Effect of Salvadora persica on resin-dentin bond stability.

BACKGROUND: The stability of resin-dentin interfaces is still highly questionable. The aim of this study was to evaluate the effect of Salvadora persica on resin-dentin bond durability. MATERIALS AND METHODS: Extracted human third molars were used to provide mid-coronal dentin, which was treated with 20% Salvadora persica extract for 1 min after acid-etching. Microtensile bond strength and interfacial nanoleakage were evaluated after 24 h and 6 months. A three-point flexure test was used to measure the stiffness of completely demineralized dentin sticks before and after treatment with Salvadora persica extract. The hydroxyproline release test was also used to measure collagen degradation by endogenous dentin proteases. Statistical analysis was performed using two-way ANOVA followed by post hoc Bonferroni test and unpaired t-test. P-values < 0.05 were considered statistically significant. RESULTS: The use of Salvadora persica as an additional primer with etch-and-rinse adhesive did not affect the immediate bond strengths and nanoleakage (p > 0.05). After 6 months, the bond strength of the control group decreased (p = 0.007), and nanoleakage increased (p = 0.006), while Salvadora persica group showed no significant difference in bond strength and nanoleakage compared to their 24 h groups (p > 0.05). Salvadora persica increased dentin stiffness and decreased collagen degradation (p < 0.001) compared to their controls. CONCLUSION: Salvadora persica extract pretreatment of acid-etched dentin preserved resin-dentin bonded interface for 6 months. CLINICAL SIGNIFICANCE: Durability of resin-dentin bonded interfaces is still highly questionable. Endogenous dentinal matrix metalloproteinases play an important role in degradation of dentinal collagen within such interfaces. Salvadora persica may preserve resin-dentin interfaces for longer periods of time contributing to greater clinical success and longevity of resin composite restorations.

Salinity alleviates arsenic stress-induced oxidative damage via antioxidative defense and metabolic adjustment in the root of the halophyte Salvadora persica.

MAIN CONCLUSION: Arsenic tolerance in the halophyte Salvadora persica is achieved by enhancing antioxidative defense and modulations of various groups of metabolites like amino acids, organic acids, sugars, sugar alcohols, and phytohormones. Salvadora persica is a facultative halophyte that thrives under high saline and arid regions of the world. In present study, we examine root metabolic responses of S. persica exposed to individual effects of high salinity (750 mM NaCl), arsenic (600 µM As), and combined treatment of salinity and arsenic (250 mM NaCl + 600 µM As) to decipher its As and salinity resistance mechanism. Our results demonstrated that NaCl supplementation reduced the levels of reactive oxygen species (ROS) under As stress. The increased activities of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GR) maintained appropriate levels of ROS [superoxide (O2•-) and hydrogen peroxide (H2O2)] under salinity and/or As stress. The metabolites like sugars, amino acids, polyphenols, and organic acids exhibited higher accumulations when salt was supplied with As. Furthermore, comparatively higher accumulations of glycine, glutamate, and cystine under combined stress of salt and As may indicate its role in glutathione and phytochelatins (PCs) synthesis in root. The levels of phytohormones such as salicylate, jasmonate, abscisic acid, and auxins were significantly increased under high As with and without salinity stress. The amino acid metabolism, glutathione metabolism, carbohydrate metabolism, tricarboxylic acid cycle (TCA cycle), phenylpropanoid biosynthesis, and phenylalanine metabolism are the most significantly altered metabolic pathways in response to NaCl and/or As stress. Our study decoded the important metabolites and metabolic pathways involved in As and/or salinity tolerance in root of the halophyte S. persica providing clues for development of salinity and As resistance crops.

The Effect of Sub-MIC of Salvadora persica on Growth and Virulence Gene Expressions in Streptococcus mutans.

Dental caries is a multifactorial infectious chronic disease caused by particular bacteria and their virulence products that causes demineralization and progressive deterioration of the dental enamel. Many studies have proven miswak to have a critical antibacterial impact, particularly on cariogenic bacteria and periodontal pathogens, in the oral cavity. This study aimed to investigate the effect of different concentrations of Salvadora persica plant extract on growth and virulence gene expressions at mRNA levels in S. mutans. A total of 191 clinical samples from tooth swabs were collected, and sub-cultured on specific medium agar identified using biochemical and molecular approaches. MIC for the extract was determined and a bacterial growth curve was made to determine the growth phases and the optimum time for adding the extract at different concentrations. RT-qPCR technique was performed, and the REST-2009 software program was used for data analysis. Out of 191 swabs from the tooth 31 isolates were identified using several biochemical and molecular tests. Several S. mutans biofilm-related virulence genes and their Ct values were produced from RT-PCR under the effect of low and high doses of Meswak concentrations. Ct values and reaction efficiency were produced in RT-qPCR by Rotorgen3000, data then were analysed by REST-2009 software. Five isolates were selected to examine the effect of the extract on the mRNA levels using qPCR after growing them with both doses of the extract for about 30hrs. Levels of virulence gene mRNA were regulated differentially in cultures with added both extract doses. The isolates produced significantly lower virulence gene mRNA levels in cultures grown with both plant extract doses. The results produced in this study here provide new insights regarding several virulence gene expressions in S. mutans at the molecular levels when grown under different concentrations of Salvadora persica plant extract.

Antimicrobial Efficacy of Glass Ionomer Cement in Incorporation with Biogenic Zingiber officinale Capped Silver-Nanobiotic, Chlorhexidine Diacetate and Lyophilized Miswak.

In the present study, Zingiber officinale is used for the synthesis of Zingiber officinale capped silver nanoparticles (ZOE-AgNPs) and compares the antimicrobial efficacy and compressive strength of conventional glass ionomer cement (GIC) combined with ZOE-AgNPs, lyophilized miswak, and chlorhexidine diacetate (CHX) against oral microbes. Five groups of the disc-shaped GIC specimens were prepared. Group A: lyophilized miswak and GIC combination, Group B: ZOE-AgNPs and GIC combinations, Group C: CHX and GIC combination, Group D: ZOE-AgNPs + CHX + GIC; Group E: Conventional GIC. Results confirmed the successful formation of ZOE-AgNPs that was monitored by UV-Vis sharp absorption spectra at 415 nm. The X-ray diffractometer (XRD) and transmission electron microscope (TEM) results revealed the formation of ZOE-AgNPs with a mean size 10.5-14.12 nm. The peaks of the Fourier transform infrared spectroscopy (FTIR) were appearing the involvement of ZOE components onto the surface of ZOE-AgNPs which played as bioreducing, and stabilizing agents. At a 24-h, one-week and three-week intervals, Group D showed the significantly highest mean inhibitory zones compared to Group A, Group B, and Group C. At microbe-level comparison, Streptococcus mutans and Staphylococcus aureus were inhibited significantly by all the specimens tested except group E when compared to Candida albicans. Group D specimens showed slightly higher (45.8 ± 5.4) mean compressive strength in comparison with other groups. The combination of GIC with ZOE-AgNPs and chlorhexidine together enhanced its antimicrobial efficacy and compressive strength compared to GIC with ZOE-AgNPs or lyophilized miswak or chlorhexidine combination alone. The present study revealed that The combination of GIC with active components of ZOE-AgNPs and chlorhexidine paves the way to lead its effective nano-dental materials applications.

Decoding first complete chloroplast genome of toothbrush tree (Salvadora persica L.): insight into genome evolution, sequence divergence and phylogenetic relationship within Brassicales.

BACKGROUND: Salvadora persica L. (Toothbrush tree - Miswak; family-Salvadoraceae) grows in the arid-land ecosystem and possesses economic and medicinal importance. The species, genus and the family have no genomic datasets available specifically on chloroplast (cp) genomics and taxonomic evolution. Herein, we have sequenced the complete chloroplast genome of S. persica for the first time and compared it with 11 related specie's cp genomes from the order Brassicales. RESULTS: The S. persica cp genome was 153,379 bp in length containing a sizeable single-copy region (LSC) of 83,818 bp which separated from the small single-copy region (SSC) of 17,683 bp by two inverted repeats (IRs) each 25,939 bp. Among these genomes, the largest cp genome size (160,600 bp) was found in M. oleifera, while in S. persica it was the smallest (153,379 bp). The cp genome of S. persica encoded 131 genes, including 37 tRNA genes, eight rRNA genes and 86 protein-coding genes. Besides, S. persica contains 27 forward, 36 tandem and 19 palindromic repeats. The S. persica cp genome had 154 SSRs with the highest number in the LSC region. Complete cp genome comparisons showed an overall high degree of sequence resemblance between S. persica and related cp genomes. Some divergence was observed in the intergenic spaces of other species. Phylogenomic analyses of 60 shared genes indicated that S. persica formed a single clade with A. tetracantha with high bootstrap values. The family Salvadoraceae is closely related to Capparaceae and Petadiplandraceae rather than to Bataceae and Koberliniacaea. CONCLUSION: The current genomic datasets provide pivotal genetic resources to determine the phylogenetic relationships, genome evolution and future genetic diversity-related studies of S. persica in complex angiosperm families.

Shear Bond Strength and Film Thickness of a Naturally Antimicrobial Modified Dental Luting Cement.

Although several natural plants and mixtures have been known and used over the centuries for their antibacterial activity, few have been thoroughly explored in the field of dentistry. Thus, the aim of this study was to enhance the antimicrobial activity of a conventional glass ionomer cement (GIC) with natural plant extracts. The effect of this alteration on the bond strength and film thickness of glass ionomer cement was evaluated and related to an 0.5% chlorohexidine modified GIC. Olive leaves (Olea europaea), Fig tree (Ficus carica), and the leaves and roots of Miswak (Salvadora persica) were used to prepare an alcoholic extract mixture. The prepared extract mixture after the evaporation of the solvent was used to modify a freeze-dried glass ionomer cement at three different extracts: water mass ratios 1:2, 1:1, and 2:1. An 0.5% chlorhexidine diacetate powder was added to a conventional GIC for the preparation of a positive control group (CHX-GIC) for comparison. The bond strength to dentine was assessed using a material-testing machine at a cross head speed of 0.5 mm/min. Failure mode was analyzed using a stereomicroscope at 12× magnification. The cement film thickness was evaluated in accordance with ISO standard 9917-1. The minimum number of samples in each group was n = 10. Statistical analysis was performed using a Kruskal-Wallis test followed by Dunn's post hoc test for pairwise comparison. There was a statistically insignificant difference between the median shear bond strength (p = 0.046) of the control group (M = 3.4 MPa), and each of the CHX-GIC (M = 1.7 MPa), and the three plant modified groups of 1:2, 1:1, 2:1 (M = 5.1, 3.2, and 4.3 MPa, respectively). The CHX-GIC group showed statistically significant lower median values compared to the three plant-modified groups. Mixed and cohesive failure modes were predominant among all the tested groups. All the tested groups (p < 0.001) met the ISO standard of having less than 25 µm film thickness, with the 2:1 group (M = 24 µm) being statistically the highest among all the other groups. The plant extracts did not alter either the shear bond strength or the film thickness of the GIC and thus might represent a promising additive to GICs.

First de novo genome specific development, characterization and validation of simple sequence repeat (SSR) markers in Genus Salvadora.

Salvadoraceae constitutes ecologically imperative desert families of 3 genera-Azima, Dobera and Salvadora. Under genus Salvadora of this family, S. oleoides is a keystone species of socio-economic and medicinal value. This species naturally grows in the arid zones but currently experiencing severe fragmentation due to land use change and reduced regeneration, which may have resulted in the depletion of genetic diversity. Hence, it is up-most important to develop genomic resources for studying the population genetics in S. oleoides. This study aims to develop robust microsatellites markers, which were not yet reported in genus Salvodora due to lack of genome sequence information. We developed novel microsatellites markers in S. oleoides using Illumina paired-end sequencing technology. In total, 14,552 simple sequence repeat (SSR) markers were successfully designed from 21,055 microsatellite repeats detected in the 13 Gb raw sequence data. Afterwards, a subset of 101 SSRs were randomly selected and validated, 94 primers were successfully amplified and 34 showed polymorphisms. These SSRs were used to estimate the measures of genetic diversity in three natural populations of state Rajasthan and Gujarat. Importantly, average number of alleles (Na), observed heterozygosity (Ho), expected heterozygosity (He), and polymorphism information content (PIC) were recorded as 2.4, 0.529, 0.357, and 0.326, respectively. Furthermore, 15 primers were evaluated in S. persica for cross-transferability, and all were successfully amplified but only eight showed polymorphisms. This study has been conducted first time for S. oleoides and pioneer among the native species of arid-zone in India.

Evaluation of the genetic diversity of wild Salvadora persica 'Arak' from Saudi Arabia.

Assessment of genetic diversity is crucial for efficient selection genotypes in plant breeding and improvement programs. Studies of genetic diversity of S. persica are rare relative to the large species diversity in Saudi Arabia, despite its valuable importance as one of the most popular medicinal plants. We investigate the genetic variability and genetic differentiation among and within wild Salvadora persica populations distributed in four regions of Saudi Arabia. Twelve sequence-related amplified polymorphism (SRAP) primers combination generated 326 alleles, with an average of 27.2 alleles per primer. All primers showed 100 polymorphism percentage, and higher PIC values exceeded 0.90. Jaccard similarity values varied between 0.04 to 0.43, with an average of 0.31, which showed a weak relationship among the accessions and their origin. Based on UGPMA and principal coordinate analysis, accessions collected from the same region showed less aggregation. Genetic diversity parameters showed that both Aflaj and Joodah populations recorded the highest mean values for the effective number of alleles (1.26 and 1.24). Shannon index and genetic heterozygosity (0.23 and 0.15 for both populations), and percent of polymorphism 45.45% for Aflaj and 43.87 for Joodah population. Most of the genetic variation was because of differences within populations (77%) and 23% among populations. SRAP markers explored the genetic diversity among and within S. persica populations. In this work, genetic diversity within populations was high, and the population structure was weak. We detected no specific geographic structure, which may reveal an active movement of plants among populations.

Evaluation of the antimicrobial activity and compressive strength of a dental cement modified using plant extract mixture.

Literature lacks sufficient data regarding addition of natural antibacterial agents to glass ionomer cement (GICs). Hence, the aim of the study was to increase the antimicrobial properties of GICs through its modification with mixture of plant extracts to be evaluated along with an 0.5% chlorohexidine-modified GIC (CHX-GIC) with regard to biological and compressive strength properties. Conventional GIC (freeze-dried version) and CHX were used. Alcoholic extract of Salvadora persica, Olea europaea, and Ficus carcia leaves were prepared using a Soxhlet extractor for 12 h. The plant extract mixture (PE) was added in three different proportions to the water used for preparation of the dental cement (Group 1:1 PE, 2:1 PE, and 1:2 PE). Specimens were then prepared and tested against the unmodified GIC (control) and the 0.5% CHX-GIC. Chemical analysis of the extract mixture was performed using Gas chromatography-mass spectrometry. Antimicrobial activity was evaluated using agar diffusion assay against Micrococcus luteus and Streptoccocus mutans. Compressive strength was evaluated according to ISO 9917-1:2007 using a Zwick testing machine at a crosshead speed of 0.5 mm/min. Antimicrobial activity against Streptoccocus mutans was significantly increased for all the extract-modified materials compared to the unmodified cement, and the highest concentration was comparable to the CHX-GIC mixture. The activity against Micrococcus luteus was also significantly increased, but only for the material with the highest extract concentration, and here the CHX-GIC group showed statistically the highest antimicrobial activity. Compressive strength results revealed that there was no statistically significant difference between the different mixtures and the control except for the highest tested concentration that showed the highest mean values. The plant extracts (PEs) enhanced the antimicrobial activity against S. mutans and also against M. luteus in the higher concentration while compressive strength was improved by addition of the PE at higher concentrations.

Alterations in lipid peroxidation, lipid profile, insulin sensitivity, and hepatic histopathological changes in diabetic rats following the treatment with Salvadora persica.

We examined the effects of various partitions of Salvadora persica extract on lipid profile (LP), lipid peroxidation, and insulin sensitivity (IS) of diabetic rats. The rats were divided into normal control, diabetic control (DC), standard, sham, and test groups. The test groups were treated with an oral dose of 200, 400, and 600 mg/kg of crude, aqueous, and ethyl acetate partition of S. persica extract. After 21 days of experiment, the fasting blood glucose (FBS), LPs, lipid peroxidation, IS, liver enzymes levels, liver histopathology, and body weight alteration were evaluated. A significant decrease in FBS and lipid profile (except HDL) were observed in rats treated with various dose of extract compared with the DC rats ( P < 0.05). Treating diabetic rats with various extracts of S. persica meaningfully decreased the level of malondialdehyde ( P < 0.05). Animals treated with various dose of aqueous extract showed better results ( P < 0.01). On the basis of used indirect indexes to determine IS, all partitions of extracts showed anti-insulin resistance effects in diabetic rats. On the basis of our statistical analyzing, treating diabetic rats with all of the three extracts of S. persica decreased the elevated levels of alanine phosphatase, aspartate aminotransferase, and alanine transferase. Also, pathological changes in the liver tissue were reduced following treatment with the S. persica. In conclusion, our results give evidence that the S. persica extract, especially aqueous partition, has a healing effect on diabetes and can be considered as an alternative therapy for this disease.

Miswak (Salvadora persica) dietary supplementation improves antioxidant status and nonspecific immunity in Nile tilapia (Oreochromis niloticus).

Nile tilapia (Oreochromis niloticus) is the most common aquaculture fish, but is exposed to various pollutants and may be susceptible to infectious diseases due to reduction in their antioxidant status and immune defense. Therefore, researchers have tried to find feed supplements of natural origin to increase the health status of fish and decrease the incidence of drug resistance. The current study was conducted to investigate the effect of dietary supplementation of Miswak (Salvadora persica, SP) on Nile tilapia. Fish were randomly allocated into four experimental groups (30 fish each); the control fish were fed on the basal commercial diet and the SP-treated groups were fed basal diet supplemented with different concentrations 0.5, 1, and 2% for 30 days. The SP supplementations had no significant effects of SP on fish growth performance traits and lipid profiles but augmented the serum protein and globulin levels. The SP significantly improved the hepatic antioxidant status through the significant decrease of malondialdehyde (MDA) and the increases of reduced glutathione (GSH) levels and the activities of total superoxide dismutase (T.SOD), catalase (CAT) and glutathione peroxidase (GPx), especially in the SP 1% group, while glutathione S-transferase (GST) activities were significantly increased due to SP in a dose-dependent manner. The same results were obtained for the mRNA expression of CAT and GPx. Regarding the nonspecific immune status of the fish kidneys, SP, especially SP1, significantly increased interleukin-1 beta (IL-1ß) and interferon-gamma (INF-γ). The data of the present study revealed the protective effect of SP on Nile tilapia health status. Therefore, SP can be considered as a promising feed additive for Nile tilapia.

The repellency and toxicity effects of essential oils from the Libyan plants Salvadora persica and Rosmarinus officinalis against nymphs of Ixodes ricinus.

Essential oils extracted from the leaves of Libyan Rosemary (Rosmarinus officinalis L.), and Miswak (Salvadora persica L.) were evaluated for their acaricidal and repellent effects on Ixodes ricinus L. nymphs (Acari: Ixodidae) using a bioassay based on an 'open filter paper method'. Rosmarinus officinalis leaf essential oil diluted to 0.5 and 1 µl/cm2 in acetone exhibited, respectively, 20 and 100% tick mortality after about 5 h of exposure. A total of 50 and 95% of I. ricinus nymphs were killed by direct contact with the oil when exposed to lethal concentrations (LC) of 0.7 µl/cm2 (LC50) and 0.95 µl/cm2 (LC95), respectively. The LC50 (0.5 µl/cm2) was reached before the end of the first 24 h of exposure time (ET), as tick mortality at 24 h was 60%. Salvadora persica leaf essential oil at 1 µl/cm2 showed a significant repellency effect against I. ricinus nymphs at 1.5 h ET. A 95% repellency was observed at a repellent concentration (RC95) of 1 µl/cm2 of S. persica, but no significant mortality was recorded at this dose of S. persica oil. Gas chromatography-mass spectrometry analyses showed that the main monoterpenes in both oils were 1,8-cineol, α-pinene, and ß-pinene, although in markedly different proportions. These results suggest that essential oils have substantial potential as alternative approaches for I. ricinus tick control.

The antiplaque/anticariogenic efficacy of Salvadora persica (Miswak) mouthrinse in comparison to that of chlorhexidine: a systematic review and meta-analysis.

BACKGROUND: The plant Salvadora persica (miswak) has a long history of use in oral hygiene. Associations between the use of Salvadora persica and decreased oral bacteria numbers and plaque scores have been reported. This systematic review and meta-analysis assessed the ability of Salvadora persica mouthrinses to reduce plaque/cariogenic bacteria, in comparison to that of chlorhexidine and/or placebo rinses. METHODS: A comprehensive literature search for clinical trials reporting the use of Salvadora persica rinses as an antibacterial and/or antiplaque agent in comparison with chlorhexidine and/or placebo rinses was conducted, with no restriction to language. MEDLINE-PubMed, Cochrane Central Register of Controlled Trials, Wiley Online Library, ScienceDirect, and Google Scholar databases were searched to include all articles published up to December 2018. Based on inclusion/exclusion criteria, data were extracted from the identified reports by two independent reviewers. The primary and secondary outcomes measured from the eligible studies were mean plaque scores and mean cariogenic bacterial counts, respectively. Risk of bias of these studies was assessed. A statistical test of homogeneity was used to determine if the results of the separate studies could be combined. Based on the chi-square test, an inconsistency coefficient was computed (I2 statistic). Sensitivity analyses using subgroups and homogeneity evaluation were conducted. RESULTS: A total of 1135 potentially eligible articles were identified, of which 19 were eventually included in the qualitative analysis whereas 18 were included in the quantitative meta-analysis. The meta-analysis showed that Salvadora persica rinses exhibited strong antiplaque effects (P < 0.00001, MD: 0.46, and 95% CI: 0.29 to 0.63). In addition, it had statistically significant anti-streptococcal (P < 0.0001, MD: -1.42, and 95% CI: -2.08 to - 0.76) and anti-lactobacilli effects (P < 0.00001, MD: -1.12, and 95% CI: -1.45 to - 0.79) when compared to placebo. However, its effects were inferior compared to those by chlorhexidine rinse (P = 0.04, MD: 0.19, and 95% CI: 0.01 to 0.37). Subgroup analyses yielded results similar to those prior to subgrouping. CONCLUSION: The use of Salvadora persica extract was associated with a significant reduction in the plaque score and cariogenic bacterial count. Although, this reduction was lower than that achieved with the gold standard chlorhexidine mouthwash, Salvadora persica-containing rinse could be considered as a suitable oral hygiene alternative for use in individuals of all ages, socioeconomic backgrounds, and health conditions especially as a long-term measure due to its efficacy, safety, availability, cost-effectiveness, and ease of use.

Antibacterial activity of Salvadora persica against oral pathogenic bacterial isolates.

OBJECTIVE: The objective was to determine the antibacterial activity of Salvadora persica extract against bacteria isolated from dental plaque of patients. MATERIALS AND METHODS: Out of 40 different clinical specimens collected from patients suffering from plaque-induced gingivitis, 12 Staphylococcus aureus and 8 Streptococcus sp. isolates were recovered. The isolates were screened for their biofilm-forming capacity using tissue culture plate (TCP), tube method (TM), and congo red agar (CRA) method. Antibacterial activity of methanolic S. persica extract as well as of commercial antimicrobials against tested isolates was performed. High-performance liquid chromatography-mass spectrometry (HPLC-MS) and gas chromatography-MS (GC-MS) analysis were performed for S. persica crude extract and its volatile oil, respectively, to determine their constituents. RESULTS: Out of 20 isolates, 80%, 85%, and 90% showed positive results using TM, CRA, and TCP, respectively. The highest antimicrobial activity of methanolic S. persica extract was observed at 200 mg/ml. HPLC-MS analysis shows many polyphenols in S. persica extract such as Chrysin-8-c-ß-D-glucopyranoside, ferulic acid, gallic acid, and stigmasterol. Chemical composition of the essential oil of S. persica was determined by GC-MS yield; a mixture of monoterpene and hydrocarbons. The major compounds were butylated hydroxytoluene followed by benzene (isothiocyanatomethyl). CONCLUSION: Methanolic extract of S. persica had significant antibacterial effect against S. aureus and Streptococcus sp. isolates, and it may be gave a good alternative method for controlling oral pathogen.

Effectiveness of root-bark extract from Salvadora persica against the growth of certain molecularly identified pathogenic bacteria.

The acetone extract from root-bark of Salvadora persica L. (Salvadoraceae), is assayed for its antibacterial activity against some bacterial pathogens. By GC/MS analysis, the main chemical components of the acetone extract were found to be benzylisothiocyanate (39.4%), and benzyl nitrile (benzeneacetonitrile) (37.9%). According the extract concentrations used, the measured inhibition zones observed were between from 13.6 to 18.6 mm, 15.3-23 mm, 13.3-18.3 mm, 13.3-18.3 mm, and 12.3-19 mm, against the isolated plant bacterial pathogens namely Agrobacterium tumefaciens, Pectobacterium atrosepticum, Enterobacter cloacae, Dickeya solani and Ralstonia solanacearum, respectively, whilst it was between 8 and 12 mm, 8-9.6 mm, 8-11.6 mm, and 8-10.3 mm against Bacillus subtilis, Sarcina lutea, Escherichia coli and Staphylococcus aureus, respectively. The minimum inhibitory concentration values of the extract were between 16 and 32 µg/mL against the growth of plant bacterial, and from 1000 to 2000 µg/mL against the growth of the human bacteria. In conclusion, the acetone extract of root-bark of S. persica showed strong antibacterial activity against the plant pathogens and some activity against the human pathogens were reported. The results suggested that using the acetone extract from root-bark of S. persica as bioactive agent against the growth of the studied plant bacterial pathogens.

Purification and characterization of cysteine protease from miswak Salvadora persica.

BACKGROUND: Generally, proteases in medicinal plants had different therapeutic effects such as anti-inflammatory effect; modulate the immune response and inhibitory effect toward tumor growth. In this study, protease was purified and characterized from miswak roots, as medicinal plant and natural toothbrush. RESULTS: Physical and chemical characterization of cysteine protease P1 were studied such as pH optimum (6.5), optimum temperature (50 °C), thermal stability (50 °C) and Km (3.3 mg azocasein/ml). The enzyme digested some proteins in the order of caseine > haemoglobin > egg albumin >gelatin > bovine serum albumin. Hg2+ had strong inhibitory effect on enzyme activity compared with other metal ions. Kinetic of inhibition for determination the type of protease was studied. Iodoactamide and p-Hydroximercuribenzaoic acid (p-HMB) caused strong inhibitory effect on enzyme activity indicating the enzyme is cysteine protease. CONCLUSIONS: The biochemical characterization of this enzyme will be display the suitable conditions for using of this enzyme in toothpaste in the future and the enzyme may be used in other applications.

Persicaline, A New Antioxidant Sulphur-Containing Imidazoline Alkaloid from Salvadora persica Roots.

Salvadora persica L. is a popular chewing stick commonly known as "miswak". During our ongoing research activities on the chemical constituents of Salvadora persica roots, which is a new sulphur-containing imidazoline alkaloid 1,3-Dibenzyl-4-(1,2,3,4-tetrahydroxy-butyl)-1,3-dihydro-imidazole-2-thione, persicaline, (1) along with five known compounds (2-6) are identified. Compounds (2, 3) were reported for the first time from the family Salvadoraeceae. The structure of the new compound was established by extensive spectroscopic data and HR-MS. The antioxidant activities of the fractions and isolates were evaluated using different in vitro methods, such as DPPH, superoxide anion and nitric oxide radicals scavenging assays. Compound (1) showed a promising antioxidant activity with IC50 0.1, 0.08, and 0.09 µM in the three assays, respectively, comparable to ascorbic acid.

Anti-plaque Efficacy of Herbal Mouthwashes Compared to Synthetic Mouthwashes in Patients Undergoing Orthodontic Treatment: A Randomised Controlled Trial.

PURPOSE: The present study compared the antiplaque effects of two herbal mouthwashes containing Salvadora persica and Azadirachta indica, respectively, with two synthetic mouthwashes containing either chlorhexidine or cetylpyridinium. MATERIALS AND METHODS: In this triple-blind, randomised controlled trial, 100 patients undergoing orthodontic treatment underwent scaling and polishing at baseline to obtain a plaque score of zero. In the first phase, they were given oral hygiene instructions and were provided with a standard toothpaste to be used twice daily for a period of three weeks. In the second phase, following scaling and polishing, they were randomly allocated to 4 groups according to 4 different types of mouthwash (A: chlorhexidine; B = cetylpyridinium; C = extracts of Salvadora persica miswak; D: extract of Azadirachta indica miswak) along with previously taught toothbrushing protocol for three more weeks. Plaque accumulation was scored three times according to the Modified Bonded Bracket Plaque Index: at the start, after the toothbrush-toothpaste trial, and at the end of mouthwash trial. The paired t-test was used to compare the pre-plaque and post-plaque indices in all groups. Analysis of mean differences of post-plaque indices between and within groups was performed using the post-hoc Tukey test. Qualitative variables were compared by Crosstab. RESULTS: Eighty participants completed the study - 63 females and 17 males. There was a statistically significant decrease in mean plaque scores after using mouthwashes in all four groups at follow-up when compared to the baseline plaque score (p = 0.001). The greatest reduction of plaque score was found in group C (extract of Salvadora persica) when compared with group A, chlorhexidine (p = 0.016). CONCLUSION: Compared to other mouthwashes, Salvadora persica miswak-based mouthwash showed a maximum reduction in the plaque scores among orthodontic patients.

Chemical effects of chewing sticks made of Salvadora persica.

OBJECTIVE: The chewing sticks are widely used in many regions of Asia and Africa as a traditional tool to maintain oral hygiene. Salvadora persica L. (S. persica), also known as Arak or Miswak, a member of the salvadoraceae family. Chewing sticks, made up from S. persica, have chemical antibacterial properties and mechanical effects. The study aimed to investigate possible chemical effects of S. persica on dental plaque, sub-gingival microbiota and gingival inflammation. In order to achieve this, we inactivated some sticks through boiling to be used as inactive, but mechanically comparable control sticks. METHODS: In a double-blinded crossover trial, 24 patients with mild-to-moderate periodontitis were randomly allocated to use either fresh activated Miswak or inactivated Miswak for 3-week period. Plaque index (PI), gingival index (GI), visual plaque index (VPI) and bleeding on probing (BOP) were evaluated before and immediately following the experimental period. Sub-gingival plaque samples were analysed using DNA-DNA hybridization technique. The study was repeated with the participants switching the type of Miswak they used after a 7-week of "washout" period. RESULTS: There were no significant differences in the microflora after using active and inactive Miswak. Sixteen species of bacteria showed an increase (P < 0.05) after the usage of inactive Miswak, as compared to pretreatment values. No species showed a similar change after the use of active Miswak. There were no significant differences between active and inactive Miswak regarding the registered clinical variables. CONCLUSION: This study has not shown any clinical effect of the chemically active Miswak, but there was a tendency towards an effect on the microflora.

Phytochemical composition, anti-biofilm and anti-quorum sensing potential of fruit, stem and leaves of Salvadora persica L. methanolic extracts.

Emergence of antibiotic resistance among pathogenic bacteria encourages us to search for new molecules as an alternative treatment. The aim of this study was to evaluate the antiquorum sensing (anti-QS) and antibiofilm potential of Salvadora persica L. methanolic extracts to prevent the infections due to Staphylococcus as an alternate to antibiotics. The methanolic extracts of S. persica L. fruit, leaves and stems was assessed for their activity in inhibiting QS-depedent phenomenon such as violacein pigment production in Chromobacterium violaceum, swarming motility of Pseudomonas aeruginosa PAO1 and biofilm formation in oral Staphylococcus strains on polymethylmetacrylate (PMMA). Methanolic fruit extract of S. persica L. showed a high degree of anti-biofilm formation on PMMA and on violacein inhibition with a percentage of reduction equal to 90% when MIC value (20 mg/ml) was used. 100 µg/ml of S. persica L. leaves exhibited inhibition in swarming motility of PAO1 at 29.17%. Because the methanolic extracts of S. persica L. demonstrated anti-QS and antibiofilm activity at very low concentrations, it could be further exploited for novel molecules to treat oral Staphylococcus infections.