In vitro evaluation of Spirulina platensis extract incorporated skin cream with its wound healing and antioxidant activities.

CONTEXT: Algae have gained importance in cosmeceutical product development due to their beneficial effects on skin health and therapeutical value with bioactive compounds. Spirulina platensis Parachas (Phormidiaceae) is renowned as a potential source of high-value chemicals and recently used in skincare products. OBJECTIVE: This study develops and evaluates skin creams incorporated with bioactive S. platensis extract. MATERIALS AND METHODS: Spirulina platensis was cultivated, the aqueous crude extract was prepared and in vitro cytotoxicity of S. platensis extract in the range of 0.001-1% concentrations for 1, 3 and 7 d on HS2 keratinocyte cells was determined. Crude extracts were incorporated in skin cream formulation at 0.01% (w/w) concentration and in vitro wound healing and genotoxicity studies were performed. Immunohistochemical staining was performed to determine the collagen activity. RESULTS: 0.1% S. platensis extract exhibited higher proliferation activity compared with the control group with 198% of cell viability after 3 d. Skin cream including 1.125% S. platensis crude extract showed enhanced wound healing effect on HS2 keratinocyte cell line and the highest HS2 cell viability % was obtained with this concentration. The micronucleus (MN) assay results indicated that S. platensis extract incorporated creams had no genotoxic effect on human peripheral blood cells. Immunohistochemical analysis showed that collagen 1 immunoreactivity was improved by increased extract concentration and it was strongly positive in cells treated with 1.125% extract incorporated skin cream. CONCLUSIONS: The cell viability, wound healing activity and genotoxicity results showed that S. platensis incorporated skin cream could be of potential value in cosmeceutical and biomedical applications.

Semi-IPN chitosan/polyvinylpyrrolidone microspheres and films: sustained release and property optimisation.

A set of chitosan-polyvinylpyrrolidone (CH-PVP) microspheres were prepared as semi-inter penetrating networks (semi-IPN) and loaded with 5-fluorouracil. In vitro release studies showed faster release for semi-IPN microspheres compared to pure CH samples, and the total release was achieved in about 20-30 days, depending on the composition. In vitro cell studies were achieved against human breast adenocarcinoma cell line cells where adsorption of cells on microspheres with a significant decrease in their number was obtained. Meanwhile, the CH-PVP films, which were prepared with the same compositions as in the microspheres, demonstrated an increase in strength from 66 to 118 MPa as the PVP content was decreased. It can be concluded that the prepared CH-PVP semi-IPN microspheres are novel promising carriers compared to pure CH microspheres since it becomes possible to adjust stability and hydrophilicity of the microspheres as well as the release rates of the drugs from the microspheres by changing the ratio of CH/PVP composition.

Metabolic activity and monoclonal antibody production of Salmonella enteritidis O and H antigen specific hybridoma cells in static culture.

The aim of the present work was to study the kinetics of two hybridomas that produce monoclonal antibody against Salmonella Enteritidis O (5A8) and H (D7) antigen. The hybridomas originated from the Ag8x653 (5A8) and Sp2/O (D7) myeloma cell lines. The relationship between the uptake of glucose and glutamine and the release of the lactate and ammonia and monoclonal antibody production into hybridoma growth were investigated in static culture with serum-containing DMEM/F:12 medium for the determination of pilot-production strategies of the hybridomas. Results showed that glucose and glutamine concentrations were reduced, with an increase in ammonia and lactate concentration in the culture medium. The hybridoma cell line 5A8 has shown lower metabolic activities compared with D7, whereas its monoclonal antibody productivity was found to be two-fold higher than the D7. MAb production by the hybridoma cell line 5A8 seems promising, considering the moderate level of productivity compared to that found in the literature.

In vitro growth stimulatory and in vivo wound healing studies on cycloartane-type saponins of Astragalus genus.

AIM OF THE STUDY: The present study was undertaken to evaluate the wound healing effects of the four chief saponins of Astragalus species [cycloastragenol (CA), astragaloside IV (AG), cyclocephaloside I (CCI) and cyclocanthoside E (CCE)]. MATERIAL AND METHODS: Effects of cell viability and proliferation of the isolated compounds were evaluated by the MTT assay on human keratinocyte. The wound healing activity was studied by using in vitro wound healing, proliferation and migration scratch assay. In order to see in vivo effectiveness of the compounds, an animal study with Sprague-Dawley male rats at the age of 12 weeks was carried out, and then the main histological outcomes were investigated to observe reepithelization, neovascularization, and presence of inflammatory cells, granulation tissue amount and maturation. RESULTS: All the compounds increased both fibroblast proliferation and migration, but the effects were much superior for CA at 1 ng/ml concentration. Among the compounds, based on the histological findings, 5% CA preparation was found to be the most remarkable in vivo wound healing agent showing greater cell density, more regularly organized dermis and more newly formed blood vessels. CONCLUSION: Results of this study indicate that the cycloartane-type saponins are the principal constituents responsible for wound healing activities of the roots of Astragalus species substantiating its use in traditional medicine.

Cytotoxic naphthoquinones from Alkanna cappadocica ( perpendicular).

In a continuing program to discover new anticancer agents from plants, especially naphthoquinones from the Alkanna genus, Alkanna cappadocica was investigated. Bioassay-guided fractionation of a dichloromethane/methanol (1:1) extract of the roots led to the isolation of four new and four known naphthoquinones. The known compounds are 11-deoxyalkannin (1), beta,beta-dimethylacrylalkannin (2), 11-O-acetylalkannin (3), and alkannin (4). The new compounds 5-O-methyl-11-deoxyalkannin (5), 8-O-methyl-11-deoxyalkannin (6), 5-O-methyl-11-O-acetylalkannin (7), and 5-O-methyl-beta,beta-dimethylacrylalkannin (8) were characterized by spectroscopic analyses (LC-ESIMS, 1D and 2D NMR). Cytotoxicity of the isolated compounds was evaluated versus 12 human cancer cell lines, HT-29, MDA-MB-231, PC-3, AU565, Hep G2, LNCaP, MCF7, HeLa, SK-BR-3, DU 145, Saos-2, and Hep 3B together with two normal cell lines, VERO and 3T3, by using the MTT assay. Compound 7 showed remarkable cytotoxicity with IC(50) values between 0.09 and 14.07 muM. It was more potent than the other compounds in six out of 12 cancer cell lines and the positive controls doxorubicin and etoposide. The mono-O-methylated alkannin derivatives and their cytotoxicities are reported for the first time.

Effect of human platelet-derived growth factor-BB on attachment of periodontal ligament cells on root surfaces.

OBJECTIVE: To evaluate the effects of platelet-derived growth factor-BB (PDGF-BB) on the attachment of human periodontal ligament cells (HPLCs) on the root surfaces demineralized with different agents. METHODS: We performed this study at Ege University, Izmir, Turkey between 2005 and 2006. Eighty root slices were subjected to one of following treatments after root planing: 1) only root planing, 2) Platelet derived growth factor-BB (PDGF-BB), 3) citric acid demineralization, 4) citric acid demineralization + PDGF-BB, 5) tetracycline hydrochloric acid (T-HCl) demineralization, 6) T-HCl demineralization + PDGF-BB, 7) ethylenediamine tetra-acetic acid (EDTA) demineralization, and 8) EDTA demineralization + PDGF-BB. Human periodontal ligament cells were seeded on the root surfaces. Following the 2-hour incubation period, the number of cells was calculated by the colorimetric assay. Three slices from each group were processed for scanning electron microscopy. The number of attached cells was tested by analysis of variance (p=0.050). RESULTS: There were no significant differences among the groups with regard to the mean number of attached cells (p=0.843), which was highest in the fourth group, and lowest in the sixth group. CONCLUSION: Root planing is the most important treatment to make the diseased root surfaces biocompatible to HPLCs adherence. Application of PDGF-BB to root surfaces demineralized with citric acid may be advocated to enhance periodontal regeneration.

Multidrug resistance in locally advanced breast cancer.

BACKGROUND: Advanced breast cancer cases can still be encountered resulting in poor prognosis. The primary treatment for these patients is chemotherapy, and multidrug resistance (MDR) is a serious obstacle in the treatment. Detecting drug resistance before first-line chemotherapy may increase the patient's survival. In this study, the role of MDR is evaluated in locally advanced breast cancer patients. METHODS: Reverse transcriptase polymerase chain reaction was used for the detection of MDR genes, ABCB1 and ABCC1. Immunohistochemistry was used for the detection of MDR proteins, P-glycoprotein (Pgp) and MDR-associated protein 1. RESULTS: Breast tissues from 25 patients both before and after chemotherapy were examined. Five patients were unresponsive to chemotherapy. Four had ABCB1 gene expression induced by chemotherapy, and Pgp positivity was detected in 9 patients after chemotherapy. Both the induction of ABCB1 gene expression (p < 0.001) and Pgp positivity (p < 0.001) during chemotherapy were significantly related with clinical response. Although 80% of the clinically unresponsive patients had ABCC1 gene expression, the relation between ABCC1 expression and clinical drug response was not significant. CONCLUSION: In locally advanced breast cancer, ABCB1 gene expression during chemotherapy contributes to clinical unresponsiveness. However, ABCC1 gene expression did not correlate strongly with the clinical response.

The effect of osteogenic medium on the adhesion of rat bone marrow stromal cell to the hydroxyapatite.

OBJECTIVE: To investigate the adhesive properties of bone marrow stromal cell (BMSC) on the hydroxyapatite (HA) particles and analyze their behavior. METHODS: The study took place in the Department of the Histology and Embryology, Celal Bayar University, Manisa and in the Department of Bioengineering, Ege University, Izmir, Turkey between 2004 and 2005. We cultured BMSC from the mature rat tibia and differentiated to the osteoblasts by osteogenic medium. The BMSCs were subcultured and were taken to the HA substrate. We measured their proliferation capacity and viability with MTT assay using the spectrophotometric method. Furthermore, we identified the osteoblast-like cells by immunohistochemical staining of osteonectin and osteocalcin and we analyzed the behavior of the cells on different sized HA particles by SEM at the end of 3 days incubation. RESULTS: Osteogenic medium caused the proliferation capacity of BMSC to speed up and the effects appeared earlier. We confirmed the osteoblastic differentiation by staining of most cells with osteoblastic markers. Subcultured cells were similarly adhesive to the HA particles and the osteogenic medium did not alter this behavior. They spread on the substrate similarly. Most of the cells demonstrated the cytoplasmic protrusion. Morphology of the cells did not change much with or without osteogenic medium. Different sizes of HA particles did not affect the adhesive properties of these cells except HA gel. The spreading and attachment ratios of the cells on HA gel were more than the others CONCLUSION: We found that there was heterogeneity in BMSC on differentiation capacity to the osteoblast, which was a sign of a subpopulation. Adhesive cells showed similar morphology and behavior under the effect of osteogenic medium. The only difference was the spreading capacity on the HA gel where cell used this substrate more effectively for adhesion.