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Curvature is an integral part of the complex in vivo tissue architecture across various length scales. Therefore, several in vitro models with a patterned curvature in different length scales have been developed to understand the role of this in cellular behavior. At the subcellular scale, wavy patterns have been reported wherein concave and convex grooves are adjacently present. However, the independent effect of continuous subcellular concave and convex shapes has not been reported, mainly owing to the limitations in fabricating such patterns. In this study, we developed continuous concave and convex grooves on polydimethylsiloxane (PDMS) using a Dracaena sanderiana (bamboo) leaf as a template. The first (negative) replica from the abaxial side of the bamboo leaf, which imparted concave grooves on PDMS, was subsequently used as a template to fabricate a positive replica of the leaf, resulting in convex grooves of the same size and arrangement as the concave grooves. We examined the influence of the groove curvature on the morphology of bone marrow-derived human mesenchymal stem cells (BM-hMSCs) and skeletal muscle cells (C2C12). BM-hMSCs and C2C12 cells aligned on both concave and convex grooves as compared to the random orientation on a flat substrate. The significant difference was observed in the morphology of both cells, in terms of area, aspect ratio, number, and length of protrusions on concave and convex patterns. We found that the number of protrusions was also dependent on the ratio of cell to pattern length scale for convex-shaped grooves but independent of length scale for concave-shaped grooves. The proliferation of BM-hMSCs was also found to be different on concave and convex shapes. Therefore, this study shows the importance of (1) convex and concave curvatures of the subcellular length scale in cellular response, (2) dependence on the ratio of cell and curvature length scale, and (3) use of natural templates for overcoming fabrication challenges.
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Adhesión Celular , Proliferación Celular , Dimetilpolisiloxanos , Células Madre Mesenquimatosas , Humanos , Proliferación Celular/efectos de los fármacos , Células Madre Mesenquimatosas/citología , Dimetilpolisiloxanos/química , Anisotropía , Hojas de la Planta/química , Propiedades de SuperficieRESUMEN
Human body temperature limits below 40°C during heat stroke or fever. The implications of prolonged exposure to the physiologically relevant temperature (40°C) on cellular mechanobiology is poorly understood. Here, we have examined the effects of heat stress (40°C for 72â h incubation) in human lung adenocarcinoma (A549), mouse melanoma (B16F10), and non-cancerous mouse origin adipose tissue cells (L929). Hyperthermia increased the level of ROS, γ-H2AX and HSP70 and decreased mitochondrial membrane potential in the cells. Heat stress impaired cell division, caused G1 arrest, induced cellular senescence, and apoptosis in all the tested cell lines. The cells incubated at 40°C for 72â h displayed a significant decrease in the f-actin level and cellular traction as compared with cells incubated at 37°C. Also, the cells showed a larger focal adhesion area and stronger adhesion at 40°C than at 37°C. The mitotic cells at 40°C were unable to round up properly and displayed retracting actin stress fibers. Hyperthermia down-regulated HDAC6, increased the acetylation level of microtubules, and perturbed the chromosome alignment in the mitotic cells at 40°C. Overexpression of HDAC6 rescued the cells from the G1 arrest and reduced the delay in cell rounding at 40°C suggesting a crucial role of HDAC6 in hyperthermia mediated responses. This study elucidates the significant role of cellular traction, focal adhesions, and cytoskeletal networks in mitotic cell rounding and chromosomal misalignment. It also highlights the significance of HDAC6 in heat-evoked senile cellular responses.
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Senescencia Celular/fisiología , Citoesqueleto/metabolismo , Puntos de Control de la Fase G1 del Ciclo Celular , Histona Desacetilasa 6/metabolismo , Hipertermia/metabolismo , Fibras de Estrés/metabolismo , Acetilación , Actinas/metabolismo , Apoptosis/fisiología , Adhesión Celular/fisiología , División Celular/fisiología , Línea Celular Tumoral , Proliferación Celular/fisiología , Forma de la Célula , Tamaño de la Célula , Puntos de Control de la Fase G1 del Ciclo Celular/genética , Puntos de Control de la Fase G1 del Ciclo Celular/fisiología , Proteínas HSP70 de Choque Térmico/metabolismo , Histona Desacetilasa 6/genética , Humanos , Hipertermia/genética , Potencial de la Membrana Mitocondrial/fisiología , Microtúbulos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Regulación hacia ArribaRESUMEN
The human immunodeficiency virus (HIV) interacts with the immune cells within the human body, where the environment is uncertain and noisy. Stochastic models can successfully encapsulate the effect of such a noisy environment compared to their deterministic counterparts. The human immune system is complex but well-coordinated with various immune cells like C D 4T cells, dendritic cells, and cytotoxic T-lymphocyte (CTL) cells, among many others. The CTL can kill the antigenic cells after its recognition. However, the efficacy of CTL in removing the infected C D 4T cells is progressively compromised in HIV-infected individuals. This paper considers a noise-induced HIV-immune cell interaction model with immune impairment. A multiplicative white noise is introduced in the infection rate parameter to represent the fluctuations around the average value of the rate parameter as a causative effect of the noise. We analyzed the deterministic and stochastic models and prescribed sufficient conditions for infection eradication and persistence. It is determined under what parametric restrictions the asymptotic solutions of the noise-induced system will be a limiting case of the deterministic solutions. Simulation results revealed that the solutions of the deterministic system either converge to a CTL-dominated interior equilibrium or a CTL-free immunodeficient equilibrium, depending on the initial values of the system. Stochastic analysis divulged that higher noise might be helpful in the infection removal process. The extinction time of infected C D 4T cells for some fixed immune impairment gradually decreases with increasing noise intensity and follows the power law.
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Infecciones por VIH , Linfocitos T Citotóxicos , Humanos , Comunicación Celular , Simulación por Computador , IncertidumbreRESUMEN
Survival and maintenance of normal physiological functions depends on continuous interaction of cells with its microenvironment. Cells sense the mechanical properties of underlying substrate by applying force and modulate their behaviour in response to the resistance offered by the substrate. Most of the studies addressing cell-substrate mechanical interactions have been carried out using elastic substrates. Since tissues within our body are viscoelastic in nature, here we explore the effect of substrate's viscoelasticity on various properties of mesenchymal stem cells. Here, we used two sets of polyacrylamide substrates having similar storage modulus (G' = 1.1-1.6 kPa) but different loss modulus (G" = 45 Pa and 300 Pa). We report that human mesenchymal stem cells spread more but apply less force on the viscoelastic substrate (substrate with higher loss modulus). We further investigated the effect of substrate viscoelasticity on the expression of other contractility-associated proteins such as focal adhesion (FA) proteins (Vinculin, Paxillin, Talin), cytoskeletal proteins (actin, mysion, intermediate filaments, and microtubules) and mechano-sensor protein Yes-Associated Protein (YAP). Our results show that substrate viscoelasticity decouples cellular traction from other known traction related phenotypes.
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Proteínas del Citoesqueleto/metabolismo , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Adhesión Celular/fisiología , Procesos de Crecimiento Celular/fisiología , Elasticidad , Humanos , Fenotipo , Propiedades de Superficie , ViscosidadRESUMEN
The world has been facing the biggest virological invasion in the form of Covid-19 pandemic since the beginning of the year 2020. In this paper, we consider a deterministic epidemic model of four compartments based on the health status of the populations of a given country to capture the disease progression. A stochastic extension of the deterministic model is further considered to capture the uncertainty or variation observed in the disease transmissibility. In the case of a deterministic system, the disease-free equilibrium will be globally asymptotically stable if the basic reproduction number is less than unity, otherwise, the disease persists. Using Lyapunov functional methods, we prove that the infected population of the stochastic system tends to zero exponentially almost surely if the basic reproduction number is less than unity. The stochastic system has no interior equilibrium, however, its asymptotic solution is shown to fluctuate around the endemic equilibrium of the deterministic system under some parametric restrictions, implying that the infection persists. A case study with the Covid-19 epidemic data of Spain is presented and various analytical results have been demonstrated. The epidemic curve in Spain clearly shows two waves of infection. The first wave was observed during March-April and the second wave started in the middle of July and not completed yet. A real-time reproduction number has been given to illustrate the epidemiological status of Spain throughout the study period. Estimated cumulative numbers of confirmed and death cases are 1,613,626 and 42,899, respectively, with case fatality rate 2.66% till the deadly virus is eliminated from Spain.
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Efficient homing of human mesenchymal stem cells (hMSCs) is likely to be dictated by a combination of physical and chemical factors present in the microenvironment. However, crosstalk between the physical and chemical cues remains incompletely understood. Here, we address this question by probing the efficiency of epidermal growth factor (EGF)-induced hMSC chemotaxis on substrates of varying stiffness (3, 30 and 600â kPa) inside a polydimethylsiloxane (PDMS) microfluidic device. Chemotactic speed was found to be the sum of a stiffness-dependent component and a chemokine concentration-dependent component. While the stiffness-dependent component scaled inversely with stiffness, the chemotactic component was independent of stiffness. Faster chemotaxis on the softest 3â kPa substrates is attributed to a combination of weaker adhesions and higher protrusion rate. While chemotaxis was mildly sensitive to contractility inhibitors, suppression of chemotaxis upon actin depolymerization demonstrates the role of actin-mediated protrusions in driving chemotaxis. In addition to highlighting the collective influence of physical and chemical cues in chemotactic migration, our results suggest that hMSC homing is more efficient on softer substrates.
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Actinas/química , Proliferación Celular/genética , Quimiotaxis/genética , Células Madre Mesenquimatosas/química , Actinas/genética , Adhesión Celular/genética , Diferenciación Celular/genética , Células Cultivadas , Microambiente Celular/efectos de los fármacos , Microambiente Celular/genética , Dimetilpolisiloxanos/química , Elasticidad/fisiología , Factor de Crecimiento Epidérmico/química , Factor de Crecimiento Epidérmico/genética , Humanos , Células Madre Mesenquimatosas/metabolismo , Multimerización de Proteína/genéticaRESUMEN
The failure of chemotherapeutic drugs in treatment of various cancers is attributed to the acquisition of drug resistance. However, the migration mechanisms of drug-resistant cancer cells remain incompletely understood. Here we address this question from a biophysical perspective by mapping the phenotypic alterations in ovarian cancer cells (OCCs) resistant to cisplatin and paclitaxel. We show that cisplatin-resistant (CisR), paclitaxel-resistant (PacR) and dual drug-resistant (i.e., resistant to both drugs) OCCs are more contractile and softer than drug-sensitive cells. Protease inhibition suppresses invasion of CisR cells but not of PacR cells, indicative of a protease-dependent mode of migration in CisR cells and a protease-independent mode of migration in PacR. Despite these differences, actomyosin contractility, mediated by the RhoA-ROCK2-Myosin II signaling pathway, regulates both modes of migration. Confined migration experiments establish the role of myosin IIA and IIB in mediating nuclear translocation and regulation of proteolytic activity. Collectively, our results highlight the importance of myosin II as a potential therapeutic target for treatment of drug-resistant ovarian cancer cells.
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Resistencia a Antineoplásicos , Miosina Tipo II/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias Ováricas/metabolismo , Línea Celular Tumoral , Femenino , Humanos , Miosina Tipo II/genética , Proteínas de Neoplasias/genética , Neoplasias Ováricas/genética , Neoplasias Ováricas/patologíaRESUMEN
Microfluidic concentration gradient generators (µ-CGGs) are critical in various biochemical assays, including cell migration, drug screening, and antimicrobial susceptibility testing. However, current µ-CGGs rely on integration with flow systems, limiting their scalability and widespread adoption owing to limited infrastructure and technical expertise. Hence, there is a need for flowless diffusional gradient generators capable of standalone operation, thereby improving throughput and usability. In this study, we model such a diffusional µ-CGG as an infinite source-sink system to capture two characteristic timescales: (i) gradient generation dictated by the diffusion timescale and (ii) stability determined by the rate of change in reservoir concentrations. Through finite-element simulations, we explored the influence of various geometric parameters such as the channel length, cross-sectional area, node and reservoir volumes, and the solute diffusivity on these timescales, along with experimental confirmation using fluorescent tracer diffusion. Our results show that while the gradient stability strongly depends on the reservoir volumes, diffusion length, and solute diffusion coefficient, they are independent of the node shape or the shape of the channel cross section. However, gradient profiles were found to be the strong functions of the diffusion length, solute diffusivity, and the geometric pattern of the microfluidic grid. Additionally, we showcased the versatility of the design by generating discrete gradient profiles and combinatorial gradients of two and three solutes, thus improving throughput in a wide range of on-chip biological assays. These findings underscore the potential of our microfluidic device as an easy-to-use, inexpensive, efficient, and high-throughput platform for various on-chip biological assays.
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With a recent amendment, India joined other countries that have removed the legislative barrier toward the use of human-relevant methods in drug development. Here, global stakeholders weigh in on the urgent need to globally harmonize the guidelines toward the standardization of microphysiological systems. We discuss a possible framework for establishing scientific confidence and regulatory approval of these methods.
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Sistemas Microfisiológicos , Políticas , Humanos , Desarrollo de MedicamentosRESUMEN
Extracellular matrices (ECM) present in our tissues play a significant role in maintaining tissue homeostasis through various physical and chemical cues such as topology, stiffness, and secretion of biochemicals. They are known to influence the behavior of resident stem cells. It is also known that ECM type and coating density on cell culture plates strongly influence in vitro cellular behavior. However, the influence of ECM protein coating on long-term mesenchymal stem cell expansion has not been studied yet. To address this gap, we cultured bone-marrow derived hMSCs for multiple passages on the tissue culture plastic plates coated with 25 µg/ml of various ECM proteins. We found that cells on plates coated with ECM proteins had much higher proliferation compared to the regular tissue culture plates. Further, gelatin-coated plates helped the cells to grow faster compared to collagen, fibronectin, and laminin coated plates. Additionally, the use of gelatin showed less size heterogeneity among the cells when expanded from passages 3 to 9 (P3 to P9). Gelatin also helped in maintaining cellular stiffness which was not observed across other ECM proteins. In summary, in this research, we have shown that gelatin which is the least expensive compared to other ECM proteins, provides a better platform for mesenchymal stem cell expansion.
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Proteínas de la Matriz Extracelular , Células Madre Mesenquimatosas , Proteínas de la Matriz Extracelular/metabolismo , Gelatina/metabolismo , Matriz Extracelular/metabolismo , Técnicas de Cultivo de Célula , Proliferación Celular , Células Cultivadas , Diferenciación CelularRESUMEN
Correction for 'Design and validation of a flowless gradient generating microfluidic device for high-throughput drug testing' by Ketaki Bachal et al., Lab Chip, 2023, 23, 261-271, https://doi.org/10.1039/D2LC00879C.
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Drug testing is a vital step in the identification of the potential efficacy of any new/existing drug and/or combinations of drugs. The conventional methods of testing the efficacy of new drugs using multiwell plates are time consuming and prone to evaporation loss and manual error. Microfluidic devices with automated generation of concentration gradients provide a promising alternative. The implementation of such microfluidic devices is still limited owing to the additional expertise and facilities required to fabricate and run these devices. Conventional microfluidic devices also need pumps, tubing, valves, and other accessories, making them bulky and non-portable. To address these problems, we have developed a method for fabricating microfluidic structures using a nonconventional technique by exploiting the Saffman-Taylor instability in lifted Hele-Shaw cells. Multi-channel structure molds with varying dimensions were fabricated by shaping ceramic polymer slurry and retaining the shape. Further using the mold thus made, polydimethyl siloxane (PDMS) devices offering static, stable, diffusion-based gradients were casted using soft lithography. We have demonstrated with COMSOL simulation, as well as using fluorescein isothiocyanate (FITC), a fluorescent dye, that the concentration gradient can be generated in this device, which remains stable for at least 5 days. Using this multichannel device, in vitro drug efficacy was validated with two drugs namely, temozolomide (TMZ) and curcumin, one FDA approved and one under research, on glioblastoma cells (U87MG). The resulting IC50 values were consistent with those reported in the literature. We have also demonstrated the possibility of conducting molecular assays post-drug testing in the device by microtubule staining after curcumin treatment on cervical cancer cells (HeLa). In summary, we have demonstrated a i) user-friendly, ii) portable, static drug testing platform that iii) does not require further accessories and can create iv) a stable gradient for a long duration. Such a device can reduce the time, manual errors, fabrication and running expenditure, and resources needed to a great extent in drug testing.
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Curcumina , Técnicas Analíticas Microfluídicas , Humanos , Curcumina/farmacología , Microfluídica/métodos , Células HeLa , Dispositivos Laboratorio en un ChipRESUMEN
Recurrent glioblastoma is highly aggressive with currently no specific treatment regime. Therefore, to identify novel therapeutic targets for recurrent GBM, we used a cellular model developed in our lab from commercially available cell line U87MG and patient-derived cultures that allows the comparison between radiation naïve (Parent) and recurrent GBM cells generated after parent cells are exposed to lethal dose of radiation. Total RNA-seq of parent and recurrent population revealed significant upregulation of cell-ECM interactions pathway in the recurrent population. These results led us to hypothesize that the physical microenvironment contributes to the aggressiveness of recurrent GBM. To verify this, we cultured parent and recurrent GBM cells on collagen-coated polyacrylamide gels mimicking the stiffness of normal brain (Young's modulus E = 0.5kPa) or tumorigenic brain (E = 10kPa) and tissue culture plastic dishes (E â¼ 1 GPa). We found that compared to parent cells, recurrent cells showed higher proliferation, invasion, migration, and resistance to EGFR inhibitor. Using orthotopic GBM mouse model and resection model, we demonstrate that recurrent cells cultured on 0.5kPa had higher in vivo tumorigenicity and recurrent disease progression than parent cells, whereas these differences were insignificant when parent and recurrent cells were cultured on plastic substrates. Furthermore, recurrent cells on 0.5kPa showed high expression of ECM proteins like Collagen, MMP2 and MMP9. These proteins were also significantly upregulated in recurrent patient biopsies. Additionally, the brain of mice injected with recurrent cells grown on 0.5kPa showed higher Young's moduli suggesting the ability of these cells to make the surrounding ECM stiffer. Total RNA-seq of parent and recurrent cells grown on plastic and 0.5kpa identified PLEKHA7 significantly upregulated specifically in recurrent cells grown on 0.5 kPa substrate. PLEKHA7 was also found to be high in recurrent GBM patient biopsies. Accordingly, PLEKHA7 knockdown reduced invasion and survival of recurrent GBM cells. Together, these data provide an in vitro model system that captures the observed in vivo and clinical behavior of recurrent GBM by mimicking mechanical microenvironment and identifies PLEKHA7 as a novel potential target for recurrent GBM.
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Neoplasias Encefálicas , Glioblastoma , Ratones , Animales , Glioblastoma/metabolismo , Proliferación Celular , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Encéfalo/patología , Colágeno/metabolismo , Línea Celular Tumoral , Microambiente TumoralRESUMEN
Although the viscoelasticity or tackiness of a pressure-sensitive adhesive gives it strength owing to energy dissipation during peeling, it also renders it nonreusable because of structural changes such as the formation of fibrils, cohesive failure, and fouling. However, an elastic layer has good structural integrity and cohesive strength but low adhesive energy. We demonstrate an effective composite adhesive in which a soft viscoelastic bulk layer is imbedded in a largely elastic thin skin layer. The composite layer is able to meet the conflicting demands of the high peel strength comparable to the viscoelastic core and the structural integrity, reusability, and antifouling properties of the elastic skin. Our model adhesive is made of poly(dimethylsiloxane), where its core and skin are created by varying the cross-linking percentage from 2 to 10%.
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Cellular alignment is important for the proper functioning of different tissues such as muscles or blood vessel walls. Hence, in tissue engineering, sufficient effort has been made to control cellular orientation and alignment. It has been shown that micro-and nanoscale anisotropic topological features on cell culture substrates can control cellular orientation. Such substrates are fabricated using various lithography techniques such as photolithography and soft lithography. Although such techniques are suitable for creating patterns in small areas to establish a proof-of-concept, patterning large areas with intricate features is an unsolved problem. In this work, we report that a replica of the groove-like anisotropic patterns of the abaxial side of aDracaena sanderiana(bamboo) leaf can be used for large-area patterning of cells. We imprinted the leaf on polydimethylsiloxane (PDMS) and characterised its surface topography using scanning electron microscopy. We further cultured bone marrow human mesenchymal cells (BM-hMSCs), skeletal muscle cells (C2C12), and neuroblastoma cells (SHSY5Y) on the patterned PDMS on which the cells orient along the direction of the grooved pattern. Further, we observed enhanced neuronal differentiation of SHSY5Y cells on biomimicked pattern compared to flat PDMS as measured by percentage of cells with neurites, neurite length and the expression of neuronal differentiation marker beta-III tubulin (TUJ1). This process is simple, frugal, and can be adopted by laboratories with resource constraints. This one-step technique to fabricate large-area anisotropic surface patterns from bamboo leaves can be used as a platform to study cellular alignment and its effect on various cellular functions, including differentiation.
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Técnicas de Cultivo de Célula , Ingeniería de Tejidos , Técnicas de Cultivo de Célula/métodos , Diferenciación Celular , Línea Celular , Células Cultivadas , Humanos , Propiedades de Superficie , Ingeniería de Tejidos/métodosRESUMEN
The Covid-19 pandemic has put the world under immeasurable stress. There is no specific drug or vaccine till now that can cure the infection or protect people from the infection of coronavirus. It is therefore prudent to use the existing resources and control strategies in an optimal way to contain the virus spread and provide the best possible treatments to the infected individuals. Use of the repurposing drugs along with the non-pharmaceutical intervention strategies may be the right way for fighting against the ongoing pandemic. It is the objective of this work to demonstrate through mathematical modelling and analysis how and to what extent such control strategies can improve the overall Covid-19 epidemic burden. The criteria for disease elimination & persistence were established through the basic reproduction number. A case study with the Indian Covid-19 epidemic data is presented to visualize and illustrate the effects of lockdown, maintaining personal hygiene & safe distancing, and repurposing drugs. It is shown that India can significantly improve the overall Covid-19 epidemic burden through the combined use of NPIs and repurposing drugs though containment of spreading is difficult without serious community participation.
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Tratamiento Farmacológico de COVID-19 , COVID-19 , SARS-CoV-2 , COVID-19/epidemiología , COVID-19/prevención & control , Control de Enfermedades Transmisibles , Reposicionamiento de Medicamentos , Humanos , Pandemias/prevención & controlRESUMEN
Microfluidic concentration gradient generators are useful in drug testing, drug screening, and other cellular applications to avoid manual errors, save time, and labor. However, expensive fabrication techniques make such devices prohibitively costly. Here, in the present work, we developed a microfluidic concentration gradient generator (µCGG) using a recently proposed non-conventional photolithography-less method. In this method, ceramic suspension fluid was shaped into a square mesh by controlling Saffman Taylor instability in a multiport lifted Hele-Shaw cell (MLHSC). Using the shaped ceramic structure as the template, µCGG was prepared by soft lithography. The concentration gradient was characterized and effect of the flow rates was studied using COMSOL simulations. The simulation result was further validated by creating a fluorescein dye (fluorescein isothiocanate) gradient in the fabricated µCGG. To demonstrate the use of this device for drug testing, we created various concentrations of an anticancer drug-curcumin-using the device and determined its inhibitory concentration on cervical cancer cell-line HeLa. We found that the IC50 of curcumin for HeLa matched well with the conventional multi-well drug testing method. This method of µCGG fabrication has multiple advantages over conventional photolithography such as: (i) the channel layout and inlet-outlet arrangements can be changed by simply wiping the ceramic fluid before it solidifies, (ii) it is cost effective, (iii) large area patterning is easily achievable, and (iv) the method is scalable. This technique can be utilized to achieve a broad range of concentration gradient to be used for various biological and non-biological applications.
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The investigation is focused on the development of a compartmentalized microfluidic device for coculturing the cells of crucial retinal cellular layers and assessing cell-to-cell interactions. A perfusion-based microfluidic co-culture device was employed and computationally validated for determining the pressure drop and fluid flow rate within the device microchannels. Fabrication was performed using PDMS polymer and coating of fibronectin and collagen facilitated adherence of the cells over the glass surface. Microfluidic device successfully supported cell proliferation, under continuous perfusion of 1 µl min-1 flow rate. The barrier integrity of this coculture was confirmed by evaluating the permeability of fluorescently labeled molecules. The coculture expressed characteristic phenotypic protein markers like recoverin, PAX6, for retinal precursor cells, and RPE65 for retinal epithelial cells. The coculture also exhibited basal expression of TNF-α under normal conditions. Differentiated photoreceptor cells positively expressed rhod inherently possess sensitivity toward violet/blue light, which was validated in R28 cells by exposure to light having a wavelength of 405 nm, which significantly decreased cell viability via increased TNF-α production and reduced rhodopsin expression. This proof-of-concept investigation proved the functionality of the retinal coculture, which may be used as an appropriate perfusion-based, preclinical tool for the evaluation of novel retinal drugs and delivery systems.
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Dispositivos Laboratorio en un Chip , Factor de Necrosis Tumoral alfa , Técnicas de Cocultivo , Retina/metabolismo , Rodopsina/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Intricate structures of natural surfaces and materials have amazed people over the ages. The unique properties of various surfaces also created interest and curiosity in researchers. In the recent past, with the advent of superior microscopy techniques, we have started to understand how these complex structures provide superior properties. With that knowledge, scientists have developed various biomimicked and bio-inspired surfaces for different non-biological applications. In the last two decades, we have also started to learn how structures of the tissue microenvironment influence cell function and behaviour, both in physiological and pathological conditions. Hence, it became essential to decipher the role and importance of structural hierarchy in the cellular context. With advances in microfabricated techniques, such complex structures were made by superimposing features of different dimensions. However, the fabricated topographies are far from matching the complexities presentin vivo. Hence, the need of biomimicking the natural surfaces for cellular applications was felt. In this review, we discuss a few examples of hierarchical surfaces found in plants, insects, and vertebrates. Such structures have been widely biomimicked for various applications but rarely studied for cell-substrate interaction and cellular response. Here, we discuss the research work wherein 2D hierarchical substrates were prepared using biomimicking to understand cellular functions such as adhesion, orientation, differentiation, and formation of spheroids. Further, we also present the status of ongoing research in mimicking 3D tissue architecture using de-cellularized plant-based and tissue/organ-based scaffolds. We will also discuss 3D printing for fabricating 2D and 3D hierarchical structures. The review will end by highlighting the various advantages and research challenges in this approach. The biomimickedin-vivolike substrate can be used to better understand cellular physiology, and for tissue engineering.
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Materiales Biomiméticos , Diferenciación Celular , Microambiente Celular , Impresión Tridimensional , Animales , Biología Celular , Células Cultivadas , Humanos , Ratones , Propiedades de Superficie , Ingeniería de Tejidos , Andamios del TejidoRESUMEN
Population extinction is a serious issue both from the theoretical and practical points of view. We explore here how environmental noise influences persistence and extinction of interacting species in presence of a pathogen even when the populations remain stable in its deterministic counterpart. Multiplicative white noise is introduced in a deterministic predator-prey-parasite system by randomly perturbing three biologically important parameters. It is revealed that the extinction criterion of species may be satisfied in multiple ways, indicating various routes to extinction, and disease eradication may be possible with the right environmental noise. Predator population cannot survive, even when its focal prey strongly persists if its growth rate is lower than some critical value, measured by half of the corresponding noise intensity. It is shown that the average extinction time of population decreases with increasing noise intensity and the probability distribution of the extinction time follows the log-normal density curve. A case study on red grouse (prey) and fox (predator) interaction in presence of the parasites trichostrongylus tenuis of grouse is presented to demonstrate that the model well fits the field data.