Isoniazid—Loaded Albumin Nanoparticles: Taguchi Optimization Method

Tuberculosis is one of the dangerous infectious diseases, killing over a million people worldwide each year. The search for new dosage forms for the treatment of drug-resistant tuberculosis is an actual task. Biocompatible polymer nanoparticles, in particular bovine serum albumin (BSA), are promising drug carriers. Nanoparticle (NP) parameters such as diameter, polydispersity, bioactive substance loading, and NP yield are very important when it comes to drug transport through the bloodstream. The most well-known and widely used first-line anti-tuberculosis drug, isoniazid (INH), is being used as a drug. BSA-INH NPs were obtained by an ethanol desolvation of an aqueous protein solution in the drug presence. The peculiarity of the method is that natural components, namely urea and cysteine, are used for the stabilization of BSA-INH NPs after desolvation. The characteristics of the obtained BSA-INH NPs are significantly affected by the concentration of protein, isoniazid, urea, and cysteine in the solution. The aim of the present study is to investigate the concentration effect of the system reacting components on the parameters of the NPs that are obtained. We have chosen the concentrations of four reacting components, i.e., BSA, isoniazid, urea, and cysteine, as controlling factors and applied the Taguchi method to analyze which concentration of each component has an important effect on BSA-INH NPs characteristics.

Autologous protein solution: a promising solution for osteoarthritis?

Osteoarthritis (OA) is a global health issue with myriad pathophysiological factors and is one of the most common causes of chronic disability in adults due to pain and altered joint function. The end stage of OA develops from a destructive inflammatory cycle, driven by the pro-inflammatory cytokines interleukin-1β (IL-1β) and tumour necrosis factor alpha (TNFα). Owing to the less predictable results of total knee arthroplasty (TKA) in younger patients presenting with knee OA, there has been a surge in research evaluating less invasive biological treatment options, one of which is autologous protein solution (APS). APS is an autologous blood derivative obtained by using a proprietary device, made of APS separator, which isolates white blood cells (WBCs) and platelets in a small volume of plasma, and APS concentrator, which further concentrates platelets, WBCs and plasma proteins, resulting in a concentrated solution with high levels of growth factors including the anti-inflammatory mediators against IL-1β and TNFα. A single intraarticular injection of APS appears to be a promising solution for treatment of early-stage OA from current evidence, the majority of which comes from preclinical studies. More clinical studies are needed before APS can be widely accepted as a treatment modality for OA. Cite this article: EFORT Open Rev 2021;6:716-726. DOI: 10.1302/2058-5241.6.200040

Characterisation of pH-shift-produced protein isolates from sago palm weevil (Rhynchophorus ferrugineus) larvae

Farm-raised sago palm weevil (Rhynchophorus ferrugineus) larvae (SPWL) can be used as a protein source for food sustainability. This study aimed to investigate the potential of pH-shift processing as a cold refinery approach to produce protein isolate from SPWL. Maximum solubility of SPWL protein was observed at pH 2.0 (acid-aided process) and pH 11.5 (alkaline-aided process). The zeta-potential of the protein solution was close to zero with the lowest solubility at pH 4.5. So, the protein precipitation was performed at this pH. Although both acid and alkaline methods yielded roughly 66% protein, their nutritional and techno-functional features differed based on the pH-shifting process. The alkaline-produced protein isolate had higher essential amino acid (EAA) content and EAA index but it was darker in colour. The acid-produced protein isolate had larger levels of umami-taste-active and functional amino acids, as well as a higher emulsifying capability.

Interfacial Crystallization and Supramolecular Self-Assembly of Spider Silk Inspired Protein at the Water-Air Interface

Macromolecular assembly into complex morphologies and architectural shapes is an area of fundamental research and technological innovation. In this work, we investigate the self-assembly process of recombinantly produced protein inspired by spider silk (spidroin). To elucidate the first steps of the assembly process, we examined highly concentrated and viscous pendant droplets of this protein in air. We show how the protein self-assembles and crystallizes at the water–air interface into a relatively thick and highly elastic skin. Using time-resolved in situ synchrotron X-ray scattering measurements during the drying process, we showed that the skin evolved to contain a high β-sheet amount over time. We also found that β-sheet formation strongly depended on protein concentration and relative humidity. These had a strong influence not only on the amount, but also on the ordering of these structures during the β-sheet formation process. We also showed how the skin around pendant droplets can serve as a reservoir for attaining liquid–liquid phase separation and coacervation from the dilute protein solution. Essentially, this study shows a new assembly route which could be optimized for the synthesis of new materials from a dilute protein solution and determine the properties of the final products.

Randomized Controlled Trial Comparing Autologous Protein Solution to Hyaluronic Acid Plus Triamcinolone for Treating Hip Osteoarthritis in Dogs

Twenty-three dogs with bilateral hip osteoarthritis were used to compare the efficacy of intra-articular injections of autologous protein solution (APS) to hyaluronic acid plus triamcinolone (HAT). Prior to treatment, owner assessments of pain and mobility were obtained using the canine brief pain inventory (CBPI) and Liverpool Osteoarthritis for Dogs (LOAD) questionnaires. Owners were also asked to list all medications used to control signs of pain associated with hip osteoarthritis (OA). In addition, objective kinetic data using a pressure sensitive walkway was used to quantify the relative weight bearing of each of the limbs (total pressure index; TPI). One hip was then selected using a random number generator for injection with HAT and the contralateral hip was injected with APS under the same sedation event. At 1-, 3-, and 6 months following injection, medication usage was recorded and dogs were re-assessed using the CBPI and LOAD questionnaires and using objective gait analysis to determine the TPI. Twenty dogs completed all aspects of the study and statistically significant (p < 0.05) improvements were noted by dog owners at every post-treatment time point in every category of pain and mobility as assessed by the CBPI and LOAD questionnaires. Only 5 dogs, compared to 14 pre-treatment, received any oral NSAID or other analgesic for the duration of the 6-month study period. The TPI, and change in TPI from baseline, were not statistically significantly different between the two treatments at any time point. These data suggest clinical efficacy of both APS and HAT, but fail to show superiority of one treatment vs. the other. The inability to detect a statistically significant difference between the two treatments could be attributable to a true lack of a difference, or a type II statistical error.

Interfacial crystallization and supramolecular self-assembly of spider silk inspired protein at the water-air interface

Abstract: Macromolecular assembly into complex morphologies and architectural shapes is an area of fundamental research and technological innovation. In this work, we investigate the self-assembly process of recombinantly produced protein inspired by spider silk (spidroin). To elucidate the first steps of the assembly process we looked into highly concentrated and viscous pendant droplets of this protein in air. We show how the protein self-assembles and crystallizes at the water-air interface into a relatively thick and highly elastic skin. Using time-resolved in-situ synchrotron x-ray scattering measurements during the drying process, we showed that the skin evolved to contain a high β-sheet amount over time. We also found that β-sheet formation strongly depended on protein concentration and relative humidity. These had a strong influence not only on the amount but also on the ordering of these structures during the β-sheet formation process. We also showed how the skin around pendant droplets can serve as a reservoir for attaining liquid-liquid phase separation and coacervation from the dilute protein solution. Essentially, this study shows a new assembly route which could be optimized for the synthesis of new materials from a dilute protein solution and determine the properties of the final products.

Interfacial crystallization and supramolecular self-assembly of spider silk inspired protein at the water-air interface

Macromolecular assembly into complex morphologies and architectural shapes is an area of fundamental research and technological innovation. In this work, we investigate the self-assembly process of recombinantly produced protein inspired by spider silk (spidroin). To elucidate the first steps of the assembly process we looked into highly concentrated and viscous pendant droplets of this protein in air. We show how the protein self-assembles and crystallizes at the water-air interface into a relatively thick and highly elastic skin. Using time-resolved in-situ synchrotron x-ray scattering measurements during the drying process, we showed that the skin evolved to contain a high β-sheet amount over time. We also found that β-sheet formation strongly depended on protein concentration and relative humidity. These had a strong influence not only on the amount but also on the ordering of these structures during the β-sheet formation process. We also showed how the skin around pendant droplets can serve as a reservoir for attaining liquid-liquid phase separation and coacervation from the dilute protein solution. Essentially, this study shows a new assembly route which could be optimized for the synthesis of new materials from a dilute protein solution and determine the properties of the final products.

Evaluation of Autologous Protein Solution Injection for Treatment of Superficial Digital Flexor Tendonitis in an Equine Model

Autologous protein solution (APS) has been used anecdotally for intralesional treatment of tendon and ligament injuries, however, its use in these injuries has never been studied in vivo. Our objective was to evaluate the effect of APS on tendon healing in an equine superficial digital flexor (SDF) tendonitis model. We hypothesized intralesional injection of APS would result in superior structural and biomechanical healing. SDF tendonitis was induced in both forelimbs of eight horses using collagenase injection. One forelimb was randomly assigned to receive an intralesional injection of APS, while the other was injected with saline. Ultrasonographic examinations were performed at weeks −1, 0, 2, 4, 8, and 12 following treatment. At 12 weeks, horses were euthanized and SDF samples harvested. Histologic evaluation, biomechanical testing, gene expression analysis, total glycosaminoglycan (GAG) and total DNA quantification were performed. Collagen type III (COL3A1) expression was significantly higher (p = 0.028) in saline treated tendon than in normal tendon. Otherwise, there were no significant differences in gene expression. There were no significant differences in histologic or ultrasonographic scores between groups. Mean total DNA content was significantly higher (p = 0.024) in saline treated tendons than normal tendons, whereas total DNA content was not significantly different between APS treated tendon and normal tendon. Elastic modulus was higher in APS treated than saline treated tendon, but the difference was not significant. Reduced expression of COL3A1 in APS treated tendon may indicate superior healing. Increased total DNA content in saline treated tendon may indicate ongoing healing processes, vs. APS treated tendons which may be in the later stages of healing. Limitations include a relatively short study period and inconsistency in size and severity of induced lesions. Intralesional injection of APS resulted in some improvements in healing characteristics.

Vapor mediation as a tool to control micro-nano scale dendritic crystallization and preferential bacterial distribution in drying respiratory droplets

Deposits of biofluid droplets on surfaces (such as respiratory droplets formed during an expiratory event fallen on surfaces) are composed of the water based salt protein solution that may also contain an infection (bacterial/viral). The final patterns of the deposit formed are dictated by the composition of the fluid and flow dynamics within the droplet. This work reports the spatio temporal, topological regulation of deposits of respiratory fluid droplets and control of motility of bacteria by tweaking flow inside droplets using non contact vapor mediated interactions. When evaporated on a glass surface, respiratory droplets form haphazard multiscale dendritic, cruciform shaped precipitates using vapor mediation as a tool to control these deposits at the level of nano, micro, millimeter scales. We morphologically control dendrite orientation, size and subsequently suppress cruciform-shaped crystals. The nucleation sites are controlled via preferential transfer of solutes in the droplets; thus, achieving control over crystal occurrence and growth dynamics. The active living matter like bacteria is also preferentially segregated with controlled motility without attenuation of its viability and pathogenesis. For the first time, we have experimentally presented a proof of concept to control the motion of live active matter like bacteria in a near nonintrusive manner. The methodology can have ramifications in biomedical applications like disease detection, controlling bacterial motility, and bacterial segregation.