The association between paraspinal muscle parameters and vertebral pedicle microstructure in patients undergoing lumbar fusion surgery
Year: 2022
Type: article
Abstract: n¼3).EV were purified from the entire secretome by differential (ultra)centrifugation.EV morphology, particle diameter and size distribution/polydispersity were assessed by transmission electron microscopy (TEM) and dynamic light scattering.Biochemical characterization included western-blot detection of CD63 and calnexin.Human AF cells isolated from IVD biopsies of disc degeneration patients were cultured with IL-1β for 48h (n¼8).Subgroups of IL-1β-stimulated AF cells were treated with entire secretome or EV from nonprimed (Control Sec) or IL-1β-primed (IL-1β Sec) MSC (n¼4).Unstimulated AF cells were used as control.Cell metabolic activity, gene expression of BAX, IL6, IL8, matrix metalloproteinase MMP1, MMP inhibitor TIMP1 and collagen type I (COL1A1), and prostaglandin (PG) E2 secretion were evaluated.Statistics: oneway ANOVA (significance, p<0.05).Results: Both EV from primed and non-primed MSC displayed typical size distribution (Fig. 1A; diameter of 105Æ53 nm, PdI of 0.30), expressed the positive EV marker CD63 and did not express calnexin, a negative EV marker (Fig. 1B).IL-1β stimulation of AF cells increased cell proliferation and metabolic activity/cell, IL6, IL8, MMP1 and TIMP1 expression (p<0.05,Fig. 1C,D) and PGE2 production versus control, while downregulating BAX and COL2A1 (p<0.0001).All treatment groups decreased BAX, IL6, IL8 and PGE2 compared to IL-1β stimulation alone of AF cells; but the secretome from primed and non-primed MSC also downregulated TIMP1 and COL1A1 (p<0.05).Interestingly, the treatment with EV from IL-1β-primed MSC upregulated TIMP1 (p<0.0001) and did not further decrease COL1A1 expression.Conclusion: Overall, the results demonstrate a potent anti-inflammatory effect of the entire secretome and EV on human AF cells.However, EV from primed MSC may have a stronger effect on AF matrix metabolism.For instance, EV encapsulated proteins may be longer protected from degradation than in the entire secretome, being beneficial for therapeutics.Investigations of the proteomic content of the EV and secretome to identify potential effector molecules/subsequent pathways is ongoing.Acknowledgment: Deutsche Wirbels€ aulenstiftung.
Source: Brain and Spine
Language: English
Subfield: Pathology and Forensic Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,033
OpenAlex ID: W4312725129