Potential novel role of the human amniotic membrane as a sustainable hemostat.

OBJECTIVES: Acute hemorrhage can cause significant morbidity and mortality arising from trauma, bleeding disorders, surgical procedures, or obstetric complications. Surgical hemostasis methods may fail to stop acute bleeding due to the complex bleeding dynamics of each bleeding type. Therefore, developing safe and effective topical hemostatic agents remains crucial. The human amniotic membrane (hAM) has established clinical evidence of effectiveness in promoting wound healing and tissue regeneration. Despite its unique biological and immunologic properties and its structural composition of established hemostatic elements, the hemostatic role of hAM has not been yet explored. The present study aimed to investigate this potential role and to describe the development protocol and characterization of hAM-derived topical hemostat. METHODS: Surface electron microscope (SEM) imaging and Fourier transform infrared (FTIR) spectroscopy were used for characterization, and mouse models with induced peritoneal and tail wound bleeding were employed to evaluate the hemostatic effectiveness using physiological studies, in comparison to a chitosan-based combat-scale hemostat. RESULTS: The hAM hemostat showed a distinctive composition by SEM and FTIR. Applying equal masses of the hAM hemostat, the commercial hemostat, or a combination reduced peritoneal wound bleeding time to averages of 108.4, 86.2, and 76.8 s, respectively, compared to the control group (300 s). Tail wound bleeding times were similarly reduced with no significant difference between the hAM and the commercial hemostat (P values = 0.29, 0.34 in peritoneal and tail wounds, respectively). Neither hemostat affected coagulation time. CONCLUSION: This study describes a simple cost-effective preparation protocol for a hAM-based hemostatic agent. The long-recognized safety, sustainability, and immunotolerance advantages of hAM can establish superiority over commercial hemostats with reported safety concerns. Robust research validation in larger-scale bleeding models is required for wider applications and severe bleeding types.

A novel long-acting antimicrobial nanomicelle spray.

Contaminated surfaces play a major role in disease transmission to humans. The vast majority of commercial disinfectants provide short-term protection of surfaces against microbial contamination. The Covid-19 pandemic has attracted attention to the importance of long-term disinfectants as they would reduce the need for staff and save time. In this study, nanoemulsions and nanomicelles containing a combination of benzalkonium chloride (BKC; a potent disinfectant and a surfactant) and benzoyl peroxide (BPO; a stable form of peroxide that is activated upon contact with lipid/membranous material) were formulated. The prepared nanoemulsion and nanomicelle formulas were of small sizes <80 nm and high positive charge >45 mV. They showed enhanced stability and prolonged antimicrobial efficacy. The antibacterial potency was evaluated in terms of long-term disinfection on surfaces as verified by repeated bacterial inoculums. Additionally, the efficacy of killing bacteria upon contact was also investigated. A nanomicelle formula (NM-3) consisting of 0.8% BPO in acetone and 2% BKC plus 1% TX-100 in distilled water (1 : 5 volume ratio) demonstrated overall surface protection over a period of 7 weeks upon a single spray application. Furthermore, its antiviral activity was tested by the embryo chick development assay. The prepared NM-3 nanoformula spray showed strong antibacterial activities against Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus as well as antiviral activities against infectious bronchitis virus due to the dual effects of BKC and BPO. The prepared NM-3 spray shows great potential as an effective solution for prolonged surface protection against multiple pathogens.

Effect of mold shape on the microstructure of gelatin sponges for tissue engineering applications.

Gelatin sponges have been used in several medical applications including tissue replacement, scaffolds, and hemostasis. Each application requires specific parameters that are tuned by the porosity of the sponges. Therefore, changes in the porosity profile of the sponges would change the sponge behavior. In this study, a gelatin solution was prepared and crosslinked with glutaraldehyde. Afterward, the solution was poured into three different mold structures with different volumes and frozen at a constant freezing rate. Each mold was investigated for its physical characteristics including swelling, degradation, porosity, crystallinity, and mechanical compression. Cube-molded gelatin sponges demonstrated high swelling capacity, degradation rate, and porosity while exhibiting low crystallinity, yield strength, and elasticity. These characteristics are suitable for hemostatic application and tissue regeneration. Therefore, it is recommended to freeze dry gelatin sponge in cuboid-shaped dimensions, for research or industry, to control the porosity and crystallinity of the sponge for the best result in biomedical applications.

Toward Scaling up the Production of Metal Oxide Nanoparticles for Application on Washable Antimicrobial Cotton Fabrics.

To examine the utilization of metal oxide nanoparticles (NPs) in different commercial products, this work focuses on the determination of cost-effective and scalable synthesis protocols. The solvothermal protocol is well-known as a scalable method but has recently been shown to lack economic feasibility. The mechanochemical method has recently been recognized to be a more economic and environmentally friendly substitute for the solvothermal method. In this study, zinc oxide nanoparticles (ZnO NPs) and copper oxide nanoparticles (CuO NPs) were synthesized using two (aqueous and organic) solvothermal (wet) methods and two (manual and automated) mechanochemical (dry) methods. The four methods were evaluated and compared. The automated mechanochemical method generated a significantly higher yield of ZnO NPs (82%) and CuO NPs (84%) using the least energy and time. However, the prepared ZnO NPs displayed higher cytotoxicity against Vero E6 cells when compared to that of CuO NPs. Because of their low cytotoxicity, CuO NPs synthesized via the automated mechanochemical method were selected for application onto cotton fabrics. Lower cytotoxicity was observed for CuO NPs treated fabrics with an IC50 of 562 mg/mL and ZnO treated fabrics with an IC50 at 23.93 mg/mL when the treated fabrics were tested against L929 fibroblast cells. Additionally, the cotton fabrics retained bactericidal and virucidal effects after four washes. Thus, the current study recommends the automated mechanochemical method as a cost-effective scalable approach for the synthesis of CuO NPs. The application of CuO NPs onto cotton fabrics generated washable antimicrobial face masks.

Multifunctional Hemostatic PVA/Chitosan Sponges Loaded with Hydroxyapatite and Ciprofloxacin.

The present study describes the development of multifunctional hemostatic sponges to control bleeding. Chitosan (Ch) and poly(vinyl alcohol) (PVA) were selected as the basic polymeric matrix [Ch/PVA] for sponges. Glycerol and citric acid were used as crosslinkers [Ch/PVA/G(Cl)] to enhance the mechanical properties of the developed sponges. Ciprofloxacin (AB) was added to the developed sponge to impart antibacterial activity. Hydroxyapatite (HA) was also added, which would make the sponge suitable for bone surgery. Among the developed sponges, the Ch/PVA/G(Cl)-HA-AB sponge demonstrated enhanced cell viability, mechanical properties, and strong antimicrobial effect against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus, in addition to platelet aggregation activity. The addition of ciprofloxacin and hydroxyapatite promotes a unique synergistic effect of antimicrobial activity and hemostasis. Thus, the present study introduces Ch/PVA/G(Cl)-HA-AB, a multifunctional hemostatic sponge that would be suitable for bone surgical applications.

Functionalized Poly(N-isopropylacrylamide)-Based Microgels in Tumor Targeting and Drug Delivery.

Over the past several decades, the development of engineered small particles as targeted and drug delivery systems (TDDS) has received great attention thanks to the possibility to overcome the limitations of classical cancer chemotherapy, including targeting incapability, nonspecific action and, consequently, systemic toxicity. Thus, this research aims at using a novel design of Poly(N-isopropylacrylamide) p(NIPAM)-based microgels to specifically target cancer cells and avoid the healthy ones, which is expected to decrease or eliminate the side effects of chemotherapeutic drugs. Smart NIPAM-based microgels were functionalized with acrylic acid and coupled to folic acid (FA), targeting the folate receptors overexpressed by cancer cells and to the chemotherapeutic drug doxorubicin (Dox). The successful conjugation of FA and Dox was demonstrated by dynamic light scattering (DLS), Fourier-transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), UV-VIS analysis, and differential scanning calorimetry (DSC). Furthermore, viability assay performed on cancer and healthy breast cells, suggested the microgels' biocompatibility and the cytotoxic effect of the conjugated drug. On the other hand, the specific tumor targeting of synthetized microgels was demonstrated by a co-cultured (healthy and cancer cells) assay monitored using confocal microscopy and flow cytometry. Results suggest successful targeting of cancer cells and drug release. These data support the use of pNIPAM-based microgels as good candidates as TDDS.

Deacetylated cellulose acetate nanofibrous dressing loaded with chitosan/propolis nanoparticles for the effective treatment of burn wounds.

Every year, about 1 out of 9 get burnt in Egypt, with a mortality rate of 37%, and they suffer from physical disfigurement and trauma. For the treatment of second-degree burns, we aim at making a smart bandage provided with control of drug release (using chitosan nanoparticles) to enhance the healing process. This bandage is composed of natural materials; namely, cellulose acetate (CA), chitosan, and propolis (bee resin) as the loaded drug. Cellulose acetate nanofibers were deacetylated by NaOH after optimizing the reaction time and the concentration of NaOH solution, and the product was confirmed with FTIR analysis. Chitosan/propolis nanoparticles were prepared by ion gelation method with size ranging from 100 to 200 nm and a polydispersity index of 0.3. Chitosan/propolis nanoparticles were preloaded in the CA solution to ensure homogeneity. Loaded deacetylated cellulose nanofibers have shown the highest hydrophobicity measured by contact angle. Cytotoxicity of propolis and chitosan/propolis nanoparticles were tested and the experimental IC50 value was about 137.5 and 116.0 µg/mL, respectively, with p-value ≤0.001. In addition, chitosan/propolis nanoparticles loaded into cellulose nanofibers showed a cell viability of 89.46% in the cell viability test. In-vivo experiments showed that after 21 days of treatment with the loaded nanofibers repairing of epithelial cells, hair follicles and sebaceous glands in the skin of the burn wound were found in albino-mice model.