Antimicrobial potential of four mica drugs and their chemical and mineralogical properties.

BACKGROUND: Mica drugs, a group of herbo-metallic traditional preparations comprising biotite mica as the major mineral ingredient, are prescribed for skin disorders and respiratory ailments and other chronic conditions in South Asian countries, particularly India and Sri Lanka. Mica-based drugs (Abhrak drugs) are subjected to unique and varied preparation procedures and the bioactivity of the drugs can be affected by drug-processing conditions, the ingredients used and the mica composition. The current study aimed to evaluate and compare, on the basis of their physical and chemical characteristics, the antimicrobial potential of two commercial mica drugs AbBb (Abhrak bhashma) and AbCh (Abhrak Chenhuram) and two mica drugs ABL1 (Abhrak Bhasma Laboratory Prepared 1) and ABL2 (Abhrak Bhasma Laboratory Prepared 2) prepared in the laboratory under different conditions. METHODS: Antimicrobial activity of all four drugs was assessed at 10 mg/ml concentration against Pseudomonas aeruginosa, Escherischia coli, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA) and Candida albicans using well diffusion assay, agar dilution assay and Miles and Misra method. Major and trace metal constituents of the drug samples were measured using atomic absorption spectrometry. Mineralogical properties, bacteria-mineral interactions, morphological changes in microbes and the surface characteristics of the drugs were determined using X-ray diffraction (XRD) analysis and scanning electron microscopy (SEM). RESULTS: The drugs ABL1, ABL2 and AbBh exhibited antimicrobial activity against only Gram-positive organisms (S. aureus and MRSA) when tested with Miles and Misra method (broth method). Mineralogical studies (XRD) revealed that biotite mica was altered into secondary clay minerals and iron oxides in the commercial drug AbCh while the other three drugs had altered mica and iron oxide phases. The essential elements (Na, K, Ca and Mg) required for microbial functions were present in varying extents in all four drugs while they were present in exceedingly high amounts in AbCh having comparatively high cation-exchange capacity, consistent with the observation that AbCh was inactive against all the microbes tested. The three drugs (ABL1, ABL2 and AbBh) showing antimicrobial activity contained comparatively high amounts of Fe, Zn and Cu that are known to display antimicrobial properties at high concentrations. SEM studies revealed that the drug particles adhered and entrapped the bacterial species, presumably modifying the physiochemical characteristics of the bacteria and eventually causing lethality. CONCLUSION: Three of the four mica drugs inhibited the tested Gram-negative bacteria and the antibacterial activity of the mica drugs depends on their constituents and the methods of preparation.

Leaf extract of Osbeckia octandra induces apoptosis in oral squamous cell carcinoma cells.

BACKGROUND: Osbeckia octandra is a plant endemic to Sri Lanka and is used in ethnomedicine for treating various diseases. However, the anti-cancer properties of O. octandra are yet to be fully investigated. In the present study, we evaluated the anti-cancer effects of O. octandra on oral cancer cells. METHODS: Human oral cancer cell lines (HSC2, YD10B, YD38, YD9, and YD32) were used in this study. BrdU incorporation, cell cycle and annexin-V/PI staining were all evaluated using flow cytometry to determine the extent to which O. octandra leaf extract inhibits cell proliferation and induces apoptosis. Cell viability and reactive oxygen species (ROS) were also measured in order to investigate the anti-cancer effects of O. octandra extracts. Western blotting was performed to detect cell cycle related protein such as cyclin d1 and cdk4, and to detect apoptosis-related proteins such as Bcl-2, Bcl-XL, Bax, Caspase-9, Cleaved caspase-3, Fas, Caspase-8, and Bid. RESULTS: Leaf extract of O. octandra reduced oral squamous cell carcinoma (OSCC) cell viability in a dose-dependent manner. Leaf extract of O. octandra has non-toxic in normal keratinocytes. Also, O. octandra extract interrupted the DNA replication via G1 phase arrests, and this effect was independent of ROS generation. In the apoptosis-related experiments, the population of annexin V-positive cells increased upon treatment with O. octandra extract. Furthermore, the expression of anti-apoptotic protein (Bcl-2 and Bcl-xL) was decreased, whereas the expression of cleaved caspase-3 protein was increased in O. octandra-treated OSCC cells. CONCLUSIONS: The results suggest that a leaf extract of O. octandra inhibited the proliferation of OSCC cells through G1 phase arrest and interrupting DNA replication. The leaf extract of O. octandra could trigger the apoptotic response via caspase 3 activation in OSCC cells. These results suggest that O. octandra has the potential to be developed as an alternative medicine for treating OSCC.

Terminalia bellirica fruit extracts: in-vitro antibacterial activity against selected multidrug-resistant bacteria, radical scavenging activity and cytotoxicity study on BHK-21 cells.

BACKGROUND: Identification of novel sources for developing new antibiotics is imperative with the emergence of antibiotic resistant bacteria. The fruits of Terminalia bellirica (Gaertn) Roxb., widely used in traditional medicine, were evaluated for antibacterial activity against multidrug-resistant (MDR) bacteria, antioxidant activity and cytotoxicity. METHODS: Twelve solvent extracts of T. bellirica fruits were prepared by direct aqueous extraction and sequential extraction with dichloromethane, methanol and water using Soxhlet, bottle-shaker and ultrasound sonicator methods. Antibacterial activity of the extracts was tested against 16 strains MDR bacteria-methicillin-resistant Staphylococcus aureus (MRSA), extended spectrum ß-lactamase (ESBL) producing Escherichia coli and MDR Acinetobacter spp., Klebsiella pneumoniae and Pseudomonas aeruginosa-and 4 control organisms, using the cut-well diffusion method. The minimum inhibitory concentration (MIC) was determined using an agar dilution method. The radical scavenging activity of six antibacterial extracts was screened against 2,2'-diphenyl-2-picrylhydrazyl (DPPH) and correlation was established between EC50 (50% effective concentration) values and the total phenolic content (TPC). Cytotoxicity was determined for the most potent antibacterial extract on baby hamster kidney (BHK-21) cells by Tryphan Blue exclusion method. Statistical analysis was carried out by one-way analysis of variance at significant level p < 0.05 using "SigmaPlot 10" and "R 3.2.0" software. RESULTS: All aqueous and methanol extracts displayed antibacterial activity (MIC 0.25-4 mg/mL) against all strains of MRSA, MDR Acinetobacter spp. and MDR P. aeruginosa. The sequential aqueous extracts (MIC, 4 mg/mL) inhibited ESBL producing-E. coli. None of the extracts exhibited activity against MDR K. pneumoniae (MIC > 5 mg/mL). The sequential methanol extract (Soxhlet) recorded high antibacterial activity and the highest DPPH radical scavenging activity (EC50, 6.99 ± 0.15 ppm) and TPC content (188.71 ± 2.12 GAE mg/g). The IC50 (50% inhibition concentration) values of the most potent antibacterial extract-the direct aqueous extract from reflux method-on BHK-21 cells were 2.62 ± 0.06 and 1.45 ± 0.08 mg/ml with 24 and 48 h exposure, respectively. CONCLUSIONS: Results indicate that T. bellirica fruit is a potential source for developing broad-spectrum antibacterial drugs against MDR bacteria, which are non-toxic to mammalian cells and impart health benefits by high antioxidant activity.