Cell Culture Tests Research Articles

Determination of Toxicity at Different Trophic Levels of Aqueous Film-Forming Foams (AFFF) Used in Fire Fighting.

Aqueous film-forming foams (AFFF), containing perfluorinated surfactants, can reach the environment. The objective of this study was to determine the ecotoxicity of AFFF, according to the type of fire to be fought (A1: 1.05g.L- 1, A2: 3.15g.L- 1 and A3: 6.30g.L- 1), to bioindicators of different trophic levels. For Artemia salina a toxic effect was observed at sample A1 (at concentrations of 100%), A2 (at concentrations above 25%) and A3 (at concentrations above 12.5%). For Lactuca sativa all samples affected the number of germinated seeds, speed and percentage of germination and root length. To the Eisenia fetida earthworm, samples A2 and A3 were considered toxic due to the percent avoidance being 70% and 100%, respectively. In Macaca mullata renal cell culture test, none of the samples were toxic by the MTT test. Therefore, it is necessary to develop methods for the safe use of AFFF by professionals.

Improving Thermosensitive Bioink Scaffold Fabrication with a Temperature-Regulated Printhead in Robot-Assisted In Situ Bioprinting System.

In situ bioprinting enables precise 3D printing inside the human body using modified bioprinters with thermosensitive bioinks such as gelatin methacrylate (GelMA). However, these devices lack refined temperature-regulated mechanisms essential for ensuring bioink viscosity, as compared to traditional bio-3D printers. Addressing this challenge, this study presents a temperature-regulated printhead designed to improve the fabrication of thermosensitive bioink scaffolds in in situ bioprinting, integrated into a UR5 robotic arm. Featuring a closed-loop system, it achieves a temperature steady error of 1 °C and a response time of approximately 1 min. The effectiveness of the printer was validated by bioprinting multilayer lattice 3D bioscaffolds. Comparisons were made with or without temperature control using different concentrations of GelMA + LAP. The deformation of the bioscaffolds under both conditions was analyzed, and cell culture tests were conducted to verify viability. Additionally, the rheology and mechanical properties of GelMA were tested. A final preliminary in situ bioprinting experiment was conducted on a model of a damaged femur to demonstrate practical application. The fabrication of this printhead is entirely open source, facilitating easy modifications to accommodate various robotic arms. We encourage readers to advance this prototype for application in increasingly complex in situ bioprinting situations, especially those utilizing thermosensitive bioinks.

In Vitro Wound-Healing Potential of Phenolic and Polysaccharide Extracts of Aloe vera Gel.

The present study aimed to conduct a comparative investigation of the biological properties of phenolic and polysaccharide extracts obtained using an ultrasound-assisted technique from Aloe vera gel and their effects on each stage of the wound healing process in in vitro experimental models. HPLC analysis showed that the phenolic extract contained aloin, ferulic, and caffeic acid, as well as quercetin dihydrate, as major compounds. Capillary zone electrophoresis indicated the prevalence of mannose and glucose in the polysaccharide extract. Cell culture testing revealed the anti-inflammatory properties of the phenolic extract at a concentration of 0.25 mg/mL through significant inhibition of pro-inflammatory cytokines-up to 28% TNF-α and 11% IL-8 secretion-in inflamed THP-1-derived macrophages, while a pro-inflammatory effect was observed at 0.5 mg/mL. The phenolic extract induced 18% stimulation of L929 fibroblast proliferation at a concentration of 0.5 mg/mL, enhanced the cell migration rate by 20%, and increased collagen type I synthesis by 18%. Moreover, the phenolic extract exhibited superior antioxidant properties by scavenging free DPPH (IC50 of 2.50 mg/mL) and ABTS (16.47 mM TE/g) radicals, and 46% inhibition of intracellular reactive oxygen species (ROS) production was achieved. The polysaccharide extract demonstrated a greater increase in collagen synthesis up to 25%, as well as antibacterial activity against Staphylococcus aureus with a bacteriostatic effect at 25 mg/mL and a bactericidal one at 50 mg/mL. All these findings indicate that the phenolic extract might be more beneficial in formulations intended for the initial phases of wound healing, such as inflammation and proliferation, while the polysaccharide extract could be more suitable for use during the remodeling stage. Moreover, they might be combined with other biomaterials, acting as efficient dressings with anti-inflammatory, antioxidant, and antibacterial properties for rapid recovery of chronic wounds.

Research on Dual-Phase Composite Forming Process and Platform Construction of Radial Gradient Long Bone Scaffold.

The structure and composition of natural bone show gradient changes. Most bone scaffolds prepared by bone tissue engineering with single materials and structures present difficulties in meeting the needs of bone defect repair. Based on the structure and composition of natural long bones, this study proposed a new bone scaffold preparation technology, the dual-phase composite forming process. Based on the composite use of multiple biomaterials, a bionic natural long bone structure bone scaffold model with bone scaffold pore structure gradient and material concentration gradient changes along the radial direction was designed. Different from the traditional method of using multiple nozzles to achieve material concentration gradient in the scaffold, the dual-phase composite forming process in this study achieved continuous 3D printing preparation of bone scaffolds with gradual material concentration gradient by controlling the speed of extruding materials from two feed barrels into a closed mixing chamber with one nozzle. Through morphological characterization and mechanical property analysis, the results showed that BS-G (radial gradient long bone scaffolds prepared by the dual-phase composite forming process) had obvious pore structure gradient changes and material concentration gradient changes, while BS-T (radial gradient long bone scaffolds prepared by printing three concentrations of material in separate regions) had a discontinuous gradient with obvious boundaries between the parts. The compressive strength of BS-G was 1.00 ± 0.19 MPa, which was higher than the compressive strength of BS-T, and the compressive strength of BS-G also met the needs of bone defect repair. The results of in vitro cell culture tests showed that BS-G had no cytotoxicity. In a Sprague-Dawley rat experimental model, blood tests and key organ sections showed no significant difference between the experimental group and the control group. The prepared BS-G was verified to have good biocompatibility and lays a foundation for the subsequent study of the bone repair effect of radial gradient long bone scaffolds in large animals.

An Investigation of In Vitro Anti-Cancer Efficacy of Dihydroartemisinin-Loaded Bovine Milk Exosomes Against Triple-Negative Breast Cancer.

Repurposing drugs offers several advantages, including reduced time and cost compared to developing new drugs from scratch. It leverages existing knowledge about drug safety, dosage, and pharmacokinetics, expediting the process of clinical trials and regulatory approval. Dihydroartemisinin (DHA) is a semi-synthetic and active metabolite of all artemisinin molecules and is FDA-approved for the treatment of malaria. Apart from having anti-malarial properties, DHA also possesses anticancer properties. However, its pharmacological actions are limited by toxicity and solubility problems. To overcome these challenges and enhance its anticancer effectiveness, we designed an exosomal formulation of DHA. We isolated exosomes from bovine milk using differential ultracentrifugation and loaded DHA using sonication. Scanning and transition electron microscopy revealed a size of roughly 100nm, with a spherical shape. Furthermore, in pH 7.4 and 5.5, the exosomes exhibited burst release followed by sustained release. Multiple in vitro cell culture tests demonstrated that Exo-DHA exhibited enhanced anticancer activity, including cytotoxicity, cellular uptake, generation of reactive oxygen species (ROS), disruption of mitochondrial membrane potential, and inhibition of colony formation. Additional evidence supporting Exo-DHA's anti-migration ability came from transwell migration and scratch assays. Based on these results, it was concluded that the anticancer efficacy of DHA was improved when loaded into bovine milk-derived exosomes. While the in vitro results are encouraging, more in vivo testing in suitable animal models and biochemical marker analysis are warranted.

Initial bone tissue reactions of hydroxyapatite/collagen-(3-glycidoxypropyl)trimethoxysilane injectable bone paste.

We have previously reported that a novel bioresorbable self-setting injectable bone paste composed of hydroxyapatite/collagen bone-like nanocomposite (HAp/Col) and (3-glycidoxypropyl)trimethoxysilane (GPTMS) was successfully prepared and was replaced with new bone within 3 months of implantation in defects created in porcine tibia. In this study, the HAp/Col-GPTMS paste was implanted into bone defects in rat tibiae to investigate the initial kinetics and bone tissue response. Even though more than 35% of GPTMS molecules should be eluted rapidly from directly injected pastes according to previously reported cell culture tests, in this study, energy-dispersive X-ray spectrometry did not detect Si (GPTMS) deposition in tissues surrounding the paste at 1 day postimplantation. Further, no abnormal inflammatory responses were observed in the surrounding tissues over the test period for both directly injected and prehardened pastes. Companying these observations with the results of the previous animal test (in which the paste was fully resorbed and was substituted with new bone), the eluted GPTMS resolved in no harm in vivo from the initial to final (completely resorbed) stages. Material resorption rates calculated from X-ray microcomputed tomography (μ-CT) images decreased with increasing in GPTMS concentration. Histological observations indicated that tartrate-resistant acid phosphatase (TRAP) active cells, (assumed to be osteoclasts), exist on the periphery of pastes. This result suggested that the paste was resorbed by osteoclasts in the same way as the HAp/Col. Since a good correlation was observed between TRAP active areas in histological sections and material resorption rate calculated from μ-CT, the TRAP activity coverage ratio offers the possibility to estimate the osteoclastic resorption ratio of materials, which are replaced with bone via bone remodeling process.

Biomimetic Bi3+ substituted borate bioactive glasses for bone tissue engineering via enhanced biological and antimicrobial activity

Biomimetic borate-based bioactive glasses (BAGs), substituted with metal oxides, serve as prominent candidates for the repair and regeneration of both hard and soft tissues. In the current study, bismuth mixed borate BAGs (60-X) B2O3–25CaO–15Na2O–XBi2O3 (X = 2, 4, 6, 8 & 10 mol.%) were synthesized using melt-quench method and systematically evaluated for their structural properties, followed by in-vitro biological performance and enhanced antimicrobial activity, using various characterization techniques and bioassays. The BAG structure was studied by Fourier transform infrared (FTIR), differential thermal analysis (DTA), and X-ray photoelectron spectroscopy (XPS) methods. The density and mechanical properties of the BAGs were found to increase with increase in bismuth content of the borate glass network. The XPS analysis confirmed the presence of Bi3+ ions embedded in the glass network. From DTA, it is noticed that the glass transition temperature (Tg) decreased and thermal stability (ΔT) values increased, with increased Bi2O3 content. Further, the bioactivity via hydroxy apatite (HAp) layer formation was confirmed by X-ray diffraction (XRD), FTIR, scanning electron microscopy- energy dispersive spectra (SEM-EDS) and Atomic force microscopy (AFM), both before and after soaking the glass samples in simulated body fluid (SBF) for 0, 7, 14, and 21 days. The results revealed that Bi3+ modified borate BAGs exhibited good HAp layer formation on their surfaces upon immersion in SBF solution. Moreover, the pH variation and degradation followed a trend, with increased in immersion time and decrease with Bi2O3 content. Lastly, in vitro cell culture tests on MG-63 osteoblast-like cells demonstrated that the as-developed BAGs were non-toxic and cytocompatible in nature; in addition, they also displayed demonstrable antibacterial activity against S. aureus and E. coli bacterial strains. Thus, the developed Bi2O3 mixed borate BAGs become suitable candidates for bone substitutes, especially for bone tissue regeneration and repair applications.

Evaluation of tribological and biological properties of Ti6Al4V coated with Si3N4/ND nanoparticles for orthopedic applications: a comprehensive analysis

In this study, the biocompatibility and tribological properties of Ti6Al4V coated with silicon nitride (Si3N4)/nanodiamond using the electrophoretic deposition method were investigated. Suspensions of various aqueous and alcoholic solutions were prepared in the presence of CTAB and SDS dispersers. The most stable suspension system for the electrophoresis process was selected (aqueous media/ SDS disperser). Four different voltages (20, 30, 40 and 50 V) were applied to study the effect of voltage on the coating property. One could find that processing with 40 V obtained the best coating. The nano-composite coating was characterized using scanning electron microscopy equipped with energy dispersive spectroscopy, mapping analysis and x-ray diffraction after the coating process. The samples were then subjected to two nanoindentation and nano-scratching tests to evaluate their tribological properties. Biocompatibility was assessed in an ex vivo environment using two cell culture tests to evaluate survival and cellular adhesion. The results showed that the hardness and modulus elasticity of the coated sample increased from 85 to 124 GPa and 1.14–3.55 GPa, respectively, compared to the non-coated sample. Additionally, the MTT test results indicated that cellular survival and proliferation of MG63 cells increased from 86% for the non-coated sample to 92% for the Ti6Al4V/Si3N4 /ND sample. These findings have implications for orthopedic implant applications.

Gallium-containing mesoporous nanoparticles influence in-vitro osteogenic and osteoclastic activity

Mesoporous silica nanoparticles were synthesized using a microemulsion-assisted sol-gel method, and calcium, gallium or a combination of both, were used as dopants. The influence of these metallic ions on the physicochemical properties of the nanoparticles was investigated by scanning and transmission electron microscopy, as well as N2 adsorption-desorption methods. The presence of calcium had a significant impact on the morphology and textural features of the nanoparticles. The addition of calcium increased the average diameter of the nanoparticles from 80 nm to 150 nm, while decreasing their specific surface area from 972 m2/g to 344 m2/g. The nanoparticles of all compositions were spheroidal, with a disordered mesoporous structure. An ion release study in cell culture medium demonstrated that gallium was released from the nanoparticles in a sustained manner. In direct contact with concentrations of up to 100 μg/mL of the nanoparticles, gallium-containing nanoparticles did not exhibit cytotoxicity towards pre-osteoblast MC3T3-E1 cells. Moreover, in vitro cell culture tests revealed that the addition of gallium to the nanoparticles enhanced osteogenic activity. Simultaneously, the nanoparticles disrupted the osteoclast differentiation of RAW 264.7 macrophage cells. These findings suggest that gallium-containing nanoparticles possess favorable physicochemical properties and biological characteristics, making them promising candidates for applications in bone tissue regeneration, particularly for unphysiological or pathological conditions such as osteoporosis.

Evaluation of Biocompatibility Properties of Dental Materials: xCELLigence® System

The toxic and biological impacts of dental materials play a pivotal role in their clinical application within dentistry. The assessment of these materials typically commences with in vitro tests upon initial development, progressing to in vivo animal experiments and clinical trials. In vitro cell culture tests afford the examination of tissue responses at the cellular level, allowing the observation of physiological activities. Moreover, these tests offer a cost-effective and time-efficient alternative to animal experiments, rendering them easily applicable and replicable. Recently, real-time cell analysis systems, such as the xCELLigence® system, have emerged as a promising substitute for traditional testing methods, potentially surpassing them in the biocompatibility evaluation of dental materials. The xCELLigence® system facilitates the concurrent observation and analysis of cells within their authentic environment, obviating the need for cell staining or marking. This review seeks to underscore the advantageous features of the xCELLigence® system, which serves to mitigate the drawbacks associated with conventional in vitro biocompatibility evaluation methods.

Fabrication of porous poly(L-lactide-co-ε-caprolactone) micropowder for microbubble effect and ultrasound-mediated drug delivery.

Biodegradable elastic poly(L-lactide-co-ε-caprolactone) (PLCL) copolymer (50:50, lactide:caprolactone molar ratio) was synthesized and porous PLCL micropowders was fabricated by a simple method involving rapid cooling of 0.1, 0.5, and 1% (wt/vol) PLCL/dioxane spray into liquid nitrogen. The physicochemical properties of the porous PLCL micropowders were examined by measuring their pore size, pore morphology, and microbead size using a scanning electron microscopy (SEM) and dye and temozolomide (TMZ)-release testing under ultrasound. Human U-87MG, glioblastoma (GBM) cell culture tests were performed to evaluate cell cytotoxicity by released drug from PLCL micropowders. In this study, the porous PLCL micropowders prepared from 1 wt%/vol% PLCL solutions showed a highly porous structure, satisfactory mechanical properties, and optimal drug release efficiency compared with those produced from 0.1 or 0.5 wt%/vol% solutions. The results of the accumulated release test with the results of the absorbance of the dye initially applied, it was confirmed that more than 80% of the added dye was trapped inside the micropowder, and clearly GBM cytotoxicity effect could be observed by the released TMZ. The drug release system using micropowders and ultrasound can be applied as a drug supply system for various diseases such as brain tumors with low drug permeability.

Targeting CD33+ Acute Myeloid Leukemia with GLK-33, a Lintuzumab-Auristatin Conjugate with a Wide Therapeutic Window.

CD33 (Siglec-3) is a cell surface receptor expressed in approximately 90% of acute myeloid leukemia (AML) blasts, making it an attractive target for therapy of AML. Although previous CD33-targeting antibody-drug conjugates (ADC) like gemtuzumab ozogamicin (GO, Mylotarg) have shown efficacy in AML treatment, they have suffered from toxicity and narrow therapeutic window. This study aimed to develop a novelADCwith improved tolerability and a wider therapeutic window. GLK-33 consists of the anti-CD33 antibody lintuzumab and eight mavg-MMAU auristatin linkerpayloads per antibody. The experimental methods included testing in cell cultures, patient-derived samples, mouse xenograft models, and rat toxicology studies. GLK-33 exhibited remarkable efficacy in reducing cell viability within CD33-positive leukemia cell lines and primary AML samples. Notably, GLK-33 demonstrated antitumor activity at single dose as low as 300 mg/kg in mice, while maintaining tolerability at single dose of 20 to 30 mg/kg in rats. In contrast with both GO and lintuzumab vedotin, GLK-33 exhibited a wide therapeutic window and activity against multidrug-resistant cells. The development of GLK-33 addresses the limitations of previous ADCs, offering a wider therapeutic window, improved tolerability, and activity against drug-resistant leukemia cells. These findings encourage further exploration of GLK-33 in AML through clinical trials.

The assessment of composition, biological properties, safety and molecular docking of corn silk (Zea mays L.) extracts from the valorization of agricultural waste products in Thailand

Corn, known as Zea mays L., is an important source of nutrition for humans and animals. Corn silk is a byproduct of corn which is typically disposed in agriculture of Thailand. Examination of the composition, biological properties, safety and molecular docking interactions of corn silk extracts was the objective of this research. The crude extracts were determined for percentage yield, phytochemical composition, antioxidative activity, enzyme inhibition activities, safety profile and active content analysis. The corn silk extract with water by reflux technique (Rcsw) had the highest percentage yield and total phenolic content at 16.23% and 23.87±3.22 mgGAE/g extract, respectively, while the corn silk extract with 50% ethanol by reflux technique (Rcs50) gave the highest total flavonoid content at 220.58±47.51 mgQE/g extract. The four selected corn silk extracts had cosmeceutical potential including antioxidant activities, tyrosinase inhibition, anti-aging activity and a non-toxic profile on HET-CAM and cell culture test. The content analysis by HPLC was the highest in the corn silk extract with water by ultrasonication (Ucsw) at 18.70±2.12 mg GA/g and 903.29±23.18 mg QE/g. The comprehension of the molecular mechanisms that affected the interaction between active ingredients and the structures of tyrosinase, collagenase and hyaluronidase has been provided via molecular docking of the extracts. The findings of this research have illustrated the potential of corn silk extracts in the advancement of innovative health care products.

Nanoencapsulation of Ruthenium Complex Ru(ThySMet): A Strategy to Improve Selective Cytotoxicity against Breast Tumor Cells in 2D and 3D Culture Models.

Ruthenium complexes have shown promise in treating many cancers, including breast cancer. Previous studies of our group have demonstrated the potential of the trans- [Ru(PPh3)2(N,N-dimethylN'-thiophenylthioureato-k2O,S)(bipy)]PF6 complex, the Ru(ThySMet), in the treatment of breast tumor cancers, both in 2D and 3D culture systems. Additionally, this complex presented low toxicity when tested in vivo. Improve the Ru(ThySMet) activity by incorporating the complex into a microemulsion (ME) and testing its in vitro effects. The ME-incorporated Ru(ThySMet) complex, Ru(ThySMet)ME, was tested for its biological effects in two- (2D) and three-dimensional (3D) cultures using different types of breast cells, MDAMB- 231, MCF-10A, 4T1.13ch5T1, HMT-3522 and Balb/C 3T3 fibroblasts. An increased selective cytotoxicity of the Ru(ThySMet)ME for tumor cells was found in 2D cell culture, compared with the original complex. This novel compound also changed the shape of tumor cells and inhibited cell migration with more specificity. Additional 3D cell culture tests using the non-neoplastic S1 and the triple-negative invasive T4-2 breast cells have shown that Ru(ThySMet)ME presented increased selective cytotoxicity for tumor cells compared with the 2D results. The morphology assay performed in 3D also revealed its ability to reduce the size of the 3D structures and increase the circularity in T4-2 cells. These results demonstrate that the Ru(ThySMet)ME is a promising strategy to increase its solubility, delivery, and bioaccumulation in target breast tumors.

Argon plasma-modified bacterial nanocellulose: Cell-specific differences in the interaction with fibroblasts and endothelial cells

Unique properties of bacterial nanocelullose (BNC), such as high purity, water retention, nanofibrillar structure and non-cytotoxicity, allow its application in regenerative medicine, particularly for active wound healing and for skin tissue engineering. However, its biological activity is not fully sufficient to be considered a good cell scaffold. This can be enhanced by physicochemical modifications, particularly plasma treatment. In this work, air-dried (AD) or lyophilized (L) BNC samples were modified in a direct current Ar+ plasma discharge (PM). BNC_AD samples showed higher surface roughness, tensile strength and Young's modulus but a lower swelling ratio than BNC_L samples. The swelling ratio of BNC_L samples further increased after PM. The samples were subjected to six-day in vitro cell culture tests with normal human dermal fibroblasts (NHDFs) or human umbilical vein endothelial cells (HUVECs), i.e. cell types important for skin defect healing. NHDFs adhered in higher numbers, by a larger cell spreading area and reached higher cell population densities on PM samples than on unmodified samples. However, in HUVECs, this effect of PM on cell growth was less pronounced or even opposite, especially on mechanically weaker and more swellable BNC_L. At the same time, both cell types preferred mechanically stronger BNC_AD for their adhesion and growth, which was more apparent in HUVECs. The expression of mRNA for markers of cell adhesion and phenotypic maturation in NHDFs (talin, vinculin, CD90) was generally similar on BNC_PM and on tissue culture polystyrene. In HUVECs, the expression of vinculin and PECAM-1 on all BNC samples was lower than on polystyrene, but the expression of KDR (VEGF receptor 2) was higher, especially on BNC_PM. These results indicate cell type-specific differences in the response to various BNC modifications, which must be properly balanced to accommodate various cell types needed for wound healing and skin tissue engineering.

Onion (Allium cepa L.) Skin Waste Valorization: Unveiling the Phenolic Profile and Biological Potential for the Creation of Bioactive Agents through Subcritical Water Extraction.

Onion skin waste (OSW), the primary non-edible byproduct from onion processing, offers a renewable source of bioactive compounds. This study aims to valorize OSW through subcritical water extraction (SWE), aligning with a circular economy and biorefinery principles. SWE was carried out at 145 °C and 50 bar for 50 min in a discontinuous reactor, producing a phenolic-rich extract (32.3 ± 2.6 mg/g) dominated by protocatechuic acid (20.3 ± 2.5 mg/g), quercetin-4'-O-glucoside (7.5 ± 0.2 mg/g), and quercetin (3.2 ± 0.6 mg/g). Additionally, the extract contains sugars (207.1 ± 20.3 mg sucrose-Eq/g), proteins (22.8 ± 1.6 mg BSA-Eq/g), and free amino acids (20.4 ± 1.2 mg arginine-Eq/g). Its phenolic richness determines its scavenging activity against ●NO and O2●- radicals and its α-glucosidase and aldose-reductase inhibition without affecting α-amylase. Notably, the extract demonstrates significant α-glucosidase inhibition (IC50 = 75.6 ± 43.5 µg/mL), surpassing acarbose (IC50 = 129.5 ± 1.0 µg/mL) in both pure enzyme and cell culture tests without showing cytotoxicity to AGS, HepG2, and Caco-2 human cell lines. The extract's bioactivity and nutritional content make it suitable for developing antioxidant and antidiabetic nutraceutical/food components, highlighting SWE's potential for OSW valorization without using organic solvents.

Cell chip device for real-time monitoring of drug release from drug-laden microparticles.

A cell chip is a microfluidic cell culture device fabricated using microchip manufacturing methods for culturing living cells in a micrometer-sized chamber to model the physiological functions of tissues and organs. It has been extensively investigated in the domain of drug transport and toxicity research. Herein, we developed a cell chip for real-time monitoring of drug release from drug carriers. The proposed system integrates three core functions: cell culture, real-time analysis, and drug delivery tests. This device was designed to be loaded with microparticles for drug release and to enable real-time drug measurement. The efficacy of the developed system was evaluated by measuring the concentration of drugs released from the microparticles prepared with poly(lactic-co-glycolic acid) (PLGA). Doxorubicin, an anticancer drug, was used as a model drug and A549 cells, a type of lung cancer cell, were simultaneously cultured to compare the drug release concentrations in the presence of cells. Furthermore, variations in cell viability with respect to the presence of drug-loaded microparticles were observed and analyzed. Notably, as the proposed system requires an extremely small number of microparticles, it affords simple implementation in a single device, thereby eliminating the need for complex accessories and instruments for analysis. Thus, the analysis process becomes more convenient and cost-efficient. Thus, the proposed method offers an easy analysis of the release behavior of various cells and drugs. The simplicity and low cost of this innovative system without sacrificing analytical precision demonstrate its potential for applications across various fields.

Clinical and Molecular Detections of Lumpy Skin Disease: Possibilities of Co-infection with Foot-and-Mouth Disease

Lumpy skin disease (LSD) and foot and mouth disease (FMD) are notable viral diseases of cattle. This report aimed to highlight the possibilities of an uncommon case of LSD and FMD co-infection. The report also presents the clinical and molecular detection of LSD virus in six crossbred calves and LSD and FMD virus co-infection in a heifer at small-scale dairy farms located in northern Ethiopia. Nasal swabs and tissue samples were collected following aseptic techniques from the six calves suspected of having LSD and a tissue sample from one heifer suspected of having LSD-FMD co-infection and submitted to the laboratory for cell culture and real-time polymerase chain reaction (PCR) tests. Different-sized, firm, painful skin nodules with necrotic centers were seen on different parts of the calves’ body. Swelling of the prescapular and prefemoral lymph nodes, conjunctivitis, and corneal cloudiness were also observed. Uniquely, one heifer was seen with erosive lesions in the oral cavity and tongue, salivation, lameness, and skin nodules. Intracytoplasmic inclusion bodies, a distinctive feature of LSD virus, and the formation of syncytia, a characteristic of FMD virus, were observed in the cell lines. The heifer was diagnosed with a rare co-infection of LSDV and FMDV based on clinical signs, cell culture, and real-time PCR test results. The other six calves were diagnosed with the LSD virus. Treatment with broad-spectrum antibiotics, local wound cleansing, and anti-inflammatory drugs was initiated. Unfortunately, the heifer with LSD-FMD co-infection died while under treatment, and just three calves with LSD were recovered. It can be concluded that vaccination of animals against both diseases and promotion of bio-security protocols in farms is more helpful than treatment, and early case reporting is also warranted to avoid losses related to the diseases.

Enhancement of in vitro antibacterial activity and bioactivity of iodine-loaded titanium by micro-scale regulation using mixed-acid treatment.

Postoperative infection and subsequent device loss are serious complications in the use of titanium dental implants and plates for jawbone reconstruction. We have previously reported that NaOH-CaCl2 -thermal-ICl3 -treated titanium (NaCaThIo) has a nano-scale surface and exhibits antibacterial activity against Staphylococcus aureus. The present study examined the surface properties of mixed-acid treated and then iodine-treated titanium (MA-NaCaThIo), and evaluated oral antibacterial activity and cytotoxicity compared with the results obtained with NaCaThIo. MA-NaCaThIo formed a surface layer with a nano-scale network structure having microscale irregularities, and both the thickness of the surface layer (1.49 ± 0.16 μm) and the average surface roughness (0.35 ± 0.03 μm) were significantly higher than those of NaCaThIo. Furthermore, MA-NaCaThIo maintained high hydrophilicity with a contact angle of 7.5 ± 1.7° even after 4 weeks, as well as improved apatite formation, iodine ion release, and antibacterial activity against Prevotella intermedia compared to NaCaThIo. Cell culture test revealed that MA-NaCaThIo exhibited no cytotoxicity against MG-63 and Vero cells, while increased cell proliferation, ALP activity and mineralization of MG-63 compared to NaCaThIo. This treated titanium is expected to be useful for the development of next-generation titanium devices having both bone-bonding and antibacterial properties.

Diagnostic evaluation of leukocyte esterase reagent strip in bedside rapid diagnosis of spontaneous bacterial peritonitis in cirrhotic patients

Background: Spontaneous bacterial peritonitis (SBP) is the most common and serious infection of liver cirrhosis patients with ascites. If not intervened with early antibiotic treatment leads to high morbidity and mortality. Thus, early diagnosis and treatment of SBP are needed for survival. Leukocyte esterase reagent can aid in early diagnosis. Aims and Objectives: This study aimed to assess the diagnostic accuracy of the leukocyte esterase dipstick test for the diagnosis of SBP in resource poor settings. Materials and Methods: This cross-sectional study was conducted during August 2018–January 2019 on patients with cirrhotic liver disease and ascites in a tertiary care medical college hospital, Puducherry. All patients underwent abdominal paracentesis, and ascitic fluid was subjected to cell counts, biochemistry tests, culture, and leukocyte esterase test (Multistrix SG8- Seimens) at 120 s and graded with five levels. Grade 3 was positive (125 polymorphonuclear leukocytes [PMNL]/mL) and grade 4 (<500/mL), which exceeds the threshold for SBP (<250 PMNL/mL). Sensitivity, specificity, positive, and negative predictive value (PPV and NPV) was calculated. Results: A total of 50 ascitic fluids were analyzed. Nine out of 50 (18%), 16 (32%), and 4 (8%) were positive for cell count method, Leucocyte esterase reagent (LER) test and culture, respectively. The sensitivity, specificity, PPV, NPV, and diagnostic accuracy for LER test were 6.25%, 91.18%, 25%, 67.39%, and 64%, respectively. Conclusion: The LER strip test is useful for SBP diagnosis in an emergency and resource poor setting. It provides good diagnostic accuracy and high NPV and it is cost effective and better bedside tool for SBP diagnosis.