce vastly different nutrient transport profiles that influence neuron survival and maturation. Here we examine the uptake of nerve growth factor (NGF) by a pheochromocytoma PC12 cell line cultured using two Campenot-like device designs, a "Standard" layout, representative of a commercial device, and a custom "Notch" layout, predicted to encourage more efficient nutrient transfer that gives rise to sustained neuron viability and extensive neurite elaboration. Exploiting in vitro culture schemes coupled with computational analyses, we identify the influence of device design geometry on the interplay between neuronal survival and maturation, gauged from morphometric assessments and the spatiotemporal distribution of NGF. Computer simulations of NGF transport within the devices revealed that the microfluidic neuron culture system is highly sensitive to change, where nutrient transport is intricately linked to device geometry and cell plating density, and premature depletion of nutrients is observed if specific design criteria are not met. This study underscores the importance of validating specific device geometries for a particular neuro-based assessment, while showcasing computational modeling as a powerful tool to achieve this goal.

Bridging the gap in peripheral nerve repair with 3D printed and bioprinted conduits.

Over the past two decades, a number of fabrication methods, including 3D printing and bioprinting, have emerged as promising technologies to bioengineer nerve conduits that closely replicate features of the native peripheral nerve, with the aim of augmenting or supplanting autologous nerve grafts. 3D printing and bioprinting offer the added advantage of rapidly creating composite peripheral nerve matrices from micron-scaled units, using an assortment of synthetic, natural and biologic materials. In this review, we explore the evolution of automated 3D manufacturing technologies for the development of peripheral nerve conduits and discuss aspects of conduit design, based on microarchitecture, material selection, cell and protein inclusion, and mechanical properties, as they are adaptable to 3D printing. Additionally, we highlight advancements in the application of bio-imaging modalities toward the fabrication of patient-specific nerve conduits. Lastly, we outline regulatory as well as clinical challenges that must be surmounted for the translation of 3D printing and bioprinting technology to the clinic. As a whole, this review addresses topics that may situate 3D manufacturing at the forefront of fabrication technologies that are exploited for the generation of future revolutionary therapies like in situ printing of peripheral nerves.

MRI compatible MS2 nanoparticles designed to cross the blood-brain-barrier: providing a path towards tinnitus treatment.

Fundamental challenges of targeting specific brain regions for treatment using pharmacotherapeutic nanoparticle (NP) carriers include circumventing the blood-brain-barrier (BBB) and tracking delivery. Angiopep-2 (AP2) has been shown to facilitate the transport of large macromolecules and synthetic nanoparticles across the BBB. Thus, conjugation of AP2 to an MS2 bacteriophage based NP should also permit transport across the BBB. We have fabricated and tested a novel MS2 capsid-based NP conjugated to the ligand AP2. The reaction efficiency was determined to be over 70%, with up to two angiopep-2 conjugated per MS2 capsid protein. When linked with a porphyrin ring, manganese (Mn2+) remained stable within MS2 and was MRI detectable. Nanoparticles were introduced intracerebroventricularly or systemically. Systemic delivery yielded dose dependent, non-toxic accumulation of NPs in the midbrain. Design of a multifunctional MRI compatible NP platform provides a significant step forward for the diagnosis and treatment of intractable brain conditions, such as tinnitus.

A drop array culture for patterning adherent mouse embryonic stem cell-derived neurospheres.

New therapeutic approaches for repairing an injured or degenerating nervous system have accelerated the development of methods to generate populations of neurons derived from various stem cell sources efficiently. Many of these methods require the generation of neurospheres. Here a simple technique is described for creating an array of adherent mouse embryonic stem cell (mESC)-derived neurospheres using a conventional plastic culture dish and a patterning template. mESC-derived neurospheres are confined to circular (4-mm diameter), gel-coated regions within an array. The adherent neurosphere arrays require 3 days to prepare from an mESC source; they can be maintained in 15 µl drops of medium, and exhibit extensive neurite elaboration after 8 days of cultivation. Additionally, the potential of treating the adherent neurospheres in selected drops of an array is demonstrated with a variety of differentiation-inducing reagents and subsequently individually analysing such neurospheres for gene expression, protein levels and morphological development. Copyright © 2016 John Wiley & Sons, Ltd.

Recent developments in multiplexing techniques for immunohistochemistry.

Methods to detect immunolabeled molecules at increasingly higher resolutions, even when present at low levels, are revolutionizing immunohistochemistry (IHC). These technologies can be valuable for the management and examination of rare patient tissue specimens, and for improved accuracy of early disease detection. The purpose of this article is to highlight recent multiplexing methods that are candidates for more prevalent use in clinical research and potential translation to the clinic. Multiplex IHC methods, which permit identification of at least 3 and up to 30 discrete antigens, have been divided into whole-section staining and spatially-patterned staining categories. Associated signal enhancement technologies that can enhance performance and throughput of multiplex IHC assays are also discussed. Each multiplex IHC technique, detailed herein, is associated with several advantages as well as tradeoffs that must be taken into consideration for proper evaluation and use of the methods.

Morphometric assessment of toxicant induced neuronal degeneration in full and restricted contact co-cultures of embryonic cortical rat neurons and astrocytes: using m-Dinitrobezene as a model neurotoxicant.

With m-Dinitrobenzene (m-DNB) as a selected model neurotoxicant, we demonstrate how to assess neurotoxicity, using morphology based measurement of neurite degeneration, in a conventional "full-contact" and a modern "restricted-contact" co-culture of rat cortical neurons and astrocytes. In the "full-contact" co-culture, neurons and astrocytes in complete physical contact are "globally" exposed to m-DNB. A newly emergent "restricted-contact" co-culture is attained with a microfluidic device that polarizes neuron somas and neurites into separate compartments, and the neurite compartment is "selectively" exposed to m-DNB. Morphometric analysis of the neuronal area revealed that m-DNB exposure produced no significant change in mean neuronal cell area in "full-contact" co-cultures, whereas a significant decrease was observed for neuron monocultures. Neurite elaboration into a neurite exclusive compartment in a compartmentalized microfluidic device, for both monocultures (no astrocytes) and "restricted" co-cultures (astrocytes touching neurites), decreased with exposure to increasing concentrations of m-DNB, but the average neurite area was higher in co-cultures. By using co-culture systems that more closely approach biological and architectural complexities, and the directionality of exposure found in the brain, this study provides a methodological foundation for unraveling the role of physical contact between astrocytes and neurons in mitigating the toxic effects of chemicals such as m-DNB.

Microfluidic device capable of medium recirculation for non-adherent cell culture.

We present a microfluidic device designed for maintenance and culture of non-adherent mammalian cells, which enables both recirculation and refreshing of medium, as well as easy harvesting of cells from the device. We demonstrate fabrication of a novel microfluidic device utilizing Braille perfusion for peristaltic fluid flow to enable switching between recirculation and refresh flow modes. Utilizing fluid flow simulations and the human promyelocytic leukemia cell line, HL-60, non-adherent cells, we demonstrate the utility of this RECIR-REFRESH device. With computer simulations, we profiled fluid flow and concentration gradients of autocrine factors and found that the geometry of the cell culture well plays a key role in cell entrapping and retaining autocrine and soluble factors. We subjected HL-60 cells, in the device, to a treatment regimen of 1.25% dimethylsulfoxide, every other day, to provoke differentiation and measured subsequent expression of CD11b on day 2 and day 4 and tumor necrosis factor-alpha (TNF-α) on day 4. Our findings display perfusion sensitive CD11b expression, but not TNF-α build-up, by day 4 of culture, with a 1:1 ratio of recirculation to refresh flow yielding the greatest increase in CD11b levels. RECIR-REFRESH facilitates programmable levels of cell differentiation in a HL-60 non-adherent cell population and can be expanded to other types of non-adherent cells such as hematopoietic stem cells.

Defined topologically-complex protein matrices to manipulate cell shape via three-dimensional fiber-like patterns.

Culturing cells in three-dimensional (3D) environments has been shown to significantly influence cell function, and may provide a more physiologically relevant environment within which to study the behavior of specific cell types. 3D tissues typically present a topologically complex fibrous adhesive environment, which is technically challenging to replicate in a controlled manner. Micropatterning technologies have provided significant insights into cell-biomaterial interactions, and can be used to create fiber-like adhesive structures, but are typically limited to flat culture systems; the methods are difficult to apply to topologically-complex surfaces. In this work, we utilize crack formation in multilayered microfabricated materials under applied strain to rapidly generate well-controlled and topologically complex 'fiber-like' adhesive protein patterns, capable of supporting cell culture and controlling cell shape on three-dimensional patterns. We first demonstrate that the features of the generated adhesive environments such as width, spacing and topology can be controlled, and that these factors influence cell morphology. The patterning technique is then applied to examine the influence of fiber structure on the nuclear morphology and actin cytoskeletal structure of cells cultured in a nanofibrous biomaterial matrix.

One-dimensional patterning of cells in silicone wells via compression-induced fracture.

We have adapted our existing compression-induced fracture technology to cell culture studies by generating linear patterns on a complex cell culture well structure rather than on simple solid constructs. We present a simple method to create one-dimensional (1D), submicron, and linear patterns of extracellular matrix on a multilayer silicone material. We identified critical design parameters necessary to optimize compression-induced fracture patterning on the wells, and applied stresses using compression Hoffman clamps. Finite-element analyses show that the incorporation of the well improves stress homogeneity (stress variation = 25%), and, thus, crack uniformity over the patterned region. Notably, a shallow well with a thick base (vs. deeper wells with thinner bases) reduces out-of-plane deflections by greater than a sixth in the cell culture region, improving clarity for optical imaging. The comparison of cellular and nuclear shape indices of a neuroblast line cultured on patterned 1D lines and unpatterned 2D surfaces reveals significant differences in cellular morphology, which could impact many cellular functions. Because 1D cell cultures recapitulate many important phenotypical traits of 3D cell cultures, our culture system offers a simple means to further study the relationship between 1D and 3D cell culture environments, without demanding expensive engineering techniques and expertise.