Clinical Study Skin Antiageing and Systemic Redox Effects

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2 Oxidative Medicine and Cellular Longevity, skin bone repair Thus they positively affected glucose and positive roles of mediators in the MCPs induced collagen. lipid metabolism in patients with type II diabetes mellitus and ATP synthesis storage as well as in sebum production. 12 improved lipid metabolism in obese people 13 and ge On these grounds we suggested that selected antioxidants. netically modified mice 14 ameliorated early alcoholic liver targeting the distinct organs tissues should be essential. injury 15 and possessed hypotensive and lipid normalising components of MCPs containing nutraceuticals for more. action in patients with primary hypertension 16 A great effective individualised and safe supplementation. majority of publications demonstrated significant wound. healing efficacy of orally administered MCPs in animal 2 Materials and Methods. models of excision and full thickness skin wounds 10 17 18. Recently collagen peptides isolated by enzymatic digestion 2 1 Patients The study enrolled a group of 41 adult healthy. from fish bovine and porcine skin as well as from chicken Caucasian volunteers of both sexes recruited from the Beauty. and bovine cartilage have drawn particular interest for the Institute on Arbat Moscow Russia staff age 37 72 years. treatment of patients with osteoarthritis Several clinical trials mean age 50 6 10 4 years 5 males and 36 females following. showed that MCPs were safe and provided an improvement the exclusion and inclusion criteria for an open single blind. in terms of pain and functions in such patients 19 From clinical study The inclusion criteria were as follows i. mechanistic point of view the oral intake of MCPs stim healthy white adult subjects of both sexes 35 75 years of age. ulated the synthesis of extracellular matrix ECM macro ii subjects with visible symptoms of aged facial skin iii. molecules such as endogenous collagen by upregulating gene subjects who agreed to interrupt any intake of antioxidant. expression of several collagen modifying enzymes involved nutraceuticals drugs for at least 1 week before and during. in posttranslational collagen modification and cross linking the entire duration of the trial and iv subjects without any. 20 Several in vitro studies have shown antioxidant prop difficulty to understand and follow the clinical investigator. erties of very low molecular weight 1 20 Da MCPs 21 22 instructions Pregnant and breastfeeding women subjects. containing proline which is a scavenger of hydroxyl radicals with allergic intolerance reactions to any component of the. Of importance for the present study MCPs are considered tested product subjects on any other nutraceutical interven. antiageing compounds because they seem to increase life tions or and therapies and subjects simultaneously engaged. span in rats by inhibiting spontaneous tumour incidence 9 in other clinical trials were excluded from the study The. possess photoprotective and immunomodulating properties participants were informed that they could interrupt clinical. 23 25 and improve eliminate signs of premature senes trial at any moment without any explanation of causative. cence of human skin 26 27 reason for their action or in case they noticed any adverse. The major concern regarding safety and clinical feasibility reaction to the tested product or had any sensation that the. of regular intake of MCPs has been raised from the well estab product intake affected their appearance negatively. lished fact that the induction of collagen synthesis mainly The protocol of the clinical trial was duly analysed and. assessed by the increased hydroxyproline levels is often approved by the Ethical Committee of the Beauty Insti. associated with oxidative stress 28 30 Moreover MCPs of tute on Arbat Moscow Russia number 11 EK 2014 All. different origin have been shown to activate innate immune recruited subjects gave their informed consent to personal. response of macrophages and neutrophils through Toll like and anamnestic data collection and biological material sam. receptor 4 which leads to NADPH oxidase NOX4 activa pling The guidelines of Helsinki Declaration for human. tion and reactive oxygen species overproduction 31 32 A experimentation were strictly followed during the conduct of. newly developed composition of MCPs with a complex of the clinical trial. essential skin targeting antioxidants that is coenzyme Q10. selenium luteolin grape skin extract demonstrated UVA 2 2 Food Supplement under Investigation Food supplement. protective effects in the preliminary in vitro experiments on containing marine collagen peptides derived from skin of. human skin biopsies 25 However the composition under deep sea fish MCPs 570 mg grape skin extract 10 mg. commercial name of CELERGEN has never been evaluated coenzyme Q10 of plant origin 10 mg luteolin 10 mg and. clinically when administered as a food supplement selenium 0 05 mg of plant origin was formulated in soft. The goal of the present clinical laboratory study was to gelatine capsules As inactive solvents refined and partly. elucidate the effects of the oral administration of CELERGEN hydrogenated soybean oil as well as small admixture of pure. on skin physiology and dermal collagen deposition in the soybean lecithin were used The product under the commer. group of healthy middle aged subjects with clinical signs cial name of CELERGEN manufacturer Laboratories Dom. of skin ageing The cutaneous clinical instrumental data Carouge Switzerland was kindly provided by Suisse Ueli. were compared with the systemic metabolic parameters of Corporation According to the manufacturer s information. collagen synthesis redox balance and energy storage For the the deep sea fish sources that is Pollachius virens Hippoglos. first time we demonstrated i remarkable improvement of sus hippoglossus and Pleuronectes platessa originated from. ageing skin physiology and structure which corresponded the French coast of the North Sea. to enhanced systemic markers of collagen synthesis ii Fish skin was homogenised in distilled water with. systemic redox balance sustained by the antioxidant com addition of complex proteases The enzymatic proteolytic. plex and iii increased systemic energy storage We also process was carried out at 40 C and pH 8 0 for 3 h after. hypothesised that moderately increased plasmatic levels of which the proteases were inactivated by short term heating. nitric oxide NO and malonyl dialdehyde MDA may play 56 C for 10 min The liquid was sterilised by Millipore. Oxidative Medicine and Cellular Longevity 3, filtration pore size 0 02 mm and spray dried to prepare PLUS technique The transepidermal water loss TEWL an. MCP powder as described in detail previously 17 33 index of skin moisture was assessed with Tewameter which. Chemical analysis by Kjeldahl assay of the powder confirmed measures the water evaporation through cutaneous levels. a 90 content of collagen peptides with moisture and ash When the skin is aged or damaged the barrier properties of. content 10 According to previous publications 10 33 the skin are affected with increased water evaporation and. 34 the molecular weight distribution of MCPs after the reduced skin hydration Sebum content was measured by the. described enzymatic digestion process was within the range SOFT PLUS sebometric probe. of 10 60 Da and MCPs were enriched in glycine glutamine. proline hydroxyproline asparagine alanine and arginine 2 5 Assessment of Ultrasound Properties of the Skin Assess. The aqueous extract of grape skin was obtained from Vitis ment of ultrasound properties of the skin was performed by. vinifera Linn fruit and contained at least 70 of polyphenols a digital ultrasound imaging system DUB CUTIS Digital. and 20 of procyanidins as per UV Vis spectrophotometry Ultraschall Bildsystem Germany which allowed determin. data The coenzyme Q10 component of plant origin was ing four parameters simultaneously epidermal and dermal. of highest purity 100 3 confirmed by both IR spec thickness and epidermal and dermal ultrasonic density The. trophotometry and high performance liquid chromatography first two parameters are indirect markers of collagen dermis. HPLC methods Food quality luteolin was extracted from and lipid epidermis synthesis and retention while the. Marigold plant petals and the extract contained 20 of second pair of parameters characterises the evenness and. luteolin and 1 of zeaxanthin evaluated by HPLC analysis order in the epidermal and dermal structures respectively. Selenium in the form of selenite according to gravimetric The elastic properties of the skin were additionally analysed. method was extracted from plant bulbs and leaves Acute by a TPM system containing elastometric sensor 22 MHz. and chronic toxicity data and documents of Certificates of which combines digital ultrasound examination with an. Analyses Security and Registration in Switzerland were duly imaging record DUB CUTIS Germany A computerised. provided by the manufacturer multifunctional diagnostic tool integrating different morpho. metric parameters epidermal thickness tone wrinkles and. 2 3 Clinical Study Design The entire trial duration was 4 elasticity for face skin biological age determination was used. months May December 2014 that is 2 months of pretreat SOFT PLUS TOP Callegari Parma Italy. ment period followed by 2 months of treatment with the. test nutraceutical administration The facial skin parameters 2 6 Reagents and Assay Kits The majority of chemical. of recruited volunteers were analysed three times at the reagents HPLC standards mediums solvents and luciferin. first visit enrollment at the second visit 2 months after luciferase for ATP assay were from Sigma Chemical Co St. the pretreatment period and at the third visit immediately Louis MO USA kits for enzyme activity assays and Griess. after the treatment period Each assessment session com reagent for nitrites nitrates determination were from Cay. prised instrumental methods for measuring skin physiology man Chemical Company Ann Arbor MI USA Manufac. parameters and ultrasound properties of the skin layers This turers of other reagents are mentioned within the respective. design allowed us to use the same subject as a control and methods. an experiment During the treatment period the volunteers. were recommended to take 2 capsules of CELERGEN a, 2 7 Redox and Oxidation Markers Studies Complete differ. day at breakfast and dinner time for 60 consecutive days. ential blood cell counts and metabolic analyses were per. At the second and third visits the participants donated. formed on fresh ethylenediaminetetraacetic acid EDTA. 20 mL of venous blood after overnight fasting and test tubes. anticoagulated venous blood of 12 hrs fasting subjects Bio. were coded by the principal clinical investigator Blood. chemical assays were performed on peripheral blood plasma. samples were routinely processed for general haematology. or red blood cells RBC either immediately ATP glu, haemoglobin content differential cell count and the rate of. tathione and coenzyme Q10 or within 72 hrs on sample. erythrocyte sedimentation and biochemistry glucose levels. aliquots stored at 80 C under argon Plasma levels of. plasma protein and lipid profiles transaminases activities. nitrites nitrates NO2 NO3 expressed as moles L were. and C reactive protein content Laboratory operators carried. measured spectrophotometrically by Griess reagent 35 Pro. out analytical determinations blindly and statistician was not. tein content was measured according to Bradford 36 using. informed which set of analyses was done in the control or. a microplate assay kit Bio Rad Hercules CA USA Total. experimental periods hence ranking this study of clinical. glutathione reduced oxidized glutathione GSH GSSG, efficacy of the nutraceutical as a single blind clinical inves.
mg g Hb levels in erythrocytes were measured by HPLC. Shimadzu Scientific Instruments Columbia MD USA, according to Reed et al 37 Total coenzyme Q10 CoQ10 H2. 2 4 Assessment of Skin Physiology Parameters Several phys CoQ10 mg L levels in plasma were quantified by HPLC. iological parameters mainly barrier properties of the facial as described previously 38 In brief 1 mL plasma sample. skin were assessed by appropriate SOFT PLUS TOP probes with adequate amount of coenzyme Q9 internal standard. with microcamera visual analysis and patented comput and 500 L acetic acid 50 solution was extracted twice. erised programs Callegari Parma Italy Skin elasticity was first with 3 5 mL and then with 2 5 mL of ethanol hexane. determined by the elastometric approach used in the SOFT mixture 2 5 vol vol with homogenisation and subsequent. 4 Oxidative Medicine and Cellular Longevity, Table 1 Subjective evaluation of the 2 month food supplement administration effects by participants 41. Number of participants,Improvement No effect Aggravation. General health conditions 21 51 20 49 0 0, Stamina muscle strength joint motility 15 36 26 64 0 0. Digestive system 0 0 41 100 0 0,Skin conditions 25 61 16 39 0 0.
centrifugation The upper phase containing hexane extract counter equipped with Wallac 1420 Software Perkin Elmer. was evaporated under nitrogen flux and then resuspended in MA USA Results were expressed as mmoles L. an adjusted amount of a methanol isopropanol 3 2 vol vol. mixture for HPLC analysis Reduced and oxidized forms of 2 9 Hydroxyproline Assay The plasma levels of free hydrox. coenzyme Q10 CoQ10 H2 and CoQ10 were quantified simul yproline Hyp and hydroxyproline in the form of oligopep. Clinical Study Skin Antiageing and Systemic Redox Effects of Supplementation with Marine Collagen Peptides and Plant Derived Antioxidants A Single Blind Case Control Clinical Study ChiaraDeLuca 1 ElenaV Mikhal chik 2 MaximV Suprun 2 MichaelPapacharalambous 3 ArseniyI Truhanov 4 andLiudmilaG Korkina 4 5

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