Calcium alginate for its use in the prevention and / or treatment of inflammation

Abstract

The present invention relates to a calcium alginate for its use in the prevention and / or treatment of inflammation.

Description

Title of the invention: Calcium alginate for its use in the prevention and / or treatment of inflammation Technical field of the invention

[0001] The present invention relates to calcium alginates for their uses in the prevention and / or treatment of inflammation. Technical background

[0002] Inflammation (or inflammatory reaction) is a natural process that generally occurs when an organism is subjected to microbial aggression or tissue damage, for example due to physical or traumatic chemical agents.

[0003] Inflammation can also result from a dysfunction of the immune system, for example in autoimmune diseases which cause the body to attack its own tissues (multiple sclerosis, polyarthritis, Crohn's disease, etc.), certain genetic diseases (epidermolysis bullosa, familial Mediterranean fever, blau syndrome, etc.) or non-genetic diseases (dermatoses, diabetes, contact dermatitis, tendinitis, etc.) and cancer.

[0004] Inflammation comprises a series of events that enable the activation of immune cells and the release of inflammatory mediators such as cytokines and prostaglandins, to clear the injured site and / or fight infection. This process promotes tissue repair and the return to homeostasis.

[0005] Inflammation can be acute or chronic.

[0006] Acute inflammation is a rapid defense mechanism of the body against external aggressions such as microbes or injuries, thanks to the action of immune cells such as neutrophils and macrophages that phagocytose pathogens. Once the aggression is controlled, the tissue is repaired and the inflammation ends.

[0007] Chronic inflammation is a prolonged response that persists when the insult is not eliminated. It involves continuous activation of immune cells such as macrophages and lymphocytes, leading to tissue damage and the formation of scar tissue, as in the case of chronic diseases or persistent infections.

[0008] Inflammation plays a role, for example, in the healing process for tissue repair, particularly of skin wounds, after an injury or trauma. Healing generally includes the following phases: - hemostasis to stop bleeding, - inflammation to eliminate pathogens and cellular debris, - proliferation to regenerate tissues and close wounds, and - remodeling to result in the formation of the final scar.

[0009] In the case of non-pathological healing, the inflammatory phase resolves rapidly; this is the case of acute inflammation. In the case of pathological healing, the inflammatory phase is prolonged and hinders healing; this is the case of chronic inflammation found in chronic wounds (diabetic wounds, venous or arterial ulcers, pressure sores, etc.).

[0010] Inflammation can be local or systemic.

[0011] Local inflammation usually occurs at a specific location in the body where there is an injury or infection.

[0012] Systemic inflammation generally affects the whole body and can be triggered by severe infections, extensive burns, autoimmune diseases, or chronic inflammatory responses.

[0013] The main cells involved in inflammation include cells derived from myeloid progenitors (neutrophils, macrophages, etc.) and lymphoid progenitors (natural killer cells and lymphocytes). Macrophages play an essential role in the regulation of inflammation. Type M1 macrophages are characterized by the secretion of pro-inflammatory factors such as TNF-α and the production of ROS, enhancing the inflammatory response. In contrast, type M2 macrophages secrete anti-inflammatory cytokines such as IL-10, promote angiogenesis, and contribute to tissue repair. Thus, the transition between type M1 and M2 macrophages regulates the shift from the inflammatory phase to the repair phase and the return to tissue homeostasis.

[0014] In summary, inflammation is essential for the body's protection and plays a role in many physiological circumstances. However, when it is not controlled, it becomes debilitating and requires therapeutic intervention. Treatment of inflammation

[0015] Pathological inflammation is classically treated with anti-inflammatory drugs to control excessive inflammation and / or prevent tissue damage. Among anti-inflammatory drugs, two main categories are distinguished: non-steroidal anti-inflammatory drugs and corticosteroids (or steroidal anti-inflammatory drugs).

[0016] However, anti-inflammatory drugs often have undesirable side effects0,2' and it is useful to diversify the therapeutic means of treating inflammation. This is what the Applicant has achieved in proposing a new means of preventing and / or treating inflammation which consists of calcium alginate. Subject of the patent

[0017] The present invention relates to calcium alginate for its use in the prevention and / or treatment of inflammation.

[0018] The calcium alginate according to the invention can be prepared in the form of a wet composition (such as gel, semi-solid emulsion, membrane or foam), in the form of a liquid composition (such as suspension or lotion) or in the form of a dry composition (powder, film, fiber, etc.).

[0019] The calcium alginate according to the invention can be formulated or incorporated into a composition, in particular a wet or dry composition, or a suitable device. Detailed description of the invention

[0020] • Calcium alginate#

[0021] The present invention relates to calcium alginate for its use in the prevention and / or treatment of inflammation.

[0022] Calcium alginate is known in the prior art to be pro-inflammatory and thus induce and accentuate an inflammatory response0'4'5'6'.

[0023] For example, the study by Gail Chan et al. analyzed the biological effect of a calcium alginate gel and concluded that it promotes the inflammatory mechanism. Indeed, the calcium alginate gel induces the secretion of interleukin I (IL-1), which is a pro-inflammatory cytokine. The authors of this study indicate that in in vitro tests, the calcium alginate gel leads to maturation of dendritic cells of the immune system and increases the secretion of inflammatory cytokines (such as IL-1). This is explained by the fact that dendritic cells require calcium for cytokine secretion. In another study, the inflammatory role of calcium is highlighted by its ability to trigger the innate immune response activated by Toll-like receptors. Adib et al.They confirm the pro-inflammatory effect of calcium alginate by showing that it activates the production of pro-inflammatory cytokines, such as TNF-α, IFN-γ factors, as well as the cytolytic activity of Natural Killer cells.

[0024] In contrast to these studies, document CN-A1-111437435, hereinafter referred to as A1, describes a hydrogel cell structure and a method for preparing such a hydrogel cell structure that would have an anti-inflammatory effect. This hydrogel cell structure comprises a hydrogel formed from calcium alginate and polyvinyl alcohol (PVA), optionally seeded with macrophages of the M2 phenotype. A1 indicates that such a hydrogel would exhibit an anti-inflammatory effect by promoting the differentiation of M1 macrophages into the M2 phenotype.

[0025] The Applicant reproduced the protocol of the second example of Al and then tested the anti-inflammatory effect of the Al hydrogel thus prepared. The protocol and the results of these tests are detailed in Example 1. It appears that, contrary to what the authors of Al claim, the results of the tests (Figures 1a to 1d) demonstrate that the Al hydrogel does not have an anti-inflammatory effect, that it is toxic to macrophages and affects their viability regardless of their phenotype ([Fig. 1a]).

[0026] Thus, calcium alginate has a pro-inflammatory effect according to the teaching of studies0,4,5,6' and according to the results of tests obtained by reproducing the tests of Al (Example 1), and would therefore not allow to control or prevent excessive inflammation, and the tissue damage and delays in healing that may result.

[0027] It is therefore very surprising that the Applicant has highlighted an anti-inflammatory action of calcium alginate and, in particular, its ability to stimulate the differentiation of pro-inflammatory macrophages M1 towards an anti-inflammatory phenotype M2.

[0028] More specifically, the Applicant has demonstrated that calcium alginate, particularly in particulate gel form, reduces both the secretion of tumor necrosis factor alpha (TNF-α) and the production of reactive oxygen species (ROS). TNF-α is produced by various cells of the immune system (such as macrophages with a pro-inflammatory M1 phenotype(7)) and ROS species are known to be pro-inflammatory0'.

[0029] Furthermore, the Applicant has demonstrated that calcium alginate, particularly in the form of a particulate gel, also increases the secretion of interleukin 10 (IL-10) which is produced by various cells of the immune system (such as by anti-inflammatory macrophages M2°') and has an anti-inflammatory effect0' (example 2).

[0030] These advantageous properties make it possible to stimulate a differentiation of pro-inflammatory macrophages M1 towards anti-inflammatory macrophages M2, and thus strengthen the anti-inflammatory action.

[0031] Consequently, calcium alginate now appears as a promising therapeutic strategy for the prevention and / or treatment of inflammation, particularly in the context of wound healing and tissue regeneration (in particular, of the skin, mucous membranes, tendons, ligaments, muscles, cartilage, viscera and / or bones).

[0032] Alginate is a polysaccharide mainly obtained from algae: kelp or fucus.

[0033] It is formed of two monomers: mannuronate or mannuronic acid (M) and guluronate or guluronic acid (G).

[0034] The general formula of alginate is as follows: [P1|GH|QH ; ..O, ? "

[0035] The proportion and distribution of these two monomers are crucial for a wide range of the physical and chemical properties of alginate; alginate is thus characterized by the mannuronic acid / guluronic acid ratio (or M / G ratio). Its chemical composition varies depending on the different species of algae, the different parts of the same plant, and is subject to seasonal changes. Nevertheless, by selecting the origin of the raw materials, it is possible to obtain a variety of alginates with consistent characteristics.

[0036] The carboxyl groups of alginate allow its bonding with cations, forming alginate salts, in particular calcium alginate.

[0037] Alginate-based products have natural intrinsic physical properties and are used in health, mainly for the treatment of gastroesophageal reflux by preventing acid reflux and for the treatment of wounds by maintaining a moist environment and absorbing exudates.

[0038] Advantageously, within the framework of the present invention, calcium alginate can be devoid of cells, such as human, animal or plant cells.

[0039] In particular, calcium alginate may be devoid of M2 and / or ML type macrophages

[0040] Calcium alginate may or may not be formulated with polyvinyl alcohol. According to a particular embodiment, calcium alginate is formulated without polyvinyl alcohol.

[0041] As mentioned above, alginate as such is a polysaccharide formed from two monomers: mannuronic acid and guluronic acid.

[0042] According to a preferred embodiment, the calcium alginate comprises mannuronic acid (M) and guluronic acid (G) in an M / G ratio greater than 0.5. Advantageously, the M / G ratio may be between 0.5 and 1.5. Preferably, the M / G ratio may be between 0.7 and 1.5. More preferably, the M / G ratio may be between 1 and 1.5. Even more preferably, the M / G ratio may be approximately 1 or 1.5. These M / G ratios, in particular, allow for improved anti-inflammatory properties.

[0043] These M / G ratios can be implemented regardless of the form of the calcium alginate of the invention, the composition and / or the device comprising the calcium alginate of the invention.

[0044] The preparation of calcium alginate according to the invention can be carried out according to methods known to those skilled in the art.

[0045] The M / G ratio can be determined by Nuclear Magnetic Resonance (NMR) spectroscopy or by the so-called gel strength method(16,17,18,19).

[0046] According to the invention, calcium alginate has a molar mass of between 60,000 and 400,000 g.mol1. Preferably, the molar mass of calcium alginate is between 100,000 and 300,000 g.mol1.

[0047] The calcium alginate may comprise a calcium content of less than 15% by weight of the total calcium alginate. Preferably, the calcium content is between 3% and 12% by weight of the total calcium alginate. Even more preferably, the calcium content by weight of the calcium alginate is between 6% and 11% by weight of the total calcium alginate.

[0048] According to a particular embodiment, calcium alginate is in particulate form.

[0049] The term “particulate” means a calcium alginate that is in the form of particles or an aggregate of particles.

[0050] Calcium alginate in particulate form comprises particles which may have an average size of between 5 pm and 2000 pm.

[0051] Particle size can be measured according to ISO 13320 by dispersing the sample in a medium and directing a laser beam through this dispersion. The light scattered at different angles is collected by detectors, and the data obtained are processed by software that applies Mie theory to calculate the particle size. The particle size distribution profile is then established by analyzing, for example, the distribution of particle diameters in the sample.

[0052] This particulate form makes it possible, in particular, to increase the anti-inflammatory action of calcium alginate, while having a homogeneous structure (or, in other words, a uniform distribution) for application in a large number of products (such as the various forms mentioned above, the composition and / or the device incorporating calcium alginate according to the invention). This is in order to guarantee uniform application and / or absorption of the calcium alginate, thus ensuring optimal efficacy of the anti-inflammatory action. • Different possible compositions comprising calcium alginate#

[0053] The present invention also relates to a composition comprising calcium alginate, as defined above, for its use in the prevention and / or treatment of inflammation.

[0054] The composition may comprise between 0.4% and 100% by weight of calcium alginate relative to the total weight of the composition.

[0055] According to one embodiment, the composition comprising calcium alginate, as defined above, is in wet form for its use in the prevention and / or treatment of inflammation; this composition is referred to hereafter as the wet composition.

[0056] The term "wet" means a mixture or structure containing a significant amount of an aqueous medium (such as water or physiological saline) or impregnated with this aqueous medium.

[0057] According to another embodiment, the composition comprising calcium alginate, as defined above, is in liquid form for its use in the prevention and / or treatment of inflammation; this composition is hereinafter referred to as the liquid composition.

[0058] The term “liquid” means a dilution of the wet composition by 10 to 20 times in an acceptable solvent.

[0059] According to another embodiment, the composition comprising calcium alginate, as defined above, is in dry form for its use in the prevention and / or treatment of inflammation; this composition is hereinafter referred to as the dry composition.

[0060] The term "dry" means a mixture or substance devoid of aqueous medium (such as water or physiological saline) or containing a minimal amount of this aqueous medium. Wet composition#:

[0061] The wet composition can be formulated as a gel, semi-solid emulsion, membrane or foam.

[0062] The term "gel" means a wet preparation (for example colloidal) containing calcium alginate according to the invention which can be diluted or dispersed in an aqueous medium (such as water or physiological saline).

[0063] A "semi-solid emulsion" is understood to be a composition comprising a lipid phase dispersed in an aqueous phase containing, for example, calcium alginate according to the invention, or vice versa. According to this embodiment, the composition may be a cream optionally comprising a fatty substance.

[0064] The term “membrane” means a generally porous solid structure comprising a material, such as calcium alginate.

[0065] The term "foam" means a structure comprising gas bubbles trapped in a solid or liquid matrix containing, for example, calcium alginate according to the invention.

[0066] The wet composition may thus include calcium alginate as defined above, and an acceptable medium.

[0067] The acceptable medium can be an aqueous medium, preferably physiologically acceptable, such as water or physiological saline.

[0068] The wet composition may comprise between 80% and 98% by weight of acceptable medium relative to the total weight of that composition. Preferably, the wet composition comprises between 90% and 98% by weight of acceptable medium relative to the total weight of that composition.

[0069] The wet composition may comprise less than 10% by weight of calcium alginate relative to the total weight of the composition. Preferably, the wet composition comprises between 0.4% and 7% by weight of calcium alginate relative to the total weight of the composition. Even more preferably, the wet composition comprises between 1% and 6% by weight of calcium alginate relative to the total weight of the composition.

[0070] The wet composition may comprise between 0.1% and 3% by weight of calcium relative to the total weight of that composition. Preferably, the wet composition may comprise between 0.2% and 2% by weight of calcium relative to the total weight of that composition.

[0071] The wet composition may include other components, such as salts, preservatives, stabilizers, active ingredients, copolymers, etc. The wet composition may include less than 10% by weight of these other components relative to the total weight of this composition.

[0072] Preferably, the calcium alginate in the wet composition is in particulate form.

[0073] Calcium alginate in particulate form comprises particles which may have an average size of between 10 µm and 1000 µm. Preferably, this size may be between 20 µm and 200 µm.

[0074] The wet composition as defined above may be in the form of calcium alginate gel.

[0075] By way of example, the preparation of the calcium alginate gel according to the invention can be carried out by mixing or dispersing a solution containing alginate and calcium in an acceptable medium. In particular, the preparation of calcium alginate gel in particulate form is well known to those skilled in the art. Calcium alginate particles can be obtained by gelling droplets of an aqueous phase containing sodium alginate in contact with a phase containing divalent cations (such as calcium), called the gelation phase. Thus, the use of calcium as a divalent cation allows the production of calcium alginate particles.

[0076] The particle size is typically defined by the droplet size before its contact with the phase containing the divalent cations. For example, for calcium alginate particles, the aqueous alginate phase can be contained in a syringe fitted with a needle. The droplets are obtained by expelling the aqueous phase through the needle. The droplet size will be defined by the needle's exit diameter, the viscosity of the aqueous phase, and the height between the needle's exit and the surface of the phase containing the calcium ions. Removing the gelling phase by filtration allows the formation of a gel consisting of calcium alginate particles.

[0077] A non-limiting example of the preparation of the calcium alginate gel according to the invention is described in Example 2. This calcium alginate gel according to the invention makes it possible to obtain an anti-inflammatory effect.

[0078] Advantageously, the preparation of the calcium alginate gel according to the invention provides a smooth and viscoelastic texture, allowing for easy and homogeneous application, ensuring good adhesion, conformability, and uniform coverage of the treated area. The wet composition according to the invention results in a creamy and soft formulation, providing a feeling of comfort and hydration.

[0079] The calcium alginate gel according to the invention can serve as a basis for the formulation of other wet composition(s), such as emulsions, foams and / or membranes. Liquid composition#:

[0080] The liquid composition can be formulated by diluting the wet composition 10 to 20 times in an acceptable solvent. The liquid composition can be formulated as a suspension or a lotion.

[0081] The acceptable solvent may be an aqueous solvent or a water-miscible solvent compatible with alginate.

[0082] The term "suspension" refers to a heterogeneous mixture comprising fine solid particles, containing, for example, calcium alginate according to the invention, dispersed in a liquid without being dissolved. By way of example, the suspension comprising calcium alginate may have a water content different from that of the calcium alginate gel.

[0083] The term "lotion" refers to an oil-in-water emulsion, particularly one with a higher aqueous phase content than in a cream. For example, a lotion containing calcium alginate may also contain an oily substance in addition to the calcium alginate suspension. The liquid composition may comprise between 98% and 99.95% by weight of an acceptable solvent relative to the total weight of the composition.

[0084] The liquid composition may comprise less than 1% by weight of calcium alginate relative to the total weight of the composition. Preferably, the liquid composition comprises between 0.02% and 0.7% by weight of calcium alginate relative to the total weight of the composition. Even more preferably, the liquid composition comprises between 0.5% and 0.6% by weight of calcium alginate relative to the total weight of the composition.

[0085] The liquid composition may comprise between 0.005% and 0.3% by weight of calcium relative to the total weight of that composition. Preferably, the wet composition may comprise between 0.01% and 0.2% by weight of calcium relative to the total weight of that composition.

[0086] The liquid composition may include other components, such as salts, preservatives, stabilizers, active ingredients, copolymers, etc. The liquid composition may include less than 1% by weight of these other components relative to the total weight of this composition. Dry composition#:

[0087] The dry composition can be formulated as a film, fiber, or powder.

[0088] The term "film" refers to a generally thin and compact solid structure comprising a material (namely, calcium alginate). The film is prepared by drying a wet calcium alginate composition according to the invention.

[0089] The term "fiber" refers to a filamentous structure (containing, for example, calcium alginate according to the invention) that is generally elongated and thin. The preparation of calcium alginate fiber is well known to those skilled in the art by extruding a sodium alginate solution through a die in a calcium chloride bath. The calcium alginate fiber according to the invention can be used for the manufacture of yarns, compresses, and wicks.

[0090] The term "powder" means a structure comprising solid particles (containing, for example, calcium alginate according to the invention) dispersed in the form of grains or fragments.

[0091] The dry composition thus comprises calcium alginate as defined above.

[0092] The dry composition may optionally comprise less than 20% by weight of aqueous medium relative to the total weight of the composition. Preferably, the dry composition may comprise between 2% and 20% by weight of aqueous medium relative to the total weight of the composition.

[0093] The dry composition may comprise more than 10% by weight of calcium alginate relative to the total weight of that composition. Preferably, the dry composition comprises between 10% and 100% by weight of calcium alginate relative to the total weight of that composition. Even more Preferably, the dry composition comprises between 30% and 90% by weight of calcium alginate relative to the total weight of this composition.

[0094] The dry composition may comprise less than 15% by weight of calcium relative to the total weight of the composition. Preferably, the dry composition comprises between 2% and 12% by weight of calcium relative to the total weight of the composition. Even more preferably, the dry composition comprises between 2.5% and 10% by weight of calcium relative to the total weight of the composition.

[0095] Preferably, the calcium alginate in the dry composition is in a particulate form whose characteristics are detailed below.

[0096] Calcium alginate in particulate form comprises particles which may have a size ranging from 5 pm to 2000 pm.

[0097] By way of example, the dry composition in powder form may be in the form of a plurality of granular particles. The particles advantageously have a size less than 200 pm, preferably a size on the order of 75 pm, which is defined by sieving or by laser diffraction measurement.

[0098] The powder in the form of a plurality of granular particles in the dry composition according to the invention can be prepared as described in patent FR-B3-2823979 filed by the Applicant.

[0099] According to one embodiment, the powder also takes the form of a plurality of cylindrical particles obtained from fibers.

[0100] The cylindrical particles can have a length between 20 pm and 2000 pm and a diameter between 5 pm and 50 pm.

[0101] The powder in the form of a plurality of cylindrical particles in the dry composition according to the invention can be prepared as described in patent EP-B1-2836243 filed by the Applicant. • Adding ingredients#

[0102] The composition according to the invention, regardless of its dry or wet form, can advantageously be associated with one or more anti-inflammatory agent(s).

[0103] For example, the anti-inflammatory agent may be chosen from a steroidal type anti-inflammatory agent (such as cortisone), an anti-steroidal type anti-inflammatory agent (such as ibuprofen), a natural active ingredient (such as curcumin) and / or ions (such as zinc).

[0104] The composition comprising calcium alginate may be associated with an ion, in particular zinc.

[0105] Zinc is a physiological trace element found within the extracellular matrix (ECM) of the dermis and epidermis. Thanks to its pharmacological properties, which have been widely studied and recognized in the literature, zinc is Used for its anti-inflammatory, antioxidant, and immunoregulatory properties: it reduces pro-inflammatory cytokines such as interleukin 6 (IL-6), interleukin 1 (IL-1), and tumor necrosis factor 30 (TNF30). Zinc contributes to maintaining skin tissue homeostasis and to skin repair / regeneration processes.

[0106] Combining zinc with calcium alginate enhances the anti-inflammatory action.

[0107] The composition may comprise less than 1% by weight of zinc relative to the total weight of the composition. Preferably, the composition comprises between 0.02% and 0.05% by weight of zinc relative to the total weight of the composition. Even more preferably, the composition comprises between 0.027% and 0.035% by weight of zinc relative to the total weight of the composition.

[0108] The composition may further include one or more ingredients selected from a buffer, a stabilizing agent (such as pectins, cellulose derivatives, xanthan gum, etc.), a preservative (such as benzalkonium chloride, potassium sorbate, sodium benzoate, tocopherol, etc.), a surfactant, a vitamin, a mineral, one or more ions (such as zinc), a peptide, proteins, any element of an extracellular matrix, a pH adjuster, an antibiotic, an antimicrobial and an antibiofilm. • Various packaging options are available, including calcium alginate #

[0109] The composition comprising calcium alginate, as defined above, can be incorporated into different types of devices such as threads, dressings, wicks, compresses, aerosols, sprays, liquid (oral or injectable) or semi-solid forms, suppositories, oral solid forms.

[0110] By way of example, dressing-type devices, such as gauze and compresses, can be selected from highly absorbent fibers, hydrofibers, alginates, hydrocellular materials, hydrocolloids, hydrogels, petrolatum-based dressings, interfaces, activated charcoal dressings, silver dressings, and hyaluronic acid-based dressings. These devices are generally used for hemostasis and wound healing. • Various possible applications#

[0111] In general, the present invention relates to calcium alginate, the composition and / or the device comprising calcium alginate according to the invention, for use in the prevention and / or treatment of any pathology having an inflammatory component.

[0112] Inflammation is generally manifested by pain, redness, edema or swelling and heat.

[0113] By way of example, inflammation may result from or be associated with various factors listed in a non-exhaustive manner: - an infection (viral, bacterial, fungal or parasitic), - trauma (e.g., physical injuries such as burns, (post-)surgical wounds, sutured wounds, etc.), - an abnormal immune reaction (such as an allergy), - chemical irritants (such as toxins, pollutants, certain industrial chemicals, etc.), - chronic conditions (such as chronic diseases, chronic wounds, oxidative stress, etc.), - cancers, and - lifestyle (such as after sports activity, smoking, alcoholism and / or stress).

[0114] The inflammation that the present invention aims to treat can occur in various organs and the method of administration and formulation of calcium alginate will be chosen according to the target organ to be treated.

[0115] In particular, the location of the inflammation may be cutaneous or extracutaneous (such as cartilaginous, mucosal, muscular, tendinous, ligamentous, visceral and / or osseous).

[0116] By way of illustration and without limitation, inflammatory skin disorders may include eczema, atopic dermatitis, urticaria, acne, seborrheic dermatitis, contact dermatitis, couperose, erythrosis, psoriasis, etc.

[0117] Inflammatory skin disorders can also be genetic and / or rare diseases, such as epidermolysis bullosa.

[0118] Extracutaneous inflammatory disorders may include, but are not limited to, rheumatoid arthritis, inflammatory bowel diseases (such as Crohn's disease), chronic diseases (such as asthma), autoimmune diseases (such as systemic lupus erythematosus), etc.

[0119] Inflammatory cartilage disorders may include, but are not limited to, osteoarthritis.

[0120] Mucosal inflammatory disorders may include, but are not limited to, bronchitis, gastritis, stomatitis, conjunctivitis, cystitis, etc.

[0121] Muscle inflammatory disorders may include, but are not limited to, various myositis (such as infectious, ossifying, etc.), fibromyalgia, etc.

[0122] Inflammatory tendon disorders may include, but are not limited to, various tendinitis (such as shoulder, wrist, etc.).

[0123] Inflammatory ligament disorders may include, but are not limited to, ligamentitis, ligament sprain, etc.

[0124] According to specific embodiments, calcium alginate, the composition and / or the device comprising calcium alginate according to the invention are used for the prevention and / or treatment of inflammation of the joints, muscles, tendons and ligaments, which may for example occur during sports practice.

[0125] The present invention further relates to calcium alginate, the composition and / or the device comprising calcium alginate according to the invention, for their use in the prevention and / or treatment of inflammation of wounds or tissue lesions to promote their healing. Tissue wounds may be cutaneous, extracutaneous, cartilaginous, mucosal, muscular, tendinous, ligamentous, visceral and / or osseous lesions.

[0126] It is also known that aging can be accompanied and exacerbated by low-level chronic and systemic inflammation(20); this latent inflammation, although not pathological, increases the risk of developing diseases and it is advantageous to prevent or limit its development.

[0127] Thus, the present invention further relates to a method for the non-therapeutic prevention and / or treatment of the inflammatory component of aging, comprising the administration of calcium alginate according to the invention, of the composition and / or of the device comprising the calcium alginate according to the invention. According to this embodiment, the present invention may more particularly relate to a cosmetic treatment method for skin aging, comprising the administration of calcium alginate according to the invention, of the composition and / or of the device comprising the calcium alginate according to the invention. • Administrative channels#

[0128] Calcium alginate, the composition and / or the device may be administered by any route, including topically, orally (or buccally), nasally, rectally or by injection.

[0129] The topical route can allow the composition according to the invention to be administered locally on the skin or mucous membranes, for example, to treat inflammatory skin conditions or to control inflammation during the healing process.

[0130] The topical route can also allow the composition according to the invention to be administered locally to certain tissues, for example, to treat inflammatory tissue conditions other than those of the skin, such as tendons, ligaments, muscles and / or bones.

[0131] The oral route can be used to treat inflammatory conditions, including inflammatory bowel diseases such as Crohn's disease, as well as other systemic inflammatory conditions such as rheumatoid arthritis.

[0132] The nasal route can be used to administer the composition according to the invention to treat inflammatory conditions of the nasal mucosa and sinuses, such as allergic rhinitis, chronic sinusitis and other inflammatory nasal conditions.

[0133] The rectal route can allow the composition according to the invention to be administered to treat inflammatory conditions affecting the rectum, colon or anus, such as ulcerative colitis, hemorrhoids and other inflammatory bowel diseases.

[0134] The injectable route allows the composition according to the invention to be administered by transcutaneous, intramuscular, or intravenous injection to treat inflammatory tissue conditions other than those of the skin, such as tendons, ligaments, muscles, viscera, and / or bones. This is intended to prevent and / or rapidly treat inflammations, such as flare-ups of rheumatoid arthritis, exacerbations of inflammatory bowel disease, or other viscera and / or bone diseases.

[0135] By way of example, the gel, cream, lotion, aerosol and / or foam can be applied directly to the wounds for local absorption and targeted action on the inflamed area.

[0136] The tablets can be administered orally.

[0137] The oral formulation can be administered when the composition is liquid and as an alternative to tablets.

[0138] As mentioned above, the injectable formulation (such as in gel form) can be administered intramuscularly, intravenously or intra-articularly to rapidly treat inflammations. Brief description of the figures

[0139] Other features and advantages of the invention will become apparent upon reading the detailed description that follows, for an understanding of which reference should be made to the accompanying figures in which:

[0140] Fig. 1a represents a histogram illustrating the effect on the viability of macrophages Ml of the hydrogel formed from calcium alginate and a polyvinyl alcohol of document Al, this hydrogel being obtained from the protocol of example 2 of Al;

[0141] Figure 1b represents a histogram illustrating the percentage of cells positive for the CD206 marker by control M1 macrophages, M1 in contact with Al hydrogel as well as by M2 macrophages;

[0142] Figure 1 represents a histogram illustrating the co-expression of CD163 and CD206 as a percentage of macrophages Ml controls, Ml in contact with Al hydrogel and M2;

[0143] Fig. 1d represents a histogram illustrating the secretion of the anti-inflammatory interleukin IL-10 by control macrophages M1, M1 in contact with Al hydrogel and M2;

[0144] Figure 2a represents a histogram illustrating the effect on the viability of macrophages Ml of calcium alginate gel in particulate form according to the invention;

[0145] Figure 2b represents a histogram illustrating the co-expression of CD163 and CD206 as a percentage of macrophages M1 controls, M1 in contact with calcium alginate gel in particulate form according to the invention and M2;

[0146] Figure 2c represents a histogram illustrating the secretion of anti-interleukin inflammatory IL-10 by macrophages M1 controls, M1 in contact with calcium alginate gel in particulate form according to the invention and M2;

[0147] Fig. 2d represents a histogram illustrating the ratio of secretion of the pro-inflammatory factor TNF-α to the secretion of the anti-inflammatory interleukin IL-10 by control macrophages M1, M1 in contact with the calcium alginate gel in particulate form according to the invention and M2;

[0148] Fig. 2e represents a histogram illustrating the production of ROS pro inflammatory by macrophages M1 controls, M1 in contact with calcium alginate gel in particulate form according to the invention and M2;

[0149] Figure 3a represents a histogram illustrating the effects on viability of macrophages Ml of two calcium alginate gels according to the invention at different M / G ratios (G1 and G2);

[0150] Figure 3b represents a histogram illustrating the co-expression of CD163 and CD206 as a percentage of M1 macrophages in contact with two calcium alginate gels according to the invention at different M / G ratios (G1 and G2) compared to control M1 and M2 macrophages;

[0151] Figure 3c represents a histogram illustrating the secretion of anti-interleukin inflammatory IL-10 by M1 macrophages in contact with two calcium alginate gels according to the invention at different M / G ratios (G1 and G2) compared to control M1 and M2 macrophages;

[0152] Fig. 3d represents a histogram illustrating the ratio of TNF-α factor secretion to interleukin IL-10 secretion by M1 macrophages in contact with two calcium alginate gels according to the invention at different M / G ratios (G1 and G2) compared to control M1 and M2 macrophages;

[0153] Figure 4 represents a histogram illustrating the TNF- factor secretion ratio compared to the secretion of interleukin IL-10 by M1 macrophages in contact with calcium alginate (G) and zinc-associated calcium alginate (G.Zn2+) gels compared to control M1 and M2 macrophages.

[0154] Example 1 - Calcium alginate according to the prior art Preparation of the hydrogel from document Al

[0155] The hydrogel formed from calcium alginate and polyvinyl alcohol described in Al was prepared from the protocol of Example 2 of Al.

[0156] This protocol is briefly summarized below:

[0157] - add an amount of 100 mg of sodium alginate (referenced as SA in Al) in a volume of 12.5 ml of deionized water, and stir magnetically until the sodium alginate dissolves completely;

[0158] - add 500 mg of PVA to 12.5 ml of water demineralized, and stir at 80°C until the PVA dissolves completely;

[0159] - mix the two solutions of sodium alginate and PVA uniformly;

[0160] - add an aqueous solution of calcium chloride of 74 µl (having a concentration of 5 mg / ml);

[0161] - Adjust the pH: one drop of sodium hydroxide (NaOH) at IM (VWR) was added to obtain a pH of 6.5 and form an intermediate hydrogel which was crosslinked with calcium ion (referenced Ca-SA / PVA hydrogel in Al) with a degree of crosslinking of 1%;

[0162] - to carry out a primary freeze-thaw cycle: the Ca-SA / PVA hydrogel was placed in a glass Petri dish, and frozen for 22h at -20°C then thawed for 2h at 25°C then a freeze-drying cycle was carried out to obtain a freeze-dried gel;

[0163] - add 500 µl of phosphate buffer (PBS) solution at pH 7.4 into the pores of the lyophilized gel so that the lyophilized gel absorbs the PBS solution and thus obtain a final hydrogel referenced (Ca-SA / PVA) 1FT+FD in Al.

[0164] Preparation of pro- and anti-inflammatory macrophages M1 and M2

[0165] PB MC cells (acronym for peripheral blood mononuclear cells) from a healthy individual (i.e. without inflammatory disorders) are isolated by centrifugation in a Ficoll solution.

[0166] Monocytes with the classic markers CD14+ / CD16- are purified.

[0167] These purified monocytes are placed in a 24-well plate (containing, for example, 400,000 cells / mL / well) and differentiated as follows:

[0168] Macrophages Ml: - monocytes are incubated with a culture medium containing 10% human serum in RPMI medium (English acronym for "Roswell Park Mondial Institut Medium") with 1% of a Penicillin and Streptomycin (PS) based antibiotic, 1% LG (acronym for "L-Glutamine") and 50 ng / mL of GM-CSF (English acronym for "Granulocyte Macrophage Colony Stimulating Factor"); - the plate in an incubator for 6 days; - from the sixth day of culture, half of the medium is replaced by another culture medium containing: 10% S VF (acronym for fetal calf serum) in an RPMI medium with 1% PS, 1% LG, 50 ng / mL of GM-CSF factor and 50 ng / mL of LPS / IFN-y activators (acronym for Lipopolysaccharide and Interferon-y), and incubated for 24h for differentiation into pro-inflammatory macrophages Ml.

[0169] M2 Macrophages: - monocytes are incubated with a culture medium containing 10% human serum in an RPMI medium with 1% PS, 1% LG and 50 ng / ml of an M-CSF factor (English acronym for "Monocyte colony-stimulating factor") to differentiate monocytes into anti-inflammatory M2 macrophages; - a plate is placed in an incubator for 6 days; - from the sixth day of culture, half of the medium is replaced with another culture medium comprising: 10% S VF in RPMI medium, 1% PS, 1% LG, 50 ng / mL of M-CSF factor, 20 ng / mL of IL-4 cytokines (acronym for interleukin 4) and IL-13 cytokines (acronym for interleukin 13), and incubated for 24h for differentiation into anti-inflammatory M2 macrophages. Results#: Effects of the hydrogel in document Al

[0170] The anti-inflammatory activity of the Al hydrogel was tested on pro-inflammatory ML macrophages

[0171] Figures 1a to 1d illustrate the effects of Al hydrogel on Ml macrophages (denoted "Al").

[0172] In particular, [Fig.la] illustrates the viability, as a percentage of living cells, of Ml macrophages in contact with Al hydrogel compared to pro-inflammatory control macrophages ML. In this figure, the viability of Ml macrophages in contact with Al hydrogel is greatly reduced compared to that of control Ml macrophages.

[0173] Thus, Al hydrogel has a toxic effect on cells, particularly on pro-inflammatory ML macrophages

[0174] Al describes a unique marker CD206 to assess the anti-inflammatory profile of macrophages.

[0175] Fig.lb illustrates the % of macrophages expressing the CD206 marker among M1 macrophages in contact with Al hydrogel and that of macrophages in control condition (M1 and M2).

[0176] In [Fig. 1b], the percentage of cells positive for the CD206 marker by control macrophages M1 and M2 is almost identical; it can be deduced that the observation of the percentage of cells positive for CD206 is not correlated with a pro- or anti-inflammatory phenotype and does not allow differentiation of an M1 profile or M2. Some studies (11,12) mention that the CD206 marker is not sufficient to indicate an anti-inflammatory state. It is therefore not a relevant marker for evaluating an anti-inflammatory effect (01,12,13,14,15).

[0177] It is known from the prior art that the co-expression of the markers CD163 and CD206 makes it possible to differentiate macrophages of profile M1 (low co-expression) from profile M2 (high co-expression)05,16).

[0178] Fig. 1a thus illustrates this CD163 / CD206 co-expression for M1 macrophages in contact with Al hydrogel and for pro- and anti-inflammatory macrophages, M1 and M2, in control condition.

[0179] In [Fig. 1e], the percentage of CD163 and CD206 co-expression in control M2 macrophages is higher than that observed in control M1 macrophages. This confirms that this CD163 and CD206 co-expression allows us to distinguish the phenotype of pro-inflammatory M1 macrophages from anti-inflammatory M2 macrophages.

[0180] Moreover, in this [Fig.1e], the Ml macrophages in contact with the Al hydrogel have a low co-expression of CD163 and CD206 compared to the M2 macrophages in the control condition, but also compared to the Ml macrophages in the control condition.

[0181] Thus, Al hydrogel does not induce the differentiation of M1 macrophages into M2 macrophages. Since the co-expression of the CD163 and CD206 markers obtained in the presence of Al hydrogel was lower than that measured for control M1 macrophages, Al hydrogel did not activate M1 macrophages into M2 macrophages. This confirms that Al hydrogel does not have an anti-inflammatory effect.

[0182] Finally, [Fig.ld] illustrates the secretion of the anti-inflammatory interleukin IL-10 by M1 macrophages in contact with Al hydrogel and by pro- and anti-inflammatory macrophages, M1 and M2, in control condition.

[0183] In [Fig.ld], the secretion of interleukin IL-10 by anti-inflammatory macrophages M2 is, as expected, much greater than that of pro-inflammatory macrophages ML. The secretion of interleukin IL-10 by macrophages Ml in contact with Al hydrogel is less than that of control macrophages Ml and M2.

[0184] Thus, Al hydrogel decreases the secretion of interleukin IL-10, even compared to control M1 macrophages. Al hydrogel did not activate M1 macrophages into M2 macrophages. This also confirms that Al hydrogel does not exhibit an anti-inflammatory effect.

[0185] Contrary to what Al claims, the results obtained and presented in figures 1a to 1d demonstrate that the Al hydrogel does not induce an M2 phenotype and does not have anti-inflammatory properties.

[0186] Example 2 - Preparation and evaluation of the effect of calcium alginate according to the invention

[0187] The anti-inflammatory effect of calcium alginate according to the invention in gel form was tested on macrophages.

[0188] Preparation of a calcium alginate gel for testing

[0189] A first calcium ion solution is prepared in 2 liters (L). This first solution contains 1% by weight of calcium chloride (CaCl2), 0.6% by weight of sodium chloride (NaCl) and 98.4% by weight of water, preferably demineralized.

[0190] Approximately 0.4% by weight of zinc gluconate was added to this first solution to obtain a zinc-enriched gel (G.Zn2+).

[0191] A second sodium alginate (AlgNa) solution is prepared in 2 liters (L). This second solution contains 2% by weight of sodium alginate, 0.9% by weight of NaCl and 97.1% by weight of water, preferably demineralized.

[0192] The second solution is sprayed through a nozzle into the first solution contained in a tank, the assembly being installed in a suitable housing and with continuous agitation in the tank using one or more magnetic stirrers. This spraying is carried out with the following parameters: - pressure in the tank of approximately 1.8 bar, - casing pressure of approximately 2.5 bar, and - spraying duration of approximately 90 minutes.

[0193] The sprayed solution is then centrifuged several times, for example three times, at a speed of about 2500 rpm for about 2 minutes. At each centrifugation, the pellet is resuspended with a quantity of vacuum-filtered demineralized water at about 0.1 µm to reach a mass of 300 g.

[0194] The pellet from the last centrifugation is sterilized by passing through a UV reactor, to obtain the calcium alginate gel at the outlet of the UV reactor.

[0195] This gel is in particulate form.

[0196] Several calcium alginate gels according to the invention have been prepared from the preparation described above, such as: - a G gel with an M / G ratio of approximately 1.5; - a G.Zn2+ gel with an M / G ratio of approximately 1.5; - a G1 gel with an M / G ratio of approximately 0.7; and - a G2 gel with an M / G ratio of approximately 1.

[0197] Evaluation of the effect of calcium alginate in gel form on pro-inflammatory macrophages Ml

[0198] Preparation of pro- and anti-inflammatory macrophages M1 and M2

[0199] PB MC cells (acronym for peripheral blood mononuclear cells) from a healthy individual (i.e., without inflammatory disorders) are isolated by Ficoll.

[0200] Monocytes with the classic markers CD14VCD16 are purified.

[0201] These purified monocytes are placed in a 24-well plate (containing, for example, 400,000 cells / mL / well) and differentiated as follows:

[0202] Macrophages Ml: - monocytes are incubated with a culture medium containing 10% human serum in RPMI medium with 1% of a Penicillin and Streptomycin-based antibiotic, 1% LG and 50 ng / ml of a GM-CSF factor; - the plate is placed in an incubator for 6 days; - From the sixth day of culture, half of the medium is replaced with another culture medium containing: 10% SVF in RPMI medium with 1% PS, 1% LG, 50 ng / mL of GM-CSF factor and 50 ng / mL of LPS / IFN-γ activators (acronym for Lipopolysaccharide and Interferon-γ) and incubated for 24 hours for differentiation into pro-inflammatory macrophages ML

[0203] M2 Macrophages: - monocytes are incubated with a culture medium containing 10% human serum in RPMI medium with 1% PS, 1% LG and 50 ng / mL of an M-CSF factor to differentiate monocytes into anti-inflammatory M2 macrophages; - the plate is placed in an incubator for 6 days; - from the sixth day of culture, half of the medium is replaced with another culture medium comprising: 10% SVF in an RPMI medium, 1% PS, 1% LG, 50 ng / mL of the M-CSF factor, 20 ng / mL of the cytokines IL-4 (acronym for interleukin 4) and the cytokines IL-13 (acronym for interleukin 13) and incubated for 24h for differentiation into anti-inflammatory M2 macrophages.

[0204] In order to test the activity of the prepared gels G, G.Zn2+, G1 and G2, a 10% dilution of each of these gels in culture medium (i.e., 100 µl in ImL) is brought into contact with pro-inflammatory macrophages ML • Characterization of the macrophage phenotype

[0205] After a 3-day culture in the presence of the different gels, the macrophages are recovered from the plate. After centrifugation and washing, extracellular labeling is performed with the following antibodies: CD163-BV421, CD206-APC and 7-aminoactinomycin D (7-AAD), then the macrophages are analyzed by flow cytometry in flux. Co-expression of CD163 and CD206 allows the identification of anti-inflammatory macrophages, and 7-AAD is used to assess cell viability. • Measurement of cytokine secretion

[0206] After a 2-day culture in contact with gels G, G.Zn2+, G1, and G2, the culture media are centrifuged to remove the different gels. After adding 50 ng / mL of LPS, these culture media are redistributed into their corresponding wells.

[0207] After an additional 24-hour culture, TNF-α and IL-10 cytokine secretions are measured in the supernatants using MACSPlex cytokine kits (Miltenyi). • Measurement of ROS species production

[0208] After a 3-day culture in contact with the G, G.Zn2+, G1 and G2 gels, the cells are recovered, centrifuged and washed.

[0209] According to the instructions for the abl 13851 (abcam) and M7514 (Thermofisher) kits, the production of ROS normalized on mitochondrial mass is measured on macrophages in contact with the different gels.

[0210] Results: Anti-inflammatory effects of calcium alginate gel on pro-inflammatory macrophages Ml

[0211] The anti-inflammatory activity of each of the gels G, G.Zn2+, G1 and G2 is tested on pro-inflammatory macrophages Ml by measuring the following parameters: - co-expression of the markers CD163 and CD206 (Figures 2b and 3b), - secretion of interleukin IL-10 (Figures 2c and 3c), - ratio of the secretion of the factors TNF-a / IL-10 ([Fig.2d], 3d, 4), - production of ROS species (Figures 2e).

[0212] These tests demonstrate the reduction of pro-inflammatory components (TNF-a, ROS) and / or the increase of anti-inflammatory components (CD 163 / CD206, IL-10) by Ml macrophages in the presence of each of the gels G, G.Zn2+, G1 and G2. • Effect of G gel on pro-inflammatory macrophages Ml

[0213] Fig. 2a evaluates the toxicity of gel G on Ml macrophages compared with control pro-inflammatory Ml macrophages.

[0214] In [Fig. 2a], the percentage of live Ml macrophages in contact with gel G is identical to that of control ML macrophages

[0215] Thus, the calcium alginate of gel G does not affect the survival of macrophages.

[0216] Fig. 2b illustrates the co-expression of CD163 and CD206 of M1 macrophages in contact with gel G compared to pro- and anti-inflammatory control macrophages, M1 and M2.

[0217] In this [Fig. 2b], the % of M2 macrophages co-expressing CD163 and CD206 is, as expected, significantly increased compared to M1 macrophages. The % of M1 macrophages co-expressing CD163 and CD206 in the presence of gel G is higher than that of control M1 macrophages.

[0218] Gel G therefore increases the co-expression of CD163 and CD206. Since this co-expression of CD163 and CD206 is a marker of anti-inflammatory M2 macrophages, gel G enabled the activation of pro-inflammatory M1 macrophages into anti-inflammatory M2 macrophages.

[0219] Fig. 2c illustrates the secretion of the anti-inflammatory interleukin IL-10 by M1 macrophages in contact with gel G and that of pro- and anti-inflammatory macrophages, M1 and M2, under control conditions.

[0220] In [Fig. 2c], the secretion of interleukin IL-10 is, as expected, significantly increased by M2 macrophages compared to control M1 macrophages. The secretion of interleukin IL-10 by M1 macrophages in the presence of gel G is greater than that of pro-inflammatory control M1 macrophages. Since interleukin IL-10 is an anti-inflammatory protein, its increase upon contact with gel G demonstrates the stimulation of M2-type anti-inflammatory activity by initially pro-inflammatory ML macrophages.

[0221] Figure 2d illustrates the ratio of TNF-α secretion to interleukin IL-10 secretion (TNF-α / IL-10) in M1 macrophages in contact with gel G and in pro- and anti-inflammatory macrophages, M1 and M2, under control conditions. As expected, this TNF-α / IL-10 ratio is significantly reduced in M2 macrophages compared to control macrophages ML. This TNF-α / IL-10 ratio in M1 macrophages in the presence of gel G is significantly reduced compared to the control M1 macrophage, further demonstrating an anti-inflammatory effect of gel G.

[0222] Fig. 2e illustrates the production of ROS species by M1 macrophages in contact with gel G and that of pro- and anti-inflammatory macrophages, M1 and M2, under control conditions.

[0223] In [Fig.2e], ROS production is, as expected, greatly reduced for M2 macrophages compared to control ML macrophages. ROS species production by M1 in the presence of G gel is also lower than that of pro-inflammatory ML macrophages. Since ROS species are pro-inflammatory, the decrease in their production in the presence of G gel corresponds to a reduction in the pro-inflammatory activity of M1 macrophages by this G gel.

[0224] All these results confirm that the calcium alginate of gel G exhibits anti-inflammatory activity. • Effect of G1 and G2 gels with different M / G ratios on pro-inflammatory macrophages Ml

[0225] Fig. 3a evaluates the toxicity of G1 and G2 gels on Ml macrophages compared with control pro-inflammatory Ml macrophages.

[0226] In [Fig.3a], the percentage of Ml macrophages in contact with each of the G1 and G2 gels is identical to that of the pro-inflammatory Ml control macrophages.

[0227] Thus, calcium alginate from gels G1 and G2 does not affect macrophage survival.

[0228] Fig. 3b illustrates the co-expression of CD163 and CD206 by M1 macrophages in contact with G1 and G2 gels compared to pro- and anti-inflammatory macrophages, M1 and M2, controls.

[0229] In this [Fig. 3b], the co-expression of CD163 and CD206 in the presence of gels G1 and G2 is greater than that of the ML control

[0230] G1 and G2 gels have the effect of increasing the co-expression of CD163 and CD206, and have allowed the differentiation of pro-inflammatory macrophages M1 into anti-inflammatory macrophages M2.

[0231] Fig. 3c illustrates the secretion of interleukin IL-10 by M1 macrophages in contact with G1 and G2 gels and that of pro- and anti-inflammatory M1 and M2 macrophages, in control condition.

[0232] In [Fig. 3c], the secretion of interleukin IL-10 by M1 macrophages in the presence of gel G2 is greater than that of the pro-inflammatory M1 control macrophages. The increased secretion of interleukin IL-10 upon contact with gel G2 induces anti-inflammatory activity by the pro-inflammatory ML macrophages.

[0233] Thus, calcium alginate in gel form (in particular G2 gel) exhibits anti-inflammatory activity.

[0234] Figure 3d illustrates the ratio of TNF-α secretion to interleukin IL-10 secretion (TNF-α / IL-10) in M1 macrophages in contact with gels G1 and G2, and in pro- and anti-inflammatory macrophages, M1 and M2, under control conditions. As expected, this TNF-α / IL-10 ratio is significantly reduced in M2 macrophages compared to control macrophages ML. This TNF-α / IL-10 ratio in M1 macrophages in the presence of gel G2 is also reduced compared to the control M1 macrophage, further demonstrating an anti-inflammatory effect of gel G2. • Effects of G and G.Zn2+ gels on pro-inflammatory M1 macrophages

[0235] Figure 4 illustrates the ratio of TNF-α secretion to interleukin IL-10 secretion (TNF-α / IL-10) in M1 macrophages in contact with G and G.Zn2+ gels, and in pro- and anti-inflammatory macrophages, M1 and M2, under control conditions. As expected, this TNF-α / IL-10 ratio is significantly decreased in M2 macrophages compared to control ML macrophages. This TNF-α / IL-10 ratio in M1 macrophages in the presence of G.Zn2+ gel is decreased compared to that in G gel. Thus, The combination of zinc with calcium alginate increases the differentiation of pro-inflammatory macrophages M1 into anti-inflammatory macrophages M2 in this G.Zn2+ gel. REFERENCES

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Claims

Demands

1. Calcium alginate for its use in the prevention and / or treatment of inflammation.

2. Calcium alginate according to claim 1, characterized in that it comprises mannuronic acid (M) and guluronic acid (G) in an M / G ratio greater than 0.5, advantageously the M / G ratio is between 0.5 and 1.5, preferably the M / G ratio is between 0.7 and 1.5, more preferably the M / G ratio is between 1 and 1.5, and even more preferably the M / G ratio is about 1 or 1.5

3. 1,J. Calcium alginate according to claim 1 or claim 2, characterized in that its molar mass is between 60,000 and 400,000 g.mol1, preferably its molar mass is between 100,000 and 300,000 g.mol1.

4. Calcium alginate according to any one of claims 1 to 3, characterized in that it comprises a calcium content of less than 15% by weight of total calcium alginate, preferably the calcium content is between 3% and 12% by weight of calcium by weight of total calcium alginate, even more preferably the calcium content is between 6% and 11% by weight of calcium by weight of total calcium alginate.

5. A composition for use in the prevention and / or treatment of inflammation, characterized in that it comprises calcium alginate according to any one of claims 1 to A

6. Composition according to claim 5, characterized in that the calcium alginate is in particulate form.

7. Composition according to claim 6, characterized in that the calcium alginate particles have an average size between 5 pm and 2000 pm.

8. A composition according to any one of claims 5 to 7, characterized in that it further comprises one or more ingredients selected from buffers, stabilizing agents, surfactants, a vitamin, a mineral, one or more ions, a peptide, proteins, any element of an extracellular matrix, a pH adjuster, an antibiotic, an antimicrobial, an antibiofilm and another anti-inflammatory.

9. Composition according to any one of claims 5 to 8, characterized in that it is incorporated into various devices such as threads, dressings, wicks, compresses, aerosols, sprays, oral or injectable liquid forms, suppositories or solid forms.

10. Composition according to any one of claims 5 to 9, characterized in that it is formulated in wet form such as in the form of a gel, semi-solid emulsion, membrane or foam.

11. Composition according to claim 10, characterized in that it comprises less than 10% by weight of calcium alginate relative to the total weight of the composition, preferably the composition comprises between 0.4% and 7% by weight of calcium alginate relative to the total weight of the composition, and even more preferably the composition comprises between 1% and 6% by weight of calcium alginate relative to the total weight of the composition.

12. Composition according to claim 10 or claim 11, characterized in that it comprises between 0.1% and 3% by weight of calcium relative to the total weight of the composition, advantageously the composition comprises between 0.2% and 2% by weight of calcium relative to the total weight of the composition.

13. Liquid composition characterized in that it is obtained from a 10- to 20-fold dilution of the wet composition according to any one of claims 10 to 12 in an acceptable solvent.

14. Composition according to any one of claims 5 to 9, characterized in that it is formulated in dry form such as in the form of film, fibre or powder.

15. Composition according to claim 14, characterized in that it comprises an amount greater than 10% by weight of calcium alginate relative to the total weight of the composition, preferably between 10% and 100% by weight of calcium alginate relative to the total weight of the composition, and even more preferably between 30% and 90% by weight of calcium alginate relative to the total weight of the composition.

16. Composition according to claim 14 or claim 15, characterized in that it comprises less than 15% by weight of calcium relative to the total weight of the composition, preferably the composition comprises between 2% and 12% by weight of calcium relative to the total weight of the composition, advantageously the composition comprises between 2.5% and 10% by weight of calcium relative to the total weight of the composition.

17. Calcium alginate according to any one of claims 1 to 4, composition and / or device comprising calcium alginate according to any one of claims 1 to 4, for use in the prevention and / or treatment of tissue inflammation, in particular of the skin, mucous membranes, tendons, ligaments, muscles, cartilage and / or bones.

18. Calcium alginate according to any one of claims 1 to 4, composition and / or device comprising calcium alginate according to any one of claims 1 to 4, for use in tissue healing, in particular wounds of the skin and / or mucous membranes, skin lesions, extracutaneous lesions, cartilage lesions, muscle lesions, bone lesions, ligamentous lesions, tendon lesions and / or visceral lesions.

19. Method for the prevention and / or non-therapeutic treatment of the chronic, low-level, non-pathological systemic inflammatory component of aging comprising the administration of calcium alginate according to any one of claims 1 to 4, of the composition and / or device comprising calcium alginate according to any one of claims 1 to 4.

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