Assessment of Antimicrobial Activity of Nanocomposites Based on Nano-Hydroxyapatite (HAP), Chitosan, and Vitamin K2.

OBJECTIVE: The objective of this study was to evaluate the antimicrobial potential of nanocomposites containing vitamin K2, hydroxyapatite nanoparticles (nHAP), and chitosan (Chito)-coated dental implants against clinically relevant microbial strains. MATERIALS AND METHODS: Four test compounds were prepared: vitamin K2 + nHAP, K2 + Chito + nHAP, vitamin K2, and vitamin K2 + Chito. Agar well diffusion test was conducted to assess the antimicrobial activity of these compounds against Staphylococcus aureus (S. aureus), Streptococcus mutans (S. mutans), Enterococcus faecalis (E. faecalis), and Candida albicans (C. albicans). Results: The vitamin K2 + nHAP nanocomposite exhibited antimicrobial activity against all tested microorganisms, with E. faecalis showing the highest sensitivity (25 mm zone of inhibition at 100 µL concentration). The K2 + Chito + nHAP nanocomposite demonstrated potent antimicrobial activity with C. albicans displaying the highest sensitivity (28 mm zone of inhibition at 100 µL concentration). Pure vitamin K2 showed limited antimicrobial activity, vitamin K2 combined with chitosan exhibited significant susceptibility to C. albicans, resulting in a substantial inhibition zone of 24 mm diameter at a concentration of 100 µL. CONCLUSION: The synergistic effects of vitamin K2 with nHAP and chitosan highlight the potential of these nanocomposites for biomedical applications. These findings contribute to the development of effective nanocomposites to address antimicrobial resistance and improve infection control in various biomedical fields.

Role of Nanoparticles in Environmental Remediation: An Insight into Heavy Metal Pollution from Dentistry.

Environmental damage is without a doubt one of the most serious issues confronting society today. As dental professionals, we must recognize that some of the procedures and techniques we have been using may pose environmental risks. The usage and discharge of heavy metals from dental set-ups pollute the environment and pose a serious threat to the ecosystem. Due to the exclusive properties of nanosized particles, nanotechnology is a booming field that is being extensively studied for the remediation of pollutants. Given that the nanoparticles have a high surface area to volume ratio and significantly greater reactivity, they have been greatly considered for environmental remediation. This review aims at identifying the heavy metal sources and their environmental impact in dentistry and provides insights into the usage of nanoparticles in environmental remediation. Although the literature on various functions of inorganic nanoparticles in environmental remediation was reviewed, the research is still confined to laboratory set-ups and there is a need for more studies on the usage of nanoparticles in environmental remediation.

Acacia catechu seed extract provokes cytotoxicity via apoptosis by intrinsic pathway in HepG2 cells.

Acacia catechu Willd (Fabaceae) is a thorny tree widely distributed in India and commonly used as traditional Ayurvedic medicine for various ailments. The current study evaluates the cytotoxic potentials of A. catechu ethanolic seed extract (ACSE) in HepG2 cells, a human hepatocellular carcinoma cell line. The HepG2 cells were treated with 0.1, 0.3, 1, 3, 10, 30, 100, 300 and 1000 µg/ml of ACSE and the cytotoxic effect was evaluated by MTT and lactate dehydrogenase (LDH) leakage assays. The IC50 of ACSE was found at 77.04 µg/ml and therefore, further studies were carried out with the concentrations of 35 and 70 µg/ml. The intracellular reactive oxygen species (ROS) generation and apoptosis-related morphological changes were evaluated. Gene expressions of Bax, Bcl-2, cytochrome C (Cyt-c), caspases-9 and 3 were analyzed by qPCR. The ACSE treatments caused LDH leakage was associated with an increased ROS generation. The increased ROS generation was associated with the downregulation of intracellular antioxidant enzyme superoxide dismutase and reduced glutathione content. AO/EB and PI staining also confirmed chromatin condensation and apoptosis. The flow cytometric analysis showed an accumulation of HepG2 cells at sub G0/G1 (apoptotic) phase upon ACSE treatments. The ACSE induced cytotoxicity and oxidative stress were related to increased apoptotic marker gene expressions such as Bax, Cyt-c, caspase-9 and 3, and decreased anti-apoptotic marker Bcl-2. The current finding suggests that ACSE has apoptosis-inducing potential via the mitochondrial pathway in HepG2 cells.

Lagerstroemia speciosa (L.) Pers. triggers oxidative stress mediated apoptosis via intrinsic mitochondrial pathway in HepG2 cells.

Hepatocellular carcinoma is the second leading cause of cancer-related mortality worldwide. Lagerstroemia speciosa Pers. (Lythraceae) commonly known as Banaba has been used in different forms in traditional medicinal systems for treating various diseases which include diabetes and obesity. In this study, we investigated the cytotoxic potential of ethanolic Banaba leaf extract (EBLE) in HepG2 cells. The phytochemical analysis of EBLE was performed by HPTLC. HepG2 cells were treated with EBLE at 25, 50, 100, and 150 µg/mL concentrations, and cytotoxicity was evaluated by MTT assay. Oxidative stress was assessed by the evaluation of lipid peroxidation, superoxide dismutase, and reduced glutathione. Apoptosis-related morphology was investigated by acridine orange and ethidium bromide (AO/EB) dual staining. Mitochondrial membrane potential (ΔΨm) was evaluated by JC-1 staining. Apoptosis-related marker genes were evaluated by qPCR. HPTLC analysis confirmed the presence of corosolic acid (12.87 µg/mg), berberine (3.19 µg/mg), and gallic acid (2.94 µg/mg) in EBLE. EBLE treatments caused significant and concentration-dependent cytotoxicity and oxidative stress in HepG2 cells. Dual staining with AO/EB confirmed membrane distortion and nuclear chromatin condensation upon EBLE treatments. JC-I staining revealed the loss of ΔΨm. Furthermore, at a molecular level, EBLE treatments interfere with Bax/Bcl-2 homeostasis and induced the pro-apoptotic marker genes such as cytochrome c, Apaf-1, and caspases 9 and 3. EBLE treatments caused cytotoxicity in HepG2 cells, and this could be due to the induction of oxidative stress and apoptosis via the intrinsic or mitochondrial pathway.

Silibinin Triggers the Mitochondrial Pathway of Apoptosis in Human Oral Squamous Carcinoma Cells.

BACKGROUND: Silibinin, a natural polyphenolic flavonoid present in seed extracts of milk thistle (Silybum marianum). It has been shown to interact with various cancer-related cell signalling pathways in preclinical models, demonstrating promising anticancer effects in vitro and in vivo. MATERIALS AND METHODS: The cytotoxic effect of silibinin was evaluated in human oral squamous carcinoma (SCC-25) cells by MTT assay. The apoptosis-related morphological changes were investigated by AO/EB dual staining. The cytochrome c, caspases-3, and -9, B-cell lymphoma-2 (Bcl-2), and B-cell associated X protein (Bax) gene expressions were analysed by PCR. RESULTS: We have shown that silibinin treatment for 24 h in SCC-25 cells induced cytotoxicity in a concentration-dependent manner. The cytotoxic potential was due to the induction of apoptosis via the release of mitochondrial cytochrome c into the cytosol and subsequent activation of caspases-3 and -9. Dual staining assay was further confirmed the induction of early apoptosis upon silibinin treatment. CONCLUSION: The results from this study show that silibinin can be considered as a promising drug candidate for the treatment of oral squamous cell carcinoma.

Evaluation of the sub-acute toxicity of Acacia catechu Willd seed extract in a Wistar albino rat model.

Acacia catechu (A. catechu) or Khair (Hindi) is used in several herbal preparations in the Ayurvedic system of medicine in India. Traditionally, this drug is beneficial against several gastrointestinal and stomach related ailments, and leprosy. The present investigation was carried out to evaluate the sub-acute oral toxicity of the ethanolic extract of A. catechu seeds in Wistar albino rats. Results obtained from the quantitative chemical analysis of A. catechu seed extract were compared with commercially available standards. A. catechu seed extract was administered orally at the doses of 250, 500 and 1000 mg/kg b.w. daily for 28 days. General behavior, bodyweight and mortality were examined during the entire study period. At the end of 28 days, hematological and biochemical parameters along with the relative organ weights were determined. It was observed that the extract did not induce death or any significant changes in the body weight, relative weight of vital organs and in hematological parameters for up to a dose of 1000 mg/kg. The oral administration of the plant extract did not produce any significant changes in the levels of glucose. In addition, there were no significant changes in the activity of both hepatotoxic and nephrotoxic marker enzymes in the serum. Oral administration of A. catechu also did not produce any significant changes in the levels of oxidative markers. Furthermore, the findings from the biochemical studies were, well corroborated with the histological findings.

ß-Sitosterol attenuates carbon tetrachloride-induced oxidative stress and chronic liver injury in rats.

Chronic liver diseases are clinically silent and responsible for significant morbidity and mortality worldwide. ß-Sitosterol (BSS), major phytosterol in plants, has a wide spectrum of protective effect against various chronic ailments. We investigated the hepatoprotective effect of BSS against carbon tetrachloride (CCl4)-induced chronic liver injury in rats. Thirty rats were divided into five groups, with six animals in each group. Group I rats served as control while groups II, III, IV, and V rats were injected intraperitoneally with CCl4 (0.2 mL/100 g b.w. in olive oil (1:1)) for 7 consecutive weeks. After 7 weeks, group II rats were left without any treatments and served as CCl4 alone group, while groups III, IV, and V rats were treated with BSS 25 and 50 mg/kg b.w. and silymarin 100 mg/kg b.w. as oral post-treatments respectively, for the next 4 weeks. At the end of the experiment, hepatotoxicity marker enzymes in serum, oxidative stress, and fibrosis marker were analyzed. CCl4 administration caused significant elevation of marker enzymes of hepatotoxicity in serum and increased lipid peroxidation and fibrosis markers such as hydroxyproline, collagen, α-smooth muscle actin, vimentin, desmin, and matrix metalloproteinases 9 in liver tissue of rats. This treatment also caused a significant diminution of intracellular enyzmic antioxidants such as SOD and CAT in the liver tissue of rats. All the above adversities were significantly mitigated by the BSS post-treatments. The results suggest that BSS could have a hepatoprotective effect against oxidative stress-mediated CLD induced by CCl4.

An eco-friendly synthesis of Enterococcus sp.-mediated gold nanoparticle induces cytotoxicity in human colorectal cancer cells.

Gold nanoparticles (AuNPs) have become frequently used materials in biotechnological and biomedical applications including cancer. They can be commonly synthesized by biological and chemical methods. In the present study, we synthesized Enterococcus-mediated AuNPs and evaluated their cytotoxicity in human colorectal cancer cell line (HT-29). AuNPs are synthesized intracellularly using Enterococcus sp. RMAA. Characterization of AuNPs has done using UV spectrophotometry and transmission electron microscope. Cytotoxicity was evaluated by MTT assay. Intercellular reactive oxygen species (ROS) expression and apoptosis-related morphology were evaluated by dichlorodihydrofluorescein diacetate and acridine orange/ethidium bromide staining via fluorescence microscopy. JC-1 staining and caspase 3 immunofluorescence expression were analyzed by confocal microscopy. Enterococcus sp. RMAA-mediated AuNPs are spherical and induced concentration-dependent cytotoxicity in HT-29 cells. AuNP treatments also induced ROS and caspase-3 expressions and reduced the mitochondrial membrane potential. Morphology related to apoptotic changes was also noticed after AuNP treatments in HT-29 cells. The present study revealed that Enterococcus-derived AuNPs induced apoptotic cell death in HT-29 cells and suggests that AuNPs could be used as a pro apoptotic agent for colon cancer treatment.

Cytotoxic potentials of silibinin assisted silver nanoparticles on human colorectal HT-29 cancer cells.

It is of interest to study the cytotoxicity of silibinin assisted silver nanoparticles in human colorectal (HT-29) cancer cells. Silver nanoparticles were synthesized using silibinin as a reducing agent. The synthesized silibinin assisted silver nanoparticles ( SSNPs) were characterized and analyzed using a transmission electron microscope and spectrophotometer. The SSNPs synthesized in this study are spherical and their size ranges from 10 to 80 nm. HT-29 cells were treated with different concentrations (2, 4, 6, 8 and 10 ng/mL) of SSNPs and cytotoxicity was evaluated. The apoptosis was using flow cytometry. p53 protein expression using western blot. SSNPs are induced a decrease in viability and increased concentration-dependent cytotoxicity in HT-29 cells. SSNPs treatment also caused apoptosis-related morphological changes. SSNPs treatments at 8 and 16 ng/ml showed a prominent apoptotic change i.e., 70.3% and 83.6% respectively, and decreased viability of HT-29 cells 20% and 11.2% respectively as compared to control cells. SSNPs treatments induced p53 expression in HT-29 cells. Data shows that SSNPs have the potential to induce apoptosis in colorectal cancer cells. This provides insights for the further evaluation of SSNPs in fighting colon cancer.

Data on known anti-virals in combating CoVid-19.

Design and development of effective anti-virals in combating CoVid-19 is a great challenge worldwide. Known drugs such as chloroquine, lopinavir, favipiravir and remdesivir are used in the management of CoVid - 19. It is known that Ivermectin and remdesivir both are effective against filoviruses, paramyxo viruses. Available data also shows that ivermectin and remedesivir repress the replication of SARS-CoV-2. Thus, we document the potential use of ivermectin and remdesivir in the management of CoVid -19.

Cytotoxic potentials of S. cumini methanolic seed kernel extract in human hepatoma HepG2 cells.

Syzygium cumini (Myrtaceae) is commonly called as Jamun or Jambolan. It has antidiabetic, anti-inflammatory, antipyretic, and antioxidant activities. Hepatocellular carcinoma is the most frequent and deadliest cancers worldwide. We investigated the cytotoxic potentials of S. cumini methanolic seed kernel extract against human hepatoma HepG2 cells. HepG2 cells were treated with 10, 20, and 40 µg/mL of seed kernel extract for 24 hours and cytotoxic analysis was performed by MTT assay. S. cumini induced apoptosis related morphological changes in HepG2 cells were analyzed by annexin V and propidium iodide double staining. Nuclear fragmentation and chromatin condensation were analyzed by Hoechst nuclear staining. Mitochondrial membrane potential (MMP) was investigated by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) staining. Protein expressions of hepatocyte nuclear factor-1α (HFN-1α) was performed using western blotting. S. cumini treatments caused a significant and a concentration-dependent increase in the cytotoxicity of HepG2 cells. S. cumini treatments increased the percentage of cells in an early and late apoptosis stage. This treatment also caused chromatin condensation and nuclear fragmentation. Further, S. cumini treatments decreased MMP and also caused a significant downregulation of HFN-1α protein expression. The present study demonstrated that S. cumini seed extract induced apoptosis in HepG2 cells through decrease in MMP and downregulation of HFN-1α.

Silibinin alleviates N-nitrosodimethylamine-induced glutathione dysregulation and hepatotoxicity in rats / 中国天然药物

The present study was designed to evaluate the hepatoprotective and antioxidant potentials of silibinin (SBN) against N-nitrosodimethylamine (DMN)-induced toxic insults in the rat liver. The liver damage was induced in Wistar albino rats by repeated administration of DMN (10 mg·kg(-1) b.w., i.p.) on 3 consecutive days per week for 3 weeks. SBN (100 mg·kg(-1) b.w., p.o.) was given daily to the DMN treated rats for two weeks. The marker enzymes of liver toxicity and second-line enzymic and non-enzymic antioxidants were evaluated in serum and liver tissues before and after SBN treatment. Histopathology of the liver was evaluated by H & E staining. The DMN treatment produced a progressive increase in all the serum marker enzymes (AST, ALT, ALP, LDH, and γ-GT), peaking on Day 21. This treatment produced highly significant decreases in all the second-line antioxidant parameters (GSH, GST, GR, GPx, and vitamins C and E). The SBN treatment significantly reversed the DMN-induced damages, towards normalcy. Histopathological studies confirmed the development of liver toxicity in DMN-treated rats, which was reversed by SBN treatment in corroboration with the aforementioned biochemical results, indicating the hepatoprotective and antioxidant properties of SBN. In conclusion, the DMN-induced degenerative changes in the liver were alleviated by SBN treatment and this protective ability may be attributed to its antioxidant, free radical scavenging, and membrane stabilizing properties.