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Gamma irradiation, which is one of the more conventional sterilization methods, was used to induce the hydrogelation of silk fibroin in this study. The physical and chemical characteristics of the irradiation-induced silk fibroin hydrogels were investigated. Silk fibroin solution with a concentration greater than 1 wt% formed hydrogel when irradiated by gamma rays at a dose of 25 or 50 kGy. The hydrogel induced by 50 kGy of radiation was more thermally stable at 80 °C than those induced by 25 kGy of radiation. When compared to the spontaneously formed hydrogels, the irradiated hydrogels contained a greater fraction of random coils and a lower fraction of β-sheets. This finding implies that gelation via gamma irradiation occurs via other processes, in addition to crystalline β–sheet formation, which is a well-established mechanism. Our observation suggests that crosslinking and chain scission via gamma irradiation could occur in parallel with the β–sheet formation. The irradiation-induced hydrogels were obtained when the solution concentration was adequate to support the radiation crosslinking of the silk fibroin chains. This work has, therefore, demonstrated that gamma irradiation can be employed as an alternative method to produce chemical-free, random coil-rich, and sterilized silk fibroin hydrogels for biomedical applications. [ABSTRACT FROM AUTHOR]

Advanced wound dressings that can deliver potent antibacterial action are still much in need, especially for treating wound infections caused by drug-resistant bacteria. In this research, we utilized electron beam (EB) irradiation to develop antibacterial hydrogel sheet dressings from poly(vinyl alcohol) (PVA) and silver nanoparticles (AgNPs) in a two-step processing and evaluated their bactericidal efficacy, as well as the AgNP release. The effect of the irradiation dose on the swelling, gel fraction, network parameters, and mechanical properties of the hydrogels was first determined to establish the optimal doses for the two-step processing. The prototypic hydrogel sheets were then formed in the first EB irradiation and served as a matrix for the AgNP synthesis by the reduction of the silver nitrate precursors during the second EB irradiation. The diffusion assay showed that the minimal inhibition concentration (MIC) of the AgNP-load hydrogels was 0.25 and 0.5 mg/cm2 against Escherichia coli and Staphylococcus aureus, respectively. At these MIC levels, the released AgNPs increased sharply before reaching the maximum, ~950 and 1800 ppb, at 24 h as analyzed by atomic absorption. Therefore, we successfully demonstrated that this two-step processing by EB irradiation provides a convenient platform to fabricate AgNP-loaded hydrogel dressings that can be further developed for wound healing. [ABSTRACT FROM AUTHOR]

The use of bioactive molecules derived from medicinal plants in wound healing has recently attracted considerable attention in both research and public interest. In this work, we demonstrated the first attempt to incorporate the extract from Thai red onion skins in hydrogel patches intended for transdermal delivery. The red onion skin extract (ROSE) was first prepared and evaluated for cytotoxicity by MTT assay with both L929 and human dermal fibroblast cells. Hydrogel patches with porous microstructure and high water content were fabricated from polyvinyl alcohol (PVA) by electron beam irradiation and characterized for their physical, mechanical, morphological, and cytocompatible properties prior to the loading of ROSE. After decontamination by electron beam irradiation, the in vitro release profile exhibited the burst release of extract from ROSE-coated hydrogel patches within 5 h, followed by the sustained release up to 48 h. Finally, evaluation of skin permeation using Franz cell setup with a newborn pig skin model showed that the permeation of ROSE from the hydrogel patch increased with time and reached the maximum of 262 µg/cm2, which was well below the cytotoxicity threshold, at 24 h. These results demonstrated that our ROSE-coated hydrogel patches could potentially be used in transdermal delivery. [ABSTRACT FROM AUTHOR]

DNA double-strand breaks (DSBs) are the most lethal form of damage to cells from irradiation. γ-H2AX (phosphorylated form of H2AX histone variant) has become one of the most reliable and sensitive biomarkers of DNA DSBs. However, the γ-H2AX foci assay still has limitations in the time consumed for manual scoring and possible variability between scorers. This study proposed a novel automated foci scoring method using a deep convolutional neural network based on a You-Only-Look-Once (YOLO) algorithm to quantify γ-H2AX foci in peripheral blood samples. FociRad, a two-stage deep learning approach, consisted of mononuclear cell (MNC) and γ-H2AX foci detections. Whole blood samples were irradiated with X-rays from a 6 MV linear accelerator at 1, 2, 4 or 6 Gy. Images were captured using confocal microscopy. Then, dose–response calibration curves were established and implemented with unseen dataset. The results of the FociRad model were comparable with manual scoring. MNC detection yielded 96.6% accuracy, 96.7% sensitivity and 96.5% specificity. γ-H2AX foci detection showed very good F1 scores (> 0.9). Implementation of calibration curve in the range of 0–4 Gy gave mean absolute difference of estimated doses less than 1 Gy compared to actual doses. In addition, the evaluation times of FociRad were very short (< 0.5 min per 100 images), while the time for manual scoring increased with the number of foci. In conclusion, FociRad was the first automated foci scoring method to use a YOLO algorithm with high detection performance and fast evaluation time, which opens the door for large-scale applications in radiation triage. [ABSTRACT FROM AUTHOR]

A quick, reliable, and reproducible biological assay to distinguish individuals with possible life-threatening risk following radiological or nuclear incidents remains a quest in biodosimetry. In this paper, we examined the use of a γ-H2AX assay as an early dose estimation for rapid triage based on both flow cytometry and image analyses. In the experiment, whole blood from 11 donors was irradiated ex vivo inside a water phantom by gamma rays from Co-60 at 0.51 Gy/min. After the lysis of red blood cells, the white blood cells were collected for immunofluorescence labeling of γ-H2AX, CD45, and nuclear stained for signal collection and visualization. Analysis by flow cytometry showed that the relative γ-H2AX intensities of lymphocytes and granulocytes increased linearly with absorbed doses from 0 to 6 Gy with a large variation among individuals observed above 2 Gy. The relative γ-H2AX intensities of lymphocytes assessed by two different laboratories were highly correlated (ICC = 0.979). Using confocal microscopic images, γ-H2AX foci were observed to be discretely distributed inside the nuclei and to increase proportionally with doses from 0 to 2 Gy, whereas large plagues of merged foci appeared at 4 and 6 Gy, resulting in the saturation of foci counts above 4 Gy. The number of total foci per cell as well as the number of foci per plane were significantly different at 0 vs 1 and 2 vs 4 Gy doses (p < 0.01). Blind tests at 0.5 Gy and 1 Gy doses showed that dose estimation by flow cytometry had a mean absolute difference of less than 0.5 Gy from the actual value. In conclusion, while flow cytometry can provide a dose estimation with an uncertainty of 0.5 Gy at doses ≤ 1 Gy, foci counting can identify merged foci that are prominent at doses ≥ 4 Gy. [ABSTRACT FROM AUTHOR]

Hydrogels are soft materials that contain high water content within their 3-dimensional structure. Such extremely hydrated environment allows hydrogels to recapitulate the structure of many native tissues inside the body. In biomedical application, hydrogels have been extensively used as biocompatible materials, drug delivery systems, and tissue-engineered scaffolds that can be designed to possess either permanent or slow-degradation properties. In this research, we applied gamma irradiation to develop transparent and conformal hydrogel sheets with sufficient mechanical strength from poly(vinyl alcohol) (PVA) and further modified the based PVA matrix with naturally-derived silk fibroin (SF) protein and silver nitrate (AgNO3) for wound healing purpose. The physical and mechanical properties of based PVA hydrogels formed at varied irradiation doses from 10 - 80 kGy were first characterized. The dose of 60 kGy was found to be optimal to process flexible and elastic PVA sheets with equilibrium degree of swelling of 1000%, gel fraction of 90%, and tensile strength of 19 kPa. To further enhance water absorption capacity, 10 - 40% (w/w) silk fibroin was added to the based PVA matrix. Based on water absorption and gel fraction data, hydrogel sheets with 8PVA:2SF formulation was selected for antibacterial test. Disc diffusion assay showed that the incorporation of 0.4 mM AgNO3 in 8PVA:2SF hydrogel sheets could inhibit the growth of Staphyllococcus aureus and Pseudomonas aeruginosa. These results demonstrated that a prototypic, antibacterial hydrogel sheet dressing composed of both synthetic and natural polymers could be developed within a single-step by gamma irradiation technique. [ABSTRACT FROM AUTHOR]

Radiolytic synthesis provides a convenient and environmentally-friendly approach to prepare metallic nanoparticles in large scale with narrow size distribution. In this report, colloidal silver nanoparticles (AgNPs) were synthesized by gamma radiation using poly(vinyl alcohol) (PVA) or silk fibroin (SF) as stabilizers and were evaluated for their antibacterial properties. The conversion of metallic silver ions to silver atoms depended on irradiation dose and stabilizer concentration as determined by UV-Vis spectrophotometry and transmission electron microscopy. The uniformly dispersed AgNPs with diameter 32.3 ± 4.40 nm were evaluated as antiseptic agents in films composed of chitosan, SF, and PVA that were processed by irradiation-induced crosslinking. Using disc diffusion assay, the films containing 432 ppm AgNPs could effectively inhibit the growth of both Staphylococcus aureus and Pseudomonas aeruginosa. Therefore, we have demonstrated in our present study that gamma radiation technique can potentially be applied in the mass production of antibacterial wound dressings. [ABSTRACT FROM AUTHOR]

Proliferation and migration of keratinocytes and fibroblasts play an important role in cutaneous wound healing, while oral mucosal squamous cell proliferation and migration are crucial for oral wound healing. In this study, the phytochemical profile of Pluchea indica branch ethanolic extract was characterized. The bioactive compound of Pluchea indica branch ethanolic extract was identified and analyzed by the validated HPLC method. The nanoparticles of P. indica branch extract were formulated by solvent displacement method to increase the solubility and the colloidal stability of the extract. The stability of the nanoparticles was investigated by using the dynamic light scattering technique. Effects of P. indica crude extract and nanoparticles on cell viability, proliferation and migration of primary epidermal keratinocytes, human dermal fibroblasts, and oral mucosal keratinocyte cells were investigated by MTT assay and scratch assay, respectively. The results showed that P. indica branch extract contained a high content of total phenolic and total flavonoids. The HPLC analysis revealed that the main compound in the extract was 4,5-O-dicaffeoylquinic acid. The cell viability of the extract and nanoparticles decreased when cells were exposed to a high concentration of extract and nanoparticles. These results demonstrate that P. indica branch extract and extract nanoparticles at specific concentrations possess in vitro wound healing activity and they may be possibly used to treat different types of wounds including dermal and oral mucosal wounds. [ABSTRACT FROM AUTHOR]

In this report, silk fibroin (SF) mats coated with silver nanoparticles (AgNPs) were manufactured as a prototypic wound dressing and evaluated for antimicrobial properties. SF was extracted from cocoons of Thai silkworms Bombyx mori (variant Nangnoi Si Sa Ket) and fabricated into nonwoven mats by electrospinning. In a one-step synthesis method, colloidal AgNPs were prepared from silver nitrate by gamma irradiation and inspected by transmission electron microscopy. Using the in vitro disc diffusion and growth-inhibition assays, AgNP-coated SF mats effectively inhibited the growth of Staphyllococus aureus and Pseudomonas aeruginosa when the coating solution containing colloidal AgNPs was 4 mM, or equivalent to 50.4 ng/cm of adsorbed AgNPs. Based on these results, the AgNP-coated SF mats can potentially be used as antimicrobial wound dressings. [ABSTRACT FROM AUTHOR]