FR2895680A1 - Non-therapeutic treatment of mammalian superficial surface of human body part, comprises disposing liquid blood fraction and complex nutritional base, mixing the mixture to obtain active liquid phase and contacting the body part - Google Patents
Non-therapeutic treatment of mammalian superficial surface of human body part, comprises disposing liquid blood fraction and complex nutritional base, mixing the mixture to obtain active liquid phase and contacting the body part Download PDFInfo
- Publication number
- FR2895680A1 FR2895680A1 FR0513459A FR0513459A FR2895680A1 FR 2895680 A1 FR2895680 A1 FR 2895680A1 FR 0513459 A FR0513459 A FR 0513459A FR 0513459 A FR0513459 A FR 0513459A FR 2895680 A1 FR2895680 A1 FR 2895680A1
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- fraction
- blood fraction
- liquid
- treated
- liquid blood
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Abstract
Description
La présente invention concerne le traitement d'une partie superficielle duThe present invention relates to the treatment of a superficial part of the
corps humain ou animal, avec une base nutritive corplexe, en milieu aqueux, autrement appelée base éco-nutritive, en abrégé BNC, telle que définie ci-après, à différentes fins, médicales, thérapeutiques, cliniques, ou cosmétiques. Par "base nutritive complexe", on entend toute composition ou formulation, en milieu aqueux, se distinguant, comme précisé ci-après, d'un milieu de culture cellulaire, en ce qu'elle exclut tout facteur de croissance cellulaire, et/ou tout extrait d'origine animale ou cellulaire, non tracés, et se rapprochant d'un milieu de culture cellulaire, en ce qu'elle permet à elle seule une culture viable in vitro de kératinocytes épidermiques, par exemple humains, en particulier pendant au moins 72 heures, avec prolifération clonale, sans assise nourricière vivante, telle que fibroblastes. De telles BNC ont été décrites dans le document WO/96/21421, qui 15 décrit leur utilisation, seule ou en combinaison avec d'autres composants, en tant que produit actif ou comme excipient. Une BNC telle que considérée selon la présente invention, ne comporte, ni dans sa composition originelle ni dans sa mise en oeuvre, de facteur de croissance d'origine cellulaire, par exemple d'EGF 20 (Epidermal Growth Factor), et/ou d'extrait biologique d'origine animale ou cellulaire Par nature, non seulement ces extraits ont une composition variable, voire indéterminée, avec parfois une origine biologique mal déterminée, et donc non traçable ou non tracée, mais également certains 25 constituants sont indéterminés quant à leur structure chimique, notamment biochimique, exacte. Une BNC telle que considérée selon la présente invention, ne comporte aucun extrait biologique tel que sérum de veau foetal (SVF), ou aucun extrait de tige pituitaire de boeuf, non traçable ou non tracée. 30 Par "tracer" ou "traçable", on entend la caractéristique selon laquelle l'origine puis le traitement d'une matière biologique peuvent être établis et contrôlés. Une BNC selon la présente invention ne comporte par exemple aucun extrait biologique cellulaire, végétal ou animal, traçable ou non, tracé ou 35 non. Une BNC considérée selon la présente invention, ne comporte pas de principe actif médicamenteux, tel qu'un antibiotique. human or animal body, with a nutrient base corplexe, in an aqueous medium, otherwise called eco-nutritive base, abbreviated BNC, as defined below, for different purposes, medical, therapeutic, clinical, or cosmetic. By "complex nutritional base" is meant any composition or formulation, in an aqueous medium, distinguished, as specified below, from a cell culture medium, in that it excludes any cell growth factor, and / or any extract of animal or cellular origin, not traced, and approaching a cell culture medium, in that it alone allows a viable in vitro culture of epidermal keratinocytes, for example human, especially during at least 72 hours, with clonal proliferation, without living feeder, such as fibroblasts. Such BNCs have been described in WO / 96/21421, which describes their use, alone or in combination with other components, as an active product or as an excipient. A BNC as considered according to the present invention has neither, in its original composition nor in its implementation, growth factor of cellular origin, for example EGF 20 (Epidermal Growth Factor), and / or Biological extract of animal or cellular origin By nature, not only these extracts have a variable composition, or even indeterminate, sometimes with a poorly determined biological origin, and therefore not traceable or not traced, but also some constituents are indeterminate as to their chemical structure, especially biochemical, exact. A BNC as considered according to the present invention does not contain any biological extract such as fetal calf serum (FCS), or any pituitary beef stem extract, which is not traceable or not traced. By "tracer" or "traceable" is meant the characteristic that the origin and then the treatment of a biological material can be established and controlled. A BNC according to the present invention does not include, for example, any biological cell, plant or animal extract, traceable or not, traced or not. A BNC considered according to the present invention does not comprise any medicinal active principle, such as an antibiotic.
De telles BNC sont constituées, outre le milieu aqueux, par une fraction d'acides aminés, dont certains essentiels, inférieure à 0,5%, préférentiellement à 0,35% en poids, par une fraction de vitamines hydrosolubles inférieure à 0,2%, et préférentiellement à 0,015% en poids, et par une fraction inorganique, dont oligo-éléments et sels métalliques, inférieure à 5%, et préférentiellement à 2% en poids, le solde de la composition étant représenté par de l'eau. Préférentiellement, une BNC selon l'invention est entièrement formulée en phase aqueuse, à partir d'entités chimiques, biochimiques et biologiques, dont la structure de chacune est identifiée, par exemple nomenclaturée ou répertoriée, en sorte que la composition chimique de la BNC est strictement définie. Une BNC selon l'invention résulte d'une action de la main de l'homme, et ne saurait être assimilée de ce fait à tout extrait naturel et/ou biologique, obtenu par exemple par fractionnement d'un matériel biologique, animal ou végétal par exemple. Conformément au document WO 96/21421, une telle base nutritive complexe a, par exemple, la composition suivante, selon Tableau 1 ci-après : 1 Tableau 1 Soit 2808,3 mg ou 0,281% en poids COMPOSANTS Eau Acides aminés L-Alanine L-Arginine HCL L-Asparagine (anhydre Acide L-aspartique .............. L-Cystéine HCl H2O Acide L -glutamique L-Glutamine Glycine ........... L-Histidine HCI. H2O L-Isoleucine L-Leucine L-Lysine HCI ................. L-Méthionine L-Phénylalanine L-Proline L-Sérine L-Thréonine L-TryptoP...h a ne L -Tyrosine 2 Na 2H20 L-Valine Concentration en mg/I q.s.p. 42,0 14,8 1754.4 50,0 6,0 Such BNCs consist, in addition to the aqueous medium, of a fraction of amino acids, some essential, less than 0.5%, preferably 0.35% by weight, with a water-soluble vitamin fraction of less than 0.2. %, and preferably to 0.015% by weight, and an inorganic fraction, including trace elements and metal salts, less than 5%, and preferably 2% by weight, the balance of the composition being represented by water. Preferably, a BNC according to the invention is entirely formulated in aqueous phase, from chemical, biochemical and biological entities, whose structure of each is identified, for example nomenclature or listed, so that the chemical composition of the BNC is strictly defined. A BNC according to the invention results from an action of the human hand, and can not be assimilated to any natural and / or biological extract, obtained for example by fractionation of a biological material, animal or plant for example. According to the document WO 96/21421, such a complex nutritive base has, for example, the following composition, according to Table 1 below: TABLE 1 Either 2808.3 mg or 0.281% by weight COMPONENTS Water Amino acids L-Alanine L -Arginine HCL L-Asparagine (anhydrous L-aspartic acid .............. L-Cysteine HCl H2O L-Glutamic Acid L-Glutamine Glycine ........... L-Histidine HCI H2O L-Isoleucine L-Leucine L-Lysine HCl ................. L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L- TryptoP ... ha ne L -Tyrosine 2 Na 2H20 L-Valine Concentration in mg / l qs 42.0 14.8 1754.4 50.0 6.0
................................................ 131,2 54 _ 13,5 10,0.. DTD: .......... ...................................... 34,6..DTD: . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126,1 24,0 ................................................ 131, 2 54 _ 13.5 10.0 .. DTD: .......... ........................... ............ 34.6..DTD:. . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126.1 24.0
............................................... 9,3 11 7 70,3 Soit 99,49mg ou 0,0099% en poids Soit 16474,2 mg à 16483,25 mg ou environ 1, 65% en poids La présente invention a pour objet une mise en oeuvre particulière d'une BNC telle que précédemment définie, permettant de nouveaux soins de la peau ou d'autres parties superficielles du corps humain ou animal, à des fins cliniques ou cosmétiques. Au premier chef, l'invention concerne une méthode de traitement, à des fins thérapeutiques, médicales, cliniques, ou cosmétiques, d'une partie superficielle du corps d'un sujet mammifère, par exemple chez l'homme, et dite ci-après partie d'intérêt. Selon cette méthode, de manière générale : a) on dispose d'une fraction sanguine liquide, par exemple d'un sérum ou d'un plasma, de la même espèce que celle du sujet mammifère à traiter, 3 Vitamines d-Biotine Acide folique Nicotinamide D-Ca Pantothénate..DTD: .......................................................................... DTD: ....................... Pyridoxine HCI Riboflavine Thiamine HCI i-Inositol Pyruvate de sodium Thymidine Adénine (HCI) Acide DL-lipoïque Composants inorganiques Chlorure de sodium KCI CuSO4.5H20 Acétate de sodium ...................................................... D-Glucose Hepes...(pipérazine)...._ Phosphoryléthanolam ine Ethanolamine Sulfate de sodium Bicarbonate de sodium FeSOd.7H20 MgC12.6H2O CaCl2.2H20 ZnSO4.7H20 (NH4)6 Mol 024 4H20 Na2SiO3.5H20 MnCl2.4H20..DTD: .......................................................... SnCl2.2H20 NH4 VO3 0,00057 300,0 (anhydre) 120,0 De 13,0 à 22,05 0.144 0,00011 b) on dispose d'une BNC, telle que définie précédemment, c) on mélange la fraction sanguine liquide avec la BNC, pour obtenir une phase liquide active de traitement local et superficiel, d) on met la partie d'intérêt du corps du sujet, à traiter, en contact, 5 avec la phase liquide active. Préférentiellement la fraction sanguine liquide est autologue, et obtenue uniquement à partir d'un échantillon sanguin du sujet mammifère à traiter, par exemple par fractionnement. Selon un premier mode d'exécution de cette méthode, la fraction 10 sanguine liquide est un sérum décomplémenté, obtenu par fractionnements successifs à partir d'un échantillon sanguin, d'abord par coagulation à température ambiante sans anti-coagulant, puis par chauffage, par exemple à 56 C pendant trente minutes ; par exemple la fraction sanguine liquide est un sérum décomplémenté, puis déplété en ses immuno-globulines, par exemple 15 par incubation avec une suspension cellulaire de kératinocytes, ensuite centrifugée avec élimination du culot cellulaire. Selon un deuxième mode d'exécution de cette méthode, la fraction sanguine liquide est un plasma. La partie d'intérêt à traiter appartient à l'épiderme du sujet 20 mammifère, et est notamment une zone dudit épiderme présentant un défaut d'aspect en raison d'un défaut naturel, ou d'une lésion tissulaire superficielle réparée, par exemple une zone cicatricielle. Cette partie superficielle d'intérêt peut résulter de toute plaie, balafre, coupure, écorchure, égratignure, entaille, éraflure, morsure, mutilation, piqûre, brûlure, contusion, escarre, ecchymose, 25 etc..., en cours de réparation, mais laissant subsister un défaut superficiel, morphologique, structurel, fonctionnel, ou d'aspect. Au deuxième chef, l'invention concerne un dispositif de soin superficiel, notamment pansement, du corps d'un sujet mammifère, par exemple chez l'homme, à des fins médicales, ou thérapeutiques, ou 30 cosmétiques, ledit dispositif comprenant : - un support comportant une face active agencée pour venir en contact avec la peau dudit sujet, et un substrat, par exemple de nature textile, susceptible de stocker, par exemple par absorption, un liquide actif de traitement, et diffuser ledit liquide actif sur ladite face active, - des moyens de maintien temporaire du support sur une partie d'intérêt superficielle, à traiter, du corps du sujet, avec ladite face active contre la peau dudit sujet, - et le cas échéant, des moyens de protection temporaire, 5 amovibles ou éliminables, de la face active dudit support. Selon l'invention, le liquide actif de traitement comprend une fraction sanguine liquide, par exemple un sérum de la même espèce mammifère que celle du sujet traité, ainsi qu'une BNC telle que précédemment définie. 10 Préférentiellement, la fraction sanguine liquide est un sérum autologue du sujet à traiter. Au troisième chef, la présente invention concerne toute composition comprenant, d'une part une fraction sanguine liquide, par exemple sérum, et d'autre part une BNC telle que précédemment définie. 15 Une telle composition peut être utilisée pour obtenir un médicament, par exemple topique. En pareil cas, préférentiellement le médicament est destiné à traiter un sujet mammifère, par exemple l'homme, et la fraction sanguine liquide est obtenue à partir de la même espèce que celle du sujet mammifère à traiter. Par 20 exemple, la fraction sanguine liquide est une fraction autologue du sujet à traiter. Par exemple, un médicament tel que défini précédemment est mis en oeuvre pour le soin des dents ou de la bouche, ou pour la pratique de la chirurgie dentaire ou parodontale. 25 Dans ces indications, un médicament selon la présente invention améliore le maintien en survie et la prise de greffes, notamment en implantologie, lors du forage et de la récupération d'os mandibulaire ou maxilaire, et de la mise en survie dudit os (au cours de l'intervention) en vue de sa réimplantation en tant que matériel vivant de comblement au niveau du 30 collet des implants. Toujours en chirurgie dentaire, ou lors de chirurgies parodontales, un médicament selon l'invention favorise la réalisation de greffes épithéliales et/ou conjonctives, et ce en vue de l'amélioration de la prise de greffons... DTD: Au quatrième chef, l'invention concerne un médicament topique comprenant une composition telle que précédemment définie, et un support adapté à l'administration superficielle dudit médicament. En pareil cas, préférentiellement, le médicament est destiné au traitement d'un sujet mammifère, et la fraction sanguine liquide est une fraction sanguine de la même espèce que celle du sujet mammifère traité. Par exemple la fraction sanguine liquide est une fraction autologue du sujet à traiter. ............................................... 9.3 11.7 70.3 Either 99.49 mg or 0.0099% by weight Or 16474.2 mg to 16483.25 mg or about 1.65% by weight The present invention relates to a particular implementation of a BNC such as than previously defined, allowing new skin care or other superficial parts of the human or animal body, for clinical or cosmetic purposes. In the first place, the invention relates to a method of treatment, for therapeutic, medical, clinical or cosmetic purposes, of a superficial part of the body of a mammalian subject, for example in humans, and hereinafter referred to as part of interest. According to this method, in general: a) a liquid blood fraction, for example a serum or a plasma, of the same species as that of the mammalian subject to be treated is available, 3 Vitamins d-Biotin Folic acid Nicotinamide D-Ca Pantothenate..DTD: ......................................... ................................. DTD: ............... ........ Pyridoxine HCI Riboflavin Thiamine HCI i-Inositol Sodium pyruvate Thymidine Adenine (HCI) DL-lipoic acid Inorganic components Sodium chloride KCI CuSO4.5H20 Sodium acetate .......... ............................................ D-Glucose Hepes .. (piperazine) Phosphorylethanolamine Ethanolamine Sodium sulphate Sodium bicarbonate FeSOd.7H2O MgC12.6H2O CaCl2.2H2O ZnSO4.7H2O (NH4) 6 Mol 024 4H2O Na2SiO3.5H2O MnCl2.4H2O..DTD: ... .................................................. ..... SnCl2.2H2O NH4 VO3 0.00057 300.0 (anhydrous) 120.0 From 13.0 to 22.05 0.144 0.00011 b) a BNC is available, such that ue defined above, c) the liquid blood fraction is mixed with the BNC, to obtain a liquid active phase of local and superficial treatment, d) the part of interest of the body of the subject, to be treated, in contact, with the active liquid phase. Preferably, the liquid blood fraction is autologous, and obtained solely from a blood sample of the mammalian subject to be treated, for example by fractionation. According to a first embodiment of this method, the liquid blood fraction is a decomplemented serum, obtained by successive fractionations from a blood sample, firstly by coagulation at room temperature without anticoagulant, then by heating, for example at 56 C for thirty minutes; for example, the liquid blood fraction is a decomplemented serum and then depleted in its immunoglobulins, for example by incubation with a cell suspension of keratinocytes, then centrifuged with removal of the cell pellet. According to a second embodiment of this method, the liquid blood fraction is a plasma. The part of interest to be treated belongs to the epidermis of the mammalian subject, and is in particular an area of said epidermis having a defect of appearance due to a natural defect, or a repaired surface tissue lesion, for example a cicatricial area. This superficial part of interest may result from any wound, scar, cut, scratch, scratch, nick, scratch, bite, mutilation, sting, burn, bruise, eschar, bruise, etc., being repaired, but leaving a superficial, morphological, structural, functional, or aspect defect. In the second head, the invention relates to a device for the superficial care, in particular dressing, of the body of a mammalian subject, for example in humans, for medical or therapeutic purposes, or cosmetics, said device comprising: support comprising an active face arranged to come into contact with the skin of said subject, and a substrate, for example of textile nature, capable of storing, for example by absorption, an active treatment liquid, and diffusing said active liquid on said active face means for temporarily holding the support on a part of superficial interest, to be treated, of the body of the subject, with said active face against the skin of said subject, and, if necessary, temporary protective means, removable or removable, the active face of said support. According to the invention, the active treatment liquid comprises a liquid blood fraction, for example a serum of the same mammalian species as that of the treated subject, and a BNC as defined above. Preferably, the liquid blood fraction is an autologous serum of the subject to be treated. Thirdly, the present invention relates to any composition comprising, firstly a liquid blood fraction, for example serum, and secondly a BNC as defined above. Such a composition can be used to obtain a drug, for example topical. In such a case, preferably the medicament is intended to treat a mammalian subject, for example the human, and the liquid blood fraction is obtained from the same species as that of the mammalian subject to be treated. For example, the liquid blood fraction is an autologous fraction of the subject to be treated. For example, a drug as defined above is used for the care of the teeth or mouth, or for the practice of dental or periodontal surgery. In these indications, a drug according to the present invention improves survival and grafting, especially in implantology, during the drilling and recovery of mandibular or maxillary bones, and the survival of said bone (at During the course of the intervention), it was reimplanted as a living filling material at the neck of the implants. Still in dental surgery, or during periodontal surgery, a drug according to the invention promotes the performance of epithelial and / or conjunctive grafts, with a view to improving the grafting of grafts ... DTD: Fourth, the invention relates to a topical medicament comprising a composition as defined above, and a carrier adapted to the superficial administration of said medicament. In such a case, preferably, the medicament is intended for the treatment of a mammalian subject, and the liquid blood fraction is a blood fraction of the same species as that of the mammalian subject treated. For example, the liquid blood fraction is an autologous fraction of the subject to be treated.
Au cinquième chef, l'invention concerne une préparation cosmétique comprenant une composition telle que précédemment définie, et un support ou véhicule cosmétiquement acceptable. En pareil cas, préférentiellement, la fraction sanguine est une fraction sanguine de la même espèce que celle du sujet mammifère traité. Par exemple, la fraction sanguine liquide est une fraction autologue du sujet à traiter. In the fifth head, the invention relates to a cosmetic preparation comprising a composition as defined above, and a cosmetically acceptable carrier or vehicle. In such a case, preferably, the blood fraction is a blood fraction of the same species as that of the mammalian subject treated. For example, the liquid blood fraction is an autologous fraction of the subject to be treated.
Au sixième chef, l'invention concerne un procédé de maintien en survie ex vivo d'un tissu ou organe d'un sujet mammifère, selon lequel on met ledit tissu ou organe en contact, par exemple par immersion, avec une composition telle que précédemment définie. En pareil cas, préférentiellement, la fraction sanguine de la composition est obtenue à partir de la même espèce que celle du sujet mammifère dont le tissu ou l'organe est prélevé, et est par exemple une fraction sanguine autologue dudit sujet mammifère. In the sixth head, the invention relates to a method for maintaining in ex vivo survival a tissue or organ of a mammalian subject, according to which said tissue or organ is brought into contact, for example by immersion, with a composition such as previously defined. In such a case, preferably, the blood fraction of the composition is obtained from the same species as that of the mammalian subject whose tissue or organ is removed, and is for example an autologous blood fraction of said mammalian subject.
Ainsi, selon l'invention, une composition telle que précédemment définie permet le maintien en survie d'expiants cutanés ou de cuirs chevelus, ainsi que d'épithélium en vue de leur greffe Grâce à la présente invention, on dispose d'une nouvelle approche du soin des parties superficielles du corps, notamment du cuir chevelu, permettant de manière générale d'entretenir ou relancer le métabolisme de greffons et des tissus superficiels comme la peau. Thus, according to the invention, a composition as defined above makes it possible to maintain skin explants or scalps, as well as epithelium, for the purpose of their grafting. Thanks to the present invention, a new approach is available. care of the superficial parts of the body, especially the scalp, which generally allows to maintain or restart the metabolism of grafts and superficial tissues such as skin.
La pertinence de la méthode selon l'invention a été évaluée, in vitro, selon les deux essais décrits ci-après : The relevance of the method according to the invention was evaluated, in vitro, according to the two tests described below:
Essai n 1 : évaluation de la croissance de kératinocytes humains normaux, cultivés pendant six jours dans une BNC supplémentée en sérum 10 humain. La BNC mise en oeuvre a la composition décrite par le tableau ci-après : 15 Tableau 2 NBRE DENOMINATION CONCENTRATION LIGNE INTERNATIONALE En mg/I NOMENCLATURE COSMETIQUE INGREDIENT (NOM INCI) 1 EAU q.s.p. (1000mI) 2 CHLORURE DE 6085 SODIUM 3 GLUTAMINE 2043.2 4 SODIUM BICARBONATE 1160 5 GLUCOSE 4500 6 ARGININE HCL 421,4 7 SODIUM ACETATE 300 8 DISODIUM PHOSPHATE 284 9 LEUCINE 131,2 SERINE 136,6 11 CHLORURE DE Mg 120,0 12 CHLORURE DE K 112 13 VALINE 70,3 14 SODIUM PYRUVATE 55 LYSINE HCL 54 16 HISTIDINE HCL 50 17 CYSTEINE HCL 42 18 ADENINE 24 19 THREONINE 24 INOSITOL 18 21 ACIDE GLUTAMIQUE 29,5 22 ASPARAGINE 29,2 23 METHIONINE 13, 5 24 TYROSINE 11,7 PHENYLALANINE 10,0 26 TRYPTOPHANE 9,3 27 ALANINE 18,1 28 GLYCINE 15,1 29 ISOLEUCINE 6,0 ACIDE ASPARTIQUE 17,3 31 SODIUM SULFATE 3, 4 NBRE DENOMINATION CONCENTRATION LIGNE INTERNATIONALE En mg/1 NOMENCLATURE COSMETIQUE INGREDIENT (NOM INCI) 32 SULFATE FERREUX 1,4 33 ACIDE FOLIQUE 1,8 34 THYMIDINE 0,73 CYANOCOBALAMINE 0,41 36 CALCIUM 1,3 PANTOTHENATE 37 THIAMINE HCL 1,3 38 ACIDE THIOCTIQUE 0,3 39 ZINC SULFATE 0,144 SODIUM SILICATE 0,142 _ 41 PYRIDOXINE HCL 1,06 42 NIACINAMIDE _ 1,04 43 RIBOFLAVIN 0,4 44 BIOTIN _ 0,02 SULFATE DE CUIVRE _ 0,003 _ AMMONIUM 0,00120 46 MOLYBDATE 47 AMMONIUM VANADATE 0,003 48 CHLORURE DE Mn _ 0,00002 49 HYALURONATE DE 700 SODIUM _ HYDROXYPROLINE 30 _ 51 PROLINE 46 52 ACIDE ASCORBIQUE 1 53 ADENOSINE 0.1 54 GUANINE 0.1 DEOXYRIBOSE _ 0.1 56 RIBOSE 0.1 57 "MILK PEPTIDE 200 COMPLEX" _ 58 CHOLINE CHLORIDE 1 59 MYO INOSITOL 2 S'agissant du composant intitulé "Milk peptide complex", ou MPC, il s'agit de peptides naturels, ou d'origine naturelle, présents dans des fractions ou extraits obtenus à partir de lait d'origine animale. Cet extrait est l'extrait dit Whey Protein (Lactis Proteinum) ayant pour n CAS 84082-51-9 et r EINECS 281-998-7, fabriqué et vendu par la firme CLR-Chemisches Laboratorium en République Fédérale d'Allemagne. Test No. 1: Evaluation of the growth of normal human keratinocytes, cultured for six days in a BNB supplemented with human serum. The BNC used has the composition described in the following table: TABLE 2 NAME DENOMINATION CONCENTRATION INTERNATIONAL LINE In mg / I COSMETIC NOMENCLATURE INGREDIENT (INCI NAME) 1 WATER q.s. (1000mI) 2 CHLORIDE OF 6085 SODIUM 3 GLUTAMINE 2043.2 4 SODIUM BICARBONATE 1160 5 GLUCOSE 4500 6 ARGININE HCL 421.4 7 SODIUM ACETATE 300 8 DISODIUM PHOSPHATE 284 9 LEUCINE 131.2 SERINE 136.6 11 CHLORIDE Mg 120.0 12 CHLORIDE DE K 112 13 VALINE 70.3 14 SODIUM PYRUVATE 55 LYSINE HCL 54 16 HISTIDINE HCL 50 17 CYSTEINE HCL 42 18 ADENINE 24 19 THREONIN 24 INOSITOL 18 21 GLUTAMIC ACID 29.5 22 ASPARAGINE 29.2 23 METHIONINE 13, 5 24 TYROSINE 11 , 7 PHENYLALANINE 10.0 26 TRYPTOPHANE 9.3 27 ALANINE 18.1 28 GLYCINE 15.1 29 ISOLEUCIN 6.0 ASPARTIC ACID 17.3 31 SODIUM SULFATE 3, 4 NO. NAME NOMENCLATURE INTERNATIONAL LINE CONCENTRATION mg / 1 COSMETIC NOMENCLATURE INGREDIENT ( INCI NAME) 32 FERROUS SULPHATE 1.4 33 FOLIC ACID 1.8 34 THYMIDINE 0.73 CYANOCOBALAMINE 0.41 36 CALCIUM 1.3 PANTOTHENATE 37 THIAMINE HCL 1.3 38 THIOCTIC ACID 0.3 39 ZINC SULFATE 0.144 SODIUM SILICATE 0.142 41 PYRIDOXINE HCL 1.06 42 NIACINAMIDE _ 1.04 43 RIBOFLAVIN 0.4 44 BIOTIN _ 0.02 COPPER SULPHATE _ 0.003 _ AM MONIUM 0.00120 46 MOLYBDATE 47 AMMONIUM VANADATE 0.003 48 CHLORIDE Mn _ 0.00002 49 HYALURONATE 700 SODIUM _ HYDROXYPROLINE 30 _ 51 PROLINE 46 52 ASCORBIC ACID 1 53 ADENOSINE 0.1 54 GUANINE 0.1 DEOXYRIBOSE _ 0.1 56 RIBOSE 0.1 57 "MILK PEPTIDE 200 COMPLEX "_ 58 CHOLINE CHLORIDE 1 59 MYO INOSITOL 2 Regarding the component called" Milk peptide complex ", or MPC, these are natural or naturally occurring peptides present in fractions or extracts obtained from milk of animal origin. This extract is the so-called Whey Protein extract (Lactis Proteinum) having for CAS No. 84082-51-9 and EINECS 281-998-7, manufactured and sold by the firm CLR-Chemisches Laboratorium in the Federal Republic of Germany.
Cet extrait présente à titre d'exemple les données analytiques suivantes : pH 5,0-7,0 (0,5% en poudre MPC dans 1-120) 2,0-6,0% Perte à la dessication (2 h à 102 C) Azote (Kjeldahl) 1,4-1,9% Protéines (Sigma Kit BCA-1) 11,0-16,0% Lactose (Boehringer Kit) 65,0ù71,0% Acide (+) Lactique (Boehringer Kit) 4,5-8,0% Cendres (1000 C) 6,0-9,0% Graisse résiduelle <0,5% Elecrophorèse (SDS-PAGE) correspond Activité biologique ùEC5o 50-500pg/ml (test de migration) II s'agit d'évaluer l'effet d'une supplémentation de la BNC en sérum humain sur les capacités de croissance in vitro de kératinocytes humains normaux. 15 L'étude est réalisée avec la BNC, dans laquelle est ajouté extemporanément du sérum humain à 3 concentrations différentes (2%, 5% et 10 %). L'étude est réalisée en comparaison avec une culture effectuée dans un milieu de culture standard de kératinocytes, le KSFM (InVitrogen), 20 supplémenté en facteurs de croissance cellulaire (EGF, Bovine Pituitary Extract ou BPE), et en comparaison avec une culture effectuée avec la même BNC dépourvue de sérum (et supplémentée en biopeptides de lait, MPC, à la concentration de 0.2 mg/ml).10 1. PROTOCOLE Récupération et préparation du sérum sanguin ; Le sérum est la partie du plasma qui reste limpide et liquide après coagulation. Il est obtenu en laissant coaguler un prélèvement sanguin recueilli 5 sur tube sec, quelques heures à température ambiante. La coagulation terminée, le sérum est récupéré à l'aide d'une pipette. II est ensuite décomplémenté par chauffage à 56 C pendant 30 min. Traitements cellulaires : Les kératinocytes humains normaux sont ensemencés à faible 10 densité en plaques 96 puits en milieu de culture standard spécifique (KSFM). Après 24h et adhésion complète, les cellules sont placées dans les différents milieux de culture étudiés - KSFM - BNC 15 - BNC + sérum humain à 2% - BNC + sérum humain à 5% - BNC + sérum humain à 10% Chaque condition est réalisée en quadruplicate. La densité cellulaire est évaluée juste avant le "switch" dans les 20 différentes conditions d'étude ( TO), puis la croissance des kératinocytes est évaluée à T 1j, T 4j et T 6 jours après le switch, par la méthode de conversion du WST-1 (lecture à 450 nm). 2. RESULTATS 25 La croissance cellulaire est objectivée par la mesure de la viabilité cellulaire à différents temps de l'expérimentation. • La figure 1 présente les résultats obtenus pour la culture des kératinocytes avec la BNC + sérum humain versus culture en milieu standard KSFM. • La figure 1 bis constitue une simplification graphique de la précédente, ne présentant que les résultats concernant la BNC + sérum humain, la condition témoin KSFM n'étant pas représentée. • On procède aussi à des observations microscopiques 5 (Grossissements x10 et x20) des kératinocytes humains normaux dans les différentes conditions d'étude, au 61ème jour de culture. > Avec la BNC (avec complexe MPC à la concentration de 0.2 mg/ml), on observe une croissance cellulaire régulière jusqu'au 6ième jour d'expérimentation. 10 > L'ajout de sérum humain entraîne une forte augmentation de la croissance cellulaire; cet effet est dose-dépendant (l'ajout de 10% de sérum humain à la BNC entraîne la croissance cellulaire de plus forte intensité cf. figure 1 bis). > Morphologiquement, les kératinocytes, en milieu avec la BNC 15 supplémentée en sérum humain, présentent un aspect proche de celui observé en milieu standard. Les cellules sont jointives, polyédriques, avec une légère tendance à s'organiser en travées cellulaires. 3. CONCLUSION Dans les conditions expérimentales ainsi définies, il apparaît que 20 l'enrichissement de la BNC en sérum humain entraîne une forte augmentation de la croissance cellulaire, et ce de manière dose-dépendante, avec un effet optimum pour concentration en sérum de 10%. Essai n 2 : études des effets d'une supplémentation en sérum ou en plasma humain sur la croissance de kératinocytes humains normaux in 25 vitro. La BNC étudiée à la même composition que celle identifiée précédemment. Il s'agit d'évaluer l'effet d'une supplémentation de la BNC en sérum ou en plasma humain sur les capacités de croissance in vitro de kératinocytes 30 humains normaux. L'étude est réalisée avec la BNC, dépourvue ou supplémentée en biopeptides de lait (complexe MPC à la concentration de 0.2 mg/ml) et dans laquelle est ajouté extemporanément du sérum ou du plasma humain à la concentration finale de 10 %. This excerpt gives as an example the following analytical data: pH 5.0-7.0 (0.5% powder MPC in 1-120) 2.0-6.0% Loss on drying (2 hr 102 C) Nitrogen (Kjeldahl) 1.4-1.9% Protein (Sigma Kit BCA-1) 11.0-16.0% Lactose (Boehringer Kit) 65.0 ± 71.0% (+) Lactic Acid (Boehringer Kit) ) 4.5-8.0% Ash (1000 C) 6.0-9.0% Residual fat <0.5% Elecrophoresis (SDS-PAGE) corresponds to biological activity at EC50 50-500pg / ml (migration test) II The aim is to evaluate the effect of BNC supplementation in human serum on the in vitro growth capacity of normal human keratinocytes. The study is carried out with BNC, in which human serum is added extemporaneously at 3 different concentrations (2%, 5% and 10%). The study is carried out in comparison with a culture carried out in a standard culture medium of keratinocytes, KSFM (InVitrogen), supplemented with cell growth factors (EGF, Bovine Pituitary Extract or BPE), and in comparison with a culture performed. with the same serum-free BNC (and supplemented with milk biopeptides, MPC, at a concentration of 0.2 mg / ml) .10 1. PROTOCOL Recovery and preparation of blood serum; Serum is the part of the plasma that remains clear and liquid after coagulation. It is obtained by letting a blood sample collected on a dry tube coagulate for a few hours at room temperature. When coagulation is complete, the serum is recovered with a pipette. It is then decomplemented by heating at 56 ° C. for 30 minutes. Cellular Treatments: Normal human keratinocytes are inoculated at low density into 96-well plates in standard specific culture medium (KSFM). After 24h and complete adhesion, the cells are placed in the various culture media studied - KSFM - BNC 15 - BNC + 2% human serum - BNC + 5% human serum - BNC + 10% human serum Each condition is carried out in quadruplicate. The cell density is evaluated just before the "switch" in the different study conditions (TO), then the keratinocyte growth is evaluated at T 1j, T 4j and T 6 days after the switch, by the conversion method. WST-1 (reading at 450 nm). 2. RESULTS Cell growth is objectified by the measurement of cell viability at different times of the experiment. • Figure 1 shows the results obtained for the culture of keratinocytes with BNC + human serum versus culture in standard medium KSFM. • Figure 1a is a graphic simplification of the previous one, presenting only the results concerning the BNC + human serum, the control condition KSFM not being represented. Microscopic observations (x10 and x20 magnifications) of normal human keratinocytes are also made under the different study conditions at the 61st day of culture. > With BNC (with MPC complex at a concentration of 0.2 mg / ml), regular cell growth is observed until the 6th day of experimentation. 10> The addition of human serum leads to a sharp increase in cell growth; this effect is dose-dependent (the addition of 10% of human serum to the BNC causes the cell growth of higher intensity see Figure 1a). Morphologically, the keratinocytes, in medium with the BNC supplemented with human serum, have an appearance similar to that observed in standard medium. The cells are contiguous, polyhedral, with a slight tendency to organize into cellular spans. 3. CONCLUSION Under the experimental conditions thus defined, it appears that the enrichment of BNC in human serum results in a large increase in cell growth, and in a dose-dependent manner, with an optimum effect for serum concentration of 10%. %. Test No. 2: Studies of the effects of serum or human plasma supplementation on the growth of normal human keratinocytes in vitro. The BNC studied in the same composition as that identified previously. The aim is to evaluate the effect of BNC supplementation in serum or human plasma on the in vitro growth capacity of normal human keratinocytes. The study is carried out with BNC, which is deprived or supplemented with milk biopeptides (MPC complex at a concentration of 0.2 mg / ml) and in which human serum or plasma is added at the final concentration of 10%.
L'étude est réalisée en comparaison avec une culture effectuée dans un milieu de culture standard de kératinocytes, le KSFM (InVitrogen), avec ou sans facteurs de croissance cellulaire (EGF, BPE) et en comparaison avec une culture effectuée dans la même BNC dépourvue de sérum ou de plasma, et supplémentée en complexe MPC à la concentration de 0.2 mg/ml. The study is carried out in comparison with a culture carried out in a standard keratinocyte culture medium, KSFM (InVitrogen), with or without cell growth factors (EGF, BPE) and in comparison with a culture carried out in the same BNC lacking serum or plasma, and supplemented with MPC complex at a concentration of 0.2 mg / ml.
1. PROTOCOLE ^ Récupération et préparation du sérum sanguin Le sérum est la partie du plasma qui reste limpide et liquide après 10 coagulation. Il est obtenu en laissant coaguler un prélèvement sanguin recueilli sur tube sec, quelques heures à température ambiante. La coagulation terminée, le sérum est récupéré à l'aide d'une pipette. II est ensuite décomplémenté par chauffage à 56 C pendant 30 min. Afin de limiter les risques d'incompatibilité entre donneurs (et de 15 rejet par réaction immunitaire), on réalise une déplétion partielle du sérum humain en ses immunoglobulines. Cette procédure consiste à faire incuber, pendant 30 minutes à 37 C, le sérum avec une suspension cellulaire de kératinocytes humains normaux (les immunoglobulines contenues dans le sérum se fixent alors sur les kératinocytes). La suspension cellulaire est 20 ensuite centrifugée 10 minutes à 1400 tours par minute et le culot cellulaire éliminé. ^ Récupération du plasma sanguin Le plasma représente la fraction liquide du sang qui constitue 55% 25 de son volume et dans laquelle baignent les autres composants majeurs du sang (globules rouges, globules blancs et thrombocytes). Dans le plasma se trouve aussi une protéine en dissolution : le fibrinogène, capable de se polymériser en fibrine. Le sérum sanguin n'est autre que du plasma débarrassé de son fibrinogène. 30 Le plasma est obtenu en récupérant le prélèvement sanguin dans un tube contenant un additif anticoagulant (tube de coagulation au citrate de sodium dans ce cas). Le plasma est récupéré après centrifugation du prélèvement sanguin pendant 15 minutes à 2000 tours par minute. ^ Traitements cellulaires Les kératinocytes humains normaux (KHNP TI) sont ensemencés à faible densité en plaques 96 puits, en milieu de culture standard spécifique (KSFM). Après 24h et adhésion complète, les cellules sont placées dans les 5 différents milieux de culture étudiés : - KSFM complet (avec facteurs de croissance : EGF et BPE) - KSFM - (déplété en facteur de croissance) - KSFM - (déplété en facteur de croissance) + sérum humain à 10% 10 - Tampon PBS + sérum humain à 10% - BNC, sans complexe MPC + sérum humain à 10% - BNC, sans complexe MPC + plasma humain à 10% - BNC, avec complexe MPC (0.2 mg/ml) - BNC, avec complexe MPC (0.2 mg/ml) + sérum humain à 10% 15 - BNC, avec complexe MPC (0.2 mg/ml) + plasma humain à 10% 1. PROTOCOL Recovery and preparation of blood serum Serum is the part of the plasma which remains clear and liquid after coagulation. It is obtained by letting a blood sample collected on a dry tube coagulate for a few hours at room temperature. When coagulation is complete, the serum is recovered with a pipette. It is then decomplemented by heating at 56 ° C. for 30 minutes. In order to limit the risks of incompatibility between donors (and rejection by immune reaction), partial depletion of human serum in its immunoglobulins is achieved. This procedure consists of incubating, for 30 minutes at 37 ° C., the serum with a cell suspension of normal human keratinocytes (the immunoglobulins contained in the serum then bind to the keratinocytes). The cell suspension is then centrifuged for 10 minutes at 1400 rpm and the cell pellet removed. Retrieving the blood plasma Plasma represents the liquid fraction of the blood which constitutes 55% of its volume and in which the other major components of the blood (red blood cells, white blood cells and thrombocytes) bathe. In the plasma is also a dissolving protein: fibrinogen, capable of polymerizing to fibrin. The blood serum is nothing but plasma freed from its fibrinogen. Plasma is obtained by collecting the blood sample in a tube containing an anticoagulant additive (sodium citrate coagulation tube in this case). The plasma is recovered after centrifugation of the blood sample for 15 minutes at 2000 rpm. Cell Treatments Normal human keratinocytes (KHNP TI) are inoculated at low density into 96-well plates in standard specific culture medium (KSFM). After 24h and complete adhesion, the cells are placed in the different culture media studied: - complete KSFM (with growth factors: EGF and BPE) - KSFM - (depleted in growth factor) - KSFM - (depleted in factor of growth) + 10% human serum 10 - PBS buffer + 10% human serum - BNC, no complex MPC + 10% human serum - BNC, no MPC complex + 10% human plasma - BNC, with MPC complex (0.2 mg / ml) - BNC, with MPC complex (0.2 mg / ml) + 10% human serum - BNC, with MPC complex (0.2 mg / ml) + 10% human plasma
Chaque condition est réalisée en quadruplicate. Each condition is done in quadruplicate.
La densité cellulaire est évaluée juste avant le "switch" dans les 20 différentes conditions d'étude (= TO) puis la croissance des kératinocytes est évaluée à T 1j, T 3j et T 6 jours après le "switch", par la méthode de conversion du WST-1 (lecture à 450 nm). The cell density is evaluated just before the "switch" in the different study conditions (= TO), then the growth of the keratinocytes is evaluated at T 1j, T 3j and T 6 days after the "switch", by the method of WST-1 conversion (reading at 450 nm).
2. RESULTATS La croissance cellulaire est objectivée par la mesure de la viabilité cellulaire à différents temps de l'expérimentation. La morphologie cellulaire est étudiée par observations microscopiques et photographiée au terme de l'expérimentation (T6j). 30 La figure 2 présente les résultats obtenus pour la culture des kératinocytes dans l'ensemble des conditions étudiées. La figure 2bis constitue une simplification graphique de la précédente, ne présentant que les résultats concernant la culture des 35 kératinocytes avec BNC +1- sérum ou plasma humain versus milieu de culture standard (KSFM complet). 25 >Milieu KSFM : la déplétion du milieu de culture standard KSFM en facteurs de croissance n'entraîne pas de ralentissement de la croissance cellulaire des kératinocytes humains normaux sur les 6 jours de culture. L'ajout de sérum humain à 10% entraîne un léger ralentissement de la croissance cellulaire (figure 3). Morphologiquement, en présence de sérum humain, les kératinocytes présentent un aspect polyédrique et s'organisent en travées cellulaires. >Tampon PBS (Ca2+, Mq2+) + sérum humain à 10% : l'ajout de sérum humain au milieu tampon PBS permet la croissance des kératinocytes humains normaux in vitro (figure 3). Néanmoins, l'observation microscopique montre des cellules présentant des signes de souffrance cellulaire. >BNC avec complexe MPC à la concentration de 0.2 rnq/ml : on observe une bonne croissance cellulaire, régulière jusqu'au hème jour d'expérimentation. D'un point de vue morphologique, les cellules présentent un aspect arrondi, régulier, identique à celui observé en conditions standard de culture. L'ajout de sérum humain à 10% stimule la croissance cellulaire, qui est de faible intensité à T3j, puis fortement accélérée, conduisant ainsi à une densité cellulaire proche de celle obtenue en milieu standard (figure 3 bis). 2. RESULTS Cell growth is objectified by the measurement of cell viability at different times of the experiment. The cellular morphology is studied by microscopic observations and photographed at the end of the experiment (T6j). Figure 2 shows the results obtained for culturing keratinocytes in all the conditions studied. Figure 2bis is a graphic simplification of the previous one, presenting only the results concerning the culture of keratinocytes with BNC + 1-serum or human plasma versus standard culture medium (complete KSFM). > KSFM medium: The depletion of the standard growth medium KSFM in growth factors does not slow down the growth of normal human keratinocytes over 6 days of culture. The addition of 10% human serum results in a slight slowing of cell growth (Figure 3). Morphologically, in the presence of human serum, the keratinocytes have a polyhedral appearance and are organized into cellular spans. > PBS buffer (Ca2 +, Mq2 +) + 10% human serum: the addition of human serum to the PBS buffer medium allows the growth of normal human keratinocytes in vitro (FIG. 3). Nevertheless, microscopic observation shows cells showing signs of cellular suffering. > BNC with MPC complex at a concentration of 0.2 mg / ml: good cell growth is observed, regular until the day of the experiment. From a morphological point of view, the cells have a rounded, regular appearance, identical to that observed under standard culture conditions. The addition of 10% human serum stimulates cell growth, which is of low intensity at T3j, then strongly accelerated, thus leading to a cell density close to that obtained in standard medium (Figure 3a).
Morphologiquement et en présence de sérum humain à 10%, les cellules présentent un aspect plus jointif. L'ajout de plasma humain à 10% stimule également la croissance cellulaire mais de façon moins intense qu'en présence de sérum humain. Morphologiquement, les cellules apparaissent plus petites et plus denses. >Milieu BNC, sans complexe MPC: l'ajout de sérum humain à 10% entraîne une forte augmentation de la croissance cellulaire, dès le 3ème jour de culture et ce, jusqu'au terme de l'expérimentation. La croissance cellulaire est identique à celle obtenue en milieu standard KSFM (figure 3bis). Morphologiquement, les cellules ont un aspect plus différencié. Morphologically and in the presence of 10% human serum, the cells have a more joined appearance. The addition of human plasma to 10% also stimulates cell growth but less intensively than in the presence of human serum. Morphologically, cells appear smaller and denser. > BNC medium, no MPC complex: the addition of 10% human serum leads to a strong increase in cell growth, from the 3rd day of culture until the end of the experiment. The cell growth is identical to that obtained in standard KSFM medium (FIG. 3bis). Morphologically, cells have a more differentiated appearance.
L'ajout de plasma humain à 10% entraîne également une stimulation de la croissance cellulaire. Celle-ci est néanmoins moins importante au terme de l'expérimentation que celle obtenue en présence de sérum humain à la même concentration. Morphologiquement, les cellules apparaissent très arrondies et sont légèrement vacuolisées. . CONCLUSION Dans les conditions expérimentales ainsi définies, il apparaît que l'enrichissement de la BNC en sérum humain à la concentration de 10% entraîne une forte augmentation de la croissance cellulaire, permettant d'obtenir une croissance et une morphologie cellulaire identiques à celles obtenues dans des conditions standard de culture (milieu KSFM). Ce résultat est obtenu aussi bien avec la BNC dépourvue de complexe MPC, qu'avec la BNC supplémentée en complexe MPC à la concentration de 0.2mg/ml. The addition of 10% human plasma also stimulates cell growth. This is nevertheless less important at the end of the experiment than that obtained in the presence of human serum at the same concentration. Morphologically, the cells appear very rounded and are slightly vacuolized. . CONCLUSION Under the experimental conditions thus defined, it appears that the enrichment of the BNC in human serum at a concentration of 10% leads to a strong increase in cell growth, making it possible to obtain a growth and a cell morphology identical to those obtained in standard culture conditions (KSFM medium). This result is obtained both with BNC deprived of MPC complex, and with BNC supplemented with MPC complex at the concentration of 0.2 mg / ml.
L'enrichissement de la BNC en plasma humain à la concentration de 10% s'accompagne également d'une augmentation de la croissance cellulaire (mais cette dernière est d'intensité plus faible qu'en présence de sérum humain à la même concentration). The enrichment of BNC in human plasma at the concentration of 10% is also accompanied by an increase in cell growth (but the latter is of lower intensity than in the presence of human serum at the same concentration).
L'ajout de sérum humain à la concentration de 10% au milieu de culture standard KSFM (sans EGF et sans BPE) ne stimule pas la croissance des kératinocytes/ qui présentent dès lors des signes de différenciation cellulaire marqués (kératinocytes organisés en travées). Cet effet peut être lié à un apport en calcium (par le sérum) trop important dans le milieu de culture, conduisant à une différenciation trop précoce des kératinocytes. The addition of human serum at the concentration of 10% in standard culture medium KSFM (without EGF and without BPE) does not stimulate the growth of keratinocytes / which therefore show marked signs of cell differentiation (keratinocytes organized in bays). This effect may be related to a calcium intake (by the serum) too important in the culture medium, leading to an early differentiation of keratinocytes.
L'ajout de sérum humain à la concentration de 10% au tampon PBS permet d'obtenir une croissance cellulaire, mais l'observation 25 morphologique des cellules montre des signes de souffrance cellulaire. 14 The addition of 10% human serum to PBS buffer provides cell growth, but morphological observation of the cells shows signs of cellular distress. 14
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WO2019211567A1 (en) * | 2018-05-04 | 2019-11-07 | Naos Institute Of Life Science | Composition comprising alpha-lipoic acid or an alpha-lipoic acid salt, a vitamin c derivative and hyaluronic acid and its use |
FR3080858A1 (en) * | 2018-05-04 | 2019-11-08 | Naos Institute Of Life Science | NON-ANIMAL CELL GROWTH FACTOR AND USE THEREOF |
US20210252054A1 (en) * | 2012-08-10 | 2021-08-19 | Aquavit Pharmaceuticals, Inc. | Vitamin supplement compositions for injection |
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FR2918876A1 (en) * | 2007-07-16 | 2009-01-23 | Oreal | USE OF GREEN LIGHT TO ACTIVATE L-AMINO ACID OXIDASE |
WO2009019381A1 (en) * | 2007-07-16 | 2009-02-12 | L'oreal | Use of green light to activate l-amino acid oxydase |
JP2010533689A (en) * | 2007-07-16 | 2010-10-28 | ロレアル | Use of green light to activate L-amino acid oxidase |
US9750675B2 (en) | 2007-07-16 | 2017-09-05 | L'oreal | Use of green light to activate L-amino acid oxidase |
US20210252054A1 (en) * | 2012-08-10 | 2021-08-19 | Aquavit Pharmaceuticals, Inc. | Vitamin supplement compositions for injection |
US11878034B2 (en) * | 2012-08-10 | 2024-01-23 | Aquavit Pharmaceuticals, Inc. | Vitamin supplement compositions for injection |
WO2019211567A1 (en) * | 2018-05-04 | 2019-11-07 | Naos Institute Of Life Science | Composition comprising alpha-lipoic acid or an alpha-lipoic acid salt, a vitamin c derivative and hyaluronic acid and its use |
FR3080858A1 (en) * | 2018-05-04 | 2019-11-08 | Naos Institute Of Life Science | NON-ANIMAL CELL GROWTH FACTOR AND USE THEREOF |
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FR2895680B1 (en) | 2011-03-18 |
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