Hydrogel composition containing cell products

Abstract

A hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropyl methylcellulose, and a plurality of mesenchymal stem cells is described herein. Further described herein is a method for treating a disease, comprising the step of administering to a patient in need thereof a composition for treatment, and a composition comprising a therapeutically effective amount of hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropyl methylcellulose, and a plurality of mesenchymal stem cells.

Description

Technical Field

[0001] [Cross - Reference to Related Applications] Claims the benefit of U.S. Provisional Patent Application No. 63 / 353,637, filed on June 20, 2022, the content of which is hereby incorporated by reference in its entirety. [Field] Provided herein are compositions comprising cell products incorporated into a hydrogel and their use for the treatment of diseases.

[0002] [Background] Mesenchymal stromal (stem) cells (MSCs) are stem cells that exhibit potent immunomodulatory properties. MSCs can affect many different immune cell subtypes of both the innate and adaptive immune systems. Multiple laboratory and clinical studies have demonstrated that MSCs can suppress the proliferation of T cells, B cells, natural killer cells, and dendritic cells in a dose - dependent manner. Additionally, MSCs have the ability to induce macrophages into a more immunotolerant (M2) phenotype characterized by alternative activation. The cytokine secretion profiles of T cells and B cells are also significantly changed by culture with MSCs into a less inflammatory and fibrosis - inducing phenotype, which may further contribute to their observed immunosuppressive properties.

[0003] [Summary] A hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropylmethylcellulose, and a plurality of mesenchymal stem cells is described herein.

[0004] A method for the treatment of a disease is further described herein, the method comprising a composition for treatment and the step of administering to a patient in need thereof a composition comprising a therapeutically effective amount of hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropylmethylcellulose, and a plurality of mesenchymal stem cells.

[0005] The foregoing and other objects, features, and advantages will become more apparent from the following detailed description, which proceeds with reference to the accompanying drawings.

Brief Description of the Drawings

[0006]

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[0007] I. Abbreviations 2D Two-dimensional 3D Three-dimensional A260 / A280 / A320 Absorbance (optical density) at 260 nm / 280 nm / 320 nm AF Amniotic fluid ANA Antinuclear antibody Arg-1 Arginase-1 BUN Blood urea nitrogen CaCl2 Calcium chloride CCL2 Chemokine (C-C motif) ligand COL1A Collagen type I alpha Cx43 Connexin 43 CXCL1 Chemokine (C-X-C motif) ligand 1 DDW Double-distilled water DPBS Dulbecco's phosphate-buffered saline without Ca and Mg ds-DNA Double-stranded DNA EVs Extracellular vesicles FN1 Fibronectin 1 GFR Glomerular filtration rate H&E Hematoxylin & eosin HA Hyaluronic acid hFF Human foreskin fibroblasts HG Hydrogel hMSC Human Mesenchymal Stem Cells HPBCD Hydroxypropyl-β-cyclodextrin HPMC Hydroxypropylmethylcellulose IC Inside the capsule IFN Interferon IL Interleukin IV Intravenous min Minute MSC Mesenchymal Stem Cells N-GAL Neutrophil gelatinase-associated lipocalin ND Not performed Os Collagen spark P Passage PAS Periodic acid-Schiff reaction PBS Phosphate Buffered Saline PO Oral RAMEB Randomly methylated β-cyclodextrin RPM Revolutions Per Minute RT Reverse Transcriptase RT-qPCR Real-Time Quantitative PCR SBECD Sulfo-butyl-ether-cyclodextrin SC Subcutaneous SLE Systemic Lupus Erythematosus TGFβ Transforming Growth Factor-β TNF-α Tumor Necrosis Factor-α UC Umbilical Cord UC-MSC Umbilical Cord-Derived Mesenchymal Stem Cells UPW Ultra-Pure Water WFI Water For Injection XTT (2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) Tetrazolium-based compound

[0008] II. Terms Unless otherwise indicated, technical terms are used in accordance with their conventional usage. Definitions of common terms in molecular biology can be found in Benjamin Lewin, Genes V, published by Oxford University Press, 1994 (ISBN 0-19-854287-9); Kendrew et al. (eds.), The Encyclopedia of Molecular Biology, published by Blackwell Science Ltd., 1994 (ISBN 0-632-02182-9); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8).

[0009] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly dictates otherwise. Further, all base sizes or amino acid sizes, and all molecular weights or molecular weight values given for nucleic acids or polypeptides are approximate values and are understood to be for illustrative purposes. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are described below. The term "comprise" means "include". The abbreviation "e.g." is derived from the Latin exempli gratia and is used herein to indicate non-limiting examples. Thus, the abbreviation "e.g." is synonymous with the term "for example".

[0010] In case of contradiction, the present specification including the definitions of the terms shall prevail. In addition, all materials, methods, and examples are for illustrative purposes only and are not intended to limit the present invention.

[0011] III. SUMMARY OF SOME EMBODIMENTS The present specification provides advantageous hydrogel-based compositions comprising MSCs, hyaluronic acid or its salts, preferably the sodium salt, and HPMC. When administered to animals in a diseased state, these compositions improve markers of inflammation and reduce markers of fibrosis, particularly in a lupus (lupus) diseased state. Such compositions can be used to treat humans, particularly humans suffering from kidney diseases. Methods of treating diseases, particularly kidney diseases associated with fibrosis and inflammation, are also provided herein. Methods of treating lupus nephritis are described herein.

[0012] The inventors have discovered that the use of a small amount of HPMC in a hyaluronate-based hydrogel composition improves the characteristics of the hydrogel and its therapeutic potential. The addition of other excipients to the hyaluronate-based hydrogel composition does not improve the hydrogel characteristics; rather, it decreases them.

[0013] When treating a disease according to an embodiment, the number of cells, preferably MSCs, per therapeutic administration to a human subject in need thereof is 0.5×10 6 ~2×10 8 cells per kg of the subject's body weight.

[0014] It is suggested that in addition to MSCs, other therapeutic cells may be administered to patients in need thereof using a formulation comprising hyaluronate and HPMC. According to an embodiment, the therapeutic cell is an epithelial cell.

[0015] In the treatment of a disease according to an embodiment, the composition is administered in an amount of 0.5 milliliters (ml) to 4.0 ml per administration. Optionally, the composition is administered in an amount of 1.0 ml per administration.

[0016] According to an embodiment, the therapies described herein are administered subcutaneously to a patient in need thereof. Alternatively, the therapy can be administered locally to the organ being treated. Optionally, the organ being treated is the kidney, and the therapy may be administered into the subcapsular space of the kidney. The administered hydrogel therapy can be administered in a single dose or repeatedly. Optionally, the hydrogel is administered once a year, twice a year, or once every two years. The hydrogels described herein have been shown to provide a paracrine effect over a long period of time.

[0017] According to an embodiment, a method of treatment is described in which the hydrogel therapy described herein is administered to a patient and then tested for the presence of an inflammatory marker, a fibrosis marker, or a disease indicator marker. After such testing, a physician may decide to re-administer the treatment based on whether or not such a marker is present.

[0018] Inflammatory diseases, particularly chronic inflammatory diseases, are caused by the action of the immune system and involve inappropriate or excessive activation of certain T cells, expression of regulatory cytokines and chemokines, loss of immune tolerance, etc. Examples of autoimmune and / or chronic inflammatory diseases are multiple sclerosis, inflammatory bowel disease (IBD), joint diseases such as rheumatoid arthritis, and systemic lupus erythematosus. Some of these diseases are rather organ / tissue specific as follows: intestine (Crohn's disease), skin (psoriasis), myelinated nerves (multiple sclerosis or MS), islets or β cells (insulin-dependent diabetes mellitus (IDDM) or type I diabetes), salivary glands (Sjögren's disease), skeletal muscle (myasthenia gravis), thyroid (Hashimoto's thyroiditis; Graves' disease), anterior chamber of the eye (uveitis), joint tissue (rheumatoid arthritis), and various cardiovascular diseases.

[0019] Progressive fibrosis is a pathological feature of many chronic inflammatory diseases and is an important cause of morbidity and mortality worldwide. Fibrosis is characterized by the accumulation of excessive extracellular matrix components (e.g., collagen, fibronectin) that form fibrous connective tissue in and around inflamed or damaged tissues. Fibrosis can cause overgrowth, sclerosis, and / or scarring that disrupts the structure of underlying organs or tissues. Controlled tissue remodeling and scarring are part of the normal wound healing process promoted by the differentiation and transformation of fibroblasts into myofibroblasts, but excessive and persistent scarring due to severe or repeated injury or dysregulation of wound healing (e.g., persistence of myofibroblasts) can ultimately lead to permanent scarring, organ dysfunction and failure, and even death.

[0020] Fibrotic changes can occur in vascular disorders (e.g., peripheral vascular disease, heart disease, brain disease) and in all major tissue and organ systems (e.g., lung, liver, kidney, heart, skin). Fibrotic disorders include a wide range of clinical manifestations, including systemic disorders such as systemic sclerosis and multiple sclerosis, and organ-specific disorders such as pulmonary, hepatic, and renal fibrosis. Non-limiting examples of fibrotic disorders or fibrotic diseases include pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, hepatic fibrosis (e.g., cirrhosis), cardiac fibrosis, endomyocardial fibrosis, vascular fibrosis (e.g., atherosclerosis, stenosis, restenosis), atrial fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis (e.g., lung), chronic kidney disease, nephrogenic systemic fibrosis, Crohn's disease, hypertrophic scar, keloid, scleroderma, systemic sclerosis (e.g., skin, lung), atherosclerotic fibrosis (e.g., knee, shoulder, other joints), Peyronie's disease, Dupuytren's contracture, adhesive capsulitis, organ transplant-related fibrosis, ischemia-related fibrosis, and the like.

[0021] Diseases of the kidney can preferably be treated by administering the hydrogel composition described herein by the subcutaneous route.

[0022] The hydrogels described herein preferably contain 0.5% to 5% by weight of hyaluronate, or a pharmaceutically acceptable salt thereof. Preferably, the salt is a sodium salt. Preferably, the amount of hyaluronate in the hydrogel is 1% to 2% hyaluronate by weight, and most preferably 1.5%.

[0023] The hydrogels described herein preferably contain 0.01% to 0.1% HPMC by weight, and most preferably 0.05% HPMC by weight.

[0024] The following examples are provided to illustrate certain features and / or embodiments. These examples should not be construed as limiting the disclosure to the specific features or embodiments described.

Example

[0025] Example 1: Preparation of a hydrogel-based cell formulation, overview. Commercially available ready-to-use hydrogels were used as is or diluted with a suitable medium as shown below. In the case of powder-based hydrogels, pharmaceutical-grade hydrogels were prepared by gradually adding the powder to water or an aqueous buffer at room temperature, and gently mixing using a magnetic stirrer to prevent the formation of bubbles until all of the powder was completely dissolved and a gel was formed.

[0026] A hydrogel-based cell formulation was prepared as follows: The hydrogel was brought to room temperature. A human mesenchymal stem cell (hMSC) suspension in medium was prepared based on the desired number of cells. hMSC sources: Promo cells catalog number C-12977 [human mesenchymal stem cells derived from adipose tissue (hMSC-AT)], umbilical cord hMSCs were purchased from OrganaBio. The medium used was MSC Growth Medium 2, Promo cells catalog number C-28009, lot 472M031; 10% supplement nutrients, (all contents), Promo cells, catalog number C-28009, lot 472M031; 1% penicillin-streptomycin solution, Biological Industries, catalog number 03-031-1B and CGM, OrganoBio. The hydrogel was added to the cell suspension in the growth medium, and the mixture was gently pipetted up and down 5 - 10 times to mix thoroughly. The mixture was then transferred to a multiwell plate. The multiwell plate was gently stirred / tilted to ensure a uniform coating on the bottom of each well.

[0027] The multiwell plate was placed in an incubator. Cell viability and proliferation were tested after 24, 48, and 72 hours.

[0028] Example 2: Preparation of hyaluronic acid-based hydrogels with or without additives. A hyaluronic acid-based hydrogel containing cells was prepared using the general process of Example 1.

[0029] Specifically, a hyaluronate-based hydrogel, HY-50 hydrogel, provided by Dechra Pharmaceuticals, (UK), was obtained and had the following components: sodium hyaluronate (17 mg), sodium chloride (7.57 mg), disodium phosphate heptahydrate (3.78 mg), sodium dihydrogen phosphate monohydrate (0.45 mg), made up to 1 ml with WFI.

[0030] To prepare a hyaluronate-based hydrogel using hydroxypropylmethylcellulose (HPMC) as an additive, HPMC was obtained from Ashland Global Specialty Chemicals (Delaware, USA) as Benecel E5. 0.1 g of HPMC was diluted to a 0.5% solution by adding it to 20 ml of PBS, incubated at 37 °C for 10 minutes, and then added to the above hyaluronate-based hydrogel at a ratio of 90% hydrogel to 10% HPMC 0.5% solution.

[0031] Example 3: Test of cell viability using XTT XTT is a test used to determine cell viability and / or proliferation under various conditions. The viability of MSC cells in the hydrogels prepared according to Example 2, with or without an HPMC additive, was tested using an XTT ((2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxyanilide) tetrazolium-based compound) Cell Viability Kit from Cell Signaling Technology (Massachusetts, USA).

[0032] Cell seeding was performed as follows: 5×10 3 cells suspended in 350 μL of hydrogel and 650 μL of medium were mixed in a 1.5 mL Eppendorf tube and vortexed for 1 second. 100 μL of the 3D hydrogel mixture was added to each well in a 96-well plate. Each sample was repeated at least 3 times and incubated at 37 °C for any desired time of 24, 48, or 72 hours.

[0033] The XTT detection solution was prepared by thawing the reagent immediately before the test and then warming it to 37 °C while gently mixing. An electron coupling solution was added to the XTT reagent (1:50 volume ratio) to prepare the XTT detection solution, and then 50 μL of the XTT detection solution was added to each well of the 96-well plate. The plate was returned to the incubator for 3 hours and the absorbance at 450 nm was read.

[0034] Cells in only the medium without hydrogel were also analyzed to determine the cell viability at each time point under the same conditions. The results of the XTT analysis of the cells in the hydrogels prepared in Example 2 with or without HPMC are shown in Table 2 below. The letter S indicates that the cell viability was the same as that of the control cells in the medium in the hydrogel. The letter L indicates that the cell viability was lower than that of the control cells in the medium in the hydrogel. The letter H indicates that the cell viability was higher than that of the control cells in the medium in the hydrogel. ND indicates that there is no available data.

[0035]

Table 1

[0036] Surprisingly, the hyaluronate-based hydrogel containing HPMC increased the cell viability compared to the medium alone.

[0037] Using other solutions, the cell viability of the hyaluronate-based gels in an amount of 10% hydrogel containing sodium alginate solution (1% sodium alginate in the solution), sulfobutylether-beta-cyclodextrin (20% in the solution), poloxamer 407 (10% in the solution), and collagen syrup (10% in the solution) was further increased. None of these agents succeeded in increasing the cell viability beyond that shown with the medium alone.

[0038] Figure 1C shows the XTT results after 24 hours as described above, and Figure 1D shows the XTT results after 48 hours. In the figures, HY-50 is the hydrogel and E5 is the reference for HPMC.

[0039] Example 4: Test of cell viability using live / dead viability / cytotoxicity assay The live / dead viability / cytotoxicity assay is based on the simultaneous quantification of live and dead cells using two fluorescent probes (Calcein AM and ethidium homodimer (EthD-1)) that measure defined parameters of cell viability, namely plasma membrane integrity and intracellular esterase activity.

[0040] Principle of the method: Live cells are characterized by the presence of intracellular esterase activity, which is determined by the enzymatic conversion of the substantially non-fluorescent cell-permeable Calcein AM to the highly fluorescent Calcein. The polyanionic dye calcein is retained almost completely within live cells, generating a strong, uniform green fluorescence within live cells (excitation ~495 nm / emission ~515 nm). EthD-1 enters cells with damaged membranes and upon binding to nucleic acids, undergoes a 40-fold fluorescence enhancement, thereby generating a bright red fluorescence in dead cells (excitation ~495 nm / emission ~635 nm). EthD-1 is excluded by the almost intact plasma membrane of live cells. The determination of cell viability depends on the physical and biochemical properties of the cells. In the following model, 5000 umbilical cord MSCs were seeded into each well of a 96-well plate.

[0041] Cell seeding into 3D hydrogel a. Add 1×10 4 cells suspended in 350 μL of hydrogel and 650 μL of medium to a 1.5 mL Eppendorf tube. b. Vortex for 1 second c. Add 100 μL of the 3D hydrogel mixture to each well in a 96-well plate. d. Incubate at 37 °C for the desired time.

[0042] Reagent preparation a. Thaw the reagents immediately before the experiment. b. Prepare the staining solution by adding 5 μL of Calcein AM (Component A) and 20 μL of ethidium homodimer-1 (Component B) to 10 mL of DPBS. * Note: Consider reducing the amount of Calcein added to epithelial cells c. Remove the medium from the cells d. Add 150 μL of the staining solution directly to the cells. e. Incubate at 20 - 25 °C for 30 minutes. Cover or wrap the plate with aluminum foil to protect from light during the incubation period. f. Perform cell imaging: Calcein AM excitation / emission - 494 / 517 nm, Ethidium Homodimer - 1 excitation / emission - 528 / 617 nm. Figure 1A shows the results of the live / dead assay after 24 hours, and Figure 1B shows the results of the live / dead assay after 48 hours, comparing cells in medium only, cells in hyaluronic acid hydrogel fortified with 10% SBECD solution as described in Examples 2 and 3 (named 10% (20% SBECD) 90% HY - 50), and cells in hyaluronic acid hydrogel fortified with 10% solution of 0.5% HPMC as described in Example 2 (named 10% (0.5% Benecel E5 hypromellose) 90% HY - 50).

[0043] As shown in the figure, the HPMC hydrogel was superior to the SBECD - based hydrogel.

[0044] Example 5: In vivo study of hydrogels containing MSCs for the treatment of lupus, particularly lupus nephritis The MRL / lpr mouse strain was generated by interbreeding four different strains of mice (LG, B6, AKR, and C3H). MRL / lpr mice are unique among lupus strains in that they produce a complete panel of lupus autoantibodies (ANA, anti - dsDNA, anti - Sm, anti - Ro, and anti - La) and have additional lupus symptoms including arthritis, encephalitis, skin rash, and vasculitis. MRL / lpr mice are widely used for the evaluation of lupus treatment candidates because of their more rapid onset and multiple disease symptoms. (Richard & Gilkeson, 2018).

[0045] The MRL / lpr mouse strain was used to determine the effect of MSCs mixed with the hydrogel composition described in Example 2 for treating lupus and various signs and symptoms thereof.

[0046] At the start of the experiment, on day -1 / 0, female MRL / lpr mice and wild-type (WT) MRL, 8 weeks old (body weight of each mouse approximately 25 - 30 g), were subjected to measurement of glomerular filtration rate (GFR), and collection of serum and urine. At 14 weeks, the MRL / lpr mice were divided into 7 groups as described in Table 3. Group 10 consisted of WT MRL mice and was used as a control. IC indicates intra-capsular administration, SC indicates subcutaneous administration, and PO indicates oral administration. The volume of the composition administered to each group was 200 microliters per administration.

[0047]

Table 2

[0048] The following clinical parameters were evaluated: Urine: Measurement of proteinuria by urine stick: twice a week (starting from week 10) Body weight: twice a week Visual inspection of lymph nodes (score swelling 0, 1, 2, 3): once a week Visual inspection of skin rash (score rash development 0, 1, 2, 3): once a week Arthritis symptoms (general overall impression): once a week Serum: Creatinine, BUN, albumin: day 0, day of sacrifice GFR: day 0, day of sacrifice

[0049] At the end of the study, the following tests were performed: Determination of pancreas size and weight, scoring, and lymph node weight

[0050] The following safety parameters were investigated: changes in body weight; food intake; occurrence in the skin, fur, eyes, mucous membranes, secretions, and excretions (e.g., diarrhea) and autonomic nervous system activity (e.g., lacrimation, salivation, piloerection, abnormal respiratory patterns). Changes in responses to walking, posture, and handling, as well as the presence of abnormal behavior, tremors, convulsions, sleep and lethargy, morbidity, and mortality. Immunomodulatory effects of MSCs in SLE (serum): anti-dsDNA and antinuclear antibody (ANA) by ELISA. Using the sera of test animals according to the manufacturer's instructions, the self-antibodies against ds-DNA in murine sera were analyzed using the CUSABIO ELISA assay (Mouse Anti-dsDNA IgG Antibody ELISA Kit).

[0051] Using the sera of test animals according to the manufacturer's instructions, the levels of antinuclear antibody (IgG) in murine sera were analyzed using the CUSABIO ELISA assay (Mouse Anti-Nuclear Antibody (IgG) ELISA Kit).

[0052] None of the treatments had a significant effect on GFR, body weight, or spleen weight. A decrease in the proteinuria score was shown in the subcutaneous hyaluronic acid hydrogel group.

[0053] RNA samples were prepared as follows to determine cytokine levels by PCR. The samples (half of the whole kidney) were stored at -80 °C. The kidney tissue was placed into a tube containing 1 metal bead and 700 μL of TRI reagent and pulverized at high speed in a pulverizer for 5 minutes. The tube was then centrifuged at 300 xg for 5 minutes. 500 μL of the upper fluid was transferred to a new tube and 500 μL of ethanol was added. Total RNA was isolated from the samples using the DirectZol miniprep kit (catalog number R2072) according to the manufacturer's protocol. The elution volume was 50 μL. The samples were stored at -80 °C.

[0054] The amount of RNA required for the synthesis of 500 ng of cDNA samples was calculated based on the RNA concentration data obtained from Nanodrop. cDNA synthesis was performed using the "High Capacity cDNA Reverse Transcription" kit from Applied Biosystems catalog number 4368814.

[0055] The relative gene expression of various cytokines / chemokines was determined by PCR and shown in the graphs of FIGS. 2B to 2G. In the graphs, "vehicle control HA-E5" indicates the use of the above hyaluronic acid, HPMC composition without cells. "HA-E5 MSCs 12M" indicates the use of 12 million cells together with the hyaluronic acid, HPMC composition. "Vehicle control HA" indicates the use of a hyaluronic acid composition (without HPMC) without cells. "HA MSCs 12M" indicates the use of a hyaluronic acid composition (without HPMC) together with 12 million cells. The marker levels indicating lupus, ANA, and ds-DNA are shown in FIGS. 2A and 2H, respectively.

[0056] In summary, MSCs subcutaneously injected with the hydrogel formulation increased the local relative expression of the anti-inflammatory cytokine IL-10 and decreased the expression of the pro-inflammatory cytokines TNFα, IL-1β, and IL-6, as well as the fibrosis markers, FN1 and COL1A. MSCs SC injected with the hydrogel compositions described herein containing HPMC are shown to be able to induce an anti-inflammatory response by a paracrine effect. Additionally, as shown in FIGS. 2A and 2H, MSCs SC injected with the hydrogel compositions described herein containing HPMC can reduce the presence of lupus-related markers, indicating the potential therapeutic value of these compositions.

[0057] According to an embodiment, a hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropylmethylcellulose, and a plurality of mesenchymal stem cells (MSCs) is described herein. Optionally, the MSCs are derived from umbilical cord or adipose tissue. Optionally, the salt of hyaluronic acid is sodium hyaluronate. Optionally, the hydrogel comprises 0.5 to 5 wt% of hyaluronic acid or a pharmaceutically acceptable salt thereof. Optionally, the hydrogel comprises 1.5 wt% of hyaluronic acid or a pharmaceutically acceptable salt thereof. Optionally, the hydrogel comprises 0.01 to 0.1 wt% of HPMC. Optionally, the hydrogel comprises 0.05 wt% of HPMC. Optionally, the composition comprises up to 6.0×10 7 cells per ml. Optionally, the composition is for use in the treatment of an inflammatory or fibrotic disease of a patient's kidney.

[0058] Furthermore, a method of treating an inflammatory or fibrotic disease of a patient's kidney is described herein, comprising administering to a patient in need thereof a therapeutically effective amount of a hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropylmethylcellulose, and a plurality of mesenchymal stem cells (MSCs). Optionally, the disease is an inflammatory disease. Optionally, the disease is a fibrotic disease. Optionally, the composition is administered by a subcutaneous or intradermal route. Optionally, the composition is administered by a subcapsular route. Optionally, the therapeutically effective amount is 0.5×10 6 to 2×10 8 cells per kilogram of the patient's body weight. Optionally, the composition is administered in a volume of 1.5 to 4 ml per administration. Optionally, the composition is administered at intervals of 6 months to 2 years. Optionally, the inflammatory disease is selected from the group consisting of CKD and lupus nephritis. Optionally, the fibrotic disease is selected from the group consisting of renal fibrosis and chronic kidney disease, and nephrogenic systemic fibrosis. Optionally, the disease is lupus nephritis.

[0059] Furthermore, a method for treating a disease selected from the group consisting of osteoarthritis, urinary incontinence, atopic dermatitis, psoriasis, or any combination thereof is described herein, the method comprising administering to a patient in need thereof a therapeutically effective amount of a hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropyl methylcellulose, and a plurality of mesenchymal stem cells (MSCs).

[0060] In one embodiment, the disease is osteoarthritis. In another embodiment, the disease is urinary incontinence. In another embodiment, the disease is atopic dermatitis. In another embodiment, the disease is psoriasis.

[0061] Furthermore, a method for treating aging is described herein, the method comprising administering to a patient in need thereof a therapeutically effective amount of a hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropyl methylcellulose, and a plurality of mesenchymal stem cells (MSCs). One of ordinary skill in the art will understand the term "aging" as the progressive decline of physiological functions necessary for survival and reproduction. In humans, aging represents the accumulation of changes in a person over time and may include physical, psychological, and social changes. In some embodiments, aging includes wrinkles, dull skin, skin discoloration, dry skin, spots and age spots, rough skin, visible pores, or any combination thereof.

[0062] Furthermore, a method for treating a wound is described herein, the method comprising administering to a patient in need thereof a therapeutically effective amount of a hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropyl methylcellulose, and a plurality of mesenchymal stem cells (MSCs). In some embodiments, the wound includes abrasions (scraping of the outer skin layer), lacerations (wounds such as fractures), contusions (bruises swollen by the accumulation of blood and dead cells under the skin), concussions (damage to the organs and tissues under the head without significant trauma), puncture wounds (trauma from a sharp object such as a knife), skin incisions, surgical wounds (intentional cutting of the skin for a surgical procedure), gunshot wounds (wounds caused by firearms), burns, chemical wounds, bites and stings, electrical wounds, ulcers, or any combination thereof.

[0063] In one embodiment, the hydrogel composition is administered by a topical route. In another embodiment, the hydrogel composition is administered by a systemic route.

[0064] Considering a number of possible embodiments to which the disclosed principles of the present invention may be applied, it will be understood that the illustrated embodiments are merely preferred examples of the present invention and should not be construed as limiting the scope of the present invention. Rather, the scope of the present invention is defined by the following claims. Accordingly, the inventors claim as their invention all that comes within the scope and spirit of these claims.

Claims

1. A hydrogel composition comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, hydroxypropyl methylcellulose, and a plurality of mesenchymal stem cells (MSCs).

2. The hydrogel composition according to claim 1, wherein the MSC is derived from the umbilical cord or fat.

3. The composition according to claim 1 or 2, wherein the salt of hyaluronic acid is sodium hyaluronate.

4. The composition according to claim 1, wherein the hydrogel contains 0.5 to 5% by weight of hyaluronic acid or a pharmaceutically acceptable salt thereof.

5. The composition according to claim 4, wherein the hydrogel contains 1.5% by weight of hyaluronic acid or a pharmaceutically acceptable salt thereof.

6. The composition according to claim 1, wherein the hydrogel contains 0.01 to 0.1% by weight of HPMC.

7. The composition according to claim 6, wherein the hydrogel contains 0.05% by weight of HPMC.

8. Up to 6.0 x 10 per 1 ml ７ The composition according to claim 1, comprising cells.

9. The composition according to claim 1, for use in the treatment of inflammatory or fibrotic disease of the kidneys of patients.

10. The composition according to claim 9, wherein the disease is an inflammatory disease.

11. The composition according to claim 9, wherein the disease is a fibrous disease.

12. The composition according to claim 9, wherein the composition is administered by a subcutaneous or intradermal route.

13. The composition according to claim 9, wherein the composition is administered via a subcosal pathway.

14. The therapeutically effective dose is 0.5 × 10 per kilogram of the patient's body weight. ６ ~2 x 10 ８ The composition according to claim 9, wherein the composition is a cell.

15. The composition according to claim 9, wherein the composition is administered in a volume of 1.5 to 4 ml per dose.

16. The composition according to claim 9, wherein the composition is administered at intervals of 6 months to 2 years.

17. The composition according to claim 10, wherein the inflammatory disease is selected from the group consisting of CKD and lupus nephritis.

18. The composition according to claim 11, wherein the fibrous disease is selected from the group consisting of renal fibrosis, chronic kidney disease, and nephrogenic systemic fibrosis.

19. The composition according to claim 17, wherein the disease is lupus nephritis.

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