Hemolysis Experiments Research Articles

Hemolytic activity of saponins from the leaves and stems of the specie Boldoa purpurascens Cav., determined by HPLC – UV: Validation of the method

This research contributes on the chemical composition of the species Boldoa purpurascens Cav. although the identification, quantification, and hemolytic activity evaluation of saponins isolated from this specie, moreover, high-performance liquid chromatography - ultraviolet detection (HPLC - UV) methodology has been developed and validated. To determine the saponin content of Boldoa purpurascens Cav. hydroalcoholic extract, a HPLC – UV was used. A Quil-A® pattern has been used to evaluate the validation parameters, such as method linearity, limits of detection and quantification, precision, accuracy, selectivity, and specificity. The research showed that each target component was well-separated under the optimal analytical settings with great selectivity. The technique was found to be precise, accurate, and repeatable. The method's linearity was determined to be robust, with an excellent determination coefficient value (R2≥0.999). The evaluation of the matrix effect revealed that each target component had a high degree of selectivity and was well-separated. It could be inferred that the established HPLC-UV methodology is accurate and useful for identifying saponins in the hydroalcoholic extract of Boldoa purpurascens Cav. The hydroalcoholic extract of Boldoa purpurascens Cav. was used to quantitatively determine the saponin content, and it was discovered that the concentration of QS 7 in the 80% hydroalcoholic extract was 1.9 x 10-2 g L-1 in the stem whereas 1.6 x 10-2 g L-1 in the leaves. The Boldoa purpurascens Cav. saponins have hemolytic activity comparable to Quil-A®, as shown by a thin layer chromatographic hemolysis experiment.

An aligned pattern sponge based on gelatin for rapid hemostasis

Post-traumatic hemorrhage is a leading cause of morbidity and mortality. However, most current hemostatic materials focus on incorporating nutritional components, with limited research addressing the impact of the material's structure on hemostasis. In this study, we developed cytocompatible and hemocompatible three-dimensional gelatin sponges with a patterned and aligned structure, designed for rapid hemostasis. The sponges were characterized by light microscope photography and scanning electron microscopy (SEM). Pattern sponges with gelatin (P-Gelatin) exhibited aligned structures on their surfaces and the inner structure. In terms of biocompatibility, MTT assay, and hemolysis experiment showed that P-Gelatin had good cytocompatibility and hemocompatibility.In vivoblood coagulation andin vivohemostasis, P-Gelatin sponges, with their aligned structure, exhibit rapid adsorption of red blood cells and platelets compared to non-patterned gelatin counterparts. This work introduces a safe and convenient patterned sponge for rapid hemostasis, especially highlighting a concept where a patterned structure can enhance the effectiveness of blood clotting, which is particularly relevant for tissue engineering.

Improvement of mechanical strength of calcium sulfate bone cement using carboxymethyl cellulose and silicon dioxide

Calcium sulfate (CS) is a commonly used filling material for bone defects; however, its mechanical strength requires further enhancement. The hydration process of CS significantly influences its mechanical properties. This study aims to improve these properties by incorporating carboxymethyl cellulose (CMC) and silicon dioxide (SiO2) to optimize the hydration process of calcium sulfate hemihydrate (CSH). After adding CMC, the content of CSH in the hydrated material significantly increased, while the amount of calcium sulfate dihydrate (CSD) decreased. In the XPS analysis, the Ca 2p peaks exhibited notable shifts following the addition of CMC and SiO2. The mechanisms by which the hydration process influences mechanical strength were further analyzed. It was found that Ca2+ ions from CSH chelated with CMC, while SiO2 formed Si-O-Ca bonds with Ca2+, both affecting the hydration process of CSH in different ways. The optimal addition amounts of CMC and SiO2 were both determined to be 1.5 %. Compared to pure CS, the mechanical properties of the experimental group increased by 520 %, rising from 4.28 MPa to 22.25 MPa. Additionally, the injectivity and erosion resistance were improved. All materials showed no cytotoxicity in hemolysis and cell experiments. In summary, this study demonstrates how the mechanical strength of CS can be enhanced by incorporating CMC and SiO2 to influence its hydration process.

Synthesis, characterisation of ZnO@PDA@Ag nanocomposite: Mechanistic interaction with BSA, photodegradation activity & in vitro cytotoxicity assay on H1299 lung cancer cell line

Despite advancements in diagnosis and therapeutic, cancer retains to be a greatest cause of fatality and economic damage to the world. Metallic nanoparticles have grabbed attention particularly in the field of medicine attributed to their noteworthy biological and catalytic properties, offering significant progress. The work aimed to create a novel biocompatible ‘Silver doped Polydopamine coated Zinc-Oxide nanocomposite’ and investigate its efficacy in treating H1299 lung cancer cells as well as water remediation. In this study, a ‘ZnO@PDA@Ag’ Nanocomposite was synthesised using co-precipitation method and was further characterised using DLS, FESEM-EDX, P-XRD, XPS, TEM and FT-IR techniques. The mechanistic interaction with blood protein, suggested strong binding interactions and notable structural changes in BSA upon exposure to the nanocomposite. The photocatalytic properties evaluated against the Rhodamine B Dye under UV–Visible light irradiation demonstrated a photodegradation of ~49.7% within 120 min for 60 μg/mL of the used nanocomposite. Herein, we present an evaluation of anticancer bioactivity of ZnO@PDA@Ag nanoparticles using MTT assay against the H1299 lung cancer cell line and its IC50 was estimated to be (42.42 ± 4) μg/mL. The cytotoxicity was enhanced through immobilization of Silver (Ag) metal on ZnO@PDA. Moreover, the hemolysis experiment was also conducted to demonstrate the biocompatibility of nanoparticle to human red blood cells. As a result, the successfully created biocompatible nanomaterial may be useful as an efficient drug-delivery system against cancer cells.

Chitosan nanoparticles of imatinib mesylate coated with TPGS for the treatment of colon cancer: In-vivo & in-vitro studies

The study aimed to develop and evaluate chitosan-based nanoparticles coated with TPGS for the targeted delivery of imatinib mesylate to colon cancer cells. Particle size and zeta potential analysis were within the acceptable range for targeting colon cancer. CS-IMT-TPGS-NPs had a significant positive zeta potential of 30.4 mV, suggesting improved cellular intake. FE-SEM and TEM demonstrated that the nanoparticles appeared spherical, smooth, and did not aggregate, with a visible TPGS coating. XRD confirmed that crystalline imatinib transitioned to an amorphous state during nano formulation. In-vitro tests on HCT-116 cells demonstrated that CS-IMT-TPGS-NPs outperformed free IMT regarding cytotoxicity, apoptosis induction, cellular uptake, and cell migration inhibition. Additionally, the nanoparticles were examined in vitro using mitochondrial membrane potential, DNA fragmentation, GAPDH relative gene expression, ROS estimation, and cell cycle analysis. The effect of therapy on expected colon-associated bacterial strains was also investigated. The biocompatibility of nanoparticles was assessed by hemolysis and platelet aggregation experiments. The anti-inflammatory impact was determined using the HET-CAM test. Non-Fickian diffusion at pH 5.5 resulted in sustained in-vitro drug release, with no initial burst. In-vivo investigations using albino Wistar rats suggest pharmacokinetic properties for produced nanoparticles, whereas histopathological examinations assess acute toxicity.

Preparation of Time-Sequential Functionalized ZnS-ZnO Film for Modulation of Interfacial Behavior of Metals in Biological Service Environments.

Blood-contact devices are prone to inflammation, endothelial dysfunction, coagulation, and the uncontrolled release of metal ions during implantation and service. Therefore, it is essential to make these multifunctional. Herein, a superhydrophobic DE@ZnS-ZnO@SA film (composed of dabigatran ester, zinc sulfite, zinc oxide, and stearic acid, respectively) is produced. The prepared film has non-adhesion and antibacterial properties, superior mechanical stability, durability, corrosion resistance, and is self-cleaning and blood-repellent. The results of the hemolysis, cytotoxicity, and other anticoagulant experiments revealed that the film had good blood compatibility, no cytotoxicity, and excellent anticoagulant properties. The film displays anticoagulant properties even after being immersed in Phosphate-Buffered Saline (PBS) for 7 days. Furthermore, the film can spontaneously release H2S gas for 90 h after soaking in an acidic environment (pH = 6) for 90 h. This property improves the acidic microenvironment of the lesion and promotes the proliferation of endothelial cells by using H2S gas. In addition, the film can inhibit the uncontrollable release of Zn2+ ions, avoiding its toxicity even when immersed in an acid environment for 35 days. This time-sequential functionalized surface has the potential to typify the future of blood-contacting scaffolds for long-lasting use.

A l-lysine-coated graphene oxide/magnetic hydroxyapatite/cubic metal-organic frameworks for targeted and pH-controlled release of doxorubicin to cancer cells

Research on biocompatible metal-organic frameworks (MOFs) has recently attracted much attention from researchers as drug delivery systems. In this study, for the first time, Cu MOFs grown on graphene oxide capped with magnetic hydroxyapatite (MOF Cu@GO/HAp-SiO2-Fe3O4), and then the surface of the resulting nanocomposites was modified with the l-lysine amino acid (Lys-MOF Cu@GO/HAp-SiO2-Fe3O4) to prepare a controlled and pH-sensitive nanocomposite for targeted delivery of doxorubicin (DOX) to MDA-MB-231 breast cancer cells and MCF-10A normal cells. Different techniques confirmed the successful preparation of the designed nanocomposites. The values of encapsulation efficiencies (EE%) for MOF Cu@GO/HAp-SiO2-Fe3O4 and Lys-MOF Cu@GO/HAp-SiO2-Fe3O4 was found to be 79.3 % and 96.5 %, respectively. The in vitro DOX release results showed that the Lys-MOF Cu@GO/HAp-SiO2-Fe3O4 has a more controlled release than that of MOF Cu@GO/HAp-SiO2-Fe3O4 due to the slow diffusion of the drug through the Lys shell. In addition, both nanocomposites showed a pH-response release behavior. The release mechanism of DOX by the designed nanocomposites also revealed that the Korsmeyer-Peppas model is the best model to describe the release of the drug. Furthermore, in vitro cytotoxicity investigation confirmed that the prepared MOF Cu@GO/HAp-SiO2-Fe3O4 and Lys-MOF Cu@GO/HAp-SiO2-Fe3O4 nanocomposite had good biocompatibility against MDA-MB-231 and MCF-10A cells. Whereas, DOX-MOF Cu@GO/HAp-SiO2-Fe3O4 and DOX-Lys-MOF Cu@GO/HAp-SiO2-Fe3O4 against MDA-MB-231 cells increased the cytotoxicity as a result of the targeting and controlled delivery of the DOX to the cancer cells. The in vitro antioxidant activity and hemolysis experiments revealed that the designed Lys-MOF Cu@GO/HAp-SiO2-Fe3O4 have excellent antioxidant activity and good haemocompatibility, respectively, as a suitable nanocomposites for drug applications. Therefore, it is suggested that the prepared nanocomposites can be good for biological applications.

Organic nanoparticles incorporated starch/carboxymethylcellulose multifunctional coating film for efficient preservation of perishable products

Most of postharvest agricultural produces are perishable due to microorganisms infections and physiological change. Herein, one kind of multifunctional coating film of SC-ECCNPs was developed by incorporating organic nanoparticles of ECCNPs into starch/carboxymethylcellulose (SC) to prolong shelf life of food with excellent performances. The SC-ECCNPs coating was prepared with starch and sodium carboxymethylcellulose as film substrate (SC) to incorporate with organic nanoparticles of ECCNPs formed by integrating epigallocatechin-3-gallate (EGCG), cysteine (Cys), and cinnamaldehyde (CA). The incorporation of ECCNPs improves the UV-resistance and physical properties of SC-ECCNPs coating and also endows it with excellent antioxidative and broad-spectrum antibacterial activity. The application possibilities of SC-ECCNPs coating were explored with strawberries and oranges as samples, validating that the SC-ECCNPs coating can prolong the shelf life of fruits at room temperature. The biosafety of the coating was further confirmed with hemolysis and MTT experiments. The SC-ECCNPs coating film was prepared with natural substrates via a simple and green method. The investigation provides an instructive way for developing advanced packaging materials with high performances.

Preparation and Characterization of Silica-Coated Sodium Alginate Hydrogel Beads and the Delivery of Curcumin

In this study, to address the defects of sodium alginate (SA), such as its susceptibility to disintegration, silica was coated on the outer layer of sodium alginate hydrogel beads in order to improve its swelling and slow-release properties. Tetraethyl orthosilicate (TEOS) was used as the hydrolyzed precursor, and the solution of silica precursor was prepared by sol-gel reaction under acidic conditions. Then SA–silica hydrogel beads prepared by ionic crosslinking method were immersed into the SiO2 precursor solution to prepare SA–silica hydrogel beads. The chemical structure and morphology of the hydrogel beads were characterized by XRD, FTIR, and SEM, and the results showed that the surface of SA–silica beads was successfully encapsulated with the outer layer of SiO2, and the surface was smooth and dense. The swelling experiments showed that the swelling performance effectively decreased with the increase of TEOS molar concentration, and the maximum swelling ratio of the hydrogel beads decreased from 41.07 to 14.3, and the time to reach the maximum swelling ratio was prolonged from 4 h to 8 h. The sustained-release experiments showed that the SA–silica hydrogel beads possessed a good pH sensitivity, and the time of sustained-release was significantly prolonged in vitro. Hemolysis and cytotoxicity experiments showed that the SA-silica hydrogel beads were biocompatible when the TEOS molar concentration was lower than 0.375 M. The SA–silica-2 hydrogel beads had good biocompatibility, swelling properties, and slow-release properties at the same time.

In silico-designed antimicrobial peptide targeting MRSA and E. coli with antibacterial and antibiofilm actions.

Antibiotic resistance is a paramount global health issue, with numerous bacterial strains continually fortifying their resistance against diverse antibiotics. This surge in resistance levels primarily stems from the overuse and misuse of antibiotics in human, animal, and environmental contexts. In this study, we advocate for exploring alternative molecules exhibiting antibacterial properties to counteract the escalating antibiotic resistance. We identified a synthetic antimicrobial peptide (AMP) by using computational search in AMP public databases and further engineering through molecular docking and dynamics. Microbiological evaluation, cytotoxicity, genotoycity, and hemolysis experiments were then performed. The designed AMP underwent rigorous testing for antibacterial and antibiofilm activities against Methicillin-Resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), representing gram-positive and gram-negative bacteria, respectively. Subsequently, the safety profile of the AMP was assessed in vitro using human fibroblast cells and a human blood sample. The selected AMP demonstrated robust antibacterial and antibiofilm efficacy against MRSA and E. coli, with an added assurance of non-cytotoxicity and non-genotoxicity towards human fibroblasts. Also, the AMP did not demonstrate any hemolytic activity. Our findings emphasize the considerable promise of the AMP as a viable alternative antibacterial agent, showcasing its potential to combat antibiotic resistance effectively.

Antimicrobial poly (1,4-butylene carbonate): Preparation, characterization, and potential applications as a material for tympanic membrane repair

Perforation of the tympanic membrane (TM) is a common condition that often requires a scaffold as a support for surgery. However, because of the external environment of the auditory canal, the scaffold could become bacterially infected and prevent the TM from healing. As a result, the perfect scaffold should have both antibacterial and biomimetic qualities. In this study, the biodegradable biomaterial poly(1,4-butylene carbonate) (PBC) films containing levofloxacin (LEV) was successfully prepared for the first time. The results showed that the hydrophilicity of the LEV/PBC film was improved after the addition of LEV, and the tensile strength was also complied with the requirements of the standard. The created antibacterial film demonstrated excellent antibacterial properties. In vitro hemolysis experiments revealed no risk of hemolysis for the new material, and the cytotoxicity study further confirmed its non-cytotoxic nature. Overall, LEV was a good component of PBC/LEV film, which is expected to be used for TM repair in the future.

Efficient Catalyst-Free One-Pot Synthesis of Polysaccharide-Polypeptide Hydrogels in Aqueous Solution.

The preparation of polysaccharide-peptide hydrogels usually involves multiple synthetic steps, thus reducing the effectiveness and practicality of these approaches. Inspired by recent discoveries in aqueous N-carboxyanhydride (NCA) ring-opening polymerization (ROP) and ring-opening polymerization-induced nanogelation, we present an aqueous one-pot strategy to prepare polysaccharide-polypeptide hydrogels. In this study, water-soluble polysaccharide carboxymethyl chitosan is used as the macromolecular initiator to prepare polysaccharide-polypeptide copolymers through the aqueous ROP of NCA. The catalyst-free approach afforded hydrogels with properties that could be controlled by adjusting the type and amount of NCA used, with the elastic modulus ranging from 50 Pa to 18000 Pa. The hydrogen bond-cross-linked hydrogel exhibited self-healing and injectable properties. Morphology characterization revealed that micelles were formed in the early stage of reaction, suggesting that the polymerization follows an aqueous ring-opening polymerization-induced self-assembly (ROPISA) mechanism and that aggregation of micelles during the reaction caused the gelation. Moreover, the hydrogels displayed high swelling ratios (>95% water content), and hemolysis and cytotoxicity experiments demonstrated that the hydrogels had excellent biocompatibility, indicating their potential in medical applications.

Inhibition of LPS-Induced Skin Inflammatory Response and Barrier Damage via MAPK/NF-κB Signaling Pathway by Houttuynia cordata Thunb Fermentation Broth.

Houttuynia cordata Thunb is rich in active substances and has excellent antioxidant and anti-inflammatory activity. Scanning electron microscopy and gel permeation chromatography were used to analyze the molecular characteristics of the fermentation broth of Houttuynia cordata Thunb obtained through fermentation with Clavispora lusitaniae (HCT-f). The molecular weight of HCT-f was 2.64265 × 105 Da, and the polydispersity coefficient was 183.10, which were higher than that of unfermented broth of Houttuynia cordata Thunb (HCT). By investigating the active substance content and in vitro antioxidant activity of HCT-f and HCT, the results indicated that HCT-f had a higher active substance content and exhibited a superior scavenging effect on 2,2-diphenyl-1-picrylhydrazyl radicals and hydroxyl radicals, with IC50 values of 11.85% and 9.01%, respectively. Our results showed that HCT-f could effectively alleviate the increase in the secretion of inflammatory factors and apoptotic factors caused by lipopolysaccharide (LPS) stimulation, and had a certain effect on repairing skin barrier damage. HCT-f could exert an anti-inflammatory effect by down-regulating signaling in the MAPK/NF-κB pathway. The results of erythrocyte hemolysis and chicken embryo experiments showed that HCT-f had a high safety profile. Therefore, this study provides a theoretical basis for the application of HCT-f as an effective ingredient in food and cosmetics.

A super hydrophilic and high strength chitosan hemostatic sponge prepared by freeze-drying and alkali treatment for rapid hemostasis

Excessive blood loss caused by natural accidents or major surgical procedures can be life-threatening and requires rapid hemorrhage management. Developing hemostatic materials with high water absorption capacity and high strength properties is still challenging. Super hydrophilic/high-strength chitosan hemostatic sponges (SHHS-CHS) were prepared by freeze-drying and alkali treatment. The freeze-drying technology formed a porous three-dimensional structure of sponges, and alkali treatment increased the hydrophilicity of sponges and the rigidity of the chitosan molecular chain, improving the hydrophilicity and mechanical properties of chitosan sponges. Characterization experiments revealed that SHHS-CHS had high water absorption capacity, rich pore structure, and excellent mechanical properties. The cytotoxicity and hemolysis experiments showed that SHHS-CHS had good biocompatibility. Compared to the commercial hemostatic gelatin sponges and chitosan hemostatic material, SHHS-CHS had better blood coagulation and red blood cell and platelet adhesion ability. Moreover, SHHS-CHS showed better hemostatic ability than gelatin sponges in rat-liver injury and rat leg vein rupture models, significantly reducing the hemostasis time and bleeding volume. In conclusion, SHHS-CHS has broad prospects for clinical application as a hemostatic material.

Magnetic biocomposite scaffold based on decellularized tendon ECM and MNP-deposited halloysite nanotubes: physicochemical, thermal, rheological, mechanical and in vitro biological evaluations

The development of new three-dimensional biomaterials with advanced versatile properties is critical to the success of tissue engineering (TE) applications. Here, (a) bioactive decellularized tendon extracellular matrix (dECM) with a sol-gel transition feature at physiological temperature, (b) halloysite nanotubes (HNT) with known mechanical properties and bioactivity, and (c) magnetic nanoparticles (MNP) with superparamagnetic and osteogenic properties were combined to develop a new scaffold that could be used in prospective bone TE applications. Deposition of MNPs on HNTs resulted in magnetic nanostructures without agglomeration of MNPs. A completely cell-free, collagen- and glycosaminoglycan- rich dECM was obtained and characterized. dECM-based scaffolds incorporated with 1%, 2% and 4% MNP-HNT were analysed for their physical, chemical, and in vitro biological properties. Fourier-transform infrared spectroscopy, x-ray powder diffractometry and vibrating sample magnetometry analyses confirmed the presence of dECM, HNT and MNP in all scaffold types. The capacity to form apatite layer upon incubation in simulated body fluid revealed that dECM-MNP-HNT is a bioactive material. Combining dECM with MNP-HNT improved the thermal stability and compressive strength of the macroporous scaffolds upto 2% MNP-HNT. In vitro cytotoxicity and hemolysis experiments showed that the scaffolds were essentially biocompatible. Human bone marrow mesenchymal stem cells adhered and proliferated well on the macroporous constructs containing 1% and 2% MNP-HNT; and remained metabolically active for at least 21 d in vitro. Collectively, the findings support the idea that magnetic nanocomposite dECM scaffolds containing MNP-HNT could be a potential template for TE applications.

DLP-printed standard tooth -colored ceramic dentures and its biocompatibility study

Currently, ceramic dentures are mainly obtained by using computer-assisted cutting block of material to obtain the shape as the removing a model of patient's mouth. Followed by immersion coloring, porcelain stacking, and sintering to obtain dentures similar to a patients' tooth, this is a large-loss and complex process. In this study, 16 standard tooth samples were successfully reduced by adding Fe2O3, Pr6O10, Er2O3, and MnO2. After the colorful ceramic powder and photosensitive resin were configured into a slurry, a close to the sample tooth dentures was successfully prepared using digital light processing (DLP). The flexural strength and compressive strength of the ceramic specimens prepared by this method reached 821.01 ± 51.72 MPa and 2.67 GPa ± 0.22 MPa. To verify the biosafety of the prepared ceramics, we performed an in vitro biological evaluation of rat fibroblasts (L929). The resulting showed that the extracts of the prepared ceramic teeth had good biosafety, and the cells could also attach to the surface of the prepared ceramics. Hemolysis experiments confirmed good blood compatibility. Thus, our results show that DLP-printed standard tooth-colored ceramic dentures have good mechanical properties and biocompatibility, and can provide a solution to patients seeking quick access to aesthetic dentures.

Preparation and Evaluation of Folate Modified PEG-PLLA Nanoparticles Loaded with Lycorine for Glioma Treatment.

Lycorine is a kind of natural active ingredient with a strong antitumor effect. In this study, folate ligand-conjugated polyethylene glycol-block-poly(l-lactide) (PEG-PLLA) nanoparticles (FA-PEG-PLLA NPs) were designed to deliver lycorine to enhance its anti-glioma activity. The successful preparation of the FA-PEG-PLLA polymer was confirmed by 1H-NMR, FT-IR, XRD, TGA, and DSC. The optimal formulation for LYC@FA-PEG-PLLA NPs was determined by response surface analysis as follows: sodium dodecyl sulfate (SDS) of 1%, carrier material of 0.03 g, organic phase volume of 3 mL, and ultrasonic power of 20%. The LYC@FA-PEG-PLLA NPs exhibited an encapsulation efficiency of 83.58% and a particle size of 49.71 nm, demonstrating good stability. Hemolysis experiments, MTT assays, and cell scratch assays revealed excellent biocompatibility of FA-PEG-PLLA and superior anti-glioma activity of LYC@FA-PEG-PLLA NPs compared to the raw drug. Additionally, cell apoptosis assays, ROS experiments, and western blot analysis demonstrated that LYC@FA-PEG-PLLA NPs contributed to cell apoptosis by inducing ROS generation and increasing the expression of NF-κB inhibitory protein IκBα. These results suggested that LYC@FA-PEG-PLLA NPs hold promise for glioma treatment.

The Osmolality and Hemolysis of High-Concentration Monoclonal Antibody Formulations.

Highly concentrated monoclonal antibody (mAb) formulations for subcutaneous administration are becoming increasingly preferred within the biopharmaceutical industry for ease of use and improved patient compliance. A common phenomenon observed in the industry is that osmolality detected via freezing-point depression (FPD) in high-concentration mAb formulations is much higher than the theoretical concentrations, yet the occurrence of this phenomenon and its possible safety issues have been rarely reported. The current study summarized theoretical osmolality of U.S. Food and Drug Administration approved high-concentration mAb formulations and evaluated effects of high osmolality on safety using hemolysis experiments for the first time. Two mAbs formulated at 150mg/mL were used as models and configured into two isotonic solutions: a, a theoretically calculated molarity in the isotonic range (H) and b, an osmolality value measured via the FPD in the isotonic range (I). The H and I formulations of each mAb were individually subjected to hemolysis experiments, and the hemolysis rates of the two formulations of the same mAb were compared. Besides, the effect of mAb concentration on osmolality detected by FPD was explored as well. The results indicated that the hemolysis rates were similar between the H and I formulations of mAbs at the same sample addition volume, and the osmolality values increased approximately linearly with the increase in mAb concentration. High osmolality for high-concentration mAb formulations would not affect product safety and the excipients could be added at relatively high levels to maintain product stability, especially for labile products.

Electrospun PCL/gelatin/arbutin nanofiber membranes as potent reactive oxygen species scavengers to accelerate cutaneous wound healing.

The presence of excessive reactive oxygen species (ROS) at a skin wound site is an important factor affecting wound healing. ROS scavenging, which regulates the ROS microenvironment, is essential for wound healing. In this study, we used novel electrospun PCL/gelatin/arbutin (PCL/G/A) nanofibrous membranes as wound dressings, with PCL/gelatin (PCL/G) as the backbone, and plant-derived arbutin (hydroquinone-β-d-glucopyranoside, ARB) as an effective antioxidant that scavenges ROS and inhibits bacterial infection in wounds. The loading of ARB increased the mechanical strength of the nanofibres, with a water vapour transmission rate of more than 2500 g/(m2 × 24 h), and the water contact angle decreased, indicating that hydrophilicity and air permeability were significantly improved. Drug release and degradation experiments showed that the nanofibre membrane controlled the drug release and exhibited favourable degradability. Haemolysis experiments showed that the PCL/G/A nanofibre membranes were biocompatible, and DPPH and ABTS+ radical scavenging experiments indicated that PCL/G/A could effectively scavenge ROS to reflect the antioxidant activity. In addition, haemostasis experiments showed that PCL/G/A had good haemostatic effects in vitro and in vivo. In vivo animal wound closure and histological staining experiments demonstrated that PCL/G/A increased collagen deposition and remodelled epithelial tissue regeneration while showing good in vivo biocompatibility and non-toxicity. In conclusion, we successfully prepared a multifunctional wound dressing, PCL/G/A, for skin wound healing and investigated its potential role in wound healing, which is beneficial for the clinical translational application of phytomedicines.

T5S1607 identified as a antibacterial FtsZ inhibitor：Virtual screening combined with bioactivity evaluation for the drug discovery

Due to antibiotic overuse, many bacteria have developed resistance, creating an urgent need for novel antimicrobial agents. It has been established that the filamentous temperature-sensitive mutant Z (FtsZ) of the bacterial cell division protein is an effective and promising antibacterial target. In this study, the optimal proteins were assessed by early recognition ability and the processed compound libraries were virtually screened using Vina. This effort resulted in the identification of 14 potentially active antimicrobial compounds. Among them, the compound T5S1607 demonstrated remarkable antibacterial efficacy against Bacillus subtilis ATCC9732 (MIC = 1 μg/mL) and Staphylococcus aureus ATC5C6538 (MIC = 4 μg/mL). Furthermore, in vitro experiments demonstrated that the selected compound T5S1607 rapidly killed bacteria and induced FtsZ protein aggregation, preventing bacterial division and leading to bacterial death. Additionally, cell toxicity and hemolysis experiments indicate that compound T5S1607 exhibits minimal toxicity to LO2 cells and shows no significant hemolytic effects on mammalian cells in vitro at the MIC concentration range. All the results indicate that compound T5S1607 is a promising antibacterial agent and a potential FtsZ inhibitor. In conclusion, this work successfully discovered FtsZ inhibitors with good activity through the virtual screening drug discovery process.