Cell-free formation of RNA granules: low complexity sequence domains form dynamic fibers within hydrogels.

Eukaryotic cells contain assemblies of RNAs and proteins termed RNA granules. Many proteins within these bodies contain KH or RRM RNA-binding domains as well as low complexity (LC) sequences of unknown function. We discovered that exposure of cell or tissue lysates to a biotinylated isoxazole (b-isox) chemical precipitated hundreds of RNA-binding proteins with significant overlap to the constituents of RNA granules. The LC sequences within these proteins are both necessary and sufficient for b-isox-mediated aggregation, and these domains can undergo a concentration-dependent phase transition to a hydrogel-like state in the absence of the chemical. X-ray diffraction and EM studies revealed the hydrogels to be composed of uniformly polymerized amyloid-like fibers. Unlike pathogenic fibers, the LC sequence-based polymers described here are dynamic and accommodate heterotypic polymerization. These observations offer a framework for understanding the function of LC sequences as well as an organizing principle for cellular structures that are not membrane bound.

A Case of Migration of a Hydrogel Spacer for Radiotherapy into the Pulmonary Artery.

A 67-year-old man was referred to our hospital for the diagnosis and treatment of prostate cancer. Multidisciplinary discussion led to intensity-modulated radiotherapy preceded by hormone therapy. Before radiotherapy, a biodegradable hydrogel spacer (HS) was placed between the prostate and rectum to reduce radiation injury risk. Three weeks postplacement, pelvic magnetic resonance imaging revealed HS migration into the pelvic vein. Subsequent whole-body contrast-enhanced computed tomography (CECT) revealed HS migration into the pulmonary artery. The patient showed no symptoms or clinical signs. Radiotherapy was completed uneventfully. Complete absorption of the migrated HS was confirmed using CECT images 5 months postplacement.

Physical Properties and Interaction With the Ocular Surface of Water-Gradient Contact Lenses.

ABSTRACT: Since the introduction of silicone hydrogel contact lenses, many silicone-hydrogel materials have been produced, including water-gradient contact lenses with a silicone hydrogel core and a thin hydrogel outer layer (e.g., delefilcon A, verofilcon A, and lehfilcon A). Their properties have been investigated in various studies assessing both the chemical-physical characteristics and the comfort, but the overall picture is not always consistent. In this study, water-gradient technology is reviewed by looking at basic physical properties both in vitro and in vivo and at the interaction with the human ocular surface. Surface and bulk dehydration, surface wetting and dewetting, shear stress, interaction with tear components and with other environmental compounds, and comfort are discussed.

Improvement of Soft Contact Lens Wettability After the Instillation of Hyaluronic Acid Eye Drops.

PURPOSE: To evaluate the effect of the topical instillation of hyaluronic acid eye drops with different viscosity on soft contact lens wettability and comfort. METHODS: A randomized and participant-masked study was performed, involving 20 participants (25.4±2.6 years). One eye wore hydrogel (ocufilcon D) contact lenses, and another eye wore silicone-hydrogel (somofilcon A) contact lenses. The in vivo wettability tear film surface quality (TFSQ) index and comfort were measured before and after the instillation of different eye drops: saline solution (control) and 0.1%, 0.2%, and 0.3% hyaluronic acid. RESULTS: Compared with saline solution, the instillation of 0.1%, 0.2%, and 0.3% hyaluronic acid improved the in vivo wettability of the hydrogel contact lenses by decreasing their TFSQ mean for 5, 10, and 30 min, respectively ( P <0.05). During silicone-hydrogel contact lens wear, the hyaluronic acid did not affect wettability because there were no changes in TFSQ mean ( P ≥0.05), but the 0.3% hyaluronic acid produced a decrease in comfort for the first 3 min ( P <0.05). CONCLUSIONS: The instillation of hyaluronic acid eye drops increased the in vivo wettability of the hydrogel contact lens, and the duration of this effect was directly related to its concentration and viscosity.

Safety of hydrogel spacers for rectal wall protection in patients with prostate cancer: A retrospective analysis of 200 consecutive cases.

OBJECTIVE: To evaluate the safety and complications of hydrogel spacer implantation. METHODS: This single-center historical cohort study retrospectively analyzed cases of hydrogel spacer implantation between October 2018 and March 2022. The survey items were the rates of possible hydrogel injection implementation, the success rate of hydrogel implantation including asymmetry, higher position, rectal wall infiltration, subcapsular injection, and other adverse events, and width created by the spacer. To investigate the learning curve, 1, 2, and 3 points were assigned to adverse event grades G1, G2, and G3, respectively. Spacer effectiveness obstruction, such as asymmetry was assigned 3 points. A Mann-Whitney U test was performed to assess statistically significant differences. RESULTS: The study included a total of 200 patients with a median (range) age of 70 (44-85) years. In 10 (5%) patients, hydrogel injection implementation was not possible. Of 190 patients who underwent hydrogel spacer placement, 168 (88%) received a satisfactory placement. The median (range) width of hydrogel spacers was 13.1 (4.4-18.7) mm. Spacer asymmetry, higher position, rectal wall infiltration, and prostate subcapsular infiltration occurred in 7 (3.7%), 5 (2.6%), 12 (6.3%), and 1 (0.5%) patients, respectively. G1 and G3 adverse events occurred in 13 (7%) and 4 (2%) patients, respectively. Practitioner #1 who performed the highest number of procedures had significantly (p = 0.04) lower total scores in group B. CONCLUSION: Spacer implantation yielded favorable outcomes with a high percentage of appropriate spacer implantation, and few major complications.

Designer matrices for intestinal stem cell and organoid culture.

Epithelial organoids recapitulate multiple aspects of real organs, making them promising models of organ development, function and disease. However, the full potential of organoids in research and therapy has remained unrealized, owing to the poorly defined animal-derived matrices in which they are grown. Here we used modular synthetic hydrogel networks to define the key extracellular matrix (ECM) parameters that govern intestinal stem cell (ISC) expansion and organoid formation, and show that separate stages of the process require different mechanical environments and ECM components. In particular, fibronectin-based adhesion was sufficient for ISC survival and proliferation. High matrix stiffness significantly enhanced ISC expansion through a yes-associated protein 1 (YAP)-dependent mechanism. ISC differentiation and organoid formation, on the other hand, required a soft matrix and laminin-based adhesion. We used these insights to build a fully defined culture system for the expansion of mouse and human ISCs. We also produced mechanically dynamic matrices that were initially optimal for ISC expansion and subsequently permissive to differentiation and intestinal organoid formation, thus creating well-defined alternatives to animal-derived matrices for the culture of mouse and human stem-cell-derived organoids. Our approach overcomes multiple limitations of current organoid cultures and greatly expands their applicability in basic and clinical research. The principles presented here can be extended to identify designer matrices that are optimal for long-term culture of other types of stem cells and organoids.

Development of an enzymatic method for the evaluation of protein deposition on contact lenses.

The aim of this study was to evaluate a new digestion method to quantify protein deposition on contact lenses. Four silicone hydrogel and one hydrogel contact lens material were incubated in lactoferrin, lysozyme, immunoglobulin A, and bovine serum albumin solutions at approximate physiological concentrations and temperature. Immobilized trypsin was used to digest the protein deposits from the contact lens surfaces. The total protein absorbed to lenses was extracted and digested using sequencing grade trypsin. The tryptic peptides were quantified using selected reaction monitoring mass spectrometry. The concentration of surface protein deposits was either lower than or the same as the total protein for all lens types and proteins. Immobilised trypsin can digest protein deposits from the surface of contact lenses. This ability to analyse the amount of protein at a contact lens surface may help in elucidating the effect of surface deposition on clinical outcomes during lens wear.

Multipurpose Lens Care Systems and Silicone Hydrogel Contact Lens Wettability: A Systematic Review.

PURPOSE: To provide a relationship between materials developed for silicone hydrogel contact lenses and multipurpose care solutions to identify improvements in wettability, for prelens noninvasive break-up time and subjective score. METHODS: This systematic review was completed according to the updated PRISMA 2020 statement recommendations and followed the explanation and elaboration guidelines. The PubMed, Web of Science, and Scopus scientific literature databases were searched from January 2000 to November 2021. RESULTS: A total of four clinical trials published between 2011 and 2017 were included in this investigation. All included studies were randomized clinical trials. The success of contact lenses is related to the comfort of their use and therefore to the stability of the tear film and the wettability of its surface. The relationship between these parameters and changes in the ocular surface and inflammatory and infectious processes has been demonstrated. CONCLUSION: Hyaluronan and propylene glycol multipurpose solution (MPS) wetting agents achieved slightly higher prelens noninvasive break-up times than poloxamine. Polyquaternium-1 achieved better wettability and patient comfort than polyhexamethylene biguanide in medium-term studies. Short-term studies did not demonstrate differences between MPSs in their effect on contact lens wettability.

Orbital and Sinonasal Complications of Hydrogel Scleral Buckle: A Case Report.

The hydrogel scleral buckle is a hydrophilic implant that is characterized by progressive expansion and can present with secondary orbital changes. The authors present a unique case of hydrogel-induced erosion of the orbital roof into the frontal sinus, with formation of a sino-orbital communication that resulted in frontal sinusitis and adjacent cerebritis. The hydrogel material is radiologically characterized as a fluid-filled hypertense T2 mass with rim enhancement and peripheral calcification. Awareness of late orbital complications from the hydrogel material is important to distinguish this entity from other mimicking orbital pathologies.

Metformin Hydrochloride Encapsulation by Alginate Strontium Hydrogel for Cartilage Regeneration by Reliving Cellular Senescence.

Cartilage lesion is a common tissue defect and is challenging in clinical practice. Trauma-induced cellular senescence could decrease the chondrocyte capability of maintaining cartilage tissue regeneration. A previous investigation showed that, by controlling the cellular senescence, the cartilage regeneration can be significantly accelerated. Based on this finding, we design a novel hydrogel, Alg/MH-Sr, that combines metformin, an established drug for inhibiting senescence, and strontium, an effective anti-inflammatory material for cartilage tissue engineering. A RT-PCR test suggests the significant inhibitory effect of the hydrogel on senescent, apoptotic, oxidative, and inflammatory genes' expression. Histological examinations demonstrate that the Alg/MH-Sr hydrogel accelerated cartilage repairment, and chondrocyte senescence was significantly inhibited. Our study demonstrates that the Alg/MH-Sr hydrogel is effective for cartilage defect treatment and provides a new clue in accelerating tissue repairment by inhibiting the senescence of cells and tissues.

An anisotropic hydrogel with electrostatic repulsion between cofacially aligned nanosheets.

Machine technology frequently puts magnetic or electrostatic repulsive forces to practical use, as in maglev trains, vehicle suspensions or non-contact bearings. In contrast, materials design overwhelmingly focuses on attractive interactions, such as in the many advanced polymer-based composites, where inorganic fillers interact with a polymer matrix to improve mechanical properties. However, articular cartilage strikingly illustrates how electrostatic repulsion can be harnessed to achieve unparalleled functional efficiency: it permits virtually frictionless mechanical motion within joints, even under high compression. Here we describe a composite hydrogel with anisotropic mechanical properties dominated by electrostatic repulsion between negatively charged unilamellar titanate nanosheets embedded within it. Crucial to the behaviour of this hydrogel is the serendipitous discovery of cofacial nanosheet alignment in aqueous colloidal dispersions subjected to a strong magnetic field, which maximizes electrostatic repulsion and thereby induces a quasi-crystalline structural ordering over macroscopic length scales and with uniformly large face-to-face nanosheet separation. We fix this transiently induced structural order by transforming the dispersion into a hydrogel using light-triggered in situ vinyl polymerization. The resultant hydrogel, containing charged inorganic structures that align cofacially in a magnetic flux, deforms easily under shear forces applied parallel to the embedded nanosheets yet resists compressive forces applied orthogonally. We anticipate that the concept of embedding anisotropic repulsive electrostatics within a composite material, inspired by articular cartilage, will open up new possibilities for developing soft materials with unusual functions.

A method for studying lipid adsorption to silicone hydrogel contact lenses.

The aim of this study was to develop an experimental methodology to measure lipid deposition with contact lenses. Contact lenses were incubated in a lipid solution. The amount and types of adsorbed lipids were assessed using mass spectrometry and confocal microscopy. The recovery of lipids from lenses varied with lipid and lens type. Most non-polar and polar lipids were desorbed from lenses during the first 5 min of extraction. Fluorescently labelled phosphatidylcholine bound within the matrix of Senofilcon A lenses but to the surface of Lotrafilcon B lenses, whereas fluorescently labelled cholesteryl ester was found throughout both lenses. The efficacy of extraction of lipids from contact lenses varies for different lipid classes and different lens materials. Differences in the amount and time of lipid desorption probably resulted from the strength of the bond between lipid and lens polymer and the depth of adsorption of lipid in the polymer.

Functional equivalence of germ plasm organizers.

The proteins Oskar (Osk) in Drosophila and Bucky ball (Buc) in zebrafish act as germ plasm organizers. Both proteins recapitulate germ plasm activities but seem to be unique to their animal groups. Here, we discover that Osk and Buc show similar activities during germ cell specification. Drosophila Osk induces additional PGCs in zebrafish. Surprisingly, Osk and Buc do not show homologous protein motifs that would explain their related function. Nonetheless, we detect that both proteins contain stretches of intrinsically disordered regions (IDRs), which seem to be involved in protein aggregation. IDRs are known to rapidly change their sequence during evolution, which might obscure biochemical interaction motifs. Indeed, we show that Buc binds to the known Oskar interactors Vasa protein and nanos mRNA indicating conserved biochemical activities. These data provide a molecular framework for two proteins with unrelated sequence but with equivalent function to assemble a conserved core-complex nucleating germ plasm.

Uptake and Release of a Multipurpose Solution Biocide (MAP-D) From Hydrogel and Silicone Hydrogel Contact Lenses Using a Radiolabel Methodology.

PURPOSE: The purpose of this study was to evaluate the uptake and release of radiolabelled myristamidopropyl dimethylamine (MAP-D) on reusable daily wear contact lenses (CLs) over 7 days. METHODS: Three silicone hydrogel (SH) CL materials (lotrafilcon B, balafilcon A, senofilcon A) and two conventional hydrogel (CH) materials (etafilcon A, omafilcon A) were tested. A short-term (experiment 1, N=4) and a longer-term (experiment 2, N=3) study was conducted. In experiment 1, the CLs were incubated in 2 mL of phosphate buffered solution (PBS) containing 14C MAP-D (5 µg/mL) for 8 hrs. The release of 14C MAP-D was measured at t=0.25, 0.5, 1, 2, 4, 8, and 24 hr in PBS. In experiment 2, the CLs were incubated in the 14C MAP-D solution for 8 hrs followed by a 16-hr release in PBS. This cycle was repeated daily for 7 days. At the end of both experiments, lenses were extracted to determine the total uptake of MAP-D. The radioactivity was measured using a beta scintillation counter. RESULTS: In experiment 1, all three SH lenses sorbed similar amounts of MAP-D (P=0.99), all of which were higher than the two CH materials (P<0.01). However, the CH materials released a greater amount of MAP-D than the SH materials (P<0.01). In experiment 2, the uptake of MAP-D in SH materials increased over 7 days, whereas the amount of MAP-D remained constant in the CH materials (P=0.99). Similar to experiment 1, the CH lenses released more MAP-D than SH lenses after 7 days (P<0.01). CONCLUSION: The SH materials absorbed greater amounts of MAP-D compared to CH materials. However, the CH materials released the greatest amount of MAP-D. Radioactive labelling of MAP-D offers a highly sensitive method of assessing the uptake and release profiles of biocides to CL materials.

Evaluation of In Vitro Wettability of Soft Contact Lenses Using Tear Supplements.

OBJECTIVE: To evaluate the interaction between tear supplements and soft contact lenses (SCLs), we measured the contact angles (CAs) on the SCLs using commercially available tear supplements. METHODS: We used four daily disposable conventional hydrogel lenses (etafilcon A, etafilcon A+ polyvinylpyrrolidone, nelfilcon A, and omafilcon A containing 2-methacryloyloxyethyl phosphorylcholine [MPC]) and four silicone hydrogel lenses (narafilcon A, senofilcon A, delefilcon A, and stenfilcon A). The CAs on the SCLs were measured using a sessile drop technique and four different types of sessile drops, including saline, artificial tears, lubricants containing 2-MPC (MPC solution), and 0.1% hyaluronate acid (HA). RESULTS: The CA values associated with the silicone hydrogel lenses were significantly (P<0.001) lower than those associated with the conventional hydrogel lenses with all four solutions. The mean CA of 0.1% HA was significantly (P<0.01) higher than that of saline. The mean CA of the MPC solution was significantly (P<0.01) lower than that of saline with the conventional hydrogel lenses but significantly (P<0.05) higher than that of saline with the silicone hydrogel lenses. CONCLUSIONS: The CAs associated with the silicone hydrogel SCLs were higher with the use of the MPC solutions and HA in vitro. The measured CAs may depend on ingredient agents, surface treatment of the CLs, and components of the tear supplements.

A Finite Element Analysis Model is Suitable for Biomechanical Analysis of Orbital Development.

ABSTRACT: The authors investigated orbital bone development in congenital microphthalmia (CM) using a three-dimensional finite element analysis model, after the orbital dimension deficiency was improved with a self-inflating hydrogel expander implant.Data of a 2-year-old male CM patient were examined. The orbital structure, eyeball, eye muscles, and self-inflating hydrogel expander were constructed according to computed tomography examination data. The effects of self-expanding spherical hydrogel at various locations in the muscle cone on orbital bone development were examined using 3-mL self-expanding expanders placed at shallow (model 1: 2 mm depth) and deep (model 2: 8 mm depth) muscle cone positions. This model simulated the hydrogel expansion process; the orbital bone biomechanics and radial displacement nephograms were obtained when the hydrogel volume increased 3, 5, 7, and 9 times and analyzed.The orbital bone biomechanics were concentrated at the medial orbital wall center, gradually spreading to the whole orbital wall. Biomechanics and radial displacement of the inferior temporal and lateral distal orbital wall region were the most significant, and were more significant in model 1 than in model 2.Finite element analysis is suitable for the biomechanical analysis of orbital development in CM. The shallow position inside CM patients' muscle cone is the optimal site for hydrogel implantation.

Tuning the Supramolecular Structure and Function of Collagen Mimetic Ionic Complementary Peptides via Electrostatic Interactions.

Collagen, the most abundant component of natural ECM, has attracted interest of scientific communities to replicate its multihierarchical self-assembling structure. Recent developments in collagen mimetic peptides were inclined toward the production of self-assembling short peptides capable of mimicking complex higher order structures with tunable mechanical properties. Here, we report for the first time, the crucial molecular design of oppositely charged collagen mimetic shortest bioactive pentapeptide sequences, as a minimalistic building block for development of next-generation biomaterials. Our rational design involves synthesis of two pentapeptides, where the fundamental molecular motif of collagen, that is, Gly-X-Y has been mutated at the central position with positively charged, lysine, and negatively charged, aspartate, residues. Depending on their overall surface charge, these peptides showed high propensity to form self-supporting hydrogel either at acidic or basic pH, which limits their biomedical applications. Interestingly, simple mixing of the two peptides was found to induce the coassembly of these designed peptides, which drives the formation of self-supporting hydrogel at physiological pH and thus enhanced the potential of exploring these peptides for biomedical purposes. This coassembly of ionic peptides was accompanied by the enhancement in the mechanical stiffness of the gels and reduction in overall zeta potential of the combined hydrogel, which provides the evidence for additional electrostatic interactions. Furthermore, the thixotropic nature of these gels offers an additional advantage of exploration of designer biomaterials as injectable gels. The nanofibers of coassembled hydrogel were found to be highly biocompatible to the fibroblast cells compared to the individual peptides, which was evident from their cytotoxicity studies. We anticipate that our rational design of ECM protein mimics in the form of short bioactive peptides will contribute significantly to the development of novel biomaterials and play a crucial role in the field of tissue engineering and regenerative medicines.

X10 expansion microscopy enables 25-nm resolution on conventional microscopes.

Expansion microscopy is a recently introduced imaging technique that achieves super-resolution through physically expanding the specimen by ~4×, after embedding into a swellable gel. The resolution attained is, correspondingly, approximately fourfold better than the diffraction limit, or ~70 nm. This is a major improvement over conventional microscopy, but still lags behind modern STED or STORM setups, whose resolution can reach 20-30 nm. We addressed this issue here by introducing an improved gel recipe that enables an expansion factor of ~10× in each dimension, which corresponds to an expansion of the sample volume by more than 1,000-fold. Our protocol, which we termed X10 microscopy, achieves a resolution of 25-30 nm on conventional epifluorescence microscopes. X10 provides multi-color images similar or even superior to those produced with more challenging methods, such as STED, STORM, and iterative expansion microscopy (iExM). X10 is therefore the cheapest and easiest option for high-quality super-resolution imaging currently available. X10 should be usable in any laboratory, irrespective of the machinery owned or of the technical knowledge.

Bio-inspired self-healing structural color hydrogel.

Biologically inspired self-healing structural color hydrogels were developed by adding a glucose oxidase (GOX)- and catalase (CAT)-filled glutaraldehyde cross-linked BSA hydrogel into methacrylated gelatin (GelMA) inverse opal scaffolds. The composite hydrogel materials with the polymerized GelMA scaffold could maintain the stability of an inverse opal structure and its resultant structural colors, whereas the protein hydrogel filler could impart self-healing capability through the reversible covalent attachment of glutaraldehyde to lysine residues of BSA and enzyme additives. A series of unprecedented structural color materials could be created by assembling and healing the elements of the composite hydrogel. In addition, as both the GelMA and the protein hydrogels were derived from organisms, the composite materials presented high biocompatibility and plasticity. These features of self-healing structural color hydrogels make them excellent functional materials for different applications.