Unveiling the potentials of hydrophilic and hydrophobic polymers in microparticle systems: Opportunities and challenges in processing techniques.

Conventional drug delivery systems are associated with various shortcomings, including low bioavailability and limited control over release. Biodegradable polymeric microparticles have emerged as versatile carriers in drug delivery systems addressing all these challenges. This comprehensive review explores the dynamic landscape of microparticles, considering the role of hydrophilic and hydrophobic materials. Within the continuously evolving domain of microparticle preparation methods, this review offers valuable insights into the latest advancements and addresses the factors influencing microencapsulation, which is pivotal for harnessing the full potential of microparticles. Exploration of the latest research in this dynamic field unlocks the possibilities of optimizing microencapsulation techniques to produce microparticles of desired characteristics and properties for different applications, which can help contribute to the ongoing evolution in the field of pharmaceutical science.

Elucidating the influence of electrostatic force on the re-arrangement of H-bonds of protein polymers in the presence of salts.

Polyampholytes/proteins have an intriguing network of hydrogen bonds (H-bonds), especially their secondary structure, which plays a crucial role in determining the conformational stability of the polymer. The changes in protein secondary structure in the protein-salt system have been extensively deciphered by researchers, yet their pathways for breakage and recreation are unknown. Understanding the mechanism of protein conformational changes towards their biological activities, like protein folding, remains one of the main challenges and requires multiscale analysis of this strongly correlated system. Herein, salts have been used to reveal the re-arrangement behavior in the H-bond network of proteins under the influence of electrostatic interactions, as the strength of electrostatic forces is much stronger than that of H-bonds. At lower salt concentrations, there are negligible changes in the secondary structures as the electrostatic forces induced by the salt ions are less. Later, the existing H-bonds break and reconstruct new H-bonds at higher salt concentrations due to the influence of the stronger electrostatic interaction induced by the large number of salt ions. Molecular dynamics (MD) simulations and FTIR studies have been used rigorously to decipher the reason behind the re-arrangement of the H-bonds within gelatin (protein). The re-arrangement in the H-bond has also been observed with time from simulations and experiments. Thus, this study could provide a fresh perspective on the conformational changes of polyampholytes/proteins and will also influence the studies of protein folding-unfolding interaction in the presence of salt ions.

Molecular interactions of acids and salts with polyampholytes.

The Hofmeister series characterizes the ability of salt anions to precipitate polyampholytes/proteins. However, the variation of protein size in the bulk solution of acids and the effect of salts on the same have not been studied well. In this article, the four acids (CH3COOH, HNO3, H2SO4, and HCl) and their effects on the hydrodynamic radius (RH) of gelatin in the bulk solution are investigated. The effects of Na salt with the same anions are also considered to draw a comparison between the interactions of acids and salts with polyampholytes. It is suggested that the interactions of polyampholytes with acids are different from those of salts. The interaction series of polyampholytes with acids with respect to the RH of the polyampholyte is CH3COO->NO3->Cl->SO42- whereas the interaction series with salts is SO42->CH3COO->Cl->NO3-. These different interactions are due to equilibration between acid dissociation and protonation of polyampholytes. Another important factor contributing to the interactions in weak acids is the fact that undissociated acid hinders the movement of dissociated acid. Experiments and simulations were performed to understand these interactions, and the results were identical in terms of the trend in RH (from the experiments) and the radius of gyration (Rg) (from the simulations). It is concluded that the valence of ions and dissociation affect the interaction in the case of acids. However, the interactions are influenced by the kosmotropic and chaotropic effect, hydration, and mobility in the case of salts.

Enhancing the Properties of Self-Healing Gelatin Alginate Hydrogels by Hofmeister Mediated Electrostatic Effect.

The cross-linker-free hydrogels have gained attention due to their lack of need for chemically modified polymers, resulting in better biocompatibility. The hydrogel properties can be enhanced by altering physical forces such as electrostatics and H-bonds. Tuning the physical interactions between polymers, salts, and plasticisers can unlock new horizons in material properties. This article examines four different salts and mixtures to determine their impact on gelatin-alginate biomaterial design. Drug release, swelling, and rheological properties are represented using a 3-D plot, and optimum samples are identified. It is concluded that kosmotropes yield better release and swelling results than chaotropes. The physical interactions of these salts with polymers are explained using DLS and FTIR/ATR studies, and these findings are corroborated with release, swelling, and rheological analyses. Another aspect of the biomaterial, self-healing property, is also considered. A 3-D plot is prepared using release kinetics, gel strength, and recovery percentage (three important factors for self-healing hydrogels). Chaotropes are identified as better candidates for self-healing behaviour. However, when considering gel strength, release, and self-healing, kosmotropes are favourable. Hence, different salts can be selected based on the desired application for hydrogels. It is also concluded that electrostatic forces hinder the formation of H-bonds between polymer chains.

Electrostatic-Dominated Conformational Fluctuations and Transition States of Phase Separation in Charge-Balanced Protein Polymer.

Hydration of the protein/polymer is the most important aspect of stability. It is well-known that salts alter the charged polymer's electrostatic forces, ultimately impacting its conformations in solution. The solvent effects lead to certain conformational fluctuations. Previous studies have shown the screening of electrostatic repulsion within the charge-imbalanced protein following charge inversion owing to counterion condensation and phase separation. This article studies conformation stability and phase separation of charge-balanced gelatin (a protein polymer at the isoelectric point) with the addition of different salts. A phenomenon has been reported where the electrostatic effect of salts results in conformational fluctuations in gelatin due to its insufficient hydrations (termed as starvation), which scales with salt concentration. This article also presents different transition states for charge-balanced proteins prior to phase separation. It is concluded that phase separation of a charge-balanced protein passes through a stable state followed by an unstable transition state, where certain unique interactions with salts occur.

Unraveling fluctuation in gelatin and monovalent salt systems: coulombic starvation.

Fluctuations play a key role in biological systems. Here, fluctuations in gelatin intensify with increasing salt concentration. We find a redistribution of hydrogen bonds in protein-salt systems due to unfulfilled hydration of the charges of gelatin and salt-ions, termed as coulombic starvation. This yielded three regions; no starvation, starvation of gelatin, and both gelatin-salt. The system reaches equilibrium with all charges being partially hydrated. This will aid in interpreting protein-metal ion interactions and designing biomaterials.

Electrospun nanofibres in drug delivery: advances in controlled release strategies.

Emerging drug-delivery systems demand a controlled or programmable or sustained release of drug molecules to improve therapeutic efficacy and patient compliance. Such systems have been heavily investigated as they offer safe, accurate, and quality treatment for numerous diseases. Amongst newly developed drug-delivery systems, electrospun nanofibres have emerged as promising drug excipients and are coming up as promising biomaterials. The inimitable characteristics of electrospun nanofibres in terms of their high surface-to-volume ratio, high porosity, easy drug encapsulation, and programmable release make them an astounding drug-delivery vehicle.

Non-prophylactic resveratrol-mediated protection of neurite integrity under chronic hypoxia is associated with reduction of Cav1.2 channel expression and calcium overloading.

Cellular hypoxia is a major cause of oxidative stress, culminating in neuronal damage in neurodegenerative diseases. Numerous ex vivo studies have implicated that hypoxia episodes leading to disruption of Ca2+ homeostasis and redox status contribute to the progression of various neuropathologies and cell death. Isolation and maintenance of primary cell culture being cost-intensive, the details of the time course relationship between Ca2+ overload, L-type Ca2+ channel function, and neurite retraction under chronic and long-term hypoxia remain undefined. In order to explore the effect of oxidative stress and Ca2+ overload on neurite length, first, we developed a 5-day-long neurite outgrowth model using N2a cell line. Second, we propose a chronic hypoxia model to investigate the modulation of the L-type Ca2+ channel (Cav1.2) and oxidative resistance gene (OXR1) expression level during the process of neurite retraction and neuronal damage over 32 h. Thirdly, we developed a framework for quantitative analysis of cytosolic Ca2+, superoxide formation, neurite length, and constriction formation in individual cells using live imaging that provides an understanding of molecular targets. Our findings suggest that an increase in cytosolic Ca2+ is a feature of an early phase of hypoxic stress. Further, we demonstrate that augmentation in the L-type channel leads to amplification in Ca2+ overload, ROS accumulation, and a reduction in neurite length during the late phase of hypoxic stress. Next, we demonstrated that non-prophylactic treatment of resveratrol leads to the reduction of calcium overloading under chronic hypoxia via lowering of L-type channel expression. Finally, we demonstrate that resveratrol-mediated reduction of Cav1.2 channel and STAT3 expression are associated with retention of neurite integrity. The proposed in vitro model assumes significance in the context of drug designing and testing that demands monitoring of neurite length and constriction formations by imaging before animal testing.

Cross-linker-free sodium alginate and gelatin hydrogels: a multiscale biomaterial design framework.

Surface functionalization and cross-linking have been adopted extensively by researchers to customize hydrogel properties, especially in the last decade. The clinical translation of such biomaterials is in a poor state due to long-term toxicity, often beyond the periphery of the short-term animal studies. We endeavor to relook at the material development strategy with all FDA-approved biopolymers in their native states, like gelatin and sodium alginate, without using any functionalization and cross-linking. The fabrication of a cross-linker-free hydrogel has remained one of the main challenges in biomaterial design and requires multiscale structuring of the hydrogels. The physical properties of these hydrogels were enhanced by plasticizers (PEG and glycerol) and a monovalent salt (NaCl). An in-depth analysis suggested that PEG forms a plasticizing layer at the sodium alginate and gelatin interface and glycerol alters the overall polymer structure. The results were further complemented by different characterization methods (scattering techniques and infrared spectroscopy) and molecular dynamics simulations. The detailed microstructural analysis surfaced the enthralling integrated swelling mechanism in gelatin chains that led to high-performing hydrogels.

Imaging Methods for the Assessment of a Complex Hydrogel as an Ocular Drug Delivery System for Glaucoma Treatment: Opportunities and Challenges in Preclinical Evaluation.

Glaucoma is one of the leading causes of loss of vision. The problems associated with the marketed formulations of anti-glaucoma drugs are low bioavailability, unwanted side effects, and low patient compliance. Hydrogels are an important class of soft materials that play a crucial role in developing an ocular drug delivery system. They assume a special significance in addressing the problems associated with the marketed formulations of eyedrops. An appropriate design of the hydrogel system capable of encapsulating single or multiple drugs for glaucoma has emerged in recent times to overcome such challenges. Although various modes of imaging play critical roles in assessing the efficacy of these formulations, evaluating hydrogels for drug permeation and retention remains challenging. Especially, the assessment of dual drugs in the hydrogel system is not straightforward due to the complexity in measuring drug penetration and retention for in vivo or ex vivo systems. There is a need to develop tools for the fabrication and validation of hydrogel-based systems that give insight into precorneal retention, biocompatibility, cellular uptake, and cell permeation. The current review highlights some of the complexities in formulating hydrogel and benchmarking technologies, including confocal laser scanning microscopy, fluorescent microscopy, slit-lamp biomicroscopy, and camera-based imaging. This review also summarizes recent evaluations of various hydrogel formulations using in vitro and in vivo models. Further the article will help researchers from various disciplines, including formulation scientists and biologists, set up preclinical protocols for evaluating polymeric hydrogels.

Government responses and COVID-19 deaths: Global evidence across multiple pandemic waves.

We provide an assessment of the impact of government closure and containment measures on deaths from COVID-19 across sequential waves of the COVID-19 pandemic globally. Daily data was collected on a range of containment and closure policies for 186 countries from January 1, 2020 until March 11th, 2021. These data were combined into an aggregate stringency index (SI) score for each country on each day (range: 0-100). Countries were divided into successive waves via a mathematical algorithm to identify peaks and troughs of disease. Within our period of analysis, 63 countries experienced at least one wave, 40 countries experienced two waves, and 10 countries saw three waves, as defined by our approach. Within each wave, regression was used to assess the relationship between the strength of government stringency and subsequent deaths related to COVID-19 with a number of controls for time and country-specific demographic, health system, and economic characteristics. Across the full period of our analysis and 113 countries, an increase of 10 points on the SI was linked to 6 percentage points (P < 0.001, 95% CI = [5%, 7%]) lower average daily deaths. In the first wave, in countries that ultimately experiences 3 waves of the pandemic to date, ten additional points on the SI resulted in lower average daily deaths by 21 percentage points (P < .001, 95% CI = [8%, 16%]). This effect was sustained in the third wave with reductions in deaths of 28 percentage points (P < .001, 95% CI = [13%, 21%]). Moreover, interaction effects show that government policies were effective in reducing deaths in all waves in all groups of countries. These findings highlight the enduring importance of non-pharmaceutical responses to COVID-19 over time.

A global panel database of pandemic policies (Oxford COVID-19 Government Response Tracker).

COVID-19 has prompted unprecedented government action around the world. We introduce the Oxford COVID-19 Government Response Tracker (OxCGRT), a dataset that addresses the need for continuously updated, readily usable and comparable information on policy measures. From 1 January 2020, the data capture government policies related to closure and containment, health and economic policy for more than 180 countries, plus several countries' subnational jurisdictions. Policy responses are recorded on ordinal or continuous scales for 19 policy areas, capturing variation in degree of response. We present two motivating applications of the data, highlighting patterns in the timing of policy adoption and subsequent policy easing and reimposition, and illustrating how the data can be combined with behavioural and epidemiological indicators. This database enables researchers and policymakers to explore the empirical effects of policy responses on the spread of COVID-19 cases and deaths, as well as on economic and social welfare.

Oral Drug Delivery: Conventional to Long Acting New-Age Designs.

The Oral route of administration forms the heartwood of the ever-growing tree of drug delivery technology. It is one of the most preferred dosage forms among patients and controlled release community. Despite the high patient compliance, the deliveries of anti-cancerous drugs, vaccines, proteins, etc. via the oral route are limited and have recorded a very low bioavailability. The oral administration must overcome the physiological barriers (low solubility, permeation and early degradation) to achieve efficient and sustained delivery. This review aims at highlighting the conventional and modern-age strategies that address some of these physiological barriers. The modern age designs include the 3D printed devices and formulations. The superiority of 3D dosage forms over conventional cargos is summarized with a focus on long-acting designs. The innovations in Pharmaceutical organizations (Lyndra, Assertio and Intec) that have taken giant steps towards commercialization of long-acting vehicles are discussed. The recent advancements made in the arena of oral peptide delivery are also highlighted. The review represents a comprehensive journey from Nano-formulations to micro-fabricated oral implants aiming at specific patient-centric designs.

Cell-to-cell variability in organelle abundance reveals mechanisms of organelle biogenesis.

How cells regulate the number of organelles is a fundamental question in cell biology. While decades of experimental work have uncovered four fundamental processes that regulate organelle biogenesis, namely, de novo synthesis, fission, fusion, and decay, a comprehensive understanding of how these processes together control organelle abundance remains elusive. Recent fluorescence microscopy experiments allow for the counting of organelles at the single-cell level. These measurements provide information about the cell-to-cell variability in organelle abundance in addition to the mean level. Motivated by such measurements, we build upon a recent study and analyze a general stochastic model of organelle biogenesis. We compute the exact analytical expressions for the probability distribution of organelle numbers, their mean, and variance across a population of single cells. It is shown that different mechanisms of organelle biogenesis lead to distinct signatures in the distribution of organelle numbers which allow us to discriminate between these various mechanisms. By comparing our theory against published data for peroxisome abundance measurements in yeast, we show that a widely believed model of peroxisome biogenesis that involves de novo synthesis, fission, and decay is inadequate in explaining the data. Also, our theory predicts bimodality in certain limits of the model. Overall, the framework developed here can be harnessed to gain mechanistic insights into the process of organelle biogenesis.

Physicochemical Response of Gelatin in a Coulombic Soup of Monovalent Salt: A Molecular Simulation and Experimental Study.

The effect of salt on the static properties of aqueous solution of gelatin is studied by molecular dynamics simulation at pH = 1.2, 7, and 10. At the isoelectric point (pH = 7), a monotonic increase in size of the polymer is obtained with the addition of sodium chloride ions. In the positive polyelectrolyte regime (pH = 1.2), collapse of gelatin is observed with increase in salt concentration. In the negative polyelectrolyte regime, we observe an interesting collapse-reexpansion behavior. This is due to the screening of repulsion between the excess charges followed by the screening of attraction of oppositely charged ions as the salt concentration is increased. This mechanism is very different from the charge inversion mechanism which causes the reexpansion in the presence of multivalent ions. The location of salt concentration corresponding to the minimum size of the chain is comparable to the theoretical estimate. The shift in the peak of radial distribution function calculated between monomers and salt ions confirms this spatial reorganization. The predictions from the simulation are verified by dynamic light scattering(DLS) and small-angle X-ray scattering (SAXS) experiments. The size of the hydrodynamic "clusters" obtained from DLS confirms the simulation predictions. Persistence length of the gelatin is calculated from SAXS to get single chain statistics, which also agrees well with the simulation results.

Piperine as a Placebo: Stability of Gelatin Capsules without a Cross-Linker.

Gelatin has been the biomaterial of choice for decades now. Its low cost, renewable, nontoxic, and biodegradable properties make it one of the most desirable materials for controlled release applications. However, the usage of gelatin is limited by its poor mechanical/thermal stability and high water solubility. Chemical cross-linkers and hydrophobic modifications of gelatin have solved this problem, but they lead to the problem of toxicity and/or a high processing cost. This research attempts to employ a nontoxic hydrophobic drug molecule to curb early degradation of gelatin in an aqueous environment. We report the design of non-cross-linked gelatin capsules with a high dissolution resistance in an aqueous medium. Piperine, a hydrophobic drug (solubility = 40 mg/L in water), was coated on the gelatin capsules to enhance its stability in an aqueous environment. The hydrophobic piperine molecules repelled the water molecules to intensify its dissolution resistance. This stabilization was used to control the release of naproxen sodium, encapsulated inside the gelatin matrix. Piperine, in this case, acts as a placebo; i.e., it has zero therapeutic effect, but its presence was necessary to control the early degradation of the gelatin matrix. The deposition of piperine was done using the solvent evaporation method where ethanol was used as the solvent. The wettability studies revealed the hydrophobic nature of the surface after the deposition of piperine, while SEM analysis showed the presence of long cylindrical (fiber-like) structures over the gelatin surface. Further investigation (FTIR/ATR and molecular dynamics) revealed that the long fiber structures were due to the crystallization of piperine over the surface of gelatin. This crystallization was triggered by the intermolecular association (hydrogen bond) of ethanol and piperine. These observations enabled us to optimize the piperine loading protocol over the gelatin capsules that helped in achieving a zero-order naproxen release for 32 h.

Sustained drug release from multi-layered sequentially crosslinked electrospun gelatin nanofiber mesh.

The aim of this study is to develop electrospun gelatin nanofibers based drug delivery carrier to achieve controlled and sustainable release of hydrophobic drug (piperine) for prolonged time. To accomplish this, we devised some strategies such as sandwiching the drug loaded gelatin nanofiber mesh with another gelatin nanofiber matrix without drug (acting as diffusion barrier), sequential crosslinking and finally, a combination of both. As fabricated multilayered electrospun nanofibers mesh was first characterized in terms of degradation study, morphology, drug-polymer interactions, thermal stability followed by studying their release kinetics in different physiological pH as per the gastrointestinal tract. Our results show that with optimized diffusional barrier support and sequential crosslinking together, a zero order sustained drug release up to 48h may be achieved with a flexibility to vary the drug loading as per the therapeutic requirements. This work lays out the possibility of systematic design of multilayer nano-fiber mesh of a biopolymer as a drug delivery vehicle for hydrophobic drugs with a desired signature of zero order release for prolonged duration.

Ionic ingress and charge-neutralization phenomena of conducting-polymer films.

Ionic ingress and diffusion through a conducting-polymer (CP) film containing embedded charges under potential and concentration gradients is studied. Electroneutrality, a common assumption in modeling of similar systems, is not justified in this case or similar diffusion-limited processes, as the timescale of ionic diffusion in the solid matrix is quite large. Counter ions therefore cannot move instantaneously for effective neutralization of excess charges. Poisson-Nernst-Planck (PNP) equations have to be solved for such cases without any simplifying assumption. Analytical solution shows the existence of a charge boundary layer, which limits and strongly influences the ionic flux. A general numerical method for solution is also developed for the dynamic modeling, analysis, and design of these types of systems. The numerical results are validated by comparison with analytical solutions as well as with some experimental results available in the literature. With this modeling framework, the basic features of controlled release of molecules across a CP film under an applied electrical potential can be explained quantitatively.