Methods and compositions for treating cartilage and disc tissue lesions
A technology of intervertebral discs and compositions, applied in the direction of bone/connective tissue cells, drug combinations, bone diseases, etc., can solve the problems of non-existing reversal or repair
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example 1
[0237]Currently, no treatment exists to prevent, stop or even delay back pain arising from disc degeneration. Previous studies have shown that linked N can act as a growth factor and stimulate proteoglycan and collagen synthesis in IVD. However, the sequences in junction N involved in regulating cellular activity are not well understood. To determine whether disc cells can proteolytically process Junction N, human disc cells were exposed to native Junction N over a 48-hour period, and as described further below, mass spectrometry analysis revealed that peptides spanning residues 1 to 8 were significantly different in AF cells than in AF cells. generated in the presence of non-NP cells. Linking N1-8 significantly induced proteoglycan production in the presence of IL-1βNP and AF cells, confirming that the biological effect is maintained in the first 8 amino acids of this peptide and indicating that this effect persists in an inflammatory environment.
[0238] Distinct effects ...
example 2
[0282] As previously demonstrated, connexin N is released from connexin by MMP cleavage. It is produced in both joints and intervertebral discs and promotes aggrecan / collagen synthesis by intervertebral disc (NP and AF) and articular cartilage (chondrocyte) cells.
[0283] Junctional N stimulation of human intervertebral discs in organ culture has been reported to increase proteoglycan synthesis 1.15 to greater than 7-fold compared to discs injected with medium alone. Proteoglycan synthesis was detectable 9 days after ligated N stimulation.
[0284] In a rabbit model of intervertebral disc degeneration, 2 weeks after L2 / 3 circular puncture, connexin injection at a concentration of 100 μg of connexin in 10 μl of saline significantly increased IVD disc height at 14 weeks after connexininfection. Aggrecan mRNA was significantly increased in AF and NP cells, especially in AF cells. Likewise, MMP-3 and ADAMTS-4 protease expression was significantly reduced. demonstrated that the...
example 3
[0289] Methodology:
[0290] Osteoarthritic (OA) cartilage was obtained from four donors with informed consent undergoing total knee arthroplasty, and OA cartilage-bone explants and OA chondrocytes were prepared from each donor. Normal human chondrocytes (PromoCell, Heidelberg, Germany) and bovine articular cartilage (12 months) were used as controls.
[0291] Explant preparation and handling
[0292] about 1cm 2 The cartilage explants were prepared from the same donor and included cartilage with cortical bone. Explants were cultured in DMEM supplemented with 10% heat-inactivated FBS. After 7 days under standard culture conditions, the explants were exposed to: IL-1β (5 ng / ml), linked N (1 μg / ml) and co-exposed to IL1β + linked N for 21 days with media changes every three days .
[0293] Tissue Processing and Analysis
[0294] A 3 mm diameter plug of cartilage was isolated from a different region of each explant. Expression of Col II, Agg, Col X and MMP-13 was assessed ...
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