Osteoblast-Based Therapy-A New Approach for Bone Repair in Osteoporosis: Pre-Clinical Setting.

BACKGROUND: Osteoporosis is a metabolic bone disease characterized by low bone density resulting in increased fracture susceptibility. This research was constructed to uncover the potential therapeutic application of osteoblasts transplantation, generated upon culturing male rat bone marrow-derived mesenchymal stem cells (BM-MSCs) in osteogenic medium (OM), OM containing gold (Au-NPs) or gold/hydroxyapatite (Au/HA-NPs) nanoparticles, in ovariectomized rats to counteract osteoporosis. METHODS: Forty rats were randomized into: (1) negative control, (2) osteoporotic rats, whereas groups (3), (4) and (5) constituted osteoporotic rats treated with osteoblasts yielded from culturing BM-MSCs in OM, OM plus Au-NPs or Au/HA-NPs, respectively. After 3 months, osterix (OSX), bone alkaline phosphatase (BALP), sclerostin (SOST) and bone sialoprotein (BSP) serum levels were assessed. In addition, gene expression levels of cathepsin K, receptor activator of nuclear factor-κb ligand (RANKL), osteoprotegerin (OPG) and RANKL/OPG ratio were evaluated using real-time PCR. Moreover, histological investigation of femur bone tissues in different groups was performed. The homing of implanted osteoblasts to the osteoporotic femur bone of rats was documented by Sex determining region Y gene detection in bone tissue. RESULTS: Our results indicated that osteoblasts infusion significantly blunted serum BALP, BSP and SOST levels, while significantly elevated OSX level. Also, they brought about significant down-regulation in gene expression levels of cathepsin K, RANKL and RANKL/OPG ratio versus untreated osteoporotic rats. Additionally, osteoblasts nidation could restore bone histoarchitecture. CONCLUSION: These findings offer scientific evidence that transplanting osteoblasts in osteoporotic rats regains the homeostasis of the bone remodeling cycle, thus providing a promising treatment strategy for primary osteoporosis.

Novel Pyrazolo[3,4-b]pyridine Derivatives: Synthesis, Characterization, Antimicrobial and Antiproliferative Profile.

Three novel series of pyridine derivatives, namely Schiff's bases, 4-thiazolidinones and azetidin-2-ones bearing pyrazolo[3,4-b]pyridine moiety, have been synthesized. The chemical structures of the synthesized compounds were characterized. The compounds were tested for their antimicrobial activity using the agar well diffusion and broth macrodilution methods. The compounds were also evaluated for their antiproliferative activity using the sulforhodamine B (SRB) assay. The majority of the tested compounds exhibited slight to high antimicrobial activity against the test microorganisms with minimum inhibitory concentrations (MICs) of 0.12-62.5 µg/mL when compared to that of 3 standard antimicrobial agents (Ampicillin, 0.007-0.03 µg/mL; Gentamicin; 0.015-0.24 µg/mL; and Amphotericin B, 0.03-0.98 µg/mL). Compound (7b) was found to be nearly as active as the standard antimicrobial drug Amphotericin B against Fusarium oxysporum fungal strain with MIC of 0.98 µg/mL. Some of the test compounds showed remarkable cytotoxic activities against Hep G2 (hepatocellular carcinoma) cells (IC50=0.0158-71.3 µM) in comparison to the standard anticancer drug doxorubicin (IC50=0.008 µM). Among the compounds tested, (5), (6a), (6b), (7b), and (10) exhibited antiproliferative potency (IC50=0.0001-0.0211 µM) that was found to be better than that of doxorubicin (IC50=0.099 µM) against MCF7 (breast adenocarcinoma) cells. In particular, (7b) displayed the highest significant antiproliferative efficacy against both Hep G2 and MCF7 cell lines showing IC50 values of 0.0158 µM and 0.0001 µM, respectively. Our findings suggest that the synthesized compounds may be promising candidates as novel antimicrobial and antiproliferative agents.

Bactericidal and Antibiotic Synergistic Effect of Nanosilver Against Methicillin-Resistant Staphylococcus aureus.

BACKGROUND: Methicillin-Resistant Staphylococcus aureus (MRSA) are bacteria responsible for several difficult-to-treat infections in humans. These strains have developed, through the process of natural selection. Infections by MRSA are more difficult to treat with standard types of antibiotics and thus more dangerous to human health. OBJECTIVES: The aim of this study was to evaluate the bactericidal and antibiotic synergistic effect of silver nanoparticles (Ag-NPs) against MRSA. MATERIALS AND METHODS: Methicillin-Resistant Staphylococcus aureus strains were isolated from clinical samples and identified, and their susceptibility was tested using the MicroScan® WalkAway-96® SI System. minimum inhibitory concentration (MIC) was determined by a microdilution method. Time kill assay was performed by exposing the MRSA isolates to different concentrations of Ag-NPs and monitoring bacterial growth, by measuring optical density at 600 nm. Tissue culture plate was used for determination of the efficacy of Ag-NPs and their combination with antibiotics in the elimination of formed biofilm. RESULTS: The MIC value of Ag-NPs against MRSA was 100 µg/mL. Methicillin-Resistant Staphylococcus aureus cells were treated with 50, 100 and 200 µg/mL of Ag-NPs and inhibited bacterial growth so that after four hours, almost all treated MRSA cells were dead. All combinations showed effectiveness against MRSA. It was observed that MRSA did not show inhibition zones with ampicillin alone. CONCLUSIONS: Silver Nanoparticles have high therapeutic activity against MRSA, thus can be suggested as an alternative or adjuvant with antibiotics for MRSA treatment. Further studies are required to understand the synergistic effect of Ag-NPs combinations and to assess the safety and efficacy of new antibiotic-Ag-NPs combinations.

Prevalence of Extended Spectrum ß-lactamase-Producing Klebsiella pneumoniae in Clinical Isolates.

BACKGROUND: Extended spectrum ß-lactamase (ESBL) are gram-negative bacteria that produce the enzyme, ß-lactamase, which can break down commonly used antibiotics, such as penicillin and cephalosporins, making infections with ESBL producing bacteria more difficult to treat. Extended spectrum ß-lactamase-producing Klebsiella pneumoniae were first reported in 1983 from Germany, and since then a steady increase in resistance against cephalosporins has been seen causing health problems. OBJECTIVES: The aim of this study was to determine the prevalence of ESBL in strains of K. pneumoniae isolated from different clinical samples. PATIENTS AND METHODS: One hundred and thirty isolates of K. pneumoniae were isolated from different clinical specimens from King Khalid hospital, Hafr Elbatin, Kingdom Saudi Arabia. These isolates were then characterized, tested for antimicrobial susceptibility and screened for ESBL production by the MicroScan WalkAway-96 SI System. Extended spectrum ß-lactamase production was confirmed by the phenotypic confirmatory disc diffusion test (PCDDT) and the double disc synergy test (DDST). RESULTS: Overall, 76.9% (100) of the isolates were resistant to cefuroxime, cefepime and cefazolin, 69.23% (90) were resistant to cefotaxime, and 46.15% (60) were resistant to cefoxitin. Extended spectrum ß-lactamase was detected in 53.8% (70) of K. pneumoniae as detected by the MicroScan "WalkAway-96" SI System and 50.07% (66) by PCDDT and 46.15% (60) by DDST. All K. pneumoniae isolates were resistant to ampicillin followed by both piperacillin and mezlocillin 92.30% (120). K. pneumoniae isolates showed high sensitivity to imipenem (15.38%) (20), followed by ertapenem, tetracycline, tigecycline pipracilline/tazobactam and amikacin (23.07%) (30). CONCLUSIONS: Our study showed that the prevalence of ESBL-producing K. pneumoniae at King Khalid Hospital was significantly high. Routine detection of ESBL-producing microorganisms is required by each of the laboratory standard detection methods to control the spread of these infections and allow a proper therapeutic strategy. For detection, the phenotypic confirmatory disc diffusion test is simple, sensitive and cost effective. However, there is a need for larger scale drug susceptibility surveillance.