Characterizing Silicone Oil-Induced Protein Aggregation with Stimulated Raman Scattering Imaging.

Particles in biopharmaceutical products present high risks due to their detrimental impacts on product quality and safety. Identification and quantification of particles in drug products are important to understand particle formation mechanisms, which can help develop control strategies for particle formation during the formulation development and manufacturing process. However, existing analytical techniques such as microflow imaging and light obscuration measurement lack the sensitivity and resolution to detect particles with sizes smaller than 2 µm. More importantly, these techniques are not able to provide chemical information to determine particle composition. In this work, we overcome these challenges by applying the stimulated Raman scattering (SRS) microscopy technique to monitor the C-H Raman stretching modes of the proteinaceous particles and silicone oil droplets formed in the prefilled syringe barrel. By comparing the relative signal intensity and spectral features of each component, most particles can be classified as protein-silicone oil aggregates. We further show that morphological features are poor indicators of particle composition. Our method has the capability to quantify aggregation in protein therapeutics with chemical and spatial information in a label-free manner, potentially allowing high throughput screening or investigation of aggregation mechanisms.

Competitive Adsorption of a Monoclonal Antibody and Nonionic Surfactant at the PDMS/Water Interface.

Interfacial adsorption of monoclonal antibodies (mAbs) can cause structural deformation and induce undesired aggregation and precipitation. Nonionic surfactants are often added to reduce interfacial adsorption of mAbs which may occur during manufacturing, storage, and/or administration. As mAbs are commonly manufactured into ready-to-use syringes coated with silicone oil to improve lubrication, it is important to understand how an mAb, nonionic surfactant, and silicone oil interact at the oil/water interface. In this work, we have coated a polydimethylsiloxane (PDMS) nanofilm onto an optically flat silicon substrate to facilitate the measurements of adsorption of a model mAb, COE-3, and a commercial nonionic surfactant, polysorbate 80 (PS-80), at the siliconized PDMS/water interface using spectroscopic ellipsometry and neutron reflection. Compared to the uncoated SiO2 surface (mimicking glass), COE-3 adsorption to the PDMS surface was substantially reduced, and the adsorbed layer was characterized by the dense but thin inner layer of 16 Å and an outer diffuse layer of 20 Å, indicating structural deformation. When PS-80 was exposed to the pre-adsorbed COE-3 surface, it removed 60 wt % of COE-3 and formed a co-adsorbed layer with a similar total thickness of 36 Å. When PS-80 was injected first or as a mixture with COE-3, it completely prevented COE-3 adsorption. These findings reveal the hydrophobic nature of the PDMS surface and confirm the inhibitory role of the nonionic surfactant in preventing COE-3 adsorption at the PDMS/water interface.

Probing Molecular Interactions of Antibody Drugs, Silicone Oil, and Surfactant at Buried Interfaces In Situ.

Antibody drugs have been rapidly developed to cure many diseases including COVID-19 infection. Silicone oil is commonly used as a lubricant coating material for devices used in the pharmaceutical industry to store and administer antibody drug formulations. However, the interaction between silicone oil and antibody molecules could lead to the adsorption, denaturation, and aggregation of antibody molecules, impacting the efficacy of antibody drugs. Here, we studied the molecular interactions between antibodies and silicone oil in situ in real time. The effect of the surfactant on such interactions was also investigated. Specifically, the adsorption dynamics of a bispecific antibody (BsAb) onto a silicone oil surface without and with different concentrations of the surfactant PS80 in antibody solutions were monitored. Also the possible lowest effective PS80 concentrations that can prevent the adsorption of BsAb as well as a monoclonal antibody (mAb) onto silicone oil were measured. It was found that different concentrations of PS80 are required for preventing the adsorption of different antibodies. Both BsAB and mAB denature on silicone oil without a surfactant. However, for a low surfactant concentration in the solution, although the surfactant could not completely prevent the antibody from adsorption, it could maintain the native structures of adsorbed BsAb and mAb antibodies on silicone oil. This is important knowledge, showing that to prevent antibody aggregation on silicone oil it is not necessary to add surfactant to a concentration high enough to completely minimize protein adsorption.

Efficient and stable electrorheological fluids based on chestnut-like cobalt hydroxide coupled with surface-functionalized carbon dots.

Intrinsically polarized electrorheological fluids (ERFs) have better thermal stability than ERFs with polar molecules, so they have a broader application prospect. However, the electrorheological efficiency of the common intrinsically polarized ERF is still lower than 1500, which is related to the poor wettability between polarized materials and the continuous phase. Carbon dots (CDs) exhibit good stability, semiconductor properties and low toxicity. We prepared biomimetic chestnut-like cobalt hydroxide coupled with surface-functionalized CD particles (Co(OH)2@CDs) by a simple hydrothermal method. Then we prepared an ERF by mixing Co(OH)2@CDs with silicone oil and studied the effect of CDs on its rheology and electrorheology properties. The synergistic effect of the lipophilic groups on the surface of CDs and the biomimetic chestnut-like structure makes Co(OH)2@CDs exhibit good wettability with silicone oil, and the optimal zero-field viscosity of Co(OH)2@CDs-ERF is only 0.46 Pa s (particle mass fraction of 40%). Exceptional electrorheological efficiency (about 10 000, shear rate 0.1 s-1, 5 kV mm-1) and dynamic shear stress stability of optimal Co(OH)2@CDs-ERF can be attributed to the dielectric enhancement of the biomimetic chestnut-like structure coupled with the semiconductor properties of CDs. In addition, Co(OH)2@CDs-ERF has excellent anti-settling performance, outstanding thermal stability and low current density.

Adsorption and Aggregation of Monoclonal Antibodies at Silicone Oil-Water Interfaces.

Monoclonal antibody (mAb) therapies are rapidly growing for the treatment of various diseases like cancer and autoimmune disorders. Many mAb drug products are sold as prefilled syringes and vials with liquid formulations. Typically, the walls of prefilled syringes are coated with silicone oil to lubricate the surfaces during use. MAbs are surface-active and adsorb to these silicone oil-solution interfaces, which is a potential source of aggregation. We studied formulations containing two different antibodies, mAb1 and mAb2, where mAb1 aggregated more when agitated in the presence of an oil-water interface. This directly correlated with differences in surface activity of the mAbs, studied with interfacial tension, surface mass adsorption, and interfacial rheology. The difference in interfacial properties between the mAbs was further reinforced in the coalescence behavior of oil droplets laden with mAbs. We also looked at the efficacy of surfactants, typically added to stabilize mAb formulations, in lowering adsorption and aggregation of mAbs at oil-water interfaces. We showed the differences between poloxamer-188 and polysorbate-20 in competing with mAbs for adsorption to interfaces and in lowering particulate and overall aggregation. Our results establish a direct correspondence between the adsorption of mAbs at oil-water interfaces and aggregation and the effect of surfactants in lowering aggregation by competitively adsorbing to these interfaces.

LABORATORY EVALUATION OF INTRAOCULAR SILICONE OIL REMOVED AFTER 23 YEARS IN SITU.

PURPOSE: To report the clinicopathologic correlation and biochemical analysis of silicone oil removed after 23 years in an eye. METHODS: A 63-years-old man with a history of HIV/AIDS and cytomegalovirus retinitis, status post retinal detachment repair with silicone oil at the age of 39 years, presented with several weeks of worse vision. He was found to have a shallow fovea-off tractional retinal detachment. After the silicone oil was removed during retinal detachment repair, it was analyzed by Fourier-transform infrared spectroscopy and gas chromatography with mass spectrometry. RESULTS: In addition to cyclic and linear silicone oil, cholesterol was found in the removed silicone oil, which was not present in unused silicone oil samples. No other chemical alterations were identified in the extracted silicone oil. CONCLUSION: Silicone oil left inside an eye over an extended period may extract lipophilic substances from adjacent tissue, with possible pathophysiologic effects. However, no other major potentially toxic substance was identified from the long-standing silicone oil sample, suggesting relative chemical stability of the tamponade agent over time.

PARTICULATE MATTER FROM SYRINGES USED FOR INTRAVITREAL INJECTIONS.

BACKGROUND: Syringes containing anti-vascular endothelial growth factor drugs to treat retinal diseases are prepared in different ways by various parties with syringe selection, preparation, and storage conditions affecting the risk of injecting particles into the vitreous. This study examines particle loads from various syringes over time. METHODS: Four syringes were studied: two plastic transfer syringes lubricated with silicone oil or oleamide, a glass syringe with baked-on silicone, and a lubricant-free polymer syringe. Syringes were rinsed with water or filled with buffer and analyzed over time; particles were quantified by flow imaging. Particle formation in a bevacizumab formulation was also characterized. RESULTS: Insulin syringes consistently showed very high particle counts. Oleamide-lubricated syringes had substantially fewer particles, but showed appreciable increases over time (leading to visible particles). Baked-on silicone glass syringes and lubricant-free polymer syringes both showed low particle levels ≥10 µm. Lubricant-free syringes showed the lowest particle levels ≥1 µm and the lowest particle levels with bevacizumab agitation. CONCLUSION: Syringes have different intrinsic particle loads which can contribute to particle loads in the delivered drug. Oleamide-lubricated transfer syringes, commonly used for bevacizumab repackaging, have time-dependent particle loads and are associated with the formation of visible particles beyond 30 days of storage.

Impact of Silicone Oil on Free Fatty Acid Particle Formation due to Polysorbate 20 Degradation.

PURPOSE: Polysorbate 20 (PS20), a commonly used surfactant in biopharmaceutical formulations, can undergo hydrolytic degradation resulting in free fatty acids (FFAs) that precipitate to form particles. This work investigates the ability for silicone oil (si-oil) coated on the interior walls of prefilled syringes (PFSs) to act as a sink for FFAs and potentially delay FFA particle formation. METHODS: Myristic acid distribution coefficient was measured in a two-phase system containing si-oil and formulation buffer at a range of aqueous conditions. An empirical model was built from these data to predict distribution coefficient based on aqueous conditions. To verify the model, PS20 was degraded using model lipases side-by-side in glass vials and PFSs while monitoring sub-visible particles. RESULTS: The empirical model demonstrates that the partitioning of myristic acid into si-oil is maximized at low pH and low PS20 concentration. The model predicts that the presence of si-oil at levels typical in PFSs provides at most an 8.5% increase in the total carrying capacity for myristic acid compared to a non-coated glass vial. The time to onset of FFA particles was equivalent between degradations performed in two PFS models coated with differing levels of silicone oil and in non-coated glass vials. CONCLUSION: Herein, we demonstrate that FFAs partition from aqueous solution into si-oil. However, the extent of the partitioning effect is not large enough to delay PS20-related FFA particle formation at typical formulation conditions (pH 5.0-7.5, 0.01% - 0.1% w/v PS20) filled in typical PFSs (<1.0 mg si-oil/mL aqueous fill).

Unique Characteristics of Intravitreal Silicone Oil on Dual-Energy Computed Tomography.

Silicone oil is used as an intravitreal injection to treat retinal detachment. This material can spread into the subarachnoid space, where it may be mistaken for acute hemorrhage on single-energy computed tomography. This report describes the appearance of intravitreal silicone oil on dual-energy computed tomography, emphasizing unique virtual monoenergetic imaging characteristics that allow for confident differentiation of silicone oil from hemorrhage as well as from other potential single-energy mimics, such as calcium and iodine.

Electrical and Magnetodielectric Properties of Magneto-Active Fabrics for Electromagnetic Shielding and Health Monitoring.

An efficient, low-cost and environmental-friendly method to fabricate magneto-active fabrics (MAFs) based on cotton fibers soaked with silicone oil and iron oxide microfibers (mFe) at mass fractions 2 wt.%, 4 wt.% and 8 wt.% is presented. It is shown that mFe induce good magnetic properties in MAFs, which are subsequently used as dielectric materials for capacitor fabrication. The electrical properties of MAFs are investigated in a static magnetic field with intensities of 0 kA/m, 160 kA/m and 320 kA/m, superimposed on a medium-frequency electric field. The influence of mFe on the electrical capacitance and dielectric loss tangent is determined, and it can be observed that the electrical conductivity, dielectric relaxation times and magnetodielectric effects are sensibly influenced by the applied magnetic and electric fields. The results indicate that the MAFs have electrical properties which could be useful for protection against electromagnetic pollution or for health monitoring.

Physical Modification of Cellulose Nanocrystals with a Synthesized Triblock Copolymer and Rheological Thickening in Silicone Oil/Grease.

This study investigates the rheological thickening effect of surface-modified cellulose nanocrystals (mCNCs) with a triblock copolymer on silicone oil. An amphipathic copolymer comprising hydrophilic methoxypolyethylene glycols and hydrophobic polydimethylsiloxane segments is synthesized and introduced in silicone oil by physical adsorption of rodlike cellulose nanocrystals (CNCs). The surface-modified copolymer acts as a "bridge" to enhance the compatibility between CNCs and base oil, which produces the hybrid oils/greases containing homogeneously dispersed mCNCs at varied loading levels of 5-20 wt %. The presence of this rigid additive remarkably increases the viscosity of silicone oil, accompanied with the transition from flowable to nonflowable behavior with a gradual increase of mCNC loading levels. Furthermore, the change of storage and loss modulus of the fabricated hybrid oils/greases indicates the formation of a three-dimensional network by the chain entanglement and interactions between the copolymer and silicone oil (at a critical loading level of 15 wt % mCNC), which promotes the rigid CNCs to act as the physical cross-linking point for the significant transition from liquid to quasi-solid state.

Enhancement of Particle Alignment Using Silicone Oil Plasticizer and Its Effects on the Field-Dependent Properties of Magnetorheological Elastomers.

The existing mold concept of fabricating magnetorheological elastomer (MRE) tends to encounter several flux issues due to magnetic flux losses inside the chamber. Therefore, this paper presents a new approach for enhancing particle alignment through MRE fabrication as a means to provide better rheological properties. A closed-loop mold, which is essentially a fully guided magnetic field inside the chamber, was designed in order to strengthen the magnetic flux during the curing process with the help of silicone oil (SO) plasticizers. The oil serves the purpose of softening the matrix. Scanning electron microscopy (SEM) was used to observe the surface morphology of the fabricated MRE samples. The field-dependent dynamic properties of the MREs were measured several ways using a rheometer, namely, strain sweep, frequency sweep, and magnetic field sweep. The analysis implied that the effectiveness of the MRE was associated with the use of the SO, and the closed-loop mold helped enhance the absolute modulus up to 0.8 MPa. The relative magnetorheological (MR) effects exhibited high values up to 646%. The high modulus properties offered by the MRE with SO are believed to be potentially useful in industry applications, particularly as vibration absorbers, which require a high range of stiffness.

Hyperbranched Silicone MDTQ Tack Promoters.

Low molecular weight, highly crosslinked silicone resins are widely used as reinforcing agents for highly transparent elastomers and adhesion/tack promoters in gels. The resins are complex mixtures and their structure / property relationships are ill defined. We report the synthesis of a library of 2, 3 and 4-fold hyperbranched polymeric oils that are comprised of linear, lightly branched or highly branched dendronic structures. Rheological examination of the fluids and tack measurements of gels filled with 10, 25 or 50% dendronic oils were made. Viscosity of the hyperbranched oils themselves was related to molecular weight, but more significantly to branch density. The properties are driven by chain entanglement. When cured into a silicone gel, less densely branched materials were more effective in improving tack than either linear oils or Me3SiO-rich, very highly branched oils of comparable molecular weight, because the latter oils underwent phase separation.

DNA-Coated Functional Oil Droplets.

Many industrial soft materials include oil-in-water (O/W) emulsions at the core of their formulations. By using tuneable interface stabilizing agents, such emulsions can self-assemble into complex structures. DNA has been used for decades as a thermoresponsive, highly specific binding agent between hard and, recently, soft colloids. Up until now, emulsion droplets functionalized with DNA had relatively low coating densities and were expensive to scale up. Here, a general O/W DNA-coating method using functional nonionic amphiphilic block copolymers, both diblock and triblock, is presented. The hydrophilic poly(ethylene glycol) ends of the surfactants are functionalized with azides, allowing for efficient, dense, and controlled coupling of dibenzocyclooctane-functionalized DNA to the polymers through a strain-promoted alkyne-azide click reaction. The protocol is readily scalable due to the triblock's commercial availability. Different production methods (ultrasonication, microfluidics, and membrane emulsification) are used with different oils (hexadecane and silicone oil) to produce functional droplets in various size ranges (submicron, ∼20 and >50 µm), showcasing the generality of the protocol. Thermoreversible submicron emulsion gels, hierarchical "raspberry" droplets, and controlled droplet release from a flat DNA-coated surface are demonstrated. The emulsion stability and polydispersity is evaluated using dynamic light scattering and optical microscopy. The generality and simplicity of the method opens up new applications in soft matter, biotechnological research, and industrial advances.

Sensitive electrochemical detection of sodium azide based on the electrocatalytic activity of the pasting liquid of a carbon paste electrode.

Sodium azide (NaN3) is highly toxic and widely used in, for example, automobile airbags and biochemical laboratories. The electrochemical detection of sodium azide on commonly used electrodes is challenging due to sluggish electron transfer, but it has been achieved using a boron-doped diamond thin-film electrode and a highly oriented pyrolytic graphite electrode. Utilizing the electrocatalytic activity of the pasting liquid of a carbon paste electrode, we developed an effective method for the electrochemical detection of sodium azide in which silicone oil was employed as the pasting liquid of the carbon paste electrode. This simple and cheap silicone-oil-based carbon paste electrode exhibited comparable sensitivity to the boron-doped diamond thin-film electrode and highly oriented pyrolytic graphite electrode. The limit of detection for sodium azide at the silicone-oil-based carbon paste electrode was found to be 0.1 µM. Recoveries from diluted human serum samples were between 97.2 and 101.3%. Graphical abstract ᅟ.

Non-invasive Oil-Based Method to Increase Topical Delivery of Nucleic Acids to Skin.

Topical delivery of nucleic acids to skin has huge prospects in developing therapeutic interventions for cutaneous disorders. In spite of initial success, clinical translation is vastly impeded by the constraints of bioavailability as well as stability in metabolically active environment of skin. Various physical and chemical methods used to overcome these limitations involve invasive procedures or compounds that compromise skin integrity. Hence, there is an increasing demand for developing safe skin penetration enhancers for efficient nucleic acid delivery to skin. Here, we demonstrate that pretreatment of skin with silicone oil can increase the transfection efficiency of non-covalently associated peptide-plasmid DNA nanocomplexes in skin ex vivo and in vivo. The method does not compromise skin integrity, as indicated by microscopic evaluation of cellular differentiation, tissue architecture, enzyme activity assessment, dye penetration tests using Franz assay, and cytotoxicity and immunogenicity analyses. Stability of nanocomplexes is not hampered on pretreatment, thereby avoiding nuclease-mediated degradation. The mechanistic insights through Fourier transform infrared (FTIR) spectroscopy reveal some alterations in the skin hydration status owing to possible occlusion effects of the enhancer. Overall, we describe a topical, non-invasive, efficient, and safe method that can be used to increase the penetration and delivery of plasmid DNA to skin for possible therapeutic applications.

Indium Nitrite (InN)-Based Ultrasensitive and Selective Ammonia Sensor Using an External Silicone Oil Filter for Medical Application.

Ammonia is an essential biomarker for noninvasive diagnosis of liver malfunction. Therefore, selective detection of ammonia is essential for medical application. Here, we demonstrate a portable device to selectively detect sub-ppm ammonia gas. The presented gas sensor is composed of a Pt coating on top of an ultrathin Indium nitrite (InN) epilayer with a lower detection limit of 0.2 ppm, at operating temperature of 200 °C, and detection time of 1 min. The sensor connected with the external filter of nonpolar 500 CS silicone oil to diagnose liver malfunction. The absorption of 0.7 ppm acetone and 0.4 ppm ammonia gas in 10 cc silicone oil is 80% (0.56 ppm) and 21.11% (0.084 ppm), respectively, with a flow rate of 10 cc/min at 25 °C. The absorption of acetone gas is 6.66-fold higher as compared to ammonia gas. The percentage variation in response for 0.7 ppm ammonia and 0.7 ppm acetone with and without silicone oil on InN sensor is 17.5% and 4%, and 22.5%, and 14% respectively. Furthermore, the percentage variation in response for 0.7 ppm ammonia gas with silicone oil on InN sensor is 4.3-fold higher than that of 0.7 ppm acetone. The results show that the InN sensor is suitable for diagnosis of liver malfunction.

Heavy and standard silicone oil: intraocular inflammation.

PURPOSE: Proliferative vitreoretinopathy in the inferior retina remains clinically challenging. Heavier-than-water intraocular tamponades have been developed to improve inferior tamponading properties, and their chemical compositions have been substantially improved over the years, in parallel with developments in vitrectomy instrumentation and surgical techniques. Herein we present an updated review of the clinical use of standard formulations and HSO, focusing on analysis of the intraocular inflammation associated with endotamponade agents, and comparison of the adverse effects of these agents on the physical and biological properties of the eye. METHODS: A detailed literature search was conducted on PubMed, EMBASE, Cochrane Library, and Google Scholar using the key words. Fifty-eight articles matched our inclusion criteria that were included in this systematic review. RESULTS: Perfluorocarbon liquids and partially fluorinated alkanes are associated with tamponade emulsification, intraocular inflammation, and rises in intraocular pressure, but these associations are not as strong when these substances are mixed with a heavy silicone oil (HSO). Two recently approved heavy silicone oil tamponades, Oxane HD and Densiron 68, are now available for use in clinical practice. While the complication spectrum of the new generation of these HSOs seems to be similar to that of conventional silicone oil tamponades, they provide better support for the inferior retina and the posterior pole. CONCLUSION: Both regular and heavy silicone oils usually yield good success rates in cases of complicated retinal detachment. Decisions as to whether to utilize heavy or regular silicone oil should be made on a case-by-case basis.

Discrete electric field mediated droplet splitting in microchannels: Fission, Cascade, and Rayleigh modes.

Numerical simulations supplemented by experiments together uncovered that strategic integration of discrete electric fields in a non-invasive manner could substantially miniaturize the droplets into smaller parts in a pressure driven oil-water flow inside microchannels. The Maxwell's stress generated from the electric field at the oil-water interface could deform, stretch, neck, pin, and disintegrate a droplet into many miniaturized daughter droplets, which eventually ushered a one-step method to form water-in-oil microemulsion employing microchannels. The interplay between electrostatic, inertial, capillary, and viscous forces led to various pathways of droplet breaking, namely, fission, cascade, or Rayleigh modes. While a localized electric field in the fission mode could split a droplet into a number of daughter droplets of smaller size, the cascade or the Rayleigh mode led to the formation of an array of miniaturized droplets when multiple electrodes generating different field intensities were ingeniously assembled around the microchannel. The droplets size and frequency could be tuned by varying the field intensity, channel diameter, electrode locations, interfacial tension, and flow ratio. The proposed methodology shows a simple methodology to transform a microdroplet into an array of miniaturized ones inside a straight microchannel for enhanced mass, energy, and momentum transfer, and higher throughput.

A Random Forest Approach for Counting Silicone Oil Droplets and Protein Particles in Antibody Formulations Using Flow Microscopy.

PURPOSE: To evaluate a random forest model that counts silicone oil droplets and non-silicone oil particles in protein formulations with large class imbalance. METHODS: In this work, we present a novel approach for automated image analysis of flow microscopy data based on random forest classification enabling rapid analysis of large data sets. The random forest approach overcomes many of the limitations of traditional classification schemes derived from simple filters or regression models. In particular, the approach does not require a priori selection of important morphology parameters. RESULTS: We analyzed silicone oil droplets and non-silicone oil particles observed in four model systems with protein concentrations of 20, 50 and 125 mg/mL. Filters based on random forests achieve higher classification accuracies when compared to regression based filters. Additionally, we showcase a procedure that allows for accurate counting of particles ≥1 µm. CONCLUSIONS: Our method is generally applicable for classification and counting of different classes of particles as long as class morphologies are differentially expressed.