Enas Elzamarany

Development of mucoadhesive microbeads using thiolated sodium alginate for intrapocket delivery of resveratrol

Resveratrol (Res), a polyphenolic phytoalexin, had shown a promising therapeutic efficacy towards treatment of periodontal disease in vitro. This work aims to develop Res microbeads with strong mucoadhesion using thiolated alginate (TA) for local treatment of periodontal pockets. TA was synthesized by conjugating sodium alginate (A) with thioglycolic acid. Product was evaluated by IR and DSC. Both A and A:TA Res microbeads with different ratios were prepared by ionotropic gelation method. Formulations were evaluated regarding their entrapment efficiency (%EE), swelling index (SI), in vitro drug release and kinetics. Selected formula was examined for its mucoadhesion by ex vivo wash-off method, surface morphology using scanning electron microscope (SEM) and stability against light. Clinical evaluation is running.Formation of TA was confirmed. %EE for all formulations ranged from 83.72 to 104.54%. Results revealed a significant lower SI for TA rich formulation (A/TA 1:1) along with slower release rate and zero-order kinetics, in addition to powerful mucoadhesion; 26% remaining of microbeads after 1h, compared to 2% for A microbeads. SEM micrographs showed a rough surface with drug precipitation. The formula maintained its %EE after 5h exposure to direct sunlight. A/TA 1:1 mucoadhesive Res microbeads could be exploited as a prolonged drug release devices for intrapocket application.

Serum Soluble Fas in Patients with Schistosomal Cor pulmonale

Background: Schistosomal cor pulmonale is considered an important pathological condition in endemic areas. Few recent studies have reported the role of apoptosis in pulmonary hypertension. Objectives: The aim of this study was to assess serum levels of soluble Fas (sFas), an inhibitor of apoptosis, in patients with schistosomal cor pulmonale as compared to patients with cor pulmonale due to chronic obstructive pulmonary disease (COPD) and normal subjects. Methods: Serum sFas was assessed in 15 men with schistosomal cor pulmonale (age 32 ± 10 years), 15 men with chronic cor pulmonale secondary to COPD and 20 healthy men, matched for age. Results: Serum levels of sFas were significantly higher in patients with schistosomal cor pulmonale (74 ± 80 U/ml) than in patients with cor pulmonale due to COPD (15 ± 10 U/ml) and normal subjects (19 ± 11 U/ml, p < 0.001 in both). In patients with schistosomal cor pulmonale, sFas was significantly higher in patients with mean pulmonary artery pressure >30 mm Hg as compared to patients with pressure ≤30 mm Hg (109 ± 97 vs. 34 ± 20 U/ml, p = 0.01). There was a significant correlation between serum sFas and the mean pulmonary artery pressure in patients with bilharzial cor pulmonale (r = 0.4, p < 0.01), but not in patients with COPD (r = 0.1, p = NS). Conclusions: Serum sFas levels are elevated in patients with schistosomal cor pulmonale and they are related to the severity of pulmonary hypertension. These findings suggest a role of apoptosis in schistosomal cor pulmonale.

Intra-arterial Infusion of Autologous Bone Marrow Mononuclear Stem Cells in Subacute Ischemic Stroke Patients

Introduction Based on many preclinical and small clinical trials, stem cells can help stroke patient with the possibility of replacing the cells and supporting the remaining cells. The aim of this study was to evaluate the safety and feasibility of bone marrow mononuclear (BMMN) stem cell transplantation in subacute ischemic stroke patients. Materials and methods Thirty-nine (n = 39) patients with subacute ischemic cerebral infarct due to large artery occlusion in the middle cerebral artery (MCA) territory were recruited. They were distributed into two groups: first group (n = 21) served as an experimental group, which received intra-arterial (IA) mononuclear stem cells (bone marrow-derived mononuclear cell), while the other group (n = 18) served as a control group. All the patients were evaluated clinically by National Institutes of Health Stroke Scale, modified Rankin Scale, Barthel Index, modified and standardized Arabic version of the Comprehensive Aphasia Test, and radiological for 12 months. Results The stem cell-treated group showed better improvement, but it was not significant when compared with the non-treated group. The volume of infarction changes at the end of the study was non-significant between both the groups. There was no, or minimal, adverse reactions in stem cell-treated group. Conclusion The study results suggest that autologous BMMN stem cell IA transplantation in subacute MCA ischemic stroke patients is safe with very minimal hazards, but no significant improvement of motor, language disturbance, or infarction volume was detected in stem cell-treated group compared with the non-treated group.

In vitro and in vivo studies of the immunomodulatory effect of Echinacea purpurea on dendritic cells

Background Extracts of Echinacea have been used traditionally for the treatment of diverse types of infections and wounds. They have become very familiar immunostimulant herbal medicine. However, the specific immunomodulatory effect of Echinacea remains to be elucidated. Aim In our study, the effect of Echinacea purpurea extract on the generation of immature DCs from monocytes was described, as well as its effect on DC differentiation. In addition, an in vivo experiment was conducted to investigate whether treatment of mice with extracts derived from E. purpurea has immunomodulatory effect on murine splenic DCs. Methods Immature DCs were generated by incubating peripheral blood monocytes with cytokine cocktail (GM-CSF + IL-4) and matured by tumor necrosis factor-α (TNF-α). The cells were randomized to 5 groups to investigate E. purpurea effect in different stages. Phenotypic analysis of cell marker CD83-expressed on DCs was performed by flow cytometry. Mice were randomly divided into 3 groups; control, E. purpurea treated and E. purpurea-TNF-α treated group. The murine splenic DCs were isolated and phenotyped for CD83 and CD11c by flow cytometry. Results Treatment of monocytes with E. purpurea prior to addition of the maturation factor TNF-α resulted in a significant decrease in the yield of DC expressing CD83. On the other hand, immature DCs generated in the culture in the presence of GM-CSF and IL-4, when treated simultaneously with E. purpurea and TNF-α, exhibited an insignificant change in the yield of CD83-expressing DCs compared with untreated control. The in vivo experiments showed that splenic DCs obtained from mice treated with E. purpurea with or without TNF-α did not exhibit significant changes in CD83 or CD11c compared with those obtained from control mice. Conclusion Our findings suggest that the immunomodulatory mechanisms of E. purpurea impact generation fate of DCs rather than differentiation stages. The results obtained in the in vivo study utilizing murine splenic DCs supported those observed in vitro.

Immunotherapeutic strategies for treatment of hepatocellular carcinoma with antigen-loaded dendritic cells: in vivo study

AbstractHepatocellular carcinoma (HCC) is one of the major health problems in the world. DCs-based vaccines are a promising immunotherapeutic strategy that aims at the optimal for induction of a specific antitumor immune response and destruction of tumor cells. The present study was conducted to investigate the immunogenic characters of whole tumor lysate-pulsed DCs vaccine and its ability to induce a specific antitumor immune response in HCC mice model. We also evaluate the effectiveness of prophylactic and therapeutic immunization strategies against HCC in mice models. Mice-derived DCs were in vitro loaded with whole tumor lysate prepared from liver tissue of HCC mice and evaluated for expression of surface maturation markers CD83 and CD86. In vivo immunization of mice with whole tumor lysate-pulsed DCs was performed in two strategies; prophylactic (pre-exposure to HCC) and therapeutic (post-exposure to HCC). Effectiveness of both protocols was investigated in terms of histopathological examination of liver sections and measurement of serum levels of immune cytokines interferon-γ (IFN-γ) and interleukin-2 (IL-2). Loading of DCs with whole tumor cell lysate exhibited a significant increase in expression of CD83 and CD86. In vivo administration of prophylactic doses of whole tumor lysate-pulsed DCs in mice before induction of HCC evokes a strong antitumor immune response presented by absence of malignant cells in liver sections and the significant increase in IFN-γ and IL-2. Data herein indicated that prophylactic vaccination with whole tumor lysate-pulsed DCs exhibited an effective antitumor immune response against HCC more than therapeutic protocol.

Inducing hepatogenic differentiation of human mesenchymal stem cells derived from umbilical cord blood

Background and aim The present work aimed to study the ability of human mesenchymal stem cells (MSCs) derived from umbilical cord blood (UCB) to transdifferentiate into hepatocytes and to assess the characterization of the transformed cells in vitro. Materials and methods MSCs were isolated from 30 UCB samples collected aseptically from completely separated placentas of full-term deliveries. The separation of MSCs was carried out from freshly isolated mononuclear cells suspensions in a primary culture for 2 weeks. MSCs were identified before induction by cytochemical stain for periodic acid-Schiff and morphology. Then, they were induced to transdifferentiate into hepatocytes by hepatogenic medium, and differentiation of hepatocytes was confirmed by morphological and functional assessments of urea production, glycogen storage, and immunocytochemistry of α-fetoprotein. Results We successfully isolated 12 MSCs units from 30 full-term UCB samples (40%). The cells showed positive staining for periodic acid-Schiff, indicating that they retain the characteristics of MSCs. The response of UCB-derived MSCs to hepatogenic medium containing hepatocyte growth factor was assessed by changes in the morphology that occurred within 2 weeks in most of the cells. The morphological changes were observed closely and the induction process was monitored by immunocytochemistry for α-fetoprotein, urea production, and glycogen storage. The results were positive in most of the induced hepatocytes, with some variations from sample to sample because of the variability in the cell count in each UCB unit used. Conclusion We concluded that MSCs in human UCB can differentiate into viable functioning hepatocytes when cultured in hepatogenic conditioned medium in vitro.