Yeast-Derived Nucleotides Enhance Fibroblast Migration and Proliferation and Provide Clinical Benefits in Atopic Dermatitis.

Nucleotides, glycosaminoglycans, and omega-3 essential fatty acids (O3s) could be used for improving skin health, although their modes of action, alone or in combination, are not yet fully understood. To gain some insight into these mechanisms, we performed two in vitro tests and one in vivo pilot trial. The effects on human dermal fibroblast proliferation and migration were evaluated with the following compounds and combinations: 0.156 mg/mL O3s, 0.0017 mg/mL hyaluronic acid (HA), 0.0004 mg/mL dermatan sulfate (DS), 0.0818 mg/mL nucleotides, and [O3s + HA + DS] and [O3s + HA + DS + nucleotides] at the same concentrations. In both in vitro assays, adding nucleotides to [O3s + HA + DS] provided significant improvements. The resulting combination [O3s + HA + DS + nucleotides] was then tested in vivo in dogs with atopic dermatitis by oral administration of a supplement providing a daily amount of 40 mg/kg nucleotides, 0.9 mg/kg HA, 0.18 mg/kg DS, 53.4 mg/kg EPA, and 7.6 mg/kg DHA. After 30 days, the pruritus visual analog scale (pVAS) score was significantly reduced, and no adverse effects were observed. In conclusion, the combination of nucleotides plus glycosaminoglycans and O3s could serve as a useful therapeutic alternative in skin health applications.

Collagen Supplementation for Joint Health: The Link between Composition and Scientific Knowledge.

Osteoarthritis (OA) is the most common joint disease, generating pain, disability, and socioeconomic costs worldwide. Currently there are no approved disease-modifying drugs for OA, and safety concerns have been identified with the chronic use of symptomatic drugs. In this context, nutritional supplements and nutraceuticals have emerged as potential alternatives. Among them, collagen is being a focus of particular interest, but under the same term different types of collagens coexist with different structures, compositions, and origins, leading to different properties and potential effects. The aim of this narrative review is to generally describe the main types of collagens currently available in marketplace, focusing on those related to joint health, describing their mechanism of action, preclinical, and clinical evidence. Native and hydrolyzed collagen are the most studied collagen types for joint health. Native collagen has a specific immune-mediated mechanism that requires the recognition of its epitopes to inhibit inflammation and tissue catabolism at articular level. Hydrolyzed collagen may contain biologically active peptides that are able to reach joint tissues and exert chondroprotective effects. Although there are preclinical and clinical studies showing the safety and efficacy of food ingredients containing both types of collagens, available research suggests a clear link between collagen chemical structure and mechanism of action.

A Novel Hyaluronic Acid Matrix Ingredient with Regenerative, Anti-Aging and Antioxidant Capacity.

Hyaluronic acid (HA) and proteoglycans (such as dermatan sulphate (DS) and chondroitin sulphate (CS)) are the main components of the extracellular matrix of the skin, along with collagen and elastin. These components decrease with age, which implies a loss of skin moisture causing wrinkles, sagging and aging. Currently, the external and internal administration of effective ingredients that can reach the epidermis and dermis is the main alternative for combating skin aging. The objective of this work was to extract, characterise and evaluate the potential of an HA matrix ingredient to support anti-aging. The HA matrix was isolated and purified from rooster comb and characterised physicochemically and molecularly. In addition, its regenerative, anti-aging and antioxidant potential and intestinal absorption were evaluated. The results show that the HA matrix is composed of 67% HA, with an average molecular weight of 1.3 MDa; 12% sulphated glycosaminoglycans, including DS and CS; 17% protein, including collagen (10.4%); and water. The in vitro evaluation of the HA matrix's biological activity showed regenerative properties in both fibroblasts and keratinocytes, as well as moisturising, anti-aging and antioxidant effects. Furthermore, the results suggest that the HA matrix could be absorbed in the intestine, implying a potential oral as well as topical use for skin care, either as an ingredient in a nutraceutical or a cosmetic product.

Effectiveness of a low-fat yoghurt supplemented with rooster comb extract on muscle strength in adults with mild knee pain and mechanisms of action on muscle regeneration.

PURPOSE: To determine the effects of the intake of low-fat yoghurt supplemented with rooster comb extract (RCE) on muscle strength. METHODS AND RESULTS: 148 subjects, with mild knee pain, participated in a randomized, placebo-controlled, double-blind, and parallel study. Muscle strength, knee effusion, and pain perception were measured. C2C12 myoblasts were used to elucidate the mechanisms of action involved. RCE improved total work and mean power in men, and also peak torque in extension by 10%. RCE reduced synovial effusion by 11.8% and pain perception by 24.6%. Both RCE and HA increased myoblast proliferation by 29%, while RCE reduced myoblast differentiation by 36.2%, suggesting a beneficial role of RCE in muscle regeneration. CONCLUSIONS: Low-fat yoghurt supplemented with RCE improved muscle strength. This effect is partially explained by muscle regeneration enhancement, reduced synovial effusion, and reduced pain perception, which could exert a beneficial clinical impact on men affected by mild knee pain.

Oral chondroitin sulfate and prebiotics for the treatment of canine Inflammatory Bowel Disease: a randomized, controlled clinical trial.

BACKGROUND: Canine inflammatory bowel disease (IBD) is a chronic enteropathy of unknown etiology, although microbiome dysbiosis, genetic susceptibility, and dietary and/or environmental factors are hypothesized to be involved in its pathogenesis. Since some of the current therapies are associated with severe side effects, novel therapeutic modalities are needed. A new oral supplement for long-term management of canine IBD containing chondroitin sulfate (CS) and prebiotics (resistant starch, ß-glucans and mannaoligosaccharides) was developed to target intestinal inflammation and oxidative stress, and restore normobiosis, without exhibiting any side effects. This double-blinded, randomized, placebo-controlled trial in dogs with IBD aims to evaluate the effects of 180 days administration of this supplement together with a hydrolyzed diet on clinical signs, intestinal histology, gut microbiota, and serum biomarkers of inflammation and oxidative stress. RESULTS: Twenty-seven client-owned biopsy-confirmed IBD dogs were included in the study, switched to the same hydrolyzed diet and classified into one of two groups: supplement and placebo. Initially, there were no significant differences between groups (p > 0.05) for any of the studied parameters. Final data analysis (supplement: n = 9; placebo: n = 10) showed a significant decrease in canine IBD activity index (CIBDAI) score in both groups after treatment (p < 0.001). After treatment, a significant decrease (1.53-fold; p < 0.01) in histologic score was seen only in the supplement group. When groups were compared, the supplement group showed significantly higher serum cholesterol (p < 0.05) and paraoxonase-1 (PON1) levels after 60 days of treatment (p < 0.01), and the placebo group showed significantly reduced serum total antioxidant capacity (TAC) levels after 120 days (p < 0.05). No significant differences were found between groups at any time point for CIBDAI, WSAVA histologic score and fecal microbiota evaluated by PCR-restriction fragment length polymorphism (PCR-RFLP). No side effects were reported in any group. CONCLUSIONS: The combined administration of the supplement with hydrolyzed diet over 180 days was safe and induced improvements in selected serum biomarkers, possibly suggesting a reduction in disease activity. This study was likely underpowered, therefore larger studies are warranted in order to demonstrate a supplemental effect to dietary treatment of this supplement on intestinal histology and CIBDAI.

A 3-Arm Randomized Trial for Achilles Tendinopathy: Eccentric Training, Eccentric Training Plus a Dietary Supplement Containing Mucopolysaccharides, or Passive Stretching Plus a Dietary Supplement Containing Mucopolysaccharides.

BACKGROUND: Tendinopathy is an overuse tendon injury that occurs in loaded tendons and results in pain and functional impairment. Although many treatments for painful tendons are described, the scientific evidence for most of the conservative and surgical treatments is not always conclusive. OBJECTIVES: This study was designed to evaluate the efficacy of 3 different interventions in patients with Achilles tendinopathy. The interventions include the combination of 2 physical therapy programs (eccentric training [EC] or passive stretching [PS]) with a supplement containing mucopolisaccharides. The efficacy of the interventions was evaluated depending on the stage of the disease. METHODS: Fifty-nine patients were randomly assigned to 1 of 3 treatment groups, and classified according to the disease stage: reactive versus degenerative tendinopathy. Treatment groups were EC; EC + a dietary supplement containing mucopolisaccharides, type I collagen, and vitamin C (MCVC); and a passive stretching program + MCVC. Patients were evaluated at baseline, 6 weeks, and 12 weeks with the Victorian Institute of Sports Assessment-Achilles questionnaire for function, a visual analog scale for pain, and ultrasound characterization for the evolution of tendon structure. RESULTS: A significant improvement in Victorian Institute of Sports Assessment-Achilles questionnaire score, pain at rest, and pain during activity were detected in all 3 treatment groups at 6 and 12 weeks' follow-up when compared with baseline. In patients with reactive tendinopathy, the reduction in pain at rest was greater in the groups who took the supplemental MCVC than in the EC alone group (P < 0.05). CONCLUSIONS: MCVC seems to be therapeutically useful for management of tendinopathies, providing some additional benefit to physical therapy. This is especially evident in early stages of the disease, when the tendon does not present severe matrix and vascular changes. CLINICALTRIALSGOV IDENTIFIER: NCT01691716.

Synergistic Effects of a Mixture of Glycosaminoglycans to Inhibit Adipogenesis and Enhance Chondrocyte Features in Multipotent Cells.

BACKGROUND/AIMS: Multipotent mesenchymal stem cells affect homeostasis of adipose and joint tissues. Factors influencing their differentiation fate are of interest for both obesity and joint problems. We studied the impact of a mixture of glycosaminoglycans (GAGs) (hyaluronic acid: dermatan sulfate 1:0.25, w/w) used in an oral supplement for joint discomfort (Oralvisc™) on the differentiation fate of multipotent cells. METHODS: Primary mouse embryo fibroblasts (MEFs) were used as a model system. Post-confluent monolayer MEF cultures non-stimulated or hormonally stimulated to adipogenesis were chronically exposed to the GAGs mixture, its individual components or vehicle. The appearance of lipid laden cells, lipid accumulation and expression of selected genes at the mRNA and protein level was assessed. RESULTS: Exposure to the GAGs mixture synergistically suppressed spontaneous adipogenesis and induced the expression of cartilage extracellular matrix proteins, aggrecan core protein, decorin and cartilage oligomeric matrix protein. Hormonally-induced adipogenesis in the presence of the GAGs mixture resulted in decreased adipogenic differentiation, down-regulation of adipogenic/lipogenic factors and genes for insulin resistance-related adipokines (resistin and retinol binding protein 4), and up-regulation of oxidative metabolism-related genes. Adipogenesis in the presence of dermatan sulfate, the minor component of the mixture, was not impaired but resulted in smaller lipid droplets and the induction of a more complete brown adipocyte-related transcriptional program in the cells in the adipose state. CONCLUSIONS: The Oralvisc™ GAGs mixture can tip the adipogenic/chondrogenic fate balance of multipotent cells away from adipogenesis while favoring chondrocyte related gene expression. The mixture and its dermatan sulfate component also have modulatory effects of interest on hormonally-induced adipogenesis and on metabolic and secretory capabilities of adipose cells.

Oral Nucleotides Only Minimally Improve 5-Fluorouracil-Induced Mucositis in Rats.

Chemotherapy-induced mucositis is characterized by inflammation and ulceration of the intestinal mucosa, compromising intestinal function. Exogenous nucleotides have been reported to repair the mucosa. The nucleotide preparation, Nucleoforce F0328 (Nucleoforce), was investigated for its potential to ameliorate intestinal mucositis in rats. Female Dark Agouti rats (n = 8/group) were gavaged once daily with Nucleoforce (175 mg/kg) or water from Days 0 to 8 and injected (i.p.) with 5-fluorouracil (5-FU; 150 mg/kg) or saline on Day 5. Histological parameters (disease severity, crypt depth, and villus height measurements) and myeloperoxidase activity were quantified. P < 0.05 was considered significant. Jejunal and ileal histological disease severity scores were significantly increased by 5-FU, compared to normal controls (P < 0.05). Nucleoforce treatment in 5-FU-injected rats significantly reduced jejunal and ileal disease severity compared to 5-FU controls (P < 0.05). In 5-FU-injected rats, jejunal and ileal villus heights and crypt depths were significantly decreased compared to 5-FU controls, with no additional Nucleoforce effect (P > 0.05). Intestinal myeloperoxidase activity was significantly elevated by 5-FU (8.8-fold), compared to normal controls (P < 0.05), which was not normalized by Nucleoforce treatment (P > 0.05). Nucleoforce only partially improved parameters associated with experimentally-induced mucositis. Future studies could investigate increased concentrations, more frequent administration, or protective microencapsulation delivery methods, to increase bioavailability.

A low-fat yoghurt supplemented with a rooster comb extract on muscle joint function in adults with mild knee pain: a randomized, double blind, parallel, placebo-controlled, clinical trial of efficacy.

Preliminary results suggested that oral-administration of rooster comb extract (RCE) rich in hyaluronic acid (HA) was associated with improved muscle strength. Following these promising results, the objective of the present study was to evaluate the effect of low-fat yoghurt supplemented with RCE rich in HA on muscle function in adults with mild knee pain; a symptom of early osteoarthritis. Participants (n = 40) received low-fat yoghurt (125 mL d(-1)) supplemented with 80 mg d(-1) of RCE and the placebo group (n = 40) consumed the same yoghurt without the RCE, in a randomized, controlled, double-blind, parallel trial over 12 weeks. Using an isokinetic dynamometer (Biodex System 4), RCE consumption, compared to control, increased the affected knee peak torque, total work and mean power at 180° s(-1), at least 11% in men (p < 0.05) with no differences in women. No dietary differences were noted. These results suggest that long-term consumption of low-fat yoghurt supplemented with RCE could be a dietary tool to improve muscle strength in men, associated with possible clinical significance. However, further studies are needed to elucidate reasons for these sex difference responses observed, and may provide further insight into muscle function.

Blood cells transcriptomics as source of potential biomarkers of articular health improvement: effects of oral intake of a rooster combs extract rich in hyaluronic acid.

The aim of the study was to explore peripheral blood gene expression as a source of biomarkers of joint health improvement related to glycosaminoglycan (GAG) intake in humans. Healthy individuals with joint discomfort were enrolled in a randomized, double-blind, placebo-controlled intervention study in humans. Subjects ate control yoghurt or yoghurt supplemented with a recently authorized novel food in Europe containing hyaluronic acid (65 %) from rooster comb (Mobilee™ as commercial name) for 90 days. Effects on functional quality-of-life parameters related to joint health were assessed. Whole-genome microarray analysis of peripheral blood samples from a subset of 20 subjects (10 placebo and 10 supplemented) collected pre- and post-intervention was performed. Mobilee™ supplementation reduced articular pain intensity and synovial effusion and improved knee muscular strength indicators as compared to placebo. About 157 coding genes were differentially expressed in blood cells between supplemented and placebo groups post-intervention, but not pre-intervention (p < 0.05; fold change ≥1.2). Among them, a reduced gene expression of glucuronidase-beta (GUSB), matrix metallopeptidase 23B (MMP23B), xylosyltransferase II (XYLT2), and heparan sulfate 6-O-sulfotransferase 1 (HS6ST1) was found in the supplemented group. Correlation analysis indicated a direct relationship between blood cell gene expression of MMP23B, involved in the breakdown of the extracellular matrix, and pain intensity, and an inverse relationship between blood cell gene expression of HS6ST1, responsible for 6-O-sulfation of heparan sulfate, and indicators of knee muscular strength. Expression levels of specific genes in blood cells, in particular genes related to GAG metabolism and extracellular matrix dynamics, are potential biomarkers of beneficial effects on articular health.

Eficacia y seguridad de un tratamiento oral a base de mucopolisacáridos, colágeno tipo i y vitamina C en pacientes con tendinopatías / Efficacy and safety of an oral treatment based on mucopolysaccharides, collagen type i and vitamin C in patients with tendinopathies

Introducción y objetivos: La tendinopatía es una lesión frecuente durante la práctica deportiva que cursa con una alteración estructural del tendón. El objetivo de este estudio fue evaluar la eficacia y la seguridad de un complemento alimentario a base de mucopolisacáridos, colágeno tipo i y vitamina C (Tendoactive(R)) sobre la evolución clínica y estructural de las tendinopatías del tendón de Aquiles, rotuliano y del epicóndilo lateral del codo. Material y métodos: Se realizó un estudio multicéntrico prospectivo, de tipo exploratorio en fase IV, abierto y no comparativo. Se incluyeron un total de 98 pacientes con tendinopatías (32 de Aquiles, 32 de rotuliano y 34 del epicóndilo lateral) que recibieron una dosis diaria de 435 mg de mucopolisacáridos, 75 mg de colágeno tipo I y 60 mg de vitamina C (equivalente a 3 cápsulas al día de Tendoactive(R)) durante 90 días consecutivos. Mensualmente se evaluó el dolor en reposo y en actividad mediante una escala visual analógica (EVA), la función articular mediante los cuestionarios VISA-A, VISA-P y PRTEE, y se caracterizó ecográficamente el tendón afectado. Resultados: En los 3 tipos de tendinopatía se registró una reducción significativa del dolor tanto en reposo como en actividad desde la primera visita de control (día 30) hasta el final del estudio (día 90). Asimismo el día 90 se detectó una mejora del 38% en VISA-A, del 46% en VISA-P y del 77% en PRTEE (p < 0,001). Simultáneamente se registró una reducción del 12% en el grosor del tendón de Aquiles, del 10% en el rotuliano y del 20% en el tendón del epicóndilo lateral (p < 0,05). Conclusiones: Los resultados del estudio indican que la administración de Tendoactive(R) es segura y eficaz para mejorar los síntomas clínicos y la evolución estructural de las tendinopatías del tendón de Aquiles, tendón rotuliano y tendón del epicóndilo lateral

Introduction and objectives: The aim of this study was to evaluate the efficacy and safety of a diet supplement containing mucopolysaccharides, collagen type I and vitamin C (Tendoactive(R)) on the clinical symptoms and tendon structure of patients with Achilles, patellar, or lateral epicondyle tendinopathy. Material and methods: Between September 2012 and February 2013, a total of 98 patients with tendinopathy were included in the study (32 of Achilles tendon, 32 of patellar tendon, and 34 of lateral epicondyle tendon). The patients received a daily dose of Tendoactive(R) containing 435 mg of mucopolysaccharides, 75 mg of collagen type I and 60 mg of vitamin C (equivalent to 2 capsules per day) for 90 consecutive days, and were followed up monthly during the study period. Clinical assessments included pain intensity, which was assessed at rest and during activity using a visual analog scale (VAS), and also a specific functional scale (VISA-A questionnaire for Achilles, VISA-P for patella, and PRTEE for the elbow). Tendon structure was analyzed using ultrasound, including measurements of cross-sectional thickness of the tendon, paratenon blurring, heteroechogenicity and hypoechogenicity levels, and neovascularization. Results: There was a significant reduction in pain at rest and during activity from the first follow-up visit (day 30) until the end of the study (day 90) for the three types of tendinopathy. The same pattern of response was observed with the functional scales. On day 90 the improvement from baseline was 38% for VISA-A, 46% for VISA-P, and 77% for PRTEE (P <0.001). Simultaneous to the clinical improvement, there was a reduction of the tendon thickness (12% in Achilles, 10% in patellar tendon and 20% in lateral epicondyle tendon; P < 0.05). Conclusions: The overall results show that Tendoactive(R) is a safe and effective treatment for improving the clinical symptoms, as well as structural evolution of injured tendons, as demonstrated in Achilles, patellar and lateral elbow epicondyle tendinopathy

Dietary nucleotide improves markers of immune response to strenuous exercise under a cold environment.

BACKGROUND: Strenuous exercise has been classically associated to immune-suppression and consequently to an increased risk of infections, especially at the upper respiratory tract. The administration of dietary nucleotides has been demonstrated useful to maintain the immune function in situations of stress and thus could be an appropriate strategy to counteract the decline of the immune function associated to strenuous exercise. The aim of the present study was to asses the impact of a specific nucleotide formulation (Inmunactive®) on the markers of immune function of athletes after a heavy exercise bout under cold conditions. METHODS: Twenty elite male taekwondo athletes were randomly divided into two groups of 10 subjects that were supplemented with placebo (P) or Inmunactive (I) at 480 mg/day during 30 days. At baseline (day 0) and after 4 wk of supplementation (day 30) each subject undertook an exhaustion exercise test using a cycloergometer. Skin temperature, core temperature, heart rate, lactate concentration and rating of perceived exertion (RPE) were recorded during the test. Blood and saliva samples were obtained before and after each exercise test for determination of blood cell concentrations, PHA-stimulated lymphocyte proliferation (PHA-LP) and salivary immunoglobulin A (SIgA). RESULTS: Exercise tests induced neutrophilia and reduction in lymphocyte blood counts on day 0 and on day 30 in both groups. However, the I group exhibited a faster recovery from the lymphopenic response than the P group, so that lymphocyte levels were higher after 150 min (P < 0.0028). Furthermore, the lymphoproliferative response was modulated by nucleotide supplementation, since it was higher in the I group on day 30 despite an almost significant (P < 0.06) exercise-evoked decrease at baseline. CONCLUSIONS: These findings suggest that supplementation with a nucleotide-based product for 4 weeks could counteract the impairment of immune function after heavy exercise.

Long-term intake of resistant starch improves colonic mucosal integrity and reduces gut apoptosis and blood immune cells.

OBJECTIVE: The potential effect of a long-term intake of resistant starch on colonic fermentation and on gut morphologic and immunologic indices of interest in bowel conditions in humans was studied in pigs. METHODS: Sixteen growing pigs were meal fed for 14 wk on a diet containing a large amount of raw potato starch (RPS; resistant starch) or corn starch (CS; digestible starch). Effects were assessed in the colon from the physicochemical properties of digesta and in the intestinal morphology, including lymphocytic infiltration, apoptosis, and proliferation activities. Hematologic and blood leukocyte cell subsets analysis were performed. RESULTS: After 97 d, the digestive content from RPS pigs was heavier than for CS pigs, producing a hypertrophy of tunica muscularis (P < 0.05). The proportion of butyrate was two-fold higher in proximal colon digesta in RPS pigs (P < 0.05). RPS-fed pigs had reduced apoptosis in the crypts, lamina propria and lymphoid nodules in the colon, and ileal Peyer's patches (P < 0.05). Fermentation of RPS reduced indices associated with damage to epithelial cells, such as crypt cell proliferation and magnesium excretion, whereas mucin sulfuration was increased, which promotes epithelial protection. The numbers of intraepithelial T cells and of blood leukocytes, neutrophils, and lymphocytes, mainly T-helper lymphocytes, were reduced in RPS pigs (P < 0.05). CONCLUSION: Long-term intake of RPS induces pronounced changes in the colonic environment, reduces damage to colonocytes, and improves mucosal integrity, reducing colonic and systemic immune reactivity, for which health benefits in inflammatory conditions are likely to be associated.

Consumption of raw potato starch increases colon length and fecal excretion of purine bases in growing pigs.

Male growing pigs were fed a diet containing 250 g/kg of native corn starch (CS; 26% amylose, 74% amylopectin) or 250 g/kg of raw potato starch (RPS), as examples of digestible starch and resistant starch (Type II), respectively. Whole-tract digestibilities of organic matter, crude protein and starch were greater in pigs fed CS than in those fed RPS through at least d 23 of the study. However, the values progressively increased in the RPS-fed pigs up to d 38, at which time the groups did not differ in organic matter and starch digestibility. The digestive tract and colonic digesta were heavier and colon length longer in pigs fed the RPS diet. Digestibility of starch in the ileum on d 38 was significantly lower in RPS-fed pigs, but rose from ileum to rectum; most starch was extensively fermented in the cecum and proximal colon. Purine base (PB) and short-chain fatty acid (SCFA) concentrations in feces initially increased and then decreased beginning on d 4 for PB and on d 21 for SCFA. PB concentration in feces was greater in pigs fed RPS than in those fed CS. In the large bowel digesta, PB and SCFA concentrations increased from the ileum to the cecum and proximal colon and then fell in the distal colon. Pigs fed the RPS diet had a higher PB concentration in the middle colonic digesta and a greater SCFA concentration in the proximal colonic digesta than the CS-fed group. Adaptation of growing pigs to supplementary RPS required 5 wk, as reflected by whole-tract digestibility and PB and SCFA fecal excretion data.