Nicoleta Radu

Biomaterials obtained from probiotic consortia of microorganisms. Potential applications in regenerative medicine

ABSTRACT The beneficial effects of metabolites resulted from the consortium of probiotic microorganisms are known since the year 220 B.C. when they were used in Asia. The effects of these probiotic consortia were brought into actuality in the last century, after second World War, by researchers of the Moscow Academy of Sciences, during the studies regarding the incidence of cancer in Russia. They have found that at residents of some areas heavily polluted with radiation, the cancer incidence it is practically non-existent. Further investigations revealed that these people were healthy, due to the consumption of a specific mixture, which contains vitamins and minerals (bioproduct), obtained through fermentation, with specific yeasts. Studies have shown that fermented drink contains vitamins from the group B (B1, B2, B3, B5, B6, B7, B9, B12), vitamins A, C, E, polyphenols, amino acids, enzymes, glucuronic acid, usnic acid, caffeine, lactic acid, malonic acid, propionic acid, acetic acid, ethanol (0.5%) and protein (15 g/L). Microbiological analysis of the species involved in the bioprocess showed the existence of the specific symbiosis between several types of microorganisms type: Schizosaccharomyces pombe, Saccharomycodes ludwigii, Brettanomyces bruxellensis, Bacterium xylinum, Bacterium xylinoides Bacterium gluconicum, Bacterium katogenum, Pichia fermentans Bacterium, Acetobacter xylinum Gluconobacter oxydans, Torulaspora, delbueckii. The studies performed in vitro with this bioproduct (mixture), have confirmed that the immunomodulatory effect is due to cultivation media, which contain flavanols and catechins.

Biostimulatory Properties of Monascus sp. Bioproducts

The therapeutic metabolites properties of microorganisms obtained from Monascus sp in solid state biosynthesis in submerged media were studied and are presented. Some products isolated from solid state biosynthesis or from submerged media, were investigated in order to establish their biological or physico - chemical properties. The primary tests performed in order to establish the antioxidant properties revealed a strong antioxidant capacity situated in the range (1420 ÷ 3700) mMol Trolox Equivalent/mg product for intracellular metabolites obtained in submerged media or in solid state biosynthesis. The immunomodulatory tests performed in vitro on a monocyte human cell line with two products isolated from solid state biosynthesis show the stimulation of human macrophage (by induction of the secretion of TNF-α and IL-6 with differentiate cell THP-1 type). The physico-chemical characteristics of investigated compounds performed by FT-IR spectroscopy show the presence of N glucosylrubropunctamine and N-glucosylmonascorubramine in biomaterials obtained by solid biosynthesis.

Therapeutic Effect of Polysaccharides from Plantago Species

Analgesic and sedative effects of polysaccharides extract separated from alcoholic media of Plantain, were performed “in vivo” on mice, using the writhing and single exploration test. Antimicrobial activity of this extract was investigated “in vitro” using the Kirby Bauer method. The analgesic and sedative effects of this fraction without antimicrobial activity were also demonstrated. Physico-chemical analyses of this extract reveal the presence of C, H, P, K, microelements and the presence of polysaccharidic chain which comprises D-xylose, L-arabinose, rhamnose, and D-galacturonic acid.

Cytotoxicity Study Regarding Some Products Derived from Monascus sp.

Several species of the genus Monascus produce citrinin, a mycotoxin, are harmful to the hepatic and renal systems. For this reason, the citotoxicity study is necessary to evaluate the toxicological effect of Monascus products obtained from solid state or submerged biosynthesis. In our study we evaluate the content of mycotoxin in some Monascus products and the cytotoxicity effect of them. The study regarding citrinin content revealed a content of 145 ppm citrinin in the product obtained by solid state biosynthesis, a maximum content. In the case in which the biomaterials are obtained in submerged media, in the intracellular materials, the citrinin concentration was 82,71 mg/L (or 0,827 mg citrinin/g wet Monascus biomass). In the case of extracellular product obtained in submerged media, the mycotoxin content fluctuate in the range (7 ÷ 24,5) mg/L. Study performed ‘‘in vitro’’ with product obtained in submerged media on fibroblast murine cell lines, reveals a cytotoxicity effect for concentration larger than 60 μg/ml.

Therapeutic Effect of Flavonoids Derived from Plantago Species

Flavonoids are widely distributed in plants fulfilling many functions including producing yellow or red/blue pigmentation in flowers and protection from attack by microbes and insects. Current research shows for these compounds an anti-allergic, and an anti-cancer activity. The objectives of the research are the investigation of other therapeutic effects of flavonoid fraction separated from alcoholic extract of Plantago sp., as following: sedative, analgesic and anti-microbial activities. The physical-chemical properties of the extract were also investigated. The results obtained indicate the presence of D-apigenin, lutelin, scutellarin, baicalein, nepetin, hispidulin, plantagoside, potassium and microelements in the flavonoids extract. The anti-inflammatory effect (study performed on mice with pletismometric method)) was meaningful after 120 minute at a dose of 400 mg/kg body. The sedative effect (study performed on mice using the writhing test) was meaningful after 30 minutes at 400 mg/kg body. The antimicrobial activities (study performed “in vitro” using Kirby Bauer method) reveal the intense bacteriostatic activity against the microorganisms like Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus sp. This research suggests that plantain may be effective in the treatment of inflammatory and microbial disease but further studies are needed.

New nanobiomaterials based on irridoidic compounds

New type of nanomaterials has been synthesized using irridoidic extract derived from Plantago sp. The irridoidic compounds were separated from Plantago lanceolata by successive extraction in aqueous media. The composition of the stable nano-emulsion used for nanomaterials synthesis has been chosen from the pseudo ternary phase diagram and the dimensions of the emulsion were confirmed by Dynamic Light Scattering measurements. The obtained nanodrops were then encapsulated in silica resulting porous core - shell particles which were characterized by Dynamic Light Scattering and electronic microscopy confirming the nanostructure of the new biomaterials.

Biological properties of nanomaterials based on irridoidic compounds

Nanomaterials obtained from irridoidic extract were used for preclinical tests performed on mice in order to establish the influence of biomaterials on the cicatrisation and diarrhoea. The results obtained revealed a positive action on the cicatrisation process, and the crude extract processed as nanopowder showed a protective action against diarrhoea disorder.

Effects of biomaterials obtained from consortia of probiotic microorganism in submerged biosynthesis on THP1 cells line

ABSTRACT Two bioproducts, clear supernatant and solid bioproduct, obtained from submerged biosynthesis, with probiotic microorganisms consortia, black tea and dextrose, were tested in order to establish their antioxidant properties and their biological activity on THP1 cells line. The chemiluminescence studies revealed antioxidant properties for clear supernatant, which ranged between 92.10% and 97.06%. The results concerning the effect generated on THP1 cell lines, showed an apoptotic effect against these cells, the percent of viable cell decreasing from 93% for untreated cells, to 54% for cells treated with liquid supernatant and to 90% for cells treated with solid bioproduct.

Zingiber officinale based bioproduct. Properties and influence on some cellulolytic and keratinolytic fungi

ABSTRACT Two biomaterials obtained from Zingiber officinale were tested in order to reveal their antioxidant and antifungal properties: bioproduct 1 obtained by rhizomes hydrodistilation and bioproduct 2 obtained by pressing rhizomes. Chemiluminescence studies, showed that the bioproduct 1 posseses a strong antioxidant activity, which ranged between 97÷99% while the bioproduct 2, has an antioxidant activity which range between 45.21%÷12.55%. The best antifungal activity were obtain against Fusarium oxysporum (DL50 = 1139 µL·L−1), Fusarium graminearum (DL50 = 1199 µL·L−1), Tricoderma viride (DL50 = 1323 µL·L−1) Aspergillus ochraceus (DL50 = 1529 µL·L−1), Sclerotinia sclerotiorum (DL50 = 1650 µL·L−1), and Botrytis alii (DL50 = 1905 µL·L−1). The most resistant strains against were Aspergillus oryzae (DL50 = 2971 µL·L−1) and Aspergillus terreus (DL50 = 5788 µL·L−1).

Influence of some spice food based bioproducts on human monocytic cells line type THP-1

ABSTRACT Two biomaterials obtained by hydrodistilation from an indigenous Satureja hortensis plant (SHEO), and from the rhizoms of Zingiber officinale (ZOEO), purchased from Romanian market, were tested in order to evaluate their effect on monocytic cells line type THP-1. The first biomaterial, (SHEO), has practically no biological effect whereas the other bioproduct, (ZOEO) has a better effect in comparison to L-NAME and etoposide. Due to a mechanism which implies cells apoptosis the quantities of nitric oxide (NO)produced by THP-1 cells are reduced under ZOEO action.