Cannabidiol adjunctive therapy for the treatment of degenerative disc disease

Abstract

Methods and compositions relating to the treatment of degenerative disc disease are provided.SOLUTION: In some embodiments, cannabidiol is utilized to enhance the therapeutic and / or regenerative activity of fibroblasts. In other embodiments, cannabidiol is utilized to augment the reparative effects of other regenerative cells, including monocytes, mesenchymal stem cells, and / or hematopoietic stem cells. In some embodiments, cannabidiol is utilized in the culture media of regenerative cells prior to administration of said cells.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] (Reference to Related Application) This application was filed on October 13, 2019, which is incorporated herein by reference in its entirety. Priority is claimed to filed U.S. Provisional Patent Application No. 62 / 914,523. (Technical field) Embodiments of the present disclosure include at least the fields of molecular biology, cell biology, and medicine. [Background technology]

[0002] Low back pain is caused, at least in most patients, by inflammatory events occurring in conjunction with lumbar disc degeneration. The relationship between inflammation and pain has been confirmed by a large number of subjects with radiologically degenerated discs. It has been established in studies showing that localized The presence of inflammation is thought to be a cause of nociception, and is associated with the presence of granulation tissue and chronic, intractable low back pain. It may be associated with disc degeneration, as exemplified by symptoms (1, 2).

[0003] Intervertebral disc degeneration continues to have a significant and growing impact worldwide, but current treatment options Treatment options do not address the underlying cause. Current treatments include bed rest, early treatment, and Nonsteroidal anti-inflammatory drugs and late-stage disc disease where pain has not improved with previous approaches These include resection, arthroplasty (joint replacement), and the injection and fixation of an artificial nucleus pulposus. The approaches mentioned above are not only unpredictable but also tend to be limited to the most advanced clinical conditions. It does nothing to change the disease process itself. Treatment may reduce the incidence of disc degeneration in adjacent discs due to changes in the biomechanical distribution of workload. results in an increase.

[0004] In recent years, both advances in biotechnology and a better understanding of the biochemical makeup and environment of the intervertebral disc have led to With growing interest in the degenerative process, there is potential for the development of novel treatments aimed directly at disc preservation. It has been shown to have a significant effect on matrix synthesis and catabolism within the disc. Certain genes have provided targets for scientists trying to shift the balance between the two. To this end, much attention in the past few years has focused on gene therapy, Efforts by the FDA have yielded promising preclinical results regarding its use in the treatment of disc degeneration ( 3) Unfortunately, none of these approaches are close to clinical implementation at the time of writing. Furthermore, in situations where disc regeneration can only be achieved through gene therapy or other interventional means, Even in these cases, the underlying processes that originally caused the degeneration are addressed to prevent recurrence. It is important to note that it must be

[0005] Currently, there are no widely available biological treatments for disc degeneration. Many different molecules with potential therapeutic benefit are being investigated. Preventing or reversing one or more aspects of these changes in the plate extracellular matrix At least four different classes of molecules may be effective in intervertebral disc repair. These include anticatabolic, mitogenic, chondrogenic morphogens and intracellular regulatory Factors included (4-6).

[0006] Intervertebral disc degeneration is characterized by the loss of proteoglycans, water, and type II collagen in the disc matrix. This includes loss of collagen. In addition, loss of high molecular weight proteoglycans and their quantification have been reported. Other changes that are more difficult to detect include collagen cross-linking, proteoglycan organization, and other changes in the matrix. The qualitative changes in the disc are less clear. This appears to be a change in the differentiated chondrocyte phenotype to a more fibrotic phenotype in these vertebrae. Combined with changes in the disc matrix, the final result is a pathologically related disc and vertebral anatomy. leading to structural changes ( 7 , 8 ).

[0007] Matrix loss is an imbalance between matrix synthesis and degradation, so increasing synthesis Increase the disc matrix by increasing the One approach is to inhibit the degradation of matrices by inhibiting degradative enzymes. The purpose of this is to prevent loss of data.

[0008] Degenerated discs have elevated levels of matrix metalloproteinases (MMPs). Within the matrix, MMP activity is usually inhibited by tissue inhibitors of MMPs (TIMPs). (9-11) Wallach et al. To determine whether 1 can increase the accumulation of matrix proteoglycans, The researchers confirmed that TIMP-1 expression in intervertebral disc cells was indeed It was found that this increased the accumulation of proteoglycans and also increased the "measured synthesis rate" of proteoglycans ( 11) Inflammatory processes are well known to be associated with degenerative disc disease. In patients with leukin-1 (12-16); TNF-α (17-20); and interleukin-1 (18-21); Several studies have shown elevated levels of proinflammatory cytokines, such as leukin-6 (21). Inflammatory cytokines activate matrix metalloproteinases, which destroy the extracellular matrix. (22,23) stimulation (17,24), and apoptosis of nucleus pulposus cells accelerate degeneration by several mechanisms, including stimulation of nitric oxide production (25, 26). The cells in degenerated discs not only produce inflammatory cytokines but also other inflammatory cytokines. They are known to be pro-inflammatory cytokines in that they produce greater amounts of inflammatory cytokines in response to cytokines. It is interesting that the hypersensitivity to steroids is due to a self-amplifying feedback loop ( 27) suggest the presence of inflammatory cytokines and extracellular matrix proteins. Effects on production are also thought to contribute in part to degeneration by reducing regenerative activity. (28) For example, in one study, researchers developed in vitro nucleus pulposus tissue. The inflammatory cytokine TNF-α (up to 50 ng / mL) was administered for 48 hours. The histological appearance, proteoglycan and collagen content, and proteoglycan Tissues were evaluated for lysine and collagen synthesis. The effect of TNF-α on NP cell gene expression was measured using Degradation was assessed by immunoblot analysis. At doses of 1 to 5 ng / mL, TNF-α was Induced multiple cellular responses, including the expression of both aggrecan and type II collagen genes. Decreased expression; decreased accumulation and overall synthesis of aggrecan and collagen; MMP-1, MM Increased expression of P-3, MMP-13, ADAM-TS4, and ADAM-TS5;ADA Induction of M-TS-dependent proteoglycan degradation. Within 48 hours, these cellular responses These results suggest that the NP tissue contained only 25% of the original proteoglycan content of the marrow. This strongly supports the ability of TNF-α to potently inhibit nuclear cell function and block regenerative capacity (2 9).

[0009] The importance of inflammatory cytokines in disc degeneration as a direct cause of pathology This is highlighted by studies showing its effect on direct pain induction as well. reported that calcium ions in dorsal root ganglion (DRG) neurons innervating the rat intervertebral disc TNF-α on the molecular model of pain: CGRP expression To investigate the effects of the inhibitor, etanercept, retrograde neuronal tracing and immunohistochemistry were used. The idea was that degeneration of the lumbar intervertebral disc was the cause of lower back pain. TNF-α in DRG neurons may be a major contributing factor to disc pain. We evaluated the effect of TNF-α inhibition on CGRP expression in the L4 / 5 disc innervating the spinal cord. The gold fluoride-labeled neurons were distributed throughout the L1–L6 DRG in all groups. The proportion of CGRP-immunoreactive neurons among the luoroGold-labeled neurons was The puncture + saline group had a 21% ± 4% success rate, the puncture + saline group had a 32% ± 7% success rate, and the puncture + etanerce group had a 32% ± 7% success rate. The percentage of CGRP-immunoreactive neurons was 23% ± 4% in the puncture + saline group. The saline group had significantly greater pulmonary embolism than the sham control and paracentesis + etanercept groups. In this model, CGRP is upregulated in DRG neurons innervating the injured disc. Direct application of etanercept into the disc immediately after disc puncture has been shown to We concluded that CGRP expression in DRG neurons innervating the disc is suppressed ( 30) Other studies have shown that in addition to TNF-α, other inflammatory cytokines such as IL-1 (3 1) has been reported to be associated with direct pain induction (32-43).

[0010] Regenerative cells, including mesenchymal stem cells, fibroblasts, hematopoietic stem cells, and even immune cells, are all The present disclosure relates to an intervertebral disc. Reduces inflammation and enhances the efficacy of regenerative cells and / or growth factors when administered to children with degenerative diseases The present invention provides the use of cannabidiol as a means of stimulating Summary of the Invention

[0011] The present disclosure is directed to methods and compositions relating to the treatment of degenerative disc disease (degenerative disc disease). Certain embodiments are directed to those who have, are suspected of having, or are at risk of having degenerative disc disease. In some embodiments, the administration of one or more compositions to an individual at risk of developing a disease is The composition administered is any form of cannabidiol for therapeutic intervention against degenerative disc disease. and in some embodiments, at least one anti-degenerative disc agent (CBD) for degenerative disc disease. This includes combining one of these with a therapeutic or preventative intervention. Therapeutic interventions therefor may include, by way of example, a therapeutically effective amount of fibroblasts.

[0012] Certain embodiments relate to the administration of cannabidiol to an individual. Cannabidiol can be administered systemically and / or locally to the body (e.g., to the intervertebral disc). The administration may induce apoptosis of cells in the nucleus pulposus, including endogenous and / or exogenous cells in the nucleus pulposus. The drug can be administered to an individual at a dose and frequency sufficient to reduce the amount of extracellular fluid in the nucleus pulposus. Inducible cells include, for example, fibroblasts, fibroblasts differentiated into notochordal cells, and / or chondrocytes. Therapeutic interventions for degenerative disc disease encompassed herein include fibroblasts differentiated into fibroblasts. Endogenous cells within the nucleus pulposus may include chondrocytes, notochordal cells, notochordal precursor cells, and prechondrocytes. Cannabidiol may be present in the tissues necessary for therapeutic efficacy. Any dosage required (50 mg to 500 mg, or a dose derivable therein) Cannabidiol may be administered to an individual daily at a dose of 100mg / day or 200mg / day (including any range). The administration may be at any frequency required for therapeutic effect, including weekly, biweekly, monthly, bimonthly, etc. Cannabidiol may include (-)-cannabidiol, or (-)-cannabidiol. Cannabidiol may comprise a penetration enhancing gel. In some embodiments, the cannabidiol may be used in a hiatal or steroidal antihistamine formulation. and / or one or more extracellular matrix molecules containing long-chain hyaluronic acid molecules. It may be administered together with a localizing composition that may be used.

[0013] Fibroblasts encompassed herein include fibroblasts that may or may not be administered to an individual. The cells may or may not be engineered. In some embodiments, fibroblasts are derived from the notochord. In some embodiments, the cells differentiate into non-fibroblastic cells, such as fibroblasts or chondrocytes. The cells may, at least in certain cases, be treated with cannabidiol, which increases immunomodulatory activity. The cells are contacted with hCG, oxytocin, or a combination thereof. 1 nM to 1 μM, or 10 nM to 100 nM per million, or any inducible concentration therein Oxytocin can be contacted with or administered to cells at any concentration within the range of 1 nM to 1 0 μM, or 100 nM to 1 μM, or any inducible concentration range therein, Fibroblasts can be contacted or administered from adipose tissue, skin tissue, bone marrow, peripheral blood, Wharton's Wharton's Jelly, placenta, fetal membranes, mobilized peripheral blood, and The tissue may be from one or more tissues selected from the group consisting of: The encapsulated fibroblasts can proliferate at a rate of 14 to 21 hours per cell proliferation. fibroblasts contained in the fibroblasts ranged from 0.1 pg to 77 pg per million fibroblast cultures. g of interleukin-1 can be secreted at 75% confluence on the surface. The fibroblast content ranges from 1 pg to 500 pg per million fibroblast cultures. FGF-1 may be secreted at 75% confluence on the surface. Fibroblasts reduce the ability of responding T cells to proliferate in a mixed lymphocyte reaction by, for example, about 20% can be substantially reduced beyond

[0014] In some embodiments, the fibroblasts are contacted with cannabidiol to induce fibroblast proliferation. Increased regenerative activity. Contact of fibroblasts with cannabidiol increases at least some In this case, it may enhance the production of IGF-1 and / or EGF-1. The compound may comprise or consist of (-)-cannabidiol. It is also possible.

[0015] The foregoing describes the features and technical advantages of the present disclosure in order that the following detailed description may be better understood. have been outlined quite broadly. Additional features and advantages form the subject of the claims herein. The concepts and specific embodiments disclosed are intended to carry out the same objectives of the present design. It will be readily apparent to those skilled in the art that the present invention may be readily utilized as a basis for modifying or designing other structures for the purpose of It should be understood that such equivalent constructions are also within the scope of the appended claims. It should be understood by those skilled in the art that this does not depart from the spirit and scope of what is described. Further objects and advantages are set forth in the novel features believed characteristic of the design disclosed herein. The following, both as to organization and method of operation, when considered in conjunction with the accompanying drawings, It will be better understood from the description below. However, each figure is for purposes of illustration and explanation. It should be expressly understood that these are provided for purposes of illustration only and are not intended as a definition of the limits of the present disclosure. do. [Brief explanation of the drawings]

[0016] For a more complete understanding of the present disclosure, reference is now made to the following descriptions taken in conjunction with the accompanying drawings, in which:

[0017] [Figure 1] Figure 1 shows the stimulation of IGF-1 from fibroblasts by CBD (cannabidiol) at various concentrations. In a grouping of three bars, the middle bar at each concentration represents 100,000 fibroblasts co-cultured with CBD at the indicated concentration. The right bar at each concentration represents 200,000 fibroblasts co-cultured with CBD at the indicated concentration. The left bar at each concentration represents 0 fibroblasts co-cultured with CBD at the indicated concentration.

[0018] [Figure 2] Figure 2 shows the stimulation of EGF-1 from fibroblasts by CBD at various concentrations. In a grouping of three bars, the middle bar at each concentration represents 100,000 fibroblasts co-cultured with CBD at the indicated concentration. The right bar at each concentration represents 200,000 fibroblasts co-cultured with CBD at the indicated concentration. The left bar at each concentration represents 0 fibroblasts co-cultured with CBD at the indicated concentration. DETAILED DESCRIPTION OF THE INVENTION

[0019] In accordance with long-standing patent law convention, the terms "a" and "an" refer to the entire disclosure of this specification, including the claims. As used herein, "one or more" means "one or more" and some embodiments of the disclosure may Consisting of or consisting essentially of one or more of the elements, method steps, and / or methods set forth Any of the methods or compositions described herein can be used in combination with any other method or composition described herein. It can be implemented in terms of methods or compositions, and different embodiments can be combined. It is intended that:

[0020] Throughout this specification, unless the context requires otherwise, the terms "comprises" and "comprises" and "comprising" refers to a listed step or element or group of steps or elements. It does not imply the inclusion of other steps or elements or groups of steps or elements. It will be understood that "consisting of" does not mean that "consisting of" means including, but not limited to, anything that follows Therefore, the term "consisting of" does not mean that the listed elements are required. "Consisting essentially of" means that the element is essential or essential and that no other element is present. is meant to include any elements listed after the phrase, and is a disclosure of the listed elements limited to other elements that do not interfere with or contribute to the activity or action specified in Thus, the term "consisting essentially of" does not mean that the listed elements are necessary or mandatory. indicates that other elements are not optional and may affect the activity or operation of the listed elements. It may or may not be present depending on whether

[0021] Throughout this specification, the terms "one embodiment," "embodiment," "particular embodiment," "related embodiment," "related" and "related" are used interchangeably. "some embodiments," "particular embodiments," "additional embodiments," or "further embodiments" Reference to a particular feature, configuration, or combination thereof described in connection with an embodiment is not intended to be limiting. or features are included in at least one embodiment of the present invention. The appearances of the foregoing phrases in various places throughout this specification do not necessarily all refer to the same embodiment. Furthermore, any particular feature, structure, or characteristic may be incorporated into one or more embodiments. can be combined in any suitable manner.

[0022] I. Cannabidiol

[0023] As used herein, "cannabidiol" or "CBD" refers to cannabidiol. cannabidiol; cannabidiol prodrug; pharmaceutically acceptable derivatives of cannabidiol ( Pharmacology of cannabidiol, cannabidiol prodrugs, and cannabidiol derivatives CBD refers to the compound 2-[3-methyl-6-(1-methylethyl)amino]-3-(2-methyl-1-methylethyl)amino]-3-(2 ... [thenyl]-2-cyclohexen-1-yl-5-pentyl-1,3-benzenediol and pharmaceutically acceptable salts, solvates, metabolites (e.g., skin metabolites) thereof, and metabolic precursors. The synthesis of CBD is described, for example, in Petilka et al., Helv.C. him. Acta, 52:1102 (1969), and Mechoulam et al., J.A. Chem. Soc., 87:3273 (1965), and these are (Incorporated herein by reference).

[0024] Certain embodiments of the present disclosure relate to the use of cannabidiol (CBD) for regenerative cell activity or is included as an adjuvant for regenerative cell therapy. In certain embodiments, cannabidiol is used in combination with other cannabidiol derivatives for the treatment of intervertebral disc degeneration or degenerative disc disease. Cannabidiol is administered systemically and / or locally. In some embodiments, the CBD may be administered in vitro or ex vivo prior to implantation. In certain embodiments, CBD is used to stimulate regenerative cells in vivo. administered intravenously, which may increase the suitability of the microenvironment to maintain regenerative cell activity and It may inhibit disease and / or directly stimulate regenerative cells to perform healing functions.

[0025] The CBD may or may not be synthetic CBD. The CBD may be purified CBD. The CBD may be derived from a plant. The CBD may be administered transdermally to the subject's upper arm and shoulder. In some embodiments, CBD is administered transdermally to the thigh or back of a subject. CBD can be administered directly to the disc, whether or not it shows signs of degeneration. In certain cases, topical administration of effective amounts of cannabidiol (CBD) can compared to oral CBD, the intensity of at least one adverse event or side effect was At least one adverse event or side effect may be gastrointestinal (GI) adverse events. At least one adverse event or side effect may affect liver function. In some embodiments, at least one adverse event may be a side effect. In some embodiments, the frequency and intensity of somnolence is an adverse event of transdermal administration. is reduced as

[0026] In some embodiments, the CBD comprises or consists of (-)-CBD. An effective amount (e.g., a therapeutically effective amount) of CBD is about 50 mg to about 500 mg per day. In some embodiments, an effective amount of CBD that can be administered to an individual is about 50 mg. In some embodiments, the individual is administered The effective amount of CBD that can be administered starts at about 50 mg daily and titrates up to about 250 mg daily. In some embodiments, an effective amount of CBD that can be administered to an individual begins at 250 mg / day. In some embodiments, an effective amount of CBD that can be administered to an individual is 500 ml daily. In some embodiments, a daily dose of 500 mg is recommended for those over 35 kg. CBD is administered in a variety of therapeutic doses, including once-daily or twice-daily administration. It may be administered as frequently as is useful for effect. In some embodiments, an effective amount of CBD may be administered in divided doses. The recommended daily dose may be 390 mg.

[0027] As used herein, the term "transdermally" refers to the administration of CBD through the skin. This means contacting the CBD with the skin of the patient or subject under conditions effective to penetrate the skin. In certain embodiments, CBD is formulated as a gel or oil. In an embodiment, the CBD is formulated as a penetration-enhancing gel. The gel contains 1% (wt / wt In some embodiments, the gel may contain 4 to 7.5% (wt / wt) CBD. In some embodiments, the gel contains 7.5% (wt / wt) CBD. In some embodiments, the transdermal preparation contains CBD (wt / wt). The CBD can be delivered by a bandage, pad, or patch. CBD can be in gel form and administered transdermally in one or two daily doses. Pharmaceutically produced as a clear permeation-enhancing gel designed to provide controlled drug delivery. CBD gel can be used in a range of 1% (wt / wt) CBD to 7.5% (wt / wt) CBD. , or any range derivable therein. CBD gels can be, for example, 4.2% (wt The CBD gel may have 7.5% (wt / wt) CBD or 7.5% (wt / wt) CBD. , by the patient or caregiver, on the patient's upper arms and shoulders, back, thighs, or any combination thereof. CBD gels can be applied topically to various skin conditions. and other conventional excipients such as suspending and dispersing agents.

[0028] CBD gel contains solubilizers, penetration enhancers, solubilizers, antioxidants, thickeners, and The CBD gel composition may contain, for example, about 0.1% of the composition. cannabidiol present in an amount of about 15% to about 9% (wt / wt) of the composition; A lower alcohol having 1 to 6 carbon atoms present in an amount of 5% (wt / wt); composition a first penetration enhancer present in an amount of about 0.1% to about 20% (wt / wt) of The amount of water may be sufficient to make a total of 100% (wt / wt). Other formulations of CBD gel are available internationally. Publication No. WO2010 / 127033, the entire contents of which are incorporated herein by reference. It will be incorporated into the specification.

[0029] In some embodiments, a cannabinoid (e.g., cannabidiol or cannabinoid) (a prodrug of Bisiol) to achieve therapeutically effective plasma concentrations in individuals However, cannabinoids, including those containing cannabinol, are delivered systemically. Oral dosage forms must overcome several obstacles to achieve therapeutically effective systemic concentrations. First, cannabinoids are generally highly lipophilic. The increased water solubility thereby limits the amount of cannabinoid available for absorption in the gastrointestinal tract. Second, cannabidiol, like other cannabinoids, is absorbed through the human gastrointestinal tract. Once absorbed, they undergo substantial first-pass metabolism. Finally, patients may wish to avoid taking oral medications. oral dosage forms do not remain in the gastrointestinal tract long enough to release the full dose and achieve therapeutic concentrations Therefore, if a patient experiences nausea or vomiting, the oral bioavailability of any product may be reduced. Therefore, taking the above into consideration, cannabinoids (pain, degenerative disc disease) and (ii) administering to a patient a cannabinoid-containing drug for the treatment of one or more medical conditions responsive to cannabinoids, including nausea, vomiting, or appetite stimulation. a therapeutically effective amount of a cannabinoid, such as a cannabinol or cannabiol prodrug, The drug is administered systemically to mammals of interest by a route of administration that does not depend on absorption from the mammal's gastrointestinal tract. It is desirable to deliver it to

[0030] One parenteral route of administration for systemic delivery of cannabidiol is transdermal administration. Therefore, the epidermis and dermis of many mammals, such as humans and guinea pigs, are active substances that penetrate the stratum corneum. It contains enzymes that can metabolize active drugs. The metabolic processes that occur produce pharmaceutically effective amounts of cannabinoids, such as cannabinol, in cannabinoids can be used to deliver them to the systemic circulation of mammals in need thereof. prodrugs (e.g., cannabidiol prodrugs), and cannabinoid prodrugs a composition comprising the compound, which can be transdermally administered to a mammal, such as a human, such that the compound is absorbed into the skin; Metabolites resulting from metabolism in the skin may be useful for the treatment of medical conditions that respond to cannabinoids. Compositions containing prodrugs of systemically available cannabinoids for the treatment of cancer are encompassed herein. Unfortunately, due to its highly lipophilic nature, cannabidiol is not readily available for consumption by humans. It is poorly absorbed through membranes such as mammalian skin. The administration of cinnabidiol to a mammal in need thereof over an adequate surface area within a reasonable time frame is However, the success of transdermal administration has been substantially limited.

[0031] In some embodiments, cannabidiol is administered to an individual locally, e.g., by administering it to the individual. Cannabidiol is administered to the intervertebral disc of the body at dosages and frequencies encompassed herein. Cannabis sativa may be administered topically to an individual at any dosage and frequency, including topical administration. Visiol may be administered in conjunction with one or more other compositions, including cell therapies encompassed herein. In some embodiments, cannabidiol is administered to the intervertebral disc in an individual. reduces apoptosis of the cells that comprise the intervertebral disc of an individual. (including notochordal cells, notochordal precursor cells, chondrocytes, and / or chondrocyte precursor cells) and / or may be exogenous cells, including any cell therapy administered to an individual.

[0032] In some embodiments, the administration of cannabidiol to an individual is in combination with other drugs that inhibit inflammation. In one embodiment, the protein, gene, and / or mRNA Administration of interleukin-1 (IL-1) receptor antagonists, including A, reduces some of the inflammation. This was done to reduce some of the symptoms, which unexpectedly synergized with the administration of cannabidiol. .

[0033] In some embodiments, a cancer with one or more therapeutic interventions for degenerative disc disease is The nabidiol or cannabidiol is administered in conjunction with a localized composition. , one or more extracellular matrix molecules containing hyaluronic acid and / or long-chain hyaluronic acid could be.

[0034] In some embodiments, cannabidiol increases regenerative activity in cells. contacted with, cultured with, or administered to cells such as fibroblasts that can Cannabidiol (including (-)-cannabidiol or (-)-cannabidio fibroblasts that have been contacted with, cultured, or administered with cannabidiol (containing cannabidiol consisting of cannabidiol) The cells may increase the production of IGF-1 and / or EGF-1. Cannabidiol containing or consisting of (-)-cannabidiol Fibroblasts contacted, cultured, or administered with a fibroblast-containing antibody (including a fibroblast-containing antibody) are included in the methods encompassed herein. Cannabidiol (including (-)-cannabidiol or (-) -Cells that have been in contact with, cultured with, or administered with cannabidiol (including cannabidiol). Fibroblasts can be administered locally to individuals, including individuals with or at risk of having degenerative disc disease. The compositions can be administered intradiscally (including intradiscally) and / or systemically.

[0035] II. Therapeutic interventions for degenerative disc disease

[0036] Certain embodiments of the present disclosure include fibroblasts, mesenchymal stem cells, hematopoietic stem cells, and tissue healing cells. Degenerative disc disease may involve a variety of cell therapies, including regenerative cells, including other cells with the ability to In certain embodiments, the present invention relates to the use of one or more therapeutic interventions for diseases. The fibroblasts may be of any autologous, allogeneic, or Fibroblasts may be derived from adipose tissue, skin tissue, bone marrow, peripheral blood, or other sources. Wharton's Jelly, placenta, fetal membranes, mobilized endometrium The tissue may be from a tissue selected from the group consisting of peripheral blood, and combinations thereof.

[0037] In certain embodiments, the method comprises the steps of: The fibroblasts containing the fibroblasts may be administered to an individual, such as an individual in need of treatment for degenerative disc disease, for example, by administering them to an intervertebral disc. Fibroblasts can be administered as a hydrogel-containing formulation. The hydrogel can be blended with, for example, batroxobin (BTX) as a gelling agent. The tissue may be composed of platelet-rich plasma (PRP) and hyaluronic acid (HA). It can be encapsulated in a PRP / HA / BTX hydrogel and contains growth medium and TGF-β In one embodiment, the cells can be cultured for up to 21 days in medium with or without 1. The hydrogel was then allowed to stand for 15 minutes, ensuring that the fibroblasts maintained high cell viability and proliferation. Jellify at 20° C. for 3 minutes at 37° C. In one embodiment, fibroblasts are used to treat degenerative disc disease. In such an embodiment, the fiber is used for intradiscal injection in a patient with The blast cells are cultured in a manner that enhances GAG production, which includes the administration of cytokines (e.g., TG This can be achieved by culturing with fibroblast-specific β- Methodologies for the expansion of mesenchymal stem cells are provided below, which are incorporated by reference. (44).

[0038] Certain embodiments involve activation of fibroblasts prior to therapeutic use and / or the administration of fibroblasts. The cells in the formulation are grown in culture alone. When grown in layers, they may exhibit typical fibroblast morphology, specifically, cells with elongated extensions. The cells may have an elongated spindle or fusiform appearance with a leading edge of the cytoplasm They may appear as larger, flattened astrocytes. A mixture of these morphologies may also be observed. The cells express fibroblast-specific markers, CD90 (Thy-1), and the 35 kDa normal cell surface glycoproteins and extracellular matrix proteins, including collagen The fibroblast dosage formulation may express proteins characteristic of fibroblasts. A suspension of autologous fibroblasts grown from each individual's own skin biopsy using a culture procedure In certain embodiments, the fibroblasts may be an autologous cell therapy product composed of It may also be used to generate other cell types for tissue repair or regeneration.

[0039] The fibroblasts encompassed herein may be derived from the skin ( by growth from a biopsy of either autologous (in the case of autologous preparations) or healthy donor skin (in the case of allogeneic preparations). Skin-derived fibroblasts are disclosed as an example and are encompassed by the present disclosure. Fibroblasts can be derived from any source or tissue. In some embodiments, fibroblasts In another embodiment, the cells are derived from a young donor. and transfected with genes that allow overcoming the Hayflick limit. Following induction of cell proliferation in culture using cell culture techniques, the technique involves the growth of cells in a liquid medium. Expansion in media such as RPMI 1640, DMEM, OPTI-MEM, or AIM-V Standard techniques for tissue culture are well known in the art and include the use of In some embodiments, dissociation involves the need to transfect the cells to allow for cell passaging. Skin tissue (dermis and epidermis layers) was collected from the ear of a subject. A biopsy can be taken from the posterior region. One embodiment disclosed for purposes of illustrating a non-limiting example. In this study, the starting material was three 3 mm punch skin biopsies collected using standard sterile practice. Biopsies are collected by a skilled artisan and dissolved in sterile phosphate buffered saline (PBS). Biopsies are shipped at 2-8°C. Refrigerated shippers are required by the manufacturer. Return to facility. In one embodiment, after arriving at the manufacturing facility, the biopsy is inspected and accepted. At the start of the process, the biopsy tissue is then subjected to enzymatic digestion. After washing, Liberase digestion enzyme solution was added without mincing, and the biopsy tissue was then washed for 3 min. Incubate at 7.0 + / - 2.0°C for 1 hour. The time for biopsy tissue digestion is determined by the culture medium. It is a critical process parameter that can affect the viability and growth rate of cells in culture. Liberase is a registered trademark of Lonza Walkersville, Inc. (Walkersville, lle, Md) and Roche Diagnostics Corp. (In Collagenase / neutral protease was obtained unformulated from San Diego, Calif. (Diamondpolis, Ind.). Alternatively, other commercially available collagenases (e.g., Serva Co. llagenase NB6 (Helidelburg, Germany) was used. After digestion, the starting growth medium (IMDM, GA, 10% fetal bovine serum (FBS)) is added. The enzyme was neutralized by centrifugation, and the cells were pelleted and added to 5.0 mL of starting growth medium. Alternatively, no centrifugation is performed and complete inactivation of the enzyme is achieved by resuspending the cells in the starting growth medium alone. For cell growth and proliferation initiation, the cell suspension was cultured in T-175 cell culture medium. Add growth initiation medium before seeding the culture flask. 75, T-150, T-185 or T-225 flasks can be used. Incubate at 37°C + / - 2.0°C, 5.0% + / - 1.0% CO2, and fresh con- Feed the paper with Pret Glow media approximately every 3-5 days. Feeding was performed by removing half of the complete growth medium and replacing it with the same volume of fresh medium. Alternatively, a complete feed can be performed. Approximately 30 days before passaging, the cells Confluence should not remain in the T-175 flask. Cell confluence is monitored throughout the process to ensure cell density is maintained. If the cell volume is greater than 40% in a T-175 flask, remove the spent medium and wash the cells. Adherent cells in flasks were released into solution by treatment with trypsin-EDTA. The cells are then trypsinized and seeded into a T-500 flask. Continue cell expansion. Alternatively, use one or two T-300 flasks, one layer cell stack, or stack (1CS), one layer cell factory (1CF) or two layer cell stack (2CS ) can be used in place of the T-500 flask.

[0040] Morphology is assessed before each passage and harvest to monitor culture purity throughout the process. Morphology compares the observed sample to a visual standard for morphological examination of cell cultures. When grown in cultured monolayers, the cells exhibit typical fibroblastic properties. Cells exhibit an elongated spindle or fusiform appearance with elongated extensions or or may appear as larger, flattened astrocytes that may have a leading edge of cytoplasm. A mixture of fibroblasts with similar shapes can also be observed. The presence of keratinocytes in the cell cultures can also be assessed. Keratinocytes appear round and irregularly shaped, and at higher confluence they They appear organized into bluestone formations. At lower confluence, keratinocytes Small colonies were observed. The cells were incubated at 37°C. 10-layer cells were cultured every 3-5 days in T-500 flasks at 5.0+ / 1.0% CO2. Cells were passaged every 5–7 days in stacks (10CS). Cells were kept in T-5 for at least 10 days before passage. 00 flask. Quality control for the safety of bulk drug substance (cells) QC) Release testing includes sterility testing and endotoxin testing. T-500 flask When the cell confluence in the medium is >95%, the cells are passaged into 10CS culture vessels. Alternatively, two 5-layer cell stacks (5CS) or 10-layer cell factories (10CF) can be used. It can be used instead of 10CS. Passage through 10CS removes spent medium and Cells are washed and treated with trypsin-EDTA to release adherent cells in the flask into solution. The cells are then transferred to 10CS. Additional complete growth medium is added. Neutralize the trypsin by adding 100 ml of PBS and transfer the cells from the T-500 flask to 2 ml of PBS containing fresh complete growth medium. Pipette the contents of the 2L bottle into a 10CS bottle and seed all layers. The cells are then incubated at 37°C + / - 2.0°C with 5.0% + / - 1.0% CO2. Feed with fresh complete grow media every 5-7 days. Cells should be kept at 4°C for 20 minutes before passage. In one embodiment, the passaged dermal fibroblasts should not remain in 10CS for more than one day. The method involves incubating proliferated fibroblasts in a protein-free medium for a certain period of time. and rendering the culture medium substantially free of immunogenic proteins present in the culture medium by Cells are harvested when cell confluence in the primary harvest 10CS is 95% or greater. The spent medium was removed, the cells were washed, and the adherent cells were treated with trypsin-EDTA to bring them into solution. The cells are released and the trypsin is neutralized by adding additional complete growth medium. The cells were collected by centrifugation, resuspended, and subjected to in-process QC testing to determine total viable cells. Cell numbers and cell viability are determined.

[0041] In some embodiments, after receiving cell count results from the primary 10CS harvest, If more cells are required, further passage into multiple cell stacks (up to four 10CS) For further passages, cells from the primary harvest are cultured in fresh complete growth medium. Add the resuspended cells to a 2 L bottle of medium containing 3 mL of PBS. Add the resuspended cells to multiple cell stacks and Incubate at 7°C + / - 2.0°C and 5.0% + / - 1.0% CO2. Once cells are confluent, harvest. Feed the cell stack as above, except that it must be at least 80% full before harvesting. The harvesting procedure is the same as that described above for the primary harvest. Mycoplasma samples from the culture medium and spent culture medium were collected and cell counts and viability were determined as described above for the primary culture medium. This method is performed as described for the collection of immunogenic proteins from animal-derived reagents. By avoiding the introduction of proteins, immunogenic proteins are reduced or eliminated. To reduce residual cell mass, cells were cryopreserved in protein-free freezing medium and then It is thawed, washed, and then pre-treated for final injection to further reduce residues. If additional drug substance is needed after harvesting and cryopreservation of cells from the passage is complete, Aliquots of frozen drug substance from cryovials are thawed and seeded into 5CS or 10CS culture vessels. Alternatively, instead of 5CS or 10CS, a 4-layer cell factory (4 CF), two 4CF, or two 5CS can be used. The vial was thawed, washed, and added to a 2 L medium bottle containing fresh complete growth medium. Culture, harvest, and cryopreserve as described above. The cell suspension should be confluent for 80 minutes before harvesting. It must be % or more.

[0042] Upon completion of culture growth, cells were harvested, washed, and then diluted to 2.2 x 10 7 cells / mL or 100,000 cells with more targets ~2.7 x 10 7 cells / mL or more Alternatively, the target can be adjusted within the formulation range to accommodate different dosages. The drug substance is Iscove's Modified Dulbecco's o's Medium (IMDM) and Profreeze-CDM™ (Lonz a, Walkerville, Md) + 7.5% dimethyl sulfoxide (DMSO) or a population of viable autologous human fibroblasts suspended in a cryopreservation medium comprising: Alternatively, a lower DMSO concentration can be used instead of 7.5%, or CryoStor ( Trademark) CS5 or CryoStor(trademark) CS10 (BioLife Solutio ns, Bothell, WA) was used instead of IMDM / Profreeze / DMSO. In addition to cell count and viability, purity / identity of drug substance may be performed and the suspension may be It must be confirmed to contain 98% or more fibroblasts. Common cellular contaminants Purity / identity assays measure the percent purity of the fibroblast population. To quantify the cell density, CD90 and CD104 (respectively, fibroblast and keratinocyte) were expressed. A fluorescently tagged antibody against CD90 (a cell surface marker for T cells) is used. hy-1) is a 35 kDa cell surface glycoprotein. The antibody has been shown to be highly specific for human fibroblasts. The integrin β4 chain is a 205 kDa transmembrane glycoprotein and interacts with the integrin α6 chain (CD49 f) to form the α6 / β4 complex. This complex is a molecular matrix of keratinocyte cells. It has been shown that ATP acts as a marker for ATP synthesis (Adams and Watt 1991).

[0043] Antibodies against the CD104 protein bind to 100% of human keratinocyte cells Cell count and viability were determined using Viacount Dye Reagent. t) and analyze the samples using the Guava PCA system. This reagent contains two dyes: one that stains all nucleated cells and one that stains all nucleated cells. It consists of membrane-permeable dyes that stain only damaged or dead cells, and membrane-impermeable dyes that stain only damaged or dead cells. The use of this dye combination allows the Guava PCA system to detect the presence of cells in the sample. To estimate the total number of cells and to determine which cells are viable, apoptotic, or This method allows for the determination of the purity / activity of autologous cultured fibroblasts. Custom developed specifically for use in determining identity.

[0044] Alternatively, use a T-175 flask (or its equivalent) or a T-500 flask (or its equivalent). or its alternatives) from spinners containing microcarriers as cell growth surfaces. Cells can be passaged onto microcarriers. They are small bead-like structures that serve as growth surfaces for ridge-dependent cells. This device is designed for optimal cell yields. Add 50 mL to 300 mL of complete growth medium to a 1 L sterile disposable spinner flask. Add sterilized microcarriers to the spinner flask. Place the culture medium in a 0.0 + / - 0.0% CO2 incubator for a short period (1-24 hours). or place on a starting plate at slow rotation (15-30 RRM) to allow cells to settle. After the attachment period is over, increase the rotation speed of the spin plate (30-120R PM). Cells should be resuspended every 1-5 days or when the medium changes color and is considered used. The cells are then sampled from the microcarriers and fed with fresh complete culture medium. The cells are collected at regular intervals by isolating and performing cell count and viability analysis. The resulting cell concentration is used to determine when to scale up the culture. Once enough cells have been produced, they are washed with PBS and microcentrifuged with trypsin-EDTA. The cells were harvested from the carriers and placed in a larger volume of microcarriers and a larger volume of complete growth medium (30 Re-inoculate the spinner flask with the appropriate amount of PBS (0 mL-2 L). Alternatively, use an existing microfluidic Add additional microcarriers and complete growth medium directly to the spinner flask containing the carrier culture. Cells can be added directly from beads without trypsinization and reseeding. Alternatively, the first T-175 or T-500 flask can be moved to the If enough cells are produced from the microcarriers, the cells can be directly transferred to a scaled-up amount of microcarriers. After the adhesion period, increase the speed of the spin plate (30-1 20 RPM). Every 1 to 5 days, or when the medium changes color and is used up, replace with new medium. Supply cells with new Complete Growth Media. Once the required number of cells for infection has been reached, wash with PBS and lyse the cells with trypsin-EDTA. The microcarriers used in the disposable spinner flasks are BioNOC II (registered trademark) (Cesco Bioengineering, Bellco Bio technology, Vineland, NJ) and FibraCel® (New Brunswick Scientific, Edison, NJ) etc. Polyblend of Cultispher-G (Percell Biolytica, A gelatin such as Cytopore® (GE Healthcare, Sweden); Healthcare, Piscataway, NJ) or 2 D MicroHex® (Nunc, Weisbaden, Germany) , Cytodex® (GE Healthcare, Piscataway, NJ), or Hy-Q Sphere® (Thermo Scientific Coated / uncoated (e.g., Fictific Hyclone, Logan, Utah) It may be made from polyethylene terephthalate.

[0045] In another embodiment, the FibraStage™ is used in place of the spinner flask device. Trademark)(New Brunswick Scientific, Edison, NJ ) or BelloCell® (Cesco Bioengineering ,Distributed by Bellco Biotechnology,Vin Using an automated bellows system such as BioNOC II (Registry: on polyblend 2D microcarriers such as FibraCel® and FibraCel® It is possible to process cells with T-175 (or alternative) or T-500 (or The cells in the flasks (or alternatives) were placed in a microcarrier containing an appropriate amount of complete growth medium. The cells are passaged into bellows bottles and placed in the system. The medium is then fed to the top and then drained to supply oxygen, this is repeated in a regular cycle. The cells are monitored, fed, washed and harvested in the same order as above. Alternatively, automated equipment can be used to process the cells. After the first passage is complete (T-175 flask or alternative), the cells are seeded into the automated device. One way is to use ACE (Automated Cellular E expansion system, which is a series of off-the-shelf or custom-built components. components to form a cell growth platform in which cells can grow without human intervention. This is a system in which cells can grow rapidly. Cells support anchorage-dependent cell adhesion. The cell tower is made up of a stack of disks that support the cell growth. After cell growth is complete, the The medium is circulated through the cells, and the cells are harvested by trypsinization.

[0046] Alternatively, the ACE system can be used to combine the cell growth surface, delivery tubing, media and reagents, and heating / cooling systems. Mechanism and computer processing for cooling, medium transfer and execution of automatic programming cycles Single lot unit barges consisting of expendable components with permanent bases housing capabilities Upon receipt, each sterile, irradiated ACE disposable unit contains The pre-filled bag is opened from its packaging and suspended, and the bag is connected via a sterile connector. Media and reagents are loaded by connecting to existing tubing. This process is as follows: Proceed as follows: a) Inside a biological safety cabinet (BSC), enzymatically digested The suspension of cells from the biopsy was placed in a "starting growth medium" containing antibiotics. The cells are then transferred from the BSC to a disposable container called a "pre-growth chamber" (a small unit at the top of the cell tower). The specimens are transferred to a permanent ACE unit already in place; b) after approximately 3 days, the specimens are transferred to a pre-propagation unit. The cells in the chamber were trypsinized and introduced into the cell tower itself, which was pre-filled with complete growth medium. Here, the "bubbling action" caused by CO2 injection causes the cells to spiral downwards. It rotates spirally and circulates the medium at a rate that causes it to settle on the face of the disk in an evenly distributed manner. c) The cells are allowed to grow for approximately 7 days, at which time they are allowed to reach confluence. Check (method unknown at time of writing) and confirm that the culture is growing. At this time, the complete growth medium is replaced with fresh complete growth medium. CGM is administered every 3-7 days. At the end of the culture period, the confluence is checked again to determine the intended Ensure that there is sufficient proliferation to possibly produce the desired amount of cells for subsequent processing. d) When the culture is sufficiently confluent, harvest it. The supernatant is then drained from the vessel. PBS (to wash the medium and remove FBS from the cells) is then added. ) is pumped into the vessel and discharged almost immediately. Trypsin-EDTA is removed from the growth surface. The cells are then pumped into a container to separate them. The trypsin / cell mixture is then pumped out of the container. The cryopreservative is pumped into the container and applied to the surface of the disc. The remaining cells are rinsed from the surface and sent to the spin separator, which also collects the cells. The cells are then collected and homogenously resuspended in the transport / injection medium. Cells are counted using an in-line automated cell counting device or via laboratory analysis for cell count and viability. A specific number of cells are counted and the appropriate Once cell concentration is reached, harvested cells are stored in a cool, dry place in a cool, dry place. It is delivered to a collection vial which can be removed.

[0047] In some embodiments, the fibroblasts may be capable of enhancing immunomodulatory activity of hCG and / or Fibroblasts are cultured or administered with oxytocin at a concentration of 1 nM to 1 μM. , or 10 nM to 100 nM, or any range of concentrations inducible therein, together with hCG Fibroblasts can be cultured at a concentration of between 1 nm and 10 μM or can be administered with hCG. Or at a concentration between 100 nM and 1 μM, or any range within which it can be induced, oxytocin The animals may be cultured with oxytocin or may be administered oxytocin. [Example]

[0048] The following examples are included to demonstrate preferred embodiments of the invention. The following techniques have been found by the inventors to work well in the practice of the present invention: and therefore can be considered to constitute a preferred mode for carrying out the invention. However, those skilled in the art will understand that, in view of the present disclosure, , and in specific embodiments disclosed and similar results can be obtained, are not intended to be construed as falling outside the spirit and scope of the present invention. It will be understood that many variations are possible without departing from the scope of the invention.

[0049] Example 1 Cannabidiol stimulates fibroblast production of IGF-1 and EGF-1 Dermal fibroblasts were obtained from Allcells, Inc. and incubated in the indicated concentrations of CBD. As shown in Figure 1, an increase in IGF-1 was observed. As expected, an increase in EGF-1 was observed. The cultures were maintained in OP medium containing 10% fetal bovine serum. The experiment was carried out in TI-MEM medium. References All publications, patents, and patent applications referenced herein are to be interpreted as though each individual publication, Any patent or patent application specifically and individually indicated to be incorporated by reference in its entirety No. 6,317,797, filed Dec. 1, 2004, and incorporated by reference in its entirety as if fully set forth herein. 1. Hwang, GJ, Suh, JS, Na, JB, Lee, HM, and Kim, NH (1997) Co ntrast enhancement pattern and frequency of previously unoperated lumbar discs o n MRI. J Magn Reson Imaging 7, 575-578 2. 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[0050] Although the present disclosure and its advantages have been described in detail, the scope of the present invention is defined by the appended claims. Various changes, substitutions, and alterations may be made herein without departing from the spirit and scope of the present design. It is understood that the scope of the present application is limited to the Any disclosures expressly incorporated herein by reference are limited to particular embodiments of the process, machine, manufacture, composition of matter, means, methods and steps. As those skilled in the art will readily understand from this disclosure, perform substantially the same function or achieve substantially the same result as the corresponding embodiment described in Any now existing or later developed process, machine, manufacture, composition of matter, or technique that accomplishes the Any stage, method, or step may be utilized in accordance with the present disclosure. The appended claims include within their scope such processes, machines, manufacture, compositions of matter, means, or methods. or steps.

Claims

1. A composition for use in the treatment of degenerative intervertebral disc disease, comprising administering cannabidiol and a therapeutically effective amount of fibroblasts to an individual.

2. The composition according to claim 1, wherein the cannabidiol and the fibroblasts are administered to the individual simultaneously or at different time points.

3. The composition according to claim 2, wherein the cannabidiol is administered before, during, or after administration of the fibroblasts.

4. The composition according to any one of claims 1 to 3, wherein the fibroblasts differentiate into notochord cells, chondrocytes, or a mixture thereof.

5. The composition according to any one of claims 1 to 3, wherein the cannabidiol is administered systemically and / or locally into the intervertebral disc of the individual.

6. The composition according to any one of claims 1 to 3, wherein the cannabidiol is administered in a dose and frequency sufficient to reduce apoptosis of cells in the nucleus pulposus.

7. The composition according to claim 6, wherein the cells in the nucleus pulposus are selected from the group consisting of chondrocytes, notochord cells, notochord progenitor cells, chondrocyte progenitor cells, and combinations thereof.

8. The composition according to claim 7, wherein the cells in the nucleus pulposus include exogenous cells.

9. The composition according to claim 7 or 8, wherein the cells in the nucleus pulposus include fibroblasts.

10. The composition according to any one of claims 1 to 3, 7 and 8, wherein the fibroblasts proliferate at a rate of 14 to 21 hours per cell proliferation.

11. The composition according to any one of claims 1 to 3, 7 and 8, wherein the fibroblasts secrete 0.1 pg to 77 pg of interleukin-1 per 1 million fibroblast cultures at 75% surface confluence, or the fibroblasts secrete 1 pg to 500 pg of FGF-1 per 1 million fibroblast cultures at 75% surface confluence.

12. The composition according to any one of claims 1 to 3, 7 and 8, wherein the fibroblasts reduce the ability of responsive T cells to proliferate in a mixed lymphocyte reaction.

13. The composition according to claim 12, wherein the reduction in proliferation includes a reduction of more than 20% compared to a control mixed lymphocyte response in which fibroblasts have not been added.

14. The composition according to any one of claims 1 to 3, 7, 8 and 13, wherein the fibroblasts are treated with human chorionic gonadotropin (hCG) to enhance immunomodulatory activity.

15. The composition according to claim 14, wherein the hCG is administered at a concentration of 1 nM to 1 μM or 10 nM to 100 nM.

16. The composition according to any one of claims 1 to 3, 7, 8, 13 and 15, wherein the fibroblasts are treated with oxytocin to enhance immunomodulatory activity.

17. The composition according to claim 16, wherein the oxytocin is administered at a concentration of 1 nM to 10 μM, or the oxytocin is administered to the cells at a concentration of 100 nM to 1 μM.

18. The composition according to any one of claims 1 to 3, 7, 8, 13, 15 and 17, wherein the cannabidiol comprises (-)-cannabidiol or consists of (-)-cannabidiol.

19. The composition according to any one of claims 1 to 3, 7, 8, 13, 15, and 17, wherein the effective amount of cannabidiol is about 50 mg to about 500 mg in total daily dose.

20. The composition according to any one of claims 1 to 3, 7, 8, 13, 15 and 17, wherein the cannabidiol is present in the gel formulation.

21. The composition according to claim 20, wherein the gel formulation comprises a penetration-enhancing gel.

22. The composition according to any one of claims 1 to 3, 7, 8, 13, 15, 17, and 21, wherein the administration of the cannabidiol and / or the fibroblasts comprises administering to the individual once a day or administering to the individual twice a day.

23. The composition according to any one of claims 1 to 3, 7, 8, 13, 15, 17 and 21, wherein the administration of the cannabidiol and / or the fibroblasts comprises administering the cannabidiol percutaneously to the individual, administering the cannabidiol intradiscally to the individual, and / or administering the cannabidiol together with the localization composition.

24. The composition according to claim 23, wherein the localization composition comprises an extracellular matrix that can act to produce sustained release.

25. The composition according to claim 24, wherein the extracellular matrix is ​​hyaluronic acid.

26. The composition according to claim 25, wherein the hyaluronic acid comprises long-chain hyaluronic acid molecules.

27. ​​The composition according to any one of claims 1 to 3, 7, 8, 13, 15, 17, 21 and 24 to 26, wherein the fibroblasts are derived from one or more tissues selected from the group consisting of adipose tissue, skin tissue, bone marrow, peripheral blood, Wharton jelly, placenta, omentum, mobilized peripheral blood, and combinations thereof.

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