Hemocompatibility Assays Research Articles

Neutralization of Naja naja venom by green and chemically synthesized titanium dioxide nanoparticles

Abstract Snake envenomation affects millions of people worldwide especially rural population in developing countries. High production cost, poor accessibility and side-effects associated with antivenom treatment call for newer approaches. Nanoparticles could provide alternative therapy due to their biocompatibility, ease of production and stability. In this study, Titanium (IV) isopropoxide was reduced to make TiO2 nanoparticles by chemical and green synthesis method utilizing potato peels and neem leaves extract. The synthesized particles were characterized by Scanning Electron Microscope, Electron Dispersive X-ray Spectroscopy and Fourier Transform Infrared Spectroscopy. The synthesized nanoparticles are spherical in shape with sizes ranging from 41-64 nm and molecules, such as polyphenols, flavonoids, and alkaloids in plant extracts act as reducing and capping agents. The nanoparticles were evaluated for in vitro venom neutralization assays against the Naja naja venom. The hemocompatibility assay showed that the TiO2 NPs are non-toxic, indicating their safe use. The TiO2 NPs were able to significantly inhibit venom induced hemolysis of RBCs and blood anticoagulation effect (p-values ≤0.000). Overall, the study suggests that TiO2 NPs could be effective in reducing the toxicity of Naja naja venom, providing a practical and cost-effective therapeutic option to be used as alternative treatments for snake envenomation.

ISO 10993-4 Compliant Hemocompatibility Evaluation of Gellan Gum Hybrid Hydrogels for Biomedical Applications.

This study examines the hemocompatibility of gellan-gum-based hybrid hydrogels, with varying gellan-gum concentrations and constant sodium alginate and silk fibroin concentrations, respectively, in accordance with ISO 10993-4 standards. While previous studies have focused on cytocompatibility, the hemocompatibility of these hydrogels remains underexplored. Hydrogels were formulated with 0.3%, 0.5%, 0.75%, and 1% gellan gum combined with 3% silk fibroin and 4.2% sodium alginate separately, using physical and ionic cross-linking. Swelling behavior was analyzed in phosphate (pH 7.4) and acetic (pH 1.2) buffers and surface morphology was examined by scanning electron microscopy (SEM). Hemocompatibility tests included complete blood count (CBC), coagulation assays, hemolysis index, erythrocyte morphology, and platelet adhesion analysis. Results showed that gellan gum-sodium alginate hydrogels exhibited faster swelling than gellan gum-silk fibroin formulations. SEM indicated smoother surfaces with sodium alginate, while silk fibroin increased roughness, further amplified by higher gellan-gum concentrations. Hemocompatibility assays confirmed normal profiles in formulations with 0.3%, 0.5%, and 0.75% gellan gum, while 1% gellan gum caused significant hemolytic and thrombogenic activity. These findings highlight the excellent hemocompatibility of gellan-gum-based hydrogels, especially the sodium alginate variants, supporting their potential in bioengineering, tissue engineering, and blood-contacting biomedical applications.

Hofmeister effect enhanced SiO2/gelatin-based hydrophobically associated hydrogels and their lubricating properties.

In recent years, hydrogel materials with suitable energy dissipation mechanisms and excellent mechanical properties have attracted much attention in tissue engineering due to their ability to mimic the natural cartilage structure. However, in cartilage tissue's regeneration and repair process, hydrogel materials should also possess satisfactory lubrication properties and biocompatibility. Therefore, preparing biocompatible low friction, high toughness hydrogels remain a challenge. In this paper, a new strategy is proposed to use gelatin, acrylamide (AM), lauryl methacrylate (LMA) and SiO2 to construct hydrophobically associated hydrogels, where gelatin was used as an emulsifier and SiO2 was used to a nano-enhanced filler. Then the Hofmeister effect enhanced SiO2/gelatin-based hydrophobically associated hydrogels were prepared by one-step immersion in ammonium sulfate solution. The results showed that the strong "salting out" effect of ammonium sulfate solution on gelatin led to further enhancement of the hydrophobic interactions between gelatin molecular chains, which significantly improved the mechanical properties and lubrication ability of the hydrogels. Furthermore, Calcein AM-PI fluorescent staining and haemolysis assays showed that the hydrogel had low cytotoxicity and good haemocompatibility, and ELISA and scratch assays confirmed the positive regulatory effect of the hydrogel on normal cell growth. The Hofmeister effect-enhanced SiO2/gelatin-based hydrophobically associated hydrogels have potential applications in articular cartilage repair.

Synthesis and evaluation of vanillin Schiff bases as potential antimicrobial agents against ESBL-producing bacteria: towards novel interventions in antimicrobial stewardship

The escalating challenge of antimicrobial resistance necessitates the development of novel antibacterial agents. In this study, a series of five vanillin Schiff bases (SB-1 to SB-5) were synthesized from vanillin and various aromatic amines. The chemical structures of these compounds were characterized using Thin Layer Chromatography (TLC), Fourier Transform Infrared Spectroscopy (FT-IR), proton nuclear magnetic resonance (1H\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$^{1}\ ext {H}$$\\end{document}-NMR), carbon-13 NMR (13C\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$^{13}\ ext {C}$$\\end{document}-NMR), and mass spectrometry techniques. Antibacterial efficacy was evaluated against strains of bacteria producing extended-spectrum beta-lactamases (ESBL), including Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae using the disc diffusion method. Cytotoxic effects were assessed through haemocompatibility and brine shrimp lethality assays. The Schiff bases demonstrated notable antibacterial activities, with SB-1, SB-2, SB-4, and SB-5 exhibiting zones of inhibition up to 16.0, 16.5, 16.6, and 15.5 mm against ESBL E. coli, respectively. SB-3 showed a maximum inhibition zone of 15.0 mm against ESBL K. pneumoniae. In cytotoxicity assays, the compounds exhibited IC50\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{50}$$\\end{document} values against red blood cells (RBCs) greater than 200 μg/mL and ranging from 45.7 to 50.5 μg/mL for the brine shrimp assay. While demonstrating potent antibacterial properties, the toxicity towards human RBCs suggests that further toxicity evaluations and structural modifications are essential for developing safer therapeutic agents based on vanillin Schiff bases.

Hydroxyapatite layer evolution on silica-based (45S10P) bioactive glass-ceramic nanoparticles doped with Zn2+ ions: Augmentation of in vitro bioactivity, antibacterial activity and textural modification for bone regeneration

Zinc-incorporated bioactive glasses (BG) and bioactive glass-ceramic (BGC) nanoparticles (BGC-NPs) represent highly adaptable, biodegradable, and bioactive materials in tissue engineering and regenerative medicine. This study delves into the synthesis, characterization and biomedical implications of Zn2+ ions-doped 45S10P spherical BGC-NPs, employing analytical techniques and in vitro assays. The resulting (Zn2++ 45S10P) BGC-NPs, prepared using a modified Strober's method, exhibited spherical morphology with excellent dispersion, as verified by TEM, FE-SEM, and DLS analyses. XRD, FTIR, and FESEM analysis revealed an augmented hydroxyapatite (HAp) layer formation with increasing zinc content. Zeta potential analysis showcased a shift from negative to positive values after immersion in simulated body fluid (SBF), indicating the bioactive potential of the developed BGC-NPs. Hemocompatibility assays indicated the biocompatibility of all BGC-NPs, demonstrating minimal hemolytic effects with incorporating Zn2+ ions below a standard threshold (<5 % lysis). Further, the migration assay revealed the potentiality of the BGC-NPs to stimulate the migration of HeLa cells. Cell viability assays employing MG-63 osteoblast-like cells highlighted enhanced cell viability over time, underscoring their non-toxic nature and potential for tissue regeneration. Antibacterial assays displayed significant inhibitory effects against Gram-negative compared to Gram-positive bacteria, underscoring their potential for anti-infection applications. Overall, the results affirm the promising prospects of Zinc incorporation into 45S10P BGC-NPs for bone tissue regeneration applications.

Identification of different extracts and phytoconstituents of Callistemon viminalis Cheel for their anti-anxiety effects based on pharmacognostic, toxicological, and pharmacological strategies

BackgroundPsychiatric disorders like depression and anxiety are global challenges, exacerbated by the limitations of synthetic medications, including addiction and toxic side effects. MethodsThis study meticulously investigated the pharmacognostic, phytochemical, toxicological, and pharmacological properties of Callistemon viminalis Cheel. Toxicological assessments, including hemocompatibility assays, LD50 studies, FOB analysis, biochemical parameters, and structural integrity of vital organs, were conducted on aqueous, methanolic, chloroform, and petroleum ether extracts of leaves and stems. Phytochemical profiling via qualitative tests and GC-MS screened extracts for molecular docking against key receptors. Categorically screened extracts were evaluated for therapeutic potential against LPS-induced anxiety in mice. ResultsToxicological evaluations on experimental animals demonstrated the safety of various extracts, evidenced by no in vitro and in vivo toxicity. GC-MS identified numerous phytochemicals that passed “Lipinski's Rule of Five.” These compounds were screened for molecular docking, revealing significant binding affinities with CB1, SERT, α2A-AR, and GABAβ2 receptors, suggesting potential therapeutic effects against anxiety. The phytoconstituents with the highest docking scores, particularly in aqueous and methanolic extracts, were further validated for their therapeutic efficacy. Preliminary analysis based on the EPM test and serum cortisol levels confirmed these extracts' superior therapeutic effectiveness. ConclusionIn conclusion, aqueous and methanolic extracts of Callistemon viminalis Cheel’s leaf and stem showed promising potential as therapeutic interventions for anxiety disorders.

EpCAM aptamer-engineered, azadiradione-loaded, chemo-photo-responsive nano-erythroliposomes for the treatment of triple-negative breast cancer

This research aimed to engineer azadiradione (AZ) and IR780-enabled, epithelial cellular adhesion molecule (EpCAM) aptamer (Apt) grafted, targeted nano-erythroliposomes (Apt/Eryt/AZ/IR780) for chemo-photo combined treatment of triple-negative breast cancer (TNBC). Physicochemical characterization revealed average particle size and polydispersity index (PDI) of 106.8±3.76 nm and 0.176 ±0.03, respectively, and zeta potential of 28.75±0.65 mV. Compared to formulations that were not NIR triggered, ∼85 % of the AZ was released in 8 h from NIR-exposed formulation. EpCAM expression analysis unequivocally demonstrated that EpCAM levels were significantly elevated in 4T1 cells compared to MDA-MB-231 cells. Cellular uptake analysis revealed strong fluorescence intensity with Apt/Eryt/AZ/IR780 with NIR exposure compared to non-aptamer formulations. Dose-dependent MTT assay (100 mM, 150 mM, and 200 mM concentration of AZ) revealed that <20 % of cells survived after treatment with Apt/Eryt/AZ/IR780 and NIR exposure. The cell viability results were further validated by employing reactive oxygen species (ROS) generation determination, mitochondrial membrane potential (MMP) inhibition, and Caspases-8 assay. In particular, 4T1 cells were found to be sensitive to AZ by decreasing MMP activity (∼17-fold reduction compared to control) and increasing Caspase-3 activity (∼544-fold enhancement compared to control). A hemocompatibility assay revealed a non-significant interaction (<0.9 %) of Apt/Eryt/AZ/IR780 with blood components. In addition, stability investigation for 30 days at 30±2 °C and 5±3 °C of Apt/Eryt/AZ/IR780. The findings revealed no significant change in average size (<8.5 %), zeta potential (<25 %), PDI (<20 %), and % of IR780 (<14 %) and AZ (<13 %) at 5±3 °C in 90 days.

Screening of the global health priority BoxⓇ reveals potential new disinfectants against the emerging multidrug-resistant pathogen Candida auris

BackgroundCandida auris has been identified by the World Health Organization as a critical pathogen due to its invasive nature, resistance to multiple drugs, and high mortality rates in hospital outbreaks. This fungus can persist on surfaces and human skin for extended periods, complicating infection control efforts. The need for effective disinfection strategies is urgent, as current disinfectants are often ineffective against C. auris biofilms. ObjectiveThe study aimed to identify potential disinfectants from a collection of 240 compounds in the Global Health Priority Box® that are effective against C. auris, particularly strains resistant to existing options. MethodsThe research employed a screening protocol using a fluconazole-resistant strain of C. auris (149/23). Antifungal activity was assessed using the microdilution method to determine Minimum Inhibitory Concentrations (MICs) and Minimum Fungicidal Concentrations (MFCs). Additional assays were conducted to evaluate biofilm inhibition, biofilm eradication, cell membrane integrity, nucleotide leakage, sorbitol protection assay, efflux pump inhibition, and hemolysis assay. ResultsTwo compounds, Hydramethylnon (MMV1577471) and Flufenerim (MMV1794206), demonstrated significant inhibitory effects against C. auris. Hydramethylnon exhibited potent antifungal activity, inhibiting up to 93 % of fungal growth with an MFC of 16 μg/mL. Flufenerim inhibited up to 58 % of fungal growth, showing fungistatic action with an MFC greater than 4 μg/mL. Biofilm inhibition tests showed that both compounds significantly inhibited biofilm formation, with increased efficacy at higher concentrations. Both compounds showed eradication rates in both stages. Furthermore, Hydramethylnon and Flufenerim did not affect cell membrane integrity or nucleotide leakage, suggesting a mode of action not reliant on disrupting these cellular components. The sorbitol protection assay revealed that neither compound caused cell wall damage. In the efflux pump inhibition assay, Hydramethylnon did not activate efflux pumps, while Flufenerim activated efflux pumps, reducing its effectiveness. Hemocompatibility assay showed safety. ConclusionThe study highlights Hydramethylnon and Flufenerim as promising candidates for further development as disinfectants, offering potential solutions to the urgent need for effective disinfection agents against C. auris. The findings underscore the value of screening compound collections to identify novel antifungal agents and understand their mechanisms of action, thereby contributing to the advancement of new disinfection strategies in healthcare settings.

Development of bio sheet using nanoparticles and the confederation of protein for casting wound

This research pioneers a sustainable approach to wound healing by developing a bio sheet for dressings. It ingeniously incorporates healing properties of reduced graphene oxide synthesized via an eco-friendly method, using phytoextracts to eliminate toxic chemicals. Collagen extracted from fish waste and fibrinogen from post-animal waste contribute to the bio sheet's green profile. The study emphasizes the global shift towards utilizing natural materials in therapeutic applications for efficient wound healing while minimizing toxic side effects. The formulation involves a simple muffle treatment for the conversion of carbon to graphene oxide, followed by a greener reduction technique to synthesize rGO. Characterization- by peaks that obtained at 255 nm, 240 nm, 280 nm, 360 nm using UV–Vis analysis were confirmed the presents of desired carbon and glycoproteins, In X-ray diffraction (2θ value) we found peaks at 24.37°, 27.84°, 17.43° which indicated the presents of rGO, Fibrinogen, and Collagen, and FT-IR methods confirms the functional groups like OH, CH2, CC, CHO and NC in the derived materials and Zeta potential distribution were done for Compounds. To add up purity and efficiency analysis via GCMS were done for proteins. The resulting bio sheet, created in a controlled environment with a gelling agent, Undergoes morphological analysis through FESEM-EDX. Additional assessments, including antioxidant, anti-inflammatory, and hemocompatibility assays, illuminate the nature of the formulated bio sheet. To addition, biosheet is analyzed for physical and mechanical properties. Further insight is gained through the analysis of surface plots of the EDX images of both the composite and bio sheet using ImageJ software. This comprehensive approach underscores the potential of sustainable and naturally derived materials in advancing wound care technologies.

Wharton's jelly stem cells delivered via a curcumin-loaded nanofibrous wound dressings improved diabetic wound healing via upregulating VEGF and IGF genes: An in vitro and in vivo study

Diabetic wounds pose an enduring clinical hurdle, marked by delayed recovery, persistent inflammation, and an elevated susceptibility to infections. Conventional treatment approaches often fall short of delivering optimal outcomes, prompting the exploration of innovative methods to enhance the healing process. Electrospun wound dressings offer superior healing, controlled drug release, enhanced cell proliferation, biocompatibility, high surface area, and antimicrobial properties. In the current study, polycaprolactone/gelatin-based nanofibrous wound dressings were developed for the delivery of Wharton's jelly stem cells and curcumin into the diabetic wounds bed. Curcumin was loaded into the polycaprolactone/gelatin solution and electrospun to produce curcumin-loaded scaffolds. In vitro experiments including scanning electron microscopy, cell viability assay, release assay, hemocompatibility assay, cell proliferation assay, and antibacterial assay were utilized to characterize the delivery system. Then, curcumin-loaded scaffolds were seeded with 30,000 Wharton's jelly stem cells and implanted into a rat model of diabetic wounds. Study showed that the scaffolds containing both Wharton's jelly stem cells and curcumin significantly improved diabetic wound closure (86.32 3.88% at the end of 14th day), augmented collagen deposition, and improved epithelial tissue formation. Gene expression studies showed that VEGF and IGF genes were significantly upregulated by the co-delivery system. Our developed system may have augmented diabetic wound healing via upregulating pro-healing genes.

Thrombogenicity of biodegradable metals

Biodegradable metals offer a promising means to ameliorate many of the long-term risks associated with vascular devices made of conventional biostable stent metals. While numerous biodegradable metal alloys have been developed and characterized in animal models, knowledge of their blood reactivity and thrombogenicity remains unknown. Metal hemocompatibility is particularly valuable because current generation drug-eluting stents pose a significant long-term thrombosis risk. In this study, four pure metals, widely used as degradable base materials (Fe, Zn, Mg, and Mo), and three alloys commonly used in cardiovascular devices [NiTi, CoCr, and stainless steel (SS)] were evaluated. This work examined how each of these metals activate platelets, coagulation factors, and inflammation using in vitro hemocompatibility assays and a clinically relevant ex vivo non-human primate arteriovenous shunt model. Testing found that while all metals promoted a downstream activation of platelets and coagulation in flowing whole blood, platelet and fibrin attachment to Mg was markedly reduced. Additionally, Fe and Mo trended toward higher platelet attachment and contact pathway activation. Overall, the results suggest that Mg may delay clot initiation, but not eliminate clot formation, indicating the importance of understanding thrombosis in Mg alloys that are currently being developed for clinical use as biodegradable stents.

Preparation of protein-stabilized Litsea cubeba essential oil nano-emulsion by ultrasonication: Bioactivity, stability, in vitro digestion, and safety evaluation

Litsea cubeba essential oil (LCEO) has garnered widespread attention due to its robust biological activity. However, challenges such as high volatility, limited water solubility, and low bioavailability impede its application. Nano-emulsion encapsulation technology offers an effective solution to these issues. In this study, we prepared litsea cubeba essential oil nano-emulsion (LCEO-NE) for the first time using whey protein (WP) as the emulsifier through an ultrasonic-assisted method, achieving high efficiency with minimal energy consumption. Transmission electron microscopy and dynamic light scattering analyses revealed that the nanoparticles were uniformly spherical, with a particle size of 183.5 ± 1.19 nm and a zeta potential of −35.5 ± 0.95 mV. Stability studies revealed that LCEO-NE exhibited excellent thermal and salt stability, maintaining its integrity for up to four weeks when stored at 4 °C and 25 °C. In vitro digestion assays confirmed the digestibility of LCEO-NE. Furthermore, evaluation of the DPPH, ABTS, and antimicrobial activities revealed that LCEO-NE displayed superior bacteriostatic and antioxidant properties compared to LCEO. Scanning electron microscopy elucidated that its bacteriostatic effect involved the disruption of bacterial microstructure. Hemocompatibility and cytotoxicity assays demonstrated the safety of LCEO-NE within the effective concentration range. This research supports the utilization of nanoparticles for encapsulating LCEO, thereby enhancing its stability and bioactivity, and consequently expanding its applications in the food and pharmaceutical industries.

Exploring the efficacy of catha edulis extract-loaded nanofibrous scaffolds seeded with bone marrow-derived stem cells for diabetic wound healing: A preclinical investigation

Objectives: The aim of this study was to investigate the potential of incorporating catha edulis extract into polycaprolacton/gelatin scaffolds using electrospinning technique for the treatment of diabetic wounds in a rat model. Methods: The in vitro characterization of the scaffolds was performed using various assays, including anti-inflammatory assay, microstructure study, DPPH radical scavenging assay, cell viability assay, hemocompatibility assay, and bacterial penetration assays. The scaffolds were then seeded with bone marrow-derived stem cells and cultured before implantation into the rat model. Results: The results of the in vitro study showed that the produced scaffolds were nanofibrous, antioxidative, and non-toxic to skin cells. In vivo study demonstrated that the stem cell and catha edulis extract-loaded scaffolds had the highest rate of wound closure and histomorphometric parameters compared to other groups. Moreover, gene expression studies showed that the developed wound dressings increased the expression of VEGF gene and reduced the expression of glutathione peroxidase gene. Conclusion: These findings suggest that the catha edulis extract-loaded polycaprolacton/gelatin scaffolds could be a promising therapeutic option for diabetic wounds.

REDV-Functionalized Recombinant Spider Silk for Next-Generation Coronary Artery Stent Coatings: Hemocompatible, Drug-Eluting, and Endothelial Cell-Specific Materials.

Coronary artery stents are life-saving devices, and millions of these devices are implanted annually to treat coronary heart disease. The current gold standard in treatment is drug-eluting stents, which are coated with a biodegradable polymer layer that elutes antiproliferative drugs to prevent restenosis due to neointimal hyperplasia. Stenting is commonly paired with systemic antiplatelet therapy to prevent stent thrombosis. Despite their clinical success, current stents have significant limitations including inducing local inflammation that drives hyperplasia; a lack of hemocompatibility that promotes thrombosis, increasing need for antiplatelet therapy; and limited endothelialization, which is a critical step in the healing process. In this research, we designed a novel material for use as a next-generation coating for drug-eluting stents that addresses the limitations described above. Specifically, we developed a recombinant spider silk material that is functionalized with an REDV cell-adhesive ligand, a peptide motif that promotes specific adhesion of endothelial cells in the cardiovascular environment. We illustrated that this REDV-modified spider silk variant [eADF4(C16)-REDV] is an endothelial-cell-specific material that can promote the formation of a near-confluent endothelium. We additionally performed hemocompatibility assays using human whole blood and demonstrated that spider silk materials exhibit excellent hemocompatibility under both static and flow conditions. Furthermore, we showed that the material displayed slow enzyme-mediated degradation. Finally, we illustrated the ability to load and release the clinically relevant drug everolimus from recombinant spider silk coatings in a quantity and at a rate similar to that of commercial devices. These results support the use of REDV-functionalized recombinant spider silk as a coating for drug-eluting stents.

Fabrication and properties of PLA/β-TCP scaffolds using liquid crystal display (LCD) photocuring 3D printing for bone tissue engineering.

Introduction: Bone defects remain a thorny challenge that clinicians have to face. At present, scaffolds prepared by 3D printing are increasingly used in the field of bone tissue repair. Polylactic acid (PLA) has good thermoplasticity, processability, biocompatibility, and biodegradability, but the PLA is brittle and has poor osteogenic performance. Beta-tricalcium phosphate (β-TCP) has good mechanical properties and osteogenic induction properties, which can make up for the drawbacks of PLA. Methods: In this study, photocurable biodegradable polylactic acid (bio-PLA) was utilized as the raw material to prepare PLA/β-TCP slurries with varying β-TCP contents (β-TCP dosage at 0%, 10%, 20%, 30%, 35% of the PLA dosage, respectively). The PLA/β-TCP scaffolds were fabricated using liquid crystal display (LCD) light-curing 3D printing technology. The characterization of the scaffolds was assessed, and the biological activity of the scaffold with the optimal compressive strength was evaluated. The biocompatibility of the scaffold was assessed through CCK-8 assays, hemocompatibility assay and live-dead staining experiments. The osteogenic differentiation capacity of the scaffold on MC3T3-E1 cells was evaluated through alizarin red staining, alkaline phosphatase (ALP) detection, immunofluorescence experiments, and RT-qPCR assays. Results: The prepared scaffold possesses a three-dimensional network structure, and with an increase in the quantity of β-TCP, more β-TCP particles adhere to the scaffold surface. The compressive strength of PLA/β-TCP scaffolds exhibits a trend of initial increase followed by decrease with an increasing amount of β-TCP, reaching a maximum value of 52.1MPa at a 10% β-TCP content. Degradation rate curve results indicate that with the passage of time, the degradation rate of the scaffold gradually increases, and the pH of the scaffold during degradation shows an alkaline tendency. Additionally, Live/dead staining and blood compatibility experiments suggest that the prepared PLA/β-TCP scaffold demonstrates excellent biocompatibility. CCK-8 experiments indicate that the PLA/β-TCP group promotes cell proliferation, and the prepared PLA/β-TCP scaffold exhibits a significant ability to enhance the osteogenic differentiation of MC3T3-E1 cells in vitro. Discussion: 3D printed LCD photocuring PLA/β-TCP scaffolds could improve surface bioactivity and lead to better osteogenesis, which may provide a unique strategy for developing bioactive implants in orthopedic applications.

Small-diameter vascular graft composing of core-shell structured micro-nanofibers loaded with heparin and VEGF for endothelialization and prevention of neointimal hyperplasia

Despite the significant progress made in recent years, clinical issues with small-diameter vascular grafts related to low mechanical strength, thrombosis, intimal hyperplasia, and insufficient endothelialization remain unresolved. This study aims to design and fabricate a core-shell fibrous small-diameter vascular graft by co-axial electrospinning process, which will mechanically and biologically meet the benchmarks for blood vessel replacement. The presented graft (PGHV) comprised polycaprolactone/gelatin (shell) loaded with heparin-VEGF and polycaprolactone (core). This study hypothesized that the shell structure of the fibers would allow rapid degradation to release heparin-VEGF, and the core would provide mechanical strength for long-term application. Physico-mechanical evaluation, in vitro biocompatibility, and hemocompatibility assays were performed to ensure safe in vivo applications. After 25 days, the PGHV group released 79.47 ± 1.54% of heparin and 86.25 ± 1.19% of VEGF, and degradation of the shell was observed but the core remained pristine. Both the control (PG) and PGHV groups demonstrated robust mechanical properties. The PGHV group showed excellent biocompatibility and hemocompatibility compared to the PG group. After four months of rat aorta implantation, PGHV exhibited smooth muscle cell regeneration and complete endothelialization with a patency rate of 100%. The novel core-shell structured graft could be pivotal in vascular tissue regeneration application.

Bioactive glass incorporated dressing matrix for rapid hemostatic action with antibacterial activity

Uncontrolled bleeding stands as a leading cause of preventable death in both civilian trauma and military battlefield scenarios. Existing hemostatic dressings like HemCon, Celox, and QuickClot, often lack instant hemostasis at the bleeding site and may have biodegradability and exothermic issues. To address the above in this communication, we have synthesized a hemostatic glass (acronym AlBG, CaO-SiO2 system, incorporated with Al2O3 and ZnO) by sol gel route and carried out detail physicochemical characterizations, e.g., XRD, FESEM, FTIR, BET and particle size analysis etc. The AlBG of particle size range 140–253 nm was incorporated into the nonwoven surgical cotton gauze to obtain AlBGscg. In vitro biological assays including cytocomtability, hemocompatibility assays of the above coated gauze was undertaken using NIH3T3 of which, the later showed optimum hemocompatibility with <6 % lysis of red blood cells. Potent antibacterial action on both gram positive and gram negative strains were obtained with significant zone of inhibition of 17 ± 0.34 mm and 16 ± 0.56 mm, respectively. Importantly, significant reduction of the clotting time (31.81 ± 0.12 % for P time and 33.3 ± 0.23 % for aPTT) compared to the control group, indicated activation of both intrinsic and extrinsic coagulation pathways. Scanning electron microscope (SEM) images exhibited an enhanced in vitro blood clot formation in case of AlBGscg in comparison to the control (uncoated surgical cotton gauze), visually. Additionally, a noticeable reduction of ∼30 % in the time required for the blood clot formation, in case of AlBGscg, was observed. The in vivo hemostatic potential of AlBGscg was evaluated in Wistar rats through femoral artery injury and was compared with commercially available surgical cotton gauze. The results demonstrated substantial reduction in the time required for hemostasis (approximately 50 % reduction), a lower amount of blood loss (7.33 % with AlBGscg compared to 22.31 % with surgical cotton gauze), and importantly, there were no incidents of re-bleeding observed with AlBGscg, highlighting its promise as a candidate for effective hemostasis.

Characterization and Hemocompatibility Assay of Microencapsulation Chitosan-Alginate Single Garlic Extract-loaded (MCA-SGE)

Characterization and Hemocompatibility Assay of Microencapsulation Chitosan-Alginate Single Garlic Extract-loaded (MCA-SGE)

Electrospun gelatin-polyvinyl alcohol-silk fibre/hydroxyapatite scaffolds and their physicochemical and biological studies

The present study is focused on the preparation of a gelatin-polyvinyl alcohol (PVA)-silk fibre (SF)/hydroxyapatite (HAP) composite. Here, HAP acted as a reinforcement in the Gelatin-PVA-SF matrix to enhance the mechanical and biological properties. The gelatin-PVA-SF/HAP (10–70 wt% of HAP was added to 10:70:20 of gelatin-PVA-SF) nanocomposite was prepared via the electrospinning method and characterised by XRD, FTIR, and SEM-EDAX techniques. The highest tensile strength (93.81 MPa) was observed for 60 wt% of HAP-added composite. To evaluate the scaffold's suitability for bone tissue engineering applications, antimicrobial activity, hemocompatibility with human blood and bioactivity studies were carried out. The zone of inhibition against the microbes is as follows: E. coli - 52 mm, S. aureus - 48 mm, C. albicans - 42 mm and hemocompatibility assay showed less hemolytic (%) 1.24% for gelatin/PVA/SF with 60 wt% of HAP composite. The results suggest that gelatin/PVA/silk fibre with 60 wt% of HAP nanocomposite could be an effective and potential candidate for bone tissue engineering applications based on the mechanical, antimicrobial activity and apatite formation ability. Cytotoxicity evaluation was carried out using L929 cells (mouse fibroblast cells) for the gelatin/PVA/silk fibre with 60 wt% of HAP nanocomposite.

Chitosan-Reinforced Gelatin Microspheres-Modified Glass Ionomer Cement (GIC): A Novel Bone Alloplast Graft Material Synthesis and an In Vivo Analysis.

Aim and objective The study aimed to assess and evaluate the efficacyof glass ionomer modified with chitosan-reinforced gelatin microspheres on bone formation. Materials and methods The study involved three groups: Group I comprised plain glass ionomer cement; Group II comprised glass ionomer cement/gelatin (70:30 wt%); in Group III, glass ionomer cement/gelatin/chitosan (70:30%) scaffold were made into discs; the gelatin microspheres were synthesized by oil emulsion method. The synthesized scaffold was subjected to the following in vitro testing, Instron Universal Testing Machine (UTM), U3000, (Instron Corporation, Norwood, Massachusetts, United States) to assess compressive strength, scanning electron microscope(SEM) examination, and biocompatibility testing using hemocompatibility assay. The material was then testedin vivo; male Wistar albino rats, a total of nine animals, were utilized for this purpose. Three animals were used in each group; a femoral defect model was the model of choice for the experiment and the animals were observed for a period of four weeks, following which the animals were sacrificed and sent for histopathological analysis. Results The compression testing was carried out using UTM; test group I was 33 MPa, test group II was 2.3 MPa, and test group III was 25.75 MPa. SEM (JSM-IT800 Schottky Field Emission NANO SEM (JEOL, Tokyo, Japan)) analysis was done to observe the porosity of the fabricated scaffold with the average measurement of0.12 ± 0.2 μm in test group II and 0.29 ± 0.4μm in test group III. Hemocompatibility reports noted 0.4-0.8% lysis for the synthesized scaffolds. Histopathology staining of the femur defects showed that group III favoured bone formation. One-way analysis of variance (ANOVA) and post hoc Bonferroni test was done on the data. The optical density values of the alizarin red stained slide showed statistical significance for group III. Conclusion In conclusion, the synthesized scaffolds are biocompatible, distribution of porosity andpore characteristics in the glass ionomer cement/gelatin/chitosan group is better than that of the glass ionomer cement/gelatin group. The glass ionomer cement/gelatin/chitosan group had better compressive strength and induced more bone formation compared to the other test group and the control. Thus, the novel glass ionomer modified with chitosan-reinforced gelatin microspheres has optimal properties to be used as a bone graft material.