Composition useful for treating diseases of motion system

Abstract

To provide a composition useful for treating or preventing diseases of a motion system of horses.SOLUTION: Provided is a composition containing buccal cells to be used for preventing or treating diseases of a motion system of horses.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to compositions useful for the treatment or prevention of disorders of the locomotor system, particularly in horses. [Background technology]

[0002] In France, the total number of horses is about 350,000, including about 28,000 racing horses. 20 to 30% of them have tendon disorders. If we consider only horse racing, this represents about 8,000 cases per year.

[0003] Conventional treatments based on anti-inflammatory drugs offer no real benefit compared to long-term natural healing with six months to a year of rest. Moreover, these treatments are essentially symptomatic and do not heal the tendon.

[0004] It has therefore been proposed to use stem cells, in particular stem cells derived from bone marrow, umbilical cord or adipose tissue, to obtain tendon healing approaching that of the original tendon.

[0005] However, adipose tissue cells appear to be less differentiated than bone marrow cells, and the authenticity of umbilical cord-derived stem cells has not really been established.

[0006] Thus, bone marrow mesenchymal stem cells are currently used as the standard treatment for equine tendonitis (Godwin et al. (2012) Equine Veterinary Journal 44:25-32).

[0007] However, bone marrow sampling is a relatively laborious procedure. Moreover, bone marrow mesenchymal stem cells are essentially validated for autologous use, which means that the cells are harvested from the animal to which they are administered, which increases the cost of the method.

[0008] It would therefore be of interest to have alternative treatments that are easier and cheaper to implement. Summary of the Invention

[0009] The present invention arises from the unexpected discovery by the inventors that administration of a composition based on heterologous gingival fibroblasts to horses suffering from tendonitis or osteochondrosis makes it possible to treat these diseases.

[0010] Advantageously, gingival fibroblasts are easily obtained and prepared, and furthermore, they are useful in terms of xenogeneic use.

[0011] Thus, the present invention relates to a composition comprising buccal cells, in particular gingival fibroblasts, for use in the prevention or treatment of a disease of the locomotor system in an individual, in particular a horse.

[0012] Furthermore, the present invention also relates to a method for preventing or treating a disease of the locomotor system in an individual, particularly a horse, comprising administering to the individual a prophylactically or therapeutically effective amount of a composition of buccal cells, particularly gingival fibroblasts.

[0013] The present invention also relates to the use of a composition comprising buccal cells, in particular gingival fibroblasts, for the preparation of a medicament for treating a disease of the locomotor system in an individual, in particular a horse. [Brief description of the drawings]

[0014] [Figure 1] Figure 1 shows ultrasound pictures of the superficial flexor tendon of four horses with tendinitis of this tendon before (D0) and 3 months after (D0+3 months) injection of a gingival fibroblast composition. The area with the tendon lesion, darker than the surroundings, is surrounded by a white circle. An injection of the composition with a syringe is performed within the lesion (arrow). After 3 months, the tendon has a homogenous appearance, indicating that the lesion has been treated. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] As intended herein, the term "comprising" has the meaning of "including" or "containing," which means that when an object "comprises" one or more elements, other elements besides those mentioned may be included in the object. In contrast, when an object is said to "consist of" one or more elements, the object is limited to the recited elements and cannot include other elements besides those mentioned.

[0016] Motor system disorders As intended herein, diseases of the locomotor system, in particular diseases of horses, refer in particular to osteoarticular diseases or diseases of the musculoskeletal system, in particular diseases of the limbs.

[0017] Diseases of the equine locomotor system are described inter alia in “Maladie des Chevaux” (1994) Institut du cheval, France Agricole Editions.

[0018] Preferably, the disease of the locomotor system according to the invention is a tendon disorder or tendon injury, in particular tendonitis.

[0019] More preferably, the disease of the locomotor system according to the invention is tendinitis of the flexor tendons, especially the superficial or deep flexor tendons of the phalanges, or tendinitis of the extensor tendons, especially the lateral or anterior phalangeal extensor tendons, especially in horses.

[0020] As intended herein, the superficial flexor tendons are also referred to as perforated tendons, and the deep tendons of the phalanges are also referred to as perforating tendons.

[0021] Also preferably, the disease of the locomotor system according to the invention is an arthropathy.

[0022] More preferably, the disease of the locomotor system according to the invention is osteochondrosis or arthrosis, especially in horses.

[0023] As intended herein, arthritis is synonymous with osteoarthritis.

[0024] Preferably, the osteochondrosis or arthropathy is associated with bone nodules, osteochondritis dissecans, or bone cysts, particularly in horses.

[0025] Also preferably, the osteochondrosis or arthropathy according to the invention is in the ball joint, the knee, the calcaneus at proximal level, the calcaneus at distal level, or in the tether of the horse.

[0026] individual Preferably, the individual according to the invention is an animal, in particular a domestic animal, more preferably selected from the group consisting of Equidae, in particular a horse, Camelidae, in particular a camel, dromedary or llama, Canidae, in particular a dog, Felidae, in particular a cat, and an ostrich.

[0027] An individual according to the invention may be male or female.

[0028] Preferably, the individual according to the invention is a horse, in particular a sports horse, more particularly a race horse, a trotting horse, a driving horse, a jumping horse, a dressage horse, a cross-country horse, an eventing horse, an endurance horse or a polo horse.

[0029] The horse according to the invention may be a foal, particularly a yearling, an adult horse or an older horse.

[0030] Preferably, the individual according to the invention is a horse suffering from lameness, in particular lameness in the front or hind legs.

[0031] composition As intended herein, the expression "buccal cells" is equivalent to "oral cavity cells" or "oral mucosa cells."

[0032] Preferably, a buccal cell according to the invention is a cell of the buccal mucosa, tongue, palate, labial mucosa, sublingual mucosa, or gingiva.

[0033] Preferably, the composition according to the invention is derived from cultures of gingival samples.

[0034] Preferably, the composition according to the invention comprises or consists of gingival fibroblasts, which are called gingival fibroblast compositions. The composition according to the invention may in particular be a culture of gingival fibroblasts, possibly a concentrate, or a suspension of gingival fibroblasts.

[0035] Procedures for obtaining, culturing and storing gingival fibroblasts are well known to those skilled in the art and are described in particular in Naveau et al. (2006) J. Periodontol. 77:238-47 and Gogly et al. (2007) Arterioscler. Thromb. Vasc. Biol. 27:1984-90, as well as in the Examples below. In particular, gingival fibroblasts can be obtained by culturing a gingival sample or biopsy, possibly after enzymatic digestion of the sample or biopsy to release the gingival fibroblasts that compose it. Thus, the cells resulting from the culture of a gingival sample or biopsy are essentially gingival fibroblasts.

[0036] In one embodiment of the invention, the gingival fibroblasts according to the invention comprise mesenchymal stem cells. As the skilled artisan will well understand, the mesenchymal stem cells are buccal mesenchymal stem cells, which means those of the oral cavity or oral mucosa, in particular the buccal mucosa, tongue, palate, labial mucosa, sublingual mucosa or gingiva.

[0037] In another embodiment of the invention, the gingival fibroblasts according to the invention are free or essentially free of mesenchymal stem cells.

[0038] Advantageously, gingival fibroblasts can be easily harvested and cultured, and furthermore, gingival fibroblasts have a fast proliferation rate.

[0039] As the skilled person will well understand, the composition according to the invention comprises a prophylactically or therapeutically effective amount of buccal cells, in particular gingival fibroblasts, according to the invention. Preferably, the composition according to the invention comprises 5 to 40 million buccal cells, in particular gingival fibroblasts, according to the invention, more preferably 10 to 30 million buccal cells, in particular gingival fibroblasts, according to the invention, even more preferably 15 to 25 million buccal cells, in particular gingival fibroblasts, according to the invention, and most preferably about 20 million buccal cells, in particular gingival fibroblasts, according to the invention. The quantification of buccal cells, in particular gingival fibroblasts, according to the invention can be carried out by any method known to the skilled person for counting mammalian cells, in particular using a cell counter.

[0040] Preferably, the buccal cells, in particular gingival fibroblasts, according to the invention are xenogeneic, meaning that they are taken from another individual, in particular another horse, than the one in which the composition is to be used. As the skilled person will well understand, the individual from whom the cells are taken and the individual to whom the cells are administered are preferably of the same species.

[0041] Also preferably, the buccal cells, in particular the gingival fibroblasts, according to the invention are autologous, meaning that they are taken from the same individual, in particular the same horse, for which the composition is to be used.

[0042] Preferably, the composition according to the invention is administered near or at the body site to be treated, more preferably at the lesion to be treated. Administration can be carried out by any method known to the skilled artisan suitable for the administration of cells. However, it is preferred that the composition is injected, in particular at the lesion to be treated. Preferably, the composition according to the invention is injected, in horses, in particular by ultrasound guidance, at the site of tendinitis to be treated, more preferably at the tendon lesion, or into or at a joint with osteochondrosis or arthropathy.

[0043] Preferably, the method of preventing or treating an individual according to the present invention comprises the steps of: - obtaining gingival fibroblasts from an individual; - culturing gingival fibroblasts; - administering the gingival fibroblasts or a culture of gingival fibroblasts to the individual or to another individual. Includes.

[0044] The invention will be further illustrated, but not limited to, by the following figures and examples. EXAMPLES

[0045] Example 1: Treatment of tendonitis Preparation of gingival fibroblasts for administration Approximately 50 mg of gingival biopsy was taken from a female horse (Horse 0) and then enzymatically digested to obtain a suspension of gingival fibroblasts. The suspension was counted and then separated into 75 cm 2 3x10 cells were cultured in a flask of 100 ml of complete medium (DMEM 4.5 g / l glucose + Glutamax (Gibco®), 20% fetal calf serum (FCS) (Gibco®) in the presence of antibiotics / antimycotics). 5 Seed gingival fibroblasts and culture in a bronchial tube at 5% CO with regular medium changes. 2 Incubate at 37° C. in ambient air until 90% confluence.

[0046] Gingival fibroblasts are then harvested. 5 150 cm 2 The cells are then seeded in 25 ml of complete medium in a flask of 1000 x g. C. Then, the cells are cultured in 5% CO with regular changes of culture medium until they reach 90% confluence. 2 Incubation is carried out at 37° C. under atmospheric conditions. This forms the first passage.

[0047] Two other passages are made. At the third passage, approximately 20 x 10 6 The gingival fibroblasts are harvested, centrifuged, washed with 10 ml of antibiotic / antimycotic-free DMEM 0% FCS medium, centrifuged, and dissolved in a volume of about 3 ml. The gingival fibroblasts are then drawn into a dosing syringe and kept at 4° C. until dosing.

[0048] Administration of gingival fibroblasts Four horses (Horses 1, 2, 3, and 4) with superficial flexor tendonitis possibly associated with lameness received injections of xenogeneic gingival fibroblasts prepared as described above. The site of tendon injury was confirmed by ultrasound examination (see figure). It is visible as a denser (darker) area compared to its surroundings.

[0049] Approximately 20×10 6 Injection of gingival fibroblasts is performed directly into the lesion (intralesional injection) with ultrasound guidance.

[0050] result Three months after the injection, an ultrasound observation of the treated area is performed (see figure). No obvious tendon pathology remains: the treated area appears homogeneous by ultrasound examination. The movement of the animal is normal.

[0051] Example 2: Treatment of osteochondrosis Gingival fibroblasts prepared as described in Example 1 are injected directly into the ball-and-socket joint of a horse with osteochondrosis of the ball-and-socket joint associated with lameness (Horse 5).

[0052] Approximately one week after injection, resolution of lameness is observed.

Claims

1. A composition comprising buccal cells for use in the prevention or treatment of a disease of the locomotor system in a horse.

2. 2. A composition for use as claimed in claim 1 in the prevention or treatment of tendonitis in horses.

3. 3. A composition for use according to claim 2 in the prevention or treatment of tendinitis of the flexor tendons, in particular the superficial or deep flexor tendons of the phalanges, or tendinitis of the extensor tendons, in particular the lateral or anterior extensor tendons of the phalanges.

4. 2. A composition for use according to claim 1 for use in the prevention or treatment of arthropathy in horses.

5. A composition for use according to claim 4 in the prevention or treatment of osteochondrosis or arthropathy.

6. 6. A composition for use according to any one of claims 1 to 5, wherein the horse has lameness.

7. 7. A composition for use according to any one of claims 1 to 6, comprising cells of the cheek, tongue, palate, labial mucosa, sublingual mucosa or gums.

8. A composition for use according to any one of claims 1 to 7, derived from cultures of gingival samples.

9. A composition for use according to any one of claims 1 to 8, comprising gingival fibroblasts.

10. 10. A composition for use according to any one of claims 1 to 9, comprising between 5 and 40 million buccal cells.

11. 11. A composition for use according to any one of claims 1 to 10, wherein the cells are xenogeneic, meaning that they are taken from another horse than the one in which the composition is to be used.

12. 11. A composition for use according to any one of claims 1 to 10, wherein the cells are autologous, meaning that they are taken from the same horse in which the composition is to be used.

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