Abeer M. El Sayed

Homing and reparative effect of intra-articular injection of autologus mesenchymal stem cells in osteoarthritic animal model

AbstractBackgroundThis work aimed to study the homing evidence and the reparative effect of mesenchymal stem cells (MSCs) in the healing process of induced osteoarthritis in experimental animal model (donkeys).MethodsTwenty-seven donkeys were equally divided into 3 groups based on the observation period after induction of arthritis (3, 6 and 9 weeks) to achieve different degrees of osteoarthritis. Each group was subdivided into three subgroups of three animals each based on the follow-up period (1, 2 and 6 months) after treatment. The induction was done through intra-articular (IA) injection of 2 ml of Amphotericin-B in both carpal joints. MSCs were harvested in a separate procedure, labeled with green fluorescent protein (GFP) using monster GFP vector and suspended in hyaluronic acid for IA injection. Treatment approaches consisted of cell-treatment using MSCs suspended in 3 ml of hyaluronic acid (HA) for the right carpal joint; and using the same amount of (HA) but without MSCs for the left contralateral carpal joint to serve as a control. Animals were assessed clinically and radiologically before and after treatment. Synovial fluid was also evaluated. Histopathologically; articular cartilage structural changes, reduction of articular cartilage matrix staining, osteophyte formation, and subchondral bone plate thickening were graded. Data was summarized using median and percentile for scores of histopathologic grading. Comparison between groups was done using non-parametric Mann Whitney test.ResultsThe reparative effect of MSCs was significant both clinically and radiologically in all treated groups (P < 0.05) compared to the control groups. Fluorescence microscopy of sections of the cell-treated joints of all animals indicated that the GFP-transduced injected cells have participated effectively in the reparative process of the damaged articular surface and have integrated within the existing articular cartilage. The cells were associated with the surface of the cartilage and, were also detected in the interior.ConclusionsHoming was confirmed by the incorporation of injected GFP-labeled MSCs within the repaired newly formed cartilage. Significant recovery proves that the use of IA injection of autologous MSCs is a viable and a practical option for treating different degrees of osteoarthritis. http://www.biomedcentral.com/1741-7015/10/44/abstract

Bioactive constituents from Harpephyllum caffrum Bernh. and Rhus coriaria L.

Background: The leaf ethanol extract of Harpephyllum caffrum Bernh. has evidenced medicinal value due to its hepatoprotective activity. It demonstrated inhibitory effects on test standard microbes approximated to 40% the potency of ofloxacin and fluconazole. The same extract evidenced in vitro cytotoxicity on human cell lines, liver carcinoma HEPG2, larynx carcinoma HEP2, and colon carcinoma HCT116 cell lines when compared to doxorubicin. Materials and Methods: Fractionation of the leaf ethanol extract led to the isolation of the polyphenols, ethyl gallate, and quercetin-3-O-rhamnoside, a hydrocarbon, hendecane, the fatty acid ester, methyl linoleate, and four triterpenoids, betulonic acid, 3-acetyl-methyl betulinate, lupenone and lupeol for the first time, in addition to the previously reported phenol acids and flavonoids, gallic acid, methyl gallate, quercetin, kaempferol, kaempferol-3-O-rhamnoside, kaempferol-3-O-galactoside, apigenin-7-O-glucoside, and quercetin-3-O-arabinoside. Results: The ethanol extract of the fruit of the genetically related Rhus coriaria L., known as sumac, afforded protocatechuic acid, isoquercitrin, and myricetin-3-O-α-L-rhamnoside from the fruits for the first time, in addition to the previously reported phenol acids and flavonoids, gallic acid, methyl gallate, kaempferol, and quercetin. Conclusion: The leaf ethanol extract of H. caffrum Bernh. exhibited variable anti-inflammatory, analgesic, and antipyretic activities, besides the hepatoprotective, in vitro cytotoxic and anti-microbial activities.

Metabolomics reveals distinct methylation reaction in MeJA elicited Nigella sativa callus via UPLC–MS and chemometrics

AbstractCell suspension cultures are now recognized as important model for studying natural products biosynthesis and functional genomics. Nevertheless, very few studies of metabolic comparisons between cell cultures (callus) and original plants have been reported, even though the biological identity of cultured cells with the normally grown plant is of great importance. In this study, an MS-based metabolomic approach was used to compare the natural products profile of intact Nigella sativa seeds versus callus . N. sativa has been used for centuries in traditional medicine for several purposes. Its phytochemical components comprise, among others, alkaloids, saponins, flavonoids and fatty acids. Ultra performance liquid chromatography coupled to ultraviolet photodiode array detection and high resolution q-TOF mass spectrometry (UPLC–PDA–MS) was utilized to analyze the secondary product metabolome of N. sativa callus, with a total of 74 metabolites including five flavonoids, 13 hydroxycinnamates, an alkaloid, saponin and 14 fatty acids. Callus maintained the capacity to produce N. sativa phenolic subclasses, with hydroxycinnamates amounting for the major secondary metabolites in callus. Alkaloids, major constituents in Nigella genus, were detected in callus though with qualitative and quantitative differences from seed tissue. Methyl jasmonate (MeJA) elicitation effect was assessed on callus with the aim of increasing secondary metabolites production. Metabolite profiles were subjected to principal component analysis and orthogonal projection to latent structures-discriminant analysis to evaluate MeJA effect. Results revealed that MeJA led to O-methylation reaction induction yielding O-feruloylquinic acid from O-caffeoylquinic acid, in addition to a methylated disaccharide. The work extends our knowledge regarding hydroxycinnamates biosynthesis, regulation and on metabolic engineering future efforts to increase its production as potential phytoalexin in N. sativa.

The Cardenolide Glycoside Acovenoside A Affords Protective Activity in Doxorubicin-Induced Cardiotoxicity in Mice

The current study aimed to investigate the protective effect of the cardenolide glycoside acovenoside A (AcoA) against doxorubicin-induced cardiotoxicity in mice. AcoA was isolated from the pericarps of Acokanthera oppositifolia to chemical homogeneity and characterized by means of one- and two-dimensional nuclear magnetic resonance spectroscopy. AcoA exhibited relatively low toxicity in mice (LD50 = 223.3 mg/kg bw). Repeated administration of doxorubicin induced cardiotoxicity manifested by reduced activity of myocardial membrane-bound ion pumps and elevated serum biomarkers of myocardial dysfunction, oxidative stress, and inflammation. Pretreatment of doxorubicin-exposed mice with AcoA (11.16 or 22.33 mg/kg bw, i.p.) for 2 weeks after 2 weeks of combined administration of AcoA and doxorubicin protected the animals dose dependently against doxorubicin-induced cardiotoxicity as indicated by normalization of the levels of different myocardial markers of oxidative stress (malondialdehyde, nitric oxide, total antioxidant capacity, and cardiac glutathione), serum myocardial diagnostic marker enzymes (serum cardiac troponin T, creatine kinase isoenzyme MB, aspartate aminotransferase, and lactate dehydrogenase), and inflammatory markers (c-reactive protein, tumor necrosis factor-α, and interleukin-6), as well as myocardial Na+/K+-ATPase activity. These effects were attributed to the negative impact of AcoA on transcription factors nuclear factor κB and interferon regulatory factor 3/7. Thus acovenoside A might act as a cardioprotective agent to prevent doxorubicin-induced cardiotoxicity.

Potential Anti-Microbial, Anti-Inflammatory and Anti-Oxidant Activities of Haplophyllum tuberculatum Growing in Libya

Ethanolic extract of the aerial parts of Haplophyllum tuberculatum demonstrated an efficient anti-fungal activity against Aspergillus fumigates, Geotricum candidum and Syncephalastrum racemosum with (MIC 0.49, 0.12 and 1.95 μg/ml). It also presented 75% potency as antibacterial agent on Staphylococcus aureus and Escherichia coli (MIC 1.95 and 15.63 μg/ml). Volatile oil of the aerial parts demonstrated significant antibacterial effect against Enterococcus faecalis and Lactobacillus acidophilus (MIC 1.95 and 0.98 μg/ml). The essential oils from aerial parts and flowers exhibited a remarkable acute anti-inflammatory activity against carrageenan induced oedema in rats 9.52 and 8.56 which were found to be comparable to the standard drug at the selected dose. The ethanolic extract of the aerial parts exhibited significance anti-oxidant activity (98%) as compared to vitamin E.

Acovenoside A Induces Mitotic Catastrophe Followed by Apoptosis in Non-Small-Cell Lung Cancer Cells

We investigated the cytotoxic potential of the cardenolide glycoside acovenoside A against non-small-cell lung cancer cells. Lung cancer is the leading cause of cancer-related mortality and the second most common cancer diagnosed. Epidemiological studies revealed a direct correlation between the regular administration of cardiac glycosides and a lower incidence of various cancers. Acovenoside A, isolated from the pericarps of Acokanthera oppositifolia, potently inhibited proliferation and induced cytotoxicity in A549 non-small-cell lung cancer cells with an IC50 of 68 ± 3 nM after 48 h of exposure. Compared to the antineoplastic agent doxorubicin, acovenoside A was more potent in inhibiting the viability of A549 cancer cells. Moreover, acovenoside A exhibited selectivity against cancer cells, being significantly less toxic to lung fibroblasts and nontoxic for peripheral blood mononuclear cells. Analysis of the cell cycle profile in acovenoside A-treated A549 cells revealed mitotic arrest, due to accumulation of the G2/M regulators cyclin B1 and CDK1, and cytokinesis failure. Furthermore, acovenoside A affected the mitochondrial membrane integrity and induced production of radical oxygen species, which resulted in induction of canonical apoptosis, manifested by caspase 3 activation and DNA fragmentation. Based on our results, acovenoside A warrants further exploration as a potential anticancer lead.

DNA fingerprinting, biological and chemical investigation of certain Yucca species

Abstract Yucca aloifolia, Y. aloifolia variegata, Y. elephantipes and Y. filamentosa were investigated. DNA sequencing was performed for the four plants and a genomic DNA fingerprint was obtained and provided. The cytotoxic activities against four human cancer cell lines were investigated. The ethanolic extracts of leaves of Y. aloifolia variegata prevailed, especially against liver cancer HepG-2 and breast cancer MCF-7. In vivo assessment of hepatoprotective activity in rats also revealed the hepatoprotective potential of the ethanolic extracts of the four plants against CCl4- induced rats’ liver damage. Qualitative and quantitative analysis of the flavonoid and phenolic content of the promising species was performed using HPLC. The analysis identified and quantified 18 flavonoids and 19 phenolic acids in the different fractions of Y. aloifolia variegata, among which the major flavonoids were hesperidin and kaemp-3-(2-p-coumaroyl) glucose and the major phenolic acids were gallic acid and protocatechuic acid.