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Effects of secreted factors in culture medium of annulus fibrosus cells on microvascular endothelial cells: elucidating the possible pathomechanisms of matrix degradation and nerve in-growth in disc degeneration.
- Source :
-
Osteoarthritis and cartilage [Osteoarthritis Cartilage] 2014 Feb; Vol. 22 (2), pp. 344-54. Date of Electronic Publication: 2013 Dec 18. - Publication Year :
- 2014
-
Abstract
- Objective: To test whether the interaction between annulus fibrosus cells (AFCs) and endothelial cells (ECs) disrupts matrix homeostasis and stimulates production of innervation mediators.<br />Methods: Human microvascular ECs were cultured in the conditioned media of AF cell culture derived from degenerated human surgical specimen. Matrix-metalloproteinases (MMPs) and platelet-derived growth factor (PDGF) of ECs of this culture were analyzed by qRT-PCR, Western, and immunofluorescence. Vascular endothelial growth factor (VEGF), Interleukin-8 (IL-8), and nerve growth factor (NGF) in the media of this cell culture were assayed by ELISA. To determine the effects of ECs on AFCs, qRT-PCR was performed to determine mRNA levels of collagen I, II and aggrecan in AFCs cultured in EC conditioned media.<br />Results: Compared to ECs cultured in naïve media, ECs exposed to AFC conditioned media expressed higher mRNA and protein levels of key biomarkers of invasive EC phenotype, MMP-2 (2×), MMP-13 (4×), and PDGF-B (1.5-2×), and NGF (24.9 ± 15.2 pg/mL vs 0 in naïve media). Treatment of AF cells with EC culture conditioned media decreased collagen type II expression two fold. Considerable quantities of pro-angiogenic factors IL-8 (396.7 ± 302.0 pg/mL) and VEGF (756.2 ± 375.9 pg/mL) were also detected in the conditioned media of untreated AF cell culture.<br />Discussion: AFCs from degenerated discs secreted factors which stimulated EC production of factors known to induce matrix degradation, angiogenesis, and innervation. IL-8 and VEGF maybe the secreted factors from AFCs which mediate a pro-angiogenic stimulus often implicated in the development of disc degeneration.<br /> (Published by Elsevier Ltd.)
- Subjects :
- Adult
Capillaries metabolism
Cell Survival
Cells, Cultured
Collagen Type II metabolism
Endothelial Cells drug effects
Female
Humans
Interleukin-8 biosynthesis
Intervertebral Disc blood supply
Intervertebral Disc Degeneration pathology
Male
Metalloproteases metabolism
Middle Aged
Neovascularization, Pathologic metabolism
Nerve Growth Factor metabolism
Platelet-Derived Growth Factor metabolism
Vascular Endothelial Growth Factor A biosynthesis
Culture Media, Conditioned pharmacology
Endothelium, Vascular metabolism
Extracellular Matrix pathology
Intervertebral Disc innervation
Intervertebral Disc Degeneration metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1522-9653
- Volume :
- 22
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Osteoarthritis and cartilage
- Publication Type :
- Academic Journal
- Accession number :
- 24361793
- Full Text :
- https://doi.org/10.1016/j.joca.2013.12.008