Preparation of Chitosan/Alginate-ellagic Acid Sustained-release Microspheres and their Inhibition of Preadipocyte Adipogenic Differentiation

Objective: In this study, chitosan/alginate-ellagic acid sustained-release microspheres were prepared, and the effect of sustained-release microspheres on preadipocyte adipogenic differentiation was analyzed. Methods: Chitosan/alginate-ellagic acid microspheres were prepared and identified by scanning electron microscopy (SEM) and infrared spectroscopy (IR). The drug release rates were measured at pH 6.8, 7.0, 7.2, 7.4 to determine sustained release of ellagic acid from microspheres. The effects of 0.1, 1, 10 mg/L chitosan/alginate-ellagic acid microsphere on 3T3-F442A preadipocyte proliferation were determined by Methyl thiazolyl tetrazolium assay (MTT), and cell morphology was checked by hematoxylin/ eosin staining (HE staining). The effect of chitosan/alginate-ellagic acid microspheres on preadipocyte adipogenic differentiation was also determined by Oil red O staining, and lipogenesis was measured by isopropanol extraction. The molecular mechanism was investigated by detecting the mRNA expression of CCAAT/enhancer binding protein alpha (C/EBPα) and peroxisome proliferatorsactivated receptor gamma (PPARγ). Results: Chitosan/alginate-ellagic acid sustained-release microspheres were successfully prepared, and the inhibition of proliferation and adipogenic differentiation of preadipocytes was found to be dosedependent. The mechanism of differentiation inhibition was found to be closely related to the expression of transcription factor C/EBPα and PPARγ. Conclusion: Chitosan/alginate can be used as a good material to prepare ellagic acid sustained-release microspheres, and these microspheres can be used for treating the obesity.

Immunological and molecular study of Chlamydia trachomatis as causative agent of abortion in Al-Muthanna province

Chlamydiasis during pregnancy should be considered a significant risk factor for adverse pregnancy outcomes in humans. 120 women who had a single or repeated abortion were selected for this study, and they were referred with a physician report for TORCH tests to determine the final diagnosis of pregnancy loss. The control were (40) healthy pregnant women with a history of a normal pregnancy. The innate immunity in abortive women was higher than normal pregnancies, that was estimated by Nitroblue Tetrazolium test done to estimate the phagocytic activity, there was a significant increase (P=0.009) in phagocytic activity in the leukocytes of abortive women which was (22%) higher than that in control. Also, the cellular immune response was higher in abortive women than that in control. Methyl Thiazolyl Tetrazolium assay was performed to estimate lymphocyte transformation index of peripheral blood leukocytes in abortive women. The results of Methyl Thiazolyl Tetrazolium assay showed a significant increase (P=0.001) in the lymphocyte transformation index in the lymphocytes of abortive women which was (27%) higher than control. The serological detection by ELISA showed that anti-C. trachomatis IgG was (14.2%), and the molecular detection by Quantitive Real Time-Polymerase Chain Reaction showed positive results (17.5%) of total abortive women. The present study demonstrated a high level of relationship between C. trachomatis and abortion among women in the study samples. The age group (20-25years) was the most susceptible to chlamydial infection and the infection was higher in recurrent miscarriages than in single miscarriage.

ANTIBACTERIAL EFFECT OF VIRGIN COCONUT OIL ON THE VIABILITY OF CHROMOGENIC BACTERIA THAT CAUSES DENTAL BLACK STAIN IN CHILDREN

Introduction: Virgin coconut oil (VCO) is a natural antibacterial agent with the ability to kill microorganisms whose cell membranes contain lipids, including Gram-positive and Gram-negative bacteria, such as Actinomyces sp. and Prevotella spp. No study on the antibacterial effects of VCO on the chromogenic bacteria that causes dental black stains.Objective: To analyze the effects of VCO, which administered in various concentrations to Actinomyces sp. and Prevotella sp.Methods: Actinomyces sp. and Prevotella sp. were isolated from the dental plaque of a child diagnosed with black stain. Each streak of bacteria was cultured on a selective medium and confirmed visually and through Gram staining. Each bacterial culture was exposed to VCO in concentrations of 12.5%, 25%, 50%, and 100%. Afterward, viability testing with a methyl-thiazolyl-tetrazolium assay was conducted, and the results were read using an Enzyme-linked immunosorbent assay reader.Results: The administration of 12.5% VCO reduced bacterial viability; however, 25% VCO significantly reduced the viability of Actinomyces sp. and 100% VCO significantly reduced the viability of Prevotella sp.Conclusion: Actinomyces sp. is more sensitive to VCO than Prevotella sp.

Sinomenine-loaded microcapsules fabricated by phase reversion emulsification-drying in liquid method: An evaluation of process parameters, characterization, and released properties

Sinomenine is a natural alkaloid with important biological activities (e.g. anti-cancer, anti-inflammatory, and anti-allergic). However, the unstability and short half-life absolutely limited its application to foods. Microencapsulation technology can offer a way to solve these issues. In this study, polylactic acid microcapsules loading sinomenine hydrochloride were fabricated by phase inversion emulsification-drying in liquid technique. The results showed that microcapsules had nice spherical shape, uniform particle size, and free flowing. The encapsulation efficiency was 89.2% and drug loading was 8.9% under the optimal conditions. In vitro release assays demonstrated that release of sinomenine from microcapsules was sustained and slow. Moreover, it was found that the sinomenine release fitted Fickian diffusion mechanism. The results of cytotoxicity study showed that sinomenine-loaded microcapsules were biocompatible. Sinomenine-loaded microcapsules could inhibit the growth of MDA-MB-231 cells using methyl thiazolyl tetrazolium assay. In summary, polylactide microcapsules exhibit excellent properties for sinomenine that can be used in drug or food industry.

Proteasome Inhibitor MG132 Enhances Sensitivity to Cisplatin on Ovarian Carcinoma Cells In Vitro and In Vivo

BackgroundPlatinum-based combination chemotherapy after surgery is considered a standard treatment; therefore, any recent drug development should be new, effective, and low toxic, and should have a synergistic effect with platinum. This study aimed to observe the growth of SKOV3 cells after treatment with cisplatin by combining with carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG132) and to investigate the effect of the relationship between MG132 and cisplatin combination.Materials and MethodsCell growth was detected by methyl thiazolyl tetrazolium assay after treatment with MG132 at 0.5, 1.5, 2.5, 3.5, and 5.0 μg/mL concentrations for 24, 48, and 72 hours; with cisplatin at 1.0, 2.0, 3.0, 4.0, and 5.0 μg/mL concentrations; and with combination with MG132 at 1.5 μg/mL for 24 hours. The apoptotic rates of cells were detected by a flow cytometer after cisplatin treatment at 1.0, 2.0, 3.0, and 4.0 μg/mL concentrations and that combined with MG132 at 1.5 μg/mL concentration for 12, 24, and 36 hours. A total of 20 BALB/c (nu/nu) female nude mice (age, 4–6 weeks; body weight, 17–19 g) were divided into 4 groups: control, MG132, cisplatin, and combination groups. The expression of Caspase3 and Beclin1 was detected by Western blot analysis and reverse transcription-polymerase chain reaction after treatment with 3.0 μg/mL of the cisplatin group and combined treatment with 1.5 μg/mL of MG132 group for 24 hours, respectively.ResultsMethyl thiazolyl tetrazolium assay demonstrated the inhibitory rates, and the flow cytometery showed that the apoptotic rates in the combination group were higher than those in the cisplatin group (P < 0.01). Western blot analysis and reverse transcription-polymerase chain reaction detected that Caspase3 and Beclin1 at a relative quantity in the combination group were higher than those in the cisplatin group (P < 0.05).ConclusionsMG132 has a synergistic antitumor effect by combining with cisplatin, and it is expected to be an effective antitumor drug for platinum-resistant refractory ovarian cancer.

Cytotoxicity Test of Brown Silk from Attacus atlas L. on Rat Smooth Muscle Cells

Natural brown silk biomaterial from silkworm Attacus atlas L. (A. atlas L.) is a biopolymer that has many advantages compared to commercial silk from Bombyx mori (B. mori). The aim of this study was to investigate the cytotoxicity of A. atlas L. silk on rat smooth muscle cells with the direct method of methyl thiazolyl tetrazolium assay compared to commercial surgical suture silk. The result showed that brown colored natural silk has low cytotoxicity as same as commercial surgical suture silk with slightly lower cell viability. It was found that A. atlas L. silk potentially can developed to be surgical suture thread as a new expectation of biopolymer material in biomedical products.