L929 Cell Line Research Articles

Evaluation of Anti-Biofilm and in Vitro Wound Healing Activity of Bacterial Cellulose Loaded with Nanoparticles and Borax

Biofilms are a severe problem for public health because of the contributing recurrence of infections. Therefore, combating biofilms is a critical issue. In our study, we loaded zinc oxide (ZnO), zinc oxide borax (ZnOBorax), zinc copper oxide (ZnCuO2) nanoparticles and borax into bacterial cellulose (BC) to impart anti-biofilm and wound healing activity. The prepared BC loaded with nanoparticles (BC–NPs) was analysed via scanning electron microscopy. The nanoparticles’ geometric structure and placement in BC fibres were observed. We evaluated the biofilm inhibition and biofilm degradation activities of the BC–NPs against some pathogens via a crystal violet (CV) assay and XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2 H-tetrazolium-5-carboxanilide) reduction assay. The effects of BC–NPs on cell proliferation and wound-healing ability were analysed in L929 cell line. BC–NPs exhibited better biofilm degradation activity than biofilm inhibition activity. According to the results of the CV assay, BC–ZnONPs, BC–Borax and BC–ZnOBoraxNPs inhibited 65.53%, 71.74% and 66.60% of biofilm formation of Staphylococcus aureus, respectively. BC–ZnCuO2NPs showed the most degradation activity on Pseudomonas aeruginosa and Listeria innocua biofilms. The XTT reduction assay results indicated a considerable reduction in the metabolic activity of the biofilms. Moreover, compared to the control group, BC loaded with borax and ZnO nanoparticle promoted cell migration without cytotoxicity.

Surface modified niosomal quercetin with cationic lipid: an appropriate drug delivery system against Pseudomonas aeruginosa Infections

The Increase in infections caused by resistant strains of Pseudomonas aeruginosa poses a formidable challenge to global healthcare systems. P. aeruginosa is capable of causing severe human infections across diverse anatomical sites, presenting considerable therapeutic obstacles due to its heightened drug resistance. Niosomal drug delivery systems offer enhanced pharmaceutical potential for loaded contents due to their desirable properties, mainly providing a controlled-release profile. This study aimed to formulate an optimized niosomal drug delivery system incorporating stearylamine (SA) to augment the anti-bacterial and anti-biofilm activities of quercetin (QCT) against both standard and clinical strains of P. aeruginosa. QCT-loaded niosome (QCT-niosome) and QCT-loaded SA- niosome (QCT-SA- niosome) were synthesized by the thin-film hydration technique, and their physicochemical characteristics were evaluated by field emission scanning electron microscopy (FE-SEM), zeta potential measurement, entrapment efficacy (EE%), and in vitro release profile. The anti-P. aeruginosa activity of synthesized niosomes was assessed using minimum inhibitory and bactericidal concentrations (MICs/MBCs) and compared with free QCT. Additionally, the minimum biofilm inhibitory and eradication concentrations (MBICs/MBECs) were carried out to analyze the ability of QCT-niosome and QCT-SA-niosome against P. aeruginosa biofilms. Furthermore, the cytotoxicity assay was conducted on the L929 mouse fibroblasts cell line to evaluate the biocompatibility of the formulated niosomes. FE-SEM analysis revealed that both synthesized niosomal formulations exhibited spherical morphology with different sizes (57.4 nm for QCT-niosome and 178.9 nm for QCT-SA-niosome). The EE% for cationic and standard niosomal formulations was reported at 75.9% and 59.6%, respectively. Both formulations showed an in vitro sustained-release profile, and QCT-SA-niosome exhibited greater stability during a 4-month storage time compared to QCT-niosome. Microbial experiments indicated that both prepared formulations had higher anti-bacterial and anti-biofilm activities than free QCT. Also, the QCT-SA-niosome exhibited greater reductions in MIC, MBC, MBIC, and MBEC values compared to the QCT-niosome at equivalent concentrations. This study supports the potential of QCT-niosome and QCT-SA-niosome as effective agents against P. aeruginosa infections, manifesting significant anti-bacterial and anti-biofilm efficacy alongside biocompatibility with L929 cell lines. Furthermore, our results suggest that optimized QCT-niosome with cationic lipids could efficiently target P. aeruginosa cells with negligible cytotoxic effect.

Antimicrobial assay and controlled drug release studies with novel eugenol imprinted p(HEMA)-bacterial cellulose nanocomposite, designed for biomedical applications

In this study, a novel bio-composite material that allow sustained release of plant derived antimicrobial compound was developed for the biomedical applications to prevent the infections caused by microorganisms resistant to commercial antimicrobials agents. With this aim, bacterial cellulose (BC)-p(HEMA) nanocomposite film that imprinted with eugenol (EU) via metal chelated monomer, MAH was prepared. Firstly, characterization studies were utilized by FTIR, SEM and BET analysis. Then antimicrobial assays, drug release studies and in vitro cytotoxicity test were performed. A significant antimicrobial effect against both Gram (+) Staphylococcus aureus and Gram (-) Escherichia coli bacteria and a yeast Candida albicans were observed even in low exposure time periods. When antimicrobial effect of EU compared with commercially used agents, both antifungal and antibacterial activity of EU were found to be higher. Then, sustained drug release studies showed that approximately 55% of EU was released up to 50 h. This result proved the achievement of the molecular imprinting for an immobilization of molecules that desired to release on an area in a long-time interval. Finally, the in vitro cytotoxicity experiment performed with the mouse L929 cell line determined that the synthesized EU-imprinted BC nanocomposite was biocompatible.

Effects of Calcination Temperature on the Synthesis of One-Pot Sol-Gelled Barium Titanate Powder and Its Performance as an Endodontic Radiopacifier.

Barium titanate (BaTiO3, BTO), conventionally used for dielectric and ferroelectric applications, has been assessed for biomedical applications, such as its utilization as a radiopacifier in mineral trioxide aggregates (MTA) for endodontic treatment. In the present study, BTO powders were prepared using the sol-gel process, followed by calcination at 400-1100 °C. The X-ray diffraction technique was then used to examine the as-prepared powders to elucidate the effect of calcination on the phase composition and crystalline size of BTO. Calcined BTO powders were then used as radiopacifiers for MTA. MTA-like cements were investigated to determine the optimal calcination temperature based on the radiopacity and diametral tensile strength (DTS). The experimental results showed that the formation of BTO phase was observed after calcination at temperatures of 600 °C and above. The calcined powders were a mixture of BaTiO3 phase with residual BaCO3 and/or Ba2TiO4 phases. The performance of MTA-like cements with BTO addition increased with increasing calcination temperature up to 1000 °C. The radiopacity, however, decreased after 7 days of simulated oral environmental storage, whereas an increase in DTS was observed. Optimal MTA-like cement was obtained by adding 40 wt.% 1000 °C-calcined BTO powder, with its resulting radiopacity and DTS at 4.83 ± 0.61 mmAl and 2.86 ± 0.33 MPa, respectively. After 7 days, the radiopacity decreased slightly to 4.69 ± 0.51 mmAl, accompanied by an increase in DTS to 3.13 ± 0.70 MPa. The optimal cement was biocompatible and verified using MG 63 and L929 cell lines, which exhibited cell viability higher than 95%.

Ethanolic extract from Sophora moorcroftiana inhibit cell proliferation and alter the mechanical properties of human cervical cancer

BackgroundCervical cancer is one of the most common gynecological malignancies. Previous studies have shown that the ethanol extract of Sophora moorcroftiana seeds (EESMS) possesses an antiproliferative effect on several tumors in vitro. Therefore, in this study, we assessed the impact of EESMS on human cervical carcinoma (HeLa) cell proliferation.MethodsThe proliferation and apoptotic effects of HeLa cells treated with EESMS were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, dual acridine orange/ethidium bromide double staining, flow cytometry, and western blotting. Single-cell level atomic force microscopy (AFM) was conducted to detect the mechanical properties of HeLa cells, and proteomics and bioinformatics methods were used to elucidate the molecular mechanisms of EESMS.ResultsEESMS treatment inhibited HeLa cell proliferation by blocking the G0/G1 phase, increasing the expression of Caspase-3 and affecting its mechanical properties, and the EESMS indicated no significant inhibitory effect on mouse fibroblasts L929 cell line. In total, 218 differentially expressed proteins were identified using two-dimensional electrophoresis, and eight differentially expressed proteins were successfully identified using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The differentially expressed proteins were involved in various cellular and biological processes.ConclusionThis study provides a perspective on how cells change through biomechanics and a further theoretical foundation for the future application of Sophora moorcroftiana as a novel low-toxicity chemotherapy medication for treating human cervical cancer.

Insights into cellular interactions of characterized Mg–Al Layered Double Hydroxide on L929 cells

Layered double hydroxides are members of an anionic clay family, characterised by unique two-dimensional layered structures and lend versatility in various applications. These biocompatible compounds have the potential to get intercalated with biological compounds and physico-chemically adsorbed onto organic molecules. Thus, making them important candidates for pharmaceutical and biomedical purposes. This study aims to synthesise, characterise and investigate the cellular toxicity interactions of Mg–Al LDH towards the mouse fibroblast L929 cell line. The Mg–Al LDH was synthesized by a meticulous process of co-precipitation followed by the hydrothermal method to ensure a well-defined and stable structure for suitable biological application. Characterisation techniques like Dynamic Light Scattering, Zeta potential, Scanning Electron Microscopy, Fourier transform infrared, and X-ray diffraction analysis were employed to provide deeper insights into the physiochemical properties and structural integrity of the synthesized Mg–Al LDH. The investigation of cellular interactions with the L929 fibroblast cell line served to assess the biocompatibility and potential cytotoxic effects of Mg–Al LDH. This was observed by assessing the morphological changes and evaluating the cytotoxic effects of Mg–Al LDH by utilising various techniques like phase contrast microscopy, fluorescent staining, and Giemsa staining. The cellular metabolic activity was assessed by MTT assay, and the subcellular lysosomal alteration was examined using the fluorescent staining method by the acridine orange staining. The dose-dependent response observed in the cellular interaction underscores the importance of dosage considerations for potential biomedical applications. By elucidating the dose-response relationship, this study contributes valuable information for the safe and effective usage of LDH in biomedical contexts.

Enhanced wound regeneration by PGS/PLA fiber dressing containing platelet-rich plasma: an in vitro study

Novel wound dressings with therapeutic effects are being continually designed to improve the wound healing process. In this study, the structural, chemical, physical, and biological properties of an electrospun poly glycerol sebacate/poly lactide acid/platelet-rich plasma (PGS/PLA-PRP) nanofibers were evaluated to determine its impacts on in vitro wound healing. Results revealed desirable cell viability in the Fibroblast (L929) and macrophage (RAW-264.7) cell lines as well as human umbilical vein endothelial cells (HUVEC). Cell migration was evident in the scratch assay (L929 cell line) so that it promoted scratch contraction to accelerate in vitro wound healing. Moreover, addition of PRP to the fiber structure led to enhanced collagen deposition (~ 2 times) in comparison with PGS/PLA scaffolds. While by addition PRP to PGS/PLA fibers not only decreased the expression levels of pro-inflammatory cytokines (IL-6 and TNF-α) in RAW-264.7 cells but also led to significantly increased levels of cytokine (IL-10) and the growth factor (TGF-β), which are related to the anti-inflammatory phase (M2 phenotype). Finally, PGS/PLA-PRP was found to induce a significant level of angiogenesis by forming branching points, loops, and tubes. Based on the results obtained, the PGS/PLA-PRP dressing developed might be a promising evolution in skin tissue engineering ensuring improved wound healing and tissue regeneration.

Evaluation of Antibiofilm, Antimicrobial, Cytotoxic and Antioxidant Effects of Some Wild Mushroom Species

In this study, ethanol extracts of some wild mushroom species, in the phylum Basidiomycota, [Armillaria mellea (Vahl) P. Kumm., Infundibulicybe geotropa (Bull.) Harmaja, Leucopaxillus gentianeus (Quél.) Kotl. and Trametes versicolor (L.) Lloyd] were tested for their antioxidant, antimicrobial, antibiofilm, and cytotoxic activities. Mushroom samples showed low antimicrobial activity on Enterococcus faecalis and Staphylococcus aureus, while biofilm inhibitory activity on test microorganisms ranging from 24.6% to 80.5%. At the end of the antioxidant activity studies A. mellea was the mushroom sample having the highest DPPH radical scavenging capacity (0.105±0.001 mg/mL) whereas T. versicolor was the one having the highest iron (III) ion reducing power (40.709±0.003 µg TE/100g). The highest polyphenol content was observed in T. versicolor (29.916±0.002 mg GAE/100g) samples, and the lowest in A. mellea (9.5±0.006 mg GAE/100g) samples. The cytotoxic effects of the samples were tested on MCF-7 and MDA-MB-231 (breast cancer) and L929 (mouse fibroblast) cell lines using the MTT method. As a result, it was observed that A. mellea and T. versicolor samples were more effective on MCF-7 cell line, A. mellea on MDA-MB-231 cell line.

Exploring the Composition and Bioactivity of Hedychium flavum Leaf and Stem Essential Oil: In Vitro Assessment of Antibacterial, Antioxidant, Cytotoxicity, and Enzyme Inhibitory Activities

Hedychium flavum is an edible, medicinal, and aromatic plant widely cultivated for its essential oil. So far, little studies have been done on its phytoconstituents and bioactivities. Hence, the research aimed to determine the chemical constituents of leaf essential oil (L-EO) and stem essential oil (S-EO) of H. flavum and first estimate their antibacterial, antioxidant, cytotoxic, and enzyme inhibitory activities. According to gas chromatography-flame ionization detector/mass spectrometer (GC-FID/MS) assay, L-EO was mainly composed of β-pinene (33.4%), α-pinene (10.4%), humulene (6.8%), β-caryophyllene (6.0%), eucalyptol (6.0%), caryophyllene oxide (5.5%), endo-borneol (3.8%), humulene epoxide II (3.7%), and D-limonene (3.2%). The predominant components of S-EO were β-pinene (17.2%), eucalyptol (9.7%), nerolidol (7.8%), α-phellandrene (6.7%), α-pinene (5.8%), β-caryophyllene (5.4%), terpinen-4-ol (4.5%), D-limonene (4.4%), p-cymene (3.7%), endo-borneol (3.5%), and α-terpineol (3.5%). For the bioactivities, L-EO and S-EO showed strong antibacterial activity against Escherichia coli, Enterococcus faecalis, and Bacillus subtilis with the minimum inhibitory concentration (MIC) of 9.77–625.00 µg/mL and the minimum bactericidal concentration (MBC) of 9.77–1250.00 µg/mL. Both L-EO and S-EO exhibited moderate antioxidant activity in the DPPH (14.52 ± 0.93 mg/mL and 4.73 ± 0.15 mg/mL, respectively) and ABTS (4.05 ± 0.75 mg/mL and 2.38 ± 0.14 mg/mL, respectively) free radical scavenging capacity. S-EO showed selective cytotoxicity on human leukemic K562 cell line (IC50 = 93.94 ± 0.91 µg/mL), which was nearly three times that of noncancer L929 cell line (IC50 = 294.49 ± 9.40 µg/mL). In enzyme inhibitory properties, L-EO exerted a potent inhibition on α-glucosidase (IC50 = 1.03 ± 0.02 mg/mL), and both essential oils had weak inhibition against cholinesterase and tyrosinase. Thus, H. flavum L-EO and S-EO possess antibacterial, antioxidant, cytotoxic, and enzyme inhibitory activities with potential for exploitation in the food, cosmetic, and pharmaceutical industries.

Antiproliferative and antimicrobial compounds from Streptomyces levis: supported by in vitro and in silico molecular docking approach

The present work describes the investigation of secondary metabolites produced by rhizospheric soil isolate Streptomyces levis with respect to partial purification, bioactivity, structural elucidation, and docking study. MIC (Minimum Inhibitory Concentration) of the fraction was found to be 16 µg/mL, 32 µg/mL, and 8 µg/mL against S. aureus, E. coli, and C. albicans respectively. The fraction exhibited an IC50 of 90.48 μg/mL and 59.10 µg/mL against L929 and HeLa cell lines respectively. Spectroscopic analysis showed abundant of Isofucosterol 3-O-[6-O-(9-Octadecanoyl)-b-D-glucopyranoside], and Viniferal. These compounds were docked against receptor molecules E6, E7, Caspase-3, and Bcl2. The in-silico results suggested that all the secondary metabolites showed higher docking scores than the standard compound 5-Fluorouracil. Molecular docking study and published literature suggested that Isofucosterol 3-O-[6-O-(9-Octadecanoyl)-b-D-glucopyranoside] inactivated viral oncoprotein E7 and arrested the cell cycle from entering in S phase. Viniferal induced apoptosis by inactivation of Bcl2.

A bi-functional nanofibrous composite membrane for wound healing applications.

Various wound dressings have been developed so far for wound healing, but most of them are ineffective in properly reestablishing the skin's structure, which increases infection risks and dehydration. Electrospun membranes are particularly interesting for wound dressing applications because they mimic the extracellular matrix of healthy skin. In this study, a potential wound healing platform capable of inducing synergistic antibacterial and antioxidation activities was developed by incorporating bio-active rosmarinic acid-hydroxyapatite hybrid (HAP-RA) with different contents (0.5, 1, and 1.5 wt.%) into the electrospun polyamide 6 (PA6) nanofibers. Then, polyethylene glycol (PEG) was introduced to the nanofibrous composite to improve the biocompatibility and biodegradability of the dressing. The results indicated that the hydrophilicity, water uptake, biodegradability, and mechanical properties of the obtained PA6/PEG/HAP-RA nanofibrous composite enhanced at 1 wt.% of HAP-RA. The nanofibrous composite had excellent antibacterial activity. The antioxidation potential of the samples was assessed in vitro. The MTT assay performed on the L929 cell line confirmed the positive effects of the nanofibrous scaffold on cell viability and proliferation. According to the results, the PA6/PEG/HAP-RA nanofibrous composite showed the desirable physiochemical and biological properties besides antibacterial and antioxidative capabilities, making it a promising candidate for further studies in wound healing applications.

Thymoquinone oxime synthesis and its effects on melanoma cells: cytotoxic, genotoxic, and apoptotic evaluation

Strong evidence supports the anticancer properties of natural plant product isolates. The cytotoxic, genotoxic, and apoptotic properties of an oxime derivative of thymoquinone (TQ) in melanoma cancer cells were investigated. The structure of TQ-Oxime was elucidated through nuclear magnetic resonance, and its effect on B16F10 and L929 cell lines was assessed using a luminometric adenosine triphosphate assay. Intracellular reactive oxygen species (iROS) were quantified via fluorometry, mitochondrial membrane potential (MMP) was assessed using flow cytometry, glutathione (GSH) levels were measured using a luminometric GSH/oxidized glutathione assay, DNA damage via comet assay, and apoptosis was detected using acridine orange/ethidium bromide staining. Concentrations (0.5-20 μM) of TQ-Oxime significantly increased cytotoxicity, DNA damage, apoptosis, and iROS, in a concentration-dependent manner compared (p < 0.001). In addition, MMP and GSH levels decreased significantly with increasing concentrations compared with the control (p < 0.001). Overall, these findings contribute to our understanding of the therapeutic potential of TQ and its derivatives in cancer treatment.

Exploring etofenamate hydrazide-hydrazone/copper(II) complexes: Synthesis, anticancer activity, carbonic anhydrase IX inhibition and docking studies

Hydrazide-hydrazone derivatives have garnered significant interest from researchers globally due to their wide range of biological activities, including antiviral, anticancer, and anti-inflammatory properties. In this study, a novel series of etofenamate hydrazide-hydrazone compounds (2a-2s) and their Cu(II) complexes (3a-3s) were designed and synthesized. The compounds were characterized using various analytical techniques such as FT-IR, 1H NMR, 13C NMR, MS, and elemental analysis. Additionally, the compound 2c and Cu(II) hydrazone complex 3a were further characterized using single X-ray crystallography. The anti-proliferative activity of the compounds was evaluated against Ishikawa human endometrial cancer cell line and non-tumour L929 cells using MTT assay. Additionally, the apoptotic potential of the compounds was investigated through caspase-3 activity, Bax and Bcl-2 gene expression analysis, and annexin-V binding. Furthermore, carbonic anhydrase IX activity and in silico studies were conducted to elucidate the mechanism of action. Overall, compound 3s demonstrated significant antiproliferative effects with an IC50 value of 0.27±0.01 µM against Ishikawa cells.

Design, synthesis and characterization of novel 1,5- and 2,5-coumarin-4-yl-methyl regioisomers of 5-pyrazol-3-yl-tetrazoles as promising anticancer and antifungal agents

A novel series of compounds was synthesized using 3-arylsydnone 1a-b. Initially, 3-arylsydnone was converted into 1-aryl-1H-pyrazole-3-carbonitriles 2a-bvia [3+2] cycloaddition with acrylonitrile by ring transformation which once again underwent [2+3] cycloaddition with sodium azide to afford 5-(1-aryl-1H-pyrazol-3-yl)-1H-tetrazoles 3a-b.These were subjected to N-alkylation with 4-bromomethyl coumarins 4c-e to afford regioisomers viz., 2,5- and 1,5-disubstituted tetrazole derivatives 5f-k and 6f-k. Single crystal X-ray analysis of compound 6f was carried out. All the synthesized compounds were characterized using various techniques including 1H NMR, 13C NMR, FT-IR, elemental analysis, and mass spectroscopy.These previously unknown compounds were screened for their anticancer activity against HCT-116 colon cancer and non-cancerous L929 cell line at a single high dose (10−5 M) concentration assay. Among the tested compounds, 6g and 5f have exhibited excellent activity with very low IC50 valuei.e.,27.85 µg/mLand 37.86 µg/mL respectively, comparable with standard Cisplatin. In the non-cancerous L929 cell line, the tested compound 6f was proven to be less toxic, and all the tested compounds exhibited toxicity at higher concentrations comparable with standard Cisplatin. Further, the in vitro antifungal activity against Candida albicans for these compounds 5f-k and 6f-k revealed potential to moderate activities compared to the standard. Moreover, docking analysis was performed to know the exact binding mode of the newly synthesized compounds with N-myristoyltransferase (PDB ID: 1IYL) from Candida albicanswhich was selected as the antifungal targets. The drug-likeness of the compound was assessed using Molinspiration, while the toxicity profile was evaluated using Protox.

Slightly degradable, naturally antibacterial hydrogel matrixes derived from polyvinyl alcohol and linear/branched-polyethyleneimine as a wound dressing material

PVA:PEI based film gels were prepared at various weight ratios, 10–100% of linear-PEI (L-PEI) and branched-PEI (B-PEI) to PVA as PVA:L-PEI or PVA:B-PEI to demonstrate the effect of PEI types on hydrolytic degradation, blood compatibility, antibacterial properties, and cytotoxicity. The hydrolytic degradation studies, done at ∼ skin pH, 5.4 and revealed that PVA:PEI-based film gels, 1:0.1 wt% are degradable with a gravimetric weight losses of16.80 ± 1.39% and 17.20 ± 1.21% up to 7 days, and decreased as the wt% of L-PEI and B-PEI were increased in the matrix. The blood compatibility assays done through hemolysis and blood clotting tests disclosed only PVA:B-PEI film gels at 1.0 mg/mL concentration were found blood compatible. The antimicrobial studies revealed that regardless of the types and weight ratio of L-PEI or B-PEI, all PVA:L-PEI and PVA:B-PEI based film gels possess a potent antibacterial activity against Pseudomonas aeruginosa, a gram-negative bacteria and Staphylococcus aureus, a gram-positive bacteria. Moreover, the cytotoxicity test results examined on the L929 cell line uncovered that only PVA:L-PEI and PVA:B-PEI even at very low concentrations of L-PEI or B-PEI, for example, PVA:L-PEI and PVA:B-PEI at 1:0.1 weight ratio film gel at 10 mg/mL show toxic effect, for example <50% cell viability suggesting lesser concentration use in biomedical applications.

Enzymatic hydrolysis of wheat gluten employing cysteine proteases from noni fruits (Morinda citrifolia L.)

Previous reports have shown that noni (Morinda citrifolia L.) puree extract (NPE) is a promising source of cysteine peptidases with potential applications in the food industry. This study expanded the applications of cysteine peptidases from noni fruit by analyzing their potential for wheat gluten hydrolysis and assessing their in vitro toxicity as a new food ingredient. NPE cysteine proteases hydrolyze both gliadins and glutenins, reaching a degree of hydrolysis of 24.40% and 31.78%, respectively. In addition, they reduced gliadin content by 96.80% after 4 h of incubation, as displayed by the R5 ELISA test kit for detecting gliadin based on the monoclonal antibody R5. The chromatographic fractionation of NPE cysteine proteases on a cation exchange column (Source 30S) resulted in an unretained peak (P1) and a retained peak (P2), which showed proteolytic activity with different electrophoretic and zymographic profiles. P1 and P2 had similar (p < 0.05) degrees of hydrolysis for gliadins (approximately 9%). P1 performed better (10.68%) than P2 (8.55%) for glutenins. NPE cysteine proteases did not exhibit genotoxicity or cytotoxicity against mouse fibroblasts (L929 cell line). These findings suggest that NPE cysteine proteases have potential for use as novel ingredients to produce gluten-free foods.

3D printed kombucha biomaterial as a tissue scaffold and L929 cell cytotoxicity assay

AbstractTissue engineering includes the construction of tissue‐organ scaffold. The advantage of three‐dimensional scaffolds over two‐dimensional scaffolds is that they provide homeostasis for a longer time. The microbial community in Symbiotic culture of bacteria and yeast (SCOBY) can be a source for kombucha (kombu tea) production. In this study, it was aimed to investigate the usage of SCOBY, which produces bacterial cellulose, as a biomaterial and 3D scaffold material. 3D printable biomaterial was obtained by partial hydrolysis of oolong tea and black tea kombucha biofilms. In order to investigate the usage of 3D kombucha biomaterial as a tissue scaffold, “L929 cell line 3D cell culture” was created and cell viability was tested in the biomaterial. At the end of the 21st day, black tea showed 51% and oolong tea 73% viability. The cytotoxicity of the materials prepared by lyophilizing oolong and black tea kombucha beverages in fibroblast cell culture was determined. Black tea IC50 value: 7.53 mg, oolong tea IC50 value is found as 6.05 mg. Fibroblast viability in 3D biomaterial + lyophilized oolong and black tea kombucha beverages, which were created using the amounts determined to these values, were investigated by cell culture Fibroblasts in lyophilized and 3D biomaterial showed viability of 58% in black tea and 78% in oolong tea at the end of the 7th day. In SEM analysis, it was concluded that fibroblast cells created adhesion to the biomaterial. 3D biomaterial from kombucha mushroom culture can be used as tissue scaffold and biomaterial.

Melanin-Ce6-loaded polydopamine nanoparticles-based enhanced phototherapy for B16 melanoma cancer cells

Melanoma is one of the most aggressive and lethal types of cancer owing to its metastatic propensity and chemoresistance property. An alternative therapeutic option is photodynamic and photothermal therapies (PDT/PTT), which employ near-infrared (NIR) light to generate heat and reactive oxygen species (ROS). As per previous reports, Melanin (Mel), and its synthetic analogs (i.e. polydopamine nanoparticles) can induce NIR light-mediated heat energy, thereby selectively targeting and ameliorating cancer cells. Similarly, chlorin e6 (Ce6) also has high ROS generation ability and antitumor activity against various types of cancer. Based on this tenet, In the current study, we have encapsulated Mel-Ce6 in a polydopamine (PDA) nanocarrier (MCP NPs) synthesized by the oxidation polymerization method. The hydrodynamic diameter of the synthesized spherical MCP NPs was 139 ± 10 nm. The MCP NPs, upon irradiation with NIR 690 nm laser for 6 min, showed photothermal efficacy of more than 50 °C. Moreover, the red fluorescence in the MCP NPs due to Ce6 can be leveraged for diagnostic purposes. Further, the MCP NPs exhibited considerable biocompatibility with the L929 cell line and exerted nearly 70% ROS-mediated cytotoxicity on the B16 melanoma cell line after the laser irradiation. Thus, the prepared MCP NPs could be a promising theranostic agent for treating the B16 melanoma cancer.

Novel Adamantane Derivatives: Synthesis, Cytotoxicity and Antimicrobial Properties

Seventeen adamantane derivatives were synthesized according to facile condensation reaction protocols. Spectral analysis (1H NMR and 13C NMR) was applied to confirm the chemical structure of the obtained substances. The synthesized compounds were tested for in vitro antimicrobial activity against a panel of Gram-positive and Gram-negative bacterial strains and towards fungi from Candida spp. Among them, four derivatives numbered 9, 14, 15 and 19 showed the highest antibacterial potential with MIC = 62.5–1000 µg/mL with respect to all Gram-positive bacteria. S. epidermidis ATCC 12228 was the most susceptible among the tested bacterial strains and C. albicans ATCC 10231 among fungi. Additionally, the cytotoxicity for three derivatives was measured with the use of the MTT test on A549, T47D, L929 and HeLa cell lines. Our cytotoxicity studies confirmed that the tested substances did not cause statistically significant changes in cell proliferation within the range of the tested doses.

Evaluation of cytocompatibility and cell proliferation of electrospun chitosan/polyvinyl alcohol/montmorillonite clay scaffold with l929 cell lines in skin regeneration activity and in silico molecular docking studies

The present investigation describes the development of a novel Chitosan/Polyvinyl Alcohol/Montmorillonite Clay (CS/PVA/MMT) scaffold by adopting an electrospinning method, and their biocompatibility was evaluated in vitro with L929 fibroblast cell line to ascertain its use in wound healing applications. The fabricated scaffold was characterized using analytical techniques. FT-IR measurement exhibited the existence of relevant functional groups and XRD implies scaffolds' amorphous nature. The scaffold's morphology and pore diameter were assessed using TEM and SEM. The pore diameter of the as-prepared scaffold was approximately 125 nm. The antimicrobial assay of the scaffold was evaluated against selected pathogens which demonstrated higher antimicrobial efficacy. The scavenging activity tested using the DPPH assay showed remarkable scavenging capability. The wound healing properties were tested through the Cytotoxicity assay conducted on the L929 assay which proved the scaffold to be a suitable material for cell proliferation. Also, a Molecular docking investigation was carried out for CS/PVA/MMT ligand using human neutrophil elastase (HNE) 1H1B protein as a receptor in the CB-Dock server. Studies conducted in silico revealed strong interaction and high binding energy ratings of CS/PVA/MMT ligand with key residues of human neutrophil elastase (HNE) 1H1B proteins that help in tissue regeneration activity.