Using osteogenic medium in the in vitro evaluation of bone biomaterials: Artefacts due to a synergistic effect.

In vitro tests using bone cells to evaluate the osteogenic potential of biomaterials usually employ the osteogenic medium (OM). The lack of correlation frequently reported between in vitro and in vivo studies in bone biomaterials, makes necessary the evaluation of the impact of osteogenic supplements on these results. This study analysed the proteomic profiles of human osteoblasts (HOb) cultured in the media with and without osteogenic agents (ascorbic acid and ß-glycerol phosphate). The cells were incubated for 1 and 7 days, on their own or in contact with Ti. The comparative Perseus analysis identified 2544 proteins whose expression was affected by osteogenic agents. We observed that the OM strongly alters protein expression profiles with a complex impact on multiple pathways associated with adhesion, immunity, oxidative stress, coagulation, angiogenesis and osteogenesis. OM-triggered changes in the HOb intracellular energy production mechanisms, with key roles in osteoblast maturation. HOb cultured with and without Ti showed enrichment in the skeletal system development function due to the OM. However, differentially expressed proteins with key regenerative functions were associated with a synergistic effect of OM and Ti. This synergy, caused by the Ti-OM interaction, could complicate the interpretation of in vitro results, highlighting the need to analyse this phenomenon in biomaterial testing.

Intake of Nutrient and Non-Nutrient Dietary Antioxidants. Contribution of Macromolecular Antioxidant Polyphenols in an Elderly Mediterranean Population.

The intake of antioxidants in the diet is a useful parameter to estimate the potential of diet to prevent chronic diseases associated with oxidative stress and ageing. The objective was to estimate the intake of nutrient and non-nutrient antioxidants associated with the dietary fiber matrix in a healthy and functionally independent population aged over 80, estimating the intake of antioxidant nutrients and including soluble low molecular weight and macromolecular polyphenols in the non-nutrient antioxidant group. Specific nutrients related to oxidative stress (copper, zinc, selenium, manganese, vitamins A, C and E) were ingested in optimal quantities according to reference values. Total intake of non-nutrient antioxidants was 2196 mg/person/day, and macromolecular polyphenols were found to be the main dietary antioxidants, contributing 71% to the total intake of phenolic compounds. The intake, metabolism and physiological effects of all nutrient and non-nutrient dietary antioxidants must therefore be taken into account when evaluating their health benefits.

By-product from decoction process of Hibiscus sabdariffa L. calyces as a source of polyphenols and dietary fiber.

BACKGROUND: Dietary fiber (DF) and antioxidant compounds are widely used as functional ingredients. The market in this field is competitive and the search for new types of quality ingredients for the food industry is intensifying. The aim of this study was to evaluate the composition and antioxidant activity of by-products generated during the decoction of calyces of four Mexican Hibiscus sabdariffa L. cultivars ('Criolla', 'China', 'Rosalis' and 'Tecoanapa') in order to assess them as a source of functional ingredients. RESULTS: Some calyx components were partially transferred to the beverage during the decoction process, while most were retained in the decoction residues. These by-products proved to be a good source of DF (407.4-457.0 g kg⁻¹ dry matter) and natural antioxidants (50.7-121.8 µmol Trolox equivalent g⁻¹ dry matter). CONCLUSION: The decoction process extracted some soluble carbohydrates, ash and some extractable polyphenols. The DF content changed in the dried residues, which could be considered as high-DF materials with a high proportion of soluble DF (∼20% of total DF) and considerable antioxidant capacity. These by-products could be used as an antioxidant DF source.

Grape antioxidant dietary fiber stimulates Lactobacillus growth in rat cecum.

UNLABELLED: The digesta is a highly active biological system where epithelial cells, microbiota, nondigestible dietary components, and a large number of metabolic products interact. The gut microbiota can be modulated by both endogenous and exogenous substrates. Undigested dietary residues are substrates for colonic microbiota and may influence gut microbial ecology. The objective of this work was to study the capacity of grape antioxidant dietary fiber (GADF), which is rich in polyphenols, to modify the bacterial profile in the cecum of rats. Male adult Wistar rats were fed for 4 wk with diets containing either cellulose or GADF as dietary fiber. The effect of GADF on bacterial growth was evaluated in vitro and on the cecal microbiota of rats using quantitative real time polymerase chain reaction (RT-PCR). The results showed that GADF intake stimulates proliferation of Lactobacillus and slightly affects the composition of Bifidobacterium species. GADF was also found to have a stimulative effect on Lactobacillus reuteri and Lactobacillus acidophilus in vitro. These findings suggest that the consumption of a diet rich in plant foods with high dietary fiber and polyphenol content may enhance the gastrointestinal health of the host through microbiota modulation. PRACTICAL APPLICATION: Grape antioxidant fiber combines nutritional and physiological properties of dietary fiber and natural antioxidants from grapes. Grape antioxidant fiber could be used as an ingredient for functional foods and as a dietary supplement to increase the intake of dietary fiber and bioactive compounds.

By-products of Opuntia ficus-indica as a source of antioxidant dietary fiber.

Dietary fiber and bioactive compounds are widely used as functional ingredients in processed foods. The market in this field is competitive and the development of new types of quality ingredients for the food industry is on the rise. Opuntia ficus-indica (cactus pear) produces edible tender stems (cladodes) and fruits with a high nutritional value in terms of minerals, protein, dietary fiber and phytochemicals; however, around 20% of fresh weight of cladodes and 45% of fresh weight of fruits are by-products. The objective of this study was therefore to determine the nutritional value of by-products obtained from cladodes and fruits from two varieties of Opuntia ficus-indica, examining their dietary fiber and natural antioxidant compound contents in order to obtain quality ingredients for functional foods and increase the added value of these by-products.

Bioactive compounds of four hot pepper varieties (Capsicum annuum L.), antioxidant capacity, and intestinal bioaccessibility.

Pepper fruits (Capsicum annuum) contain a wide array of phytochemicals with well-known antioxidant properties. Since bioactive compounds depend on their bioavailability to exert beneficial effects, it was crucial to estimate the extent of release from the food matrix and thus their bioaccessibility. Accordingly, we determined the individual carotenoid and phenolic content as well as the antioxidant properties of four red hot dried cultivars (Capsicum annuum L.) of high consumption in Mexico and estimated the extent of intestinal bioaccessibility of carotenoids with significance in human health, beta-carotene, beta-cryptoxanthin, and zeaxanthin, using an in vitro gastrointestinal model. Hot dried peppers at ripe stage had a high content of bioactive compounds that exhibited significant antioxidant properties (26-80 micromol trolox equivalents/g of dry matter), such as polyphenols (>2000 mg/100 g of dry matter) and carotenoids (95-437 mg/100 g of dry matter), which were partially bioaccessible. The amount released from the food matrix by the action of digestive enzymes was about 75% for total polyphenols, up to 49% for both beta-carotene and zeaxanthin, and up to 41% for beta-cryptoxanthin. The results suggest that from 50 to 80% of these carotenoids could reach the colon to be potentially fermented or could remain unavailable.

Stimulatory role of grape pomace polyphenols on Lactobacillus acidophilus growth.

Grape pomace is a natural product rich in dietary fiber and polyphenols. A major part of dietary polyphenols is not absorbed in the small intestine and can interact with colonic microbiota. The influence of grape polyphenols on Lactobacillus acidophilus CECT 903 growth was investigated through agar diffusion assays and cultures in liquid media. Grape phenolic extracts and some standards of phenolic compounds (caffeic acid, gallic acid, tannic acid, catechin, epicatechin, and quercetin) were assayed. All phenolic compounds tested did not exert an inhibitory effect on L. acidophilus growth at a maximum concentration of 5000 microg/disk in agar diffusion assays. In addition, a stimulatory trend in bacterial growth was observed in both grape phenolic extracts and tannic acid. The major finding was that grape pomace phenolic extract (1mg/mL) induced a significant biomass increase of L. acidophilus grown in liquid culture media. Further research into the interaction between phenolic compounds and other intestinal bacteria, as well as healthy consequences, is required.

Determination of beta-carotene and lutein available from green leafy vegetables by an in vitro digestion and colonic fermentation method.

Green leafy vegetables (Spinacea oleracea, Cnidoscolus aconitifolius, and Solanum americanum) contain a high amount of beta-carotene (27-52 mg/100 g of dry sample) and lutein (140-193 mg/100 g of dry sample). The amount of beta-carotene and lutein released from the food matrix by the action of digestive enzymes ranged from 22 to 67% and from 27 to 77%, respectively. There was a significant correlation between the enzymatic release of carotenoids (lutein + beta-carotene) and the content of Klason lignin, nonstarch polysaccharides, and resistant protein. The carotenoids released by the in vitro colonic fermentation ranged from 2 to 11%, and part of them (0.251-4.03 mg/100 g of original dry sample) remained intact in the fermentation media and could be potentially absorbed in the colon. A significant part of carotenoids seems to be unavailable in the intestinal tract (16% in S. oleracea to 58% in C. aconitifolius).

Molecular and microbiological analysis of caecal microbiota in rats fed with diets supplemented either with prebiotics or probiotics.

The potential health-improving effects of both a prebiotic and a probiotic infant formula have been evaluated in a rat model. Two groups of 10 rats were fed with either prebiotics containing fructo-oligosaccharides or probiotics containing viable Bifidobacterium lactis and Streptococcus thermophilus. The composition of their caecal microbiota was analyzed both by classical plate count of the main bacterial groups and by PCR amplification of a V3 fragment of 16S rRNA genes and denaturing gradient gel electrophoresis (DGGE). Both diets induced a significant reduction of clostridia and Bacteroides spp. compared to a control diet, whereas prebiotics were also able to reduce the number of coliforms and to increase the presence of bifidobacteria. DGGE analysis showed a significant increase of 16S rRNA gene fragments in rats fed with either probotics or prebiotics. Nineteen bands were sequenced and most of them showed similarity to cultured bacteria. Detection of Bifidobacterium spp. by this technique using genus-specific primers only permitted these bacteria to be detected in prebiotics-fed rats, whereas the use of Lactobacillus group-specific primers gave similar results in rats fed with any diet, in agreement with the plate count results.