COMPOSITIONS AND METHODS FOR REDUCING INFLAMMATION
The present invention is directed to a method of producing compositions derived from culturing cells under hypoxic conditions on a biocompatible surface in vitro. The culturing method produces both an extracellular matrix composition and a conditioned culture medium composition, which may be used separately or in combination to obtain physiologically acceptable compositions useful in a variety of medical and therapeutic applications.
1 . A method for reducing inflammation in a subject comprising administering to the subject a conditioned culture medium (CCM) composition or an extracellular matrix (ECM) composition, wherein the CCM and ECM compositions are produced by the method comprising culturing fibroblast cells on microcarrier beads or a three-dimensional surface under hypoxic conditions of 1-5% oxygen, thereby producing multipotent stem cells which produce a CCM and an ECM fraction, thereby reducing inflammation.
2 . The method of claim 1 , wherein the inflammation is associated with bone degeneration, spinal disk degeneration, osteoarthritis, pruritis, skin inflammation, psoriasis, multiple sclerosis, rheumatoid arthritis, osteoarthritis, systemic lupus erythematosus, Hashimoto's thyroidis, myasthenia gravis, diabetes type I or II, asthma, inflammatory lung injury, inflammatory liver injury, inflammatory glomerular injury, atopic dermatitis, allergic contact dermatitis, irritant contact dermatitis, seborrhoeic dermatitis, Sjoegren's syndrome, keratoconjunctivitis, uveitis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, an inflammatory disease of the joints, skin, or muscle, acute or chronic idiopathic inflammatory arthritis, myositis, a demyelinating disease, chronic obstructive pulmonary disease, interstitial lung disease, interstitial nephritis or chronic active hepatitis.
3 . The method of claim 2 , wherein the inflammation is associated with bone degeneration, spinal disk degeneration or osteoarthritis.
4 . The method of claim 1 , wherein enzyme and protease activity in the anulus fibrosus (AF) and nucleus pulposus (NP) are decreased.
5 . The method of claim 1 , wherein the expression of IL-1, IL-6 and/or TNF in anulus fibrosus (AF) tissue is decreased following administration of the CCM or the ECM.
6 . The method of claim 1 , wherein the expression of ACAN, MMP3 and/or ADAMTS4 is increased in AF tissue following administration of the CCM or the ECM
7 . The method of claim 1 , wherein the expression of TNF and/or MMP3 in nucleus pulposus (NP) tissue is decreased following administration of the CCM or the ECM.
8 . The method of claim 1 , wherein the expression of ADAMTS4 and/or ACAN in NP tissue are increased following administration of the CCM or the ECM.
9 . The method of claim 1 , wherein aggrecan is produced.
10 . The method of claim 1 , wherein the fibroblast cells are neonatal fibroblast cells.
11 . A method of treating intervertebrate disc degeneration (IDD) in a subject comprising administering to the subject a cell conditioned media (CCM) composition and/or an extracellular matrix (ECM) composition, wherein the CCM and ECM compositions are produced by the method comprising culturing fibroblast cells on microcarrier beads or a three-dimensional surface under hypoxic conditions of 1-5% oxygen, thereby producing multipotent stem cells which produce a CCM and an ECM fraction, thereby treating IDD.
12 . The method of claim 11 , wherein the expression of IL-1, IL-6 and/or TNF in anulus fibrosus (AF) tissue is decreased following administration of the CCM or the ECM.
13 . The method of claim 11 , wherein the expression of ACAN, MMP3 and/or ADAMTS4 is increased in AF tissue following administration of the CCM or the ECM.
14 . The method of claim 11 , wherein the expression of TNF and/or MMP3 in nucleus pulposus (NP) tissue is decreased following administration of the CCM or the ECM.
15 . The method of claim 11 , wherein the expression of ADAMTS4 and/or ACAN in nucleus pulposus (NP) tissue is increased following administration of the CCM or the ECM.
16 . The method of claim 11 , wherein the fibroblast cells are neonatal fibroblast cells.
17 . The method of claim 11 , wherein there is an increase in disc space and/or disc height.
18 . The method of claim 11 , wherein there is a decrease in disc degeneration.
19 . The method of claim 11 , wherein aggrecan is produced.
20 . The method of claim 11 , wherein hyaline cartilage and/or fibrous tissue is regenerated in the disc.
21 . An anti-inflammatory composition comprising a CCM composition or an ECM composition produced by the method comprising culturing fibroblast cells on microcarrier beads or a three-dimensional surface under hypoxic conditions of 1-5% oxygen, thereby producing multipotent stem cells which produce a soluble and a non-soluble fraction and a pharmaceutically acceptable carrier.