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

被引:67
作者
Moon, H. J. [1 ,2 ]
Yurube, T. [1 ,3 ]
Lozito, T. P. [4 ]
Pohl, P. [1 ]
Hartman, R. A. [1 ]
Sowa, G. A. [1 ,5 ]
Kang, J. D. [1 ]
Vo, N. V. [1 ]
机构
[1] Univ Pittsburgh, Sch Med, Dept Orthopaed Surg, Ferguson Lab Orthopaed Res, Pittsburgh, PA 15261 USA
[2] Korea Univ, Dept Neurosurg, Coll Med, Seoul, South Korea
[3] Kobe Univ, Grad Sch Med, Dept Orthopaed Surg, Kobe, Hyogo 6500017, Japan
[4] Univ Pittsburgh, Sch Med, Ctr Cellular & Mol Engn, Dept Orthopaed Surg, Pittsburgh, PA 15219 USA
[5] Univ Pittsburgh, Dept Phys Med & Rehabil, Pittsburgh, PA 15261 USA
关键词
Intervertebral disc degeneration; Angiogenesis; Tip cells; Annulus fibrosus; Endothelial cells; Metalloproteinases; HUMAN INTERVERTEBRAL DISC; OSTEOCHONDRAL JUNCTION; PDGF-BB; METALLOPROTEINASES; ANGIOGENESIS; COLLAGEN; EXPRESSION; AGE; ACTIVATION; LOCALIZATION;
D O I
10.1016/j.joca.2013.12.008
中图分类号
R826.8 [整形外科学]; R782.2 [口腔颌面部整形外科学]; R726.2 [小儿整形外科学]; R62 [整形外科学(修复外科学)];
学科分类号
100224 [整形外科学];
摘要
Objective: To test whether the interaction between annulus fibrosus cells (AFCs) and endothelial cells (ECs) disrupts matrix homeostasis and stimulates production of innervation mediators. 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. Results: Compared to ECs cultured in na ve media, ECs exposed to AFC conditioned media expressed higher mRNA and protein levels of key biomarkers of invasive EC phenotype, MMP-2 (2x), MMP-13 (4x), and PDGF-B (1.5-2 x), and NGF (24.9 15.2 pg/mL vs 0 in na ve media). Treatment of AF cells with EC culture conditioned media decreased collagen type II expression two fold. Considerable quantities of proangiogenic 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. 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. Published by Elsevier Ltd on behalf of Osteoarthritis Research Society International.
引用
收藏
页码:344 / 354
页数:11
相关论文
共 50 条
[1]
'Stress' distributions inside intervertebral discs - The effects of age and degeneration [J].
Adams, MA ;
McNally, DS ;
Dolan, P .
JOURNAL OF BONE AND JOINT SURGERY-BRITISH VOLUME, 1996, 78B (06) :965-972
[2]
Sustained loading generates stress concentrations in lumbar intervertebral discs [J].
Adams, MA ;
McMillan, DW ;
Green, TP ;
Dolan, P .
SPINE, 1996, 21 (04) :434-438
[3]
HMEC-1 - ESTABLISHMENT OF AN IMMORTALIZED HUMAN MICROVASCULAR ENDOTHELIAL-CELL LINE [J].
ADES, EW ;
CANDAL, FJ ;
SWERLICK, RA ;
GEORGE, VG ;
SUMMERS, S ;
BOSSE, DC ;
LAWLEY, TJ .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 1992, 99 (06) :683-690
[4]
Matrix metalloproteinase expression levels suggest distinct enzyme roles during lumbar disc herniation and degeneration [J].
Bachmeier, Beatrice E. ;
Nerlich, Andreas ;
Mittermaier, Norbert ;
Weiler, Christoph ;
Lumenta, Christianto ;
Wuertz, Karin ;
Boos, Norbert .
EUROPEAN SPINE JOURNAL, 2009, 18 (11) :1573-1586
[5]
The relationship between disc degeneration, low back pain, and human pain genetics [J].
Cheung, Kenneth M. C. .
SPINE JOURNAL, 2010, 10 (11) :958-960
[6]
Matrix metalloproteinases in the human intervertebral disc: Role in disc degeneration and scoliosis [J].
Crean, JKG ;
Roberts, S ;
Jaffray, DC ;
Eisenstein, SM ;
Duance, VC .
SPINE, 1997, 22 (24) :2877-2884
[7]
Davis GE, 2001, J CELL SCI, V114, P917
[8]
QUANTITATIVE-ANALYSIS OF TYPE-1 COLLAGEN AND TYPE-2 COLLAGEN IN HUMAN INTERVERTEBRAL DISKS AT VARIOUS AGES [J].
EYRE, DR ;
MUIR, H .
BIOCHIMICA ET BIOPHYSICA ACTA, 1977, 492 (01) :29-42
[9]
Nerve ingrowth into diseased intervertebral disc in chronic back pain [J].
Freemont, AJ ;
Peacock, TE ;
Goupille, P ;
Hoyland, JA ;
OBrien, J ;
Jayson, MIV .
LANCET, 1997, 350 (9072) :178-181
[10]
Nerve growth factor expression and innervation of the painful intervertebral disc [J].
Freemont, AJ ;
Watkins, A ;
Le Maitre, C ;
Baird, P ;
Jeziorska, M ;
Knight, MTN ;
Ross, ERS ;
O'Brien, JP ;
Hoyland, JA .
JOURNAL OF PATHOLOGY, 2002, 197 (03) :286-292