Intradiscal application of rhBMP-7 does not induce regeneration in a canine model of spontaneous intervertebral disc degenerationReportar como inadecuado




Intradiscal application of rhBMP-7 does not induce regeneration in a canine model of spontaneous intervertebral disc degeneration - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Arthritis Research and Therapy

, 17:137

First Online: 27 May 2015Received: 16 December 2014Accepted: 16 April 2015

Abstract

IntroductionStrategies for biological repair and regeneration of the intervertebral disc IVD by cell and tissue engineering are promising, but few have made it into a clinical setting.
Recombinant human bone morphogenetic protein 7 rhBMP-7 has been shown to stimulate matrix production by IVD cells in vitro and in vivo in animal models of induced IVD degeneration.
The aim of this study was to determine the most effective dose of an intradiscal injection of rhBMP-7 in a spontaneous canine IVD degeneration model for translation into clinical application for patients with low back pain.

MethodsCanine nucleus pulposus cells NPCs were cultured with rhBMP-7 to assess the anabolic effect of rhBMP-7 in vitro, and samples were evaluated for glycosaminoglycan GAG and DNA content, histology, and matrix-related gene expression.
Three different dosages of rhBMP-7 2.5 μg, 25 μg, and 250 μg were injected in vivo into early degenerated IVDs of canines, which were followed up for six months by magnetic resonance imaging T2-weighted images, T1rho and T2 maps.
Post-mortem, the effects of rhBMP-7 were determined by radiography, computed tomography, and macroscopy, and by histological, biochemical GAG, DNA, and collagen, and biomolecular analyses of IVD tissue.

ResultsIn vitro, rhBMP-7 stimulated matrix production of canine NPCs as GAG deposition was enhanced, DNA content was maintained, and gene expression levels of ACAN and COL2A1 were significantly upregulated.
Despite the wide dose range of rhBMP-7 2.5 to 250 μg administered in vivo, no regenerative effects were observed at the IVD level.
Instead, extensive extradiscal bone formation was noticed after intradiscal injection of 25 μg and 250 μg of rhBMP-7.

ConclusionsAn intradiscal bolus injection of 2.5 μg, 25 μg, and 250 μg rhBMP-7 showed no regenerative effects in a spontaneous canine IVD degeneration model.
In contrast, intradiscal injection of 250 μg rhBMP-7, and to a lesser extent 25 μg rhBMP-7, resulted in extensive extradiscal bone formation, indicating that a bolus injection of rhBMP-7 alone cannot be used for treatment of IVD degeneration in human or canine patients.

AbbreviationsACANaggrecan

ADAMTS5a disintegrin and metalloproteinase with thrombospondin motifs 5

AFannulus fibrosus

AICAkaike Information Criterion

ALPalkaline phosphatase

ATDC5AT805 teratocarcinoma derived chondrogenic cells

BAXB-cell lymphoma 2-associated X

BCL-2B-cell lymphoma 2

bFGFbasic fibroblast growth factor

BLASTBasic Local Alignment Search Tool

BMPbone morphogenetic protein

BMPR1ABMP receptor 1A

BMPR1BBMP receptor 1B

BMPR2BMP receptor 2

CASP3caspase 3

CCND1cyclin-D1

COL1A1collagen type 1 alpha 1

COL2A1collagen type 2 alpha 1

Ctcomputed tomography

Ctcycle threshold

DECAnimal Experiments Committee Dutch: Dierexperimentencommissie

DHIdisc height index

DMMBdimethylmethylene blue

DMSOdimethyl sulfoxide

ECMextracellular matrix

FBSfetal bovine serum

FOVfield of view

GAGglycosaminoglycan

GAPDHglyceraldehyde 3-phosphate dehydrogenase

hg-DMEMhigh-glucose Dulbecco’s modified Eagle’s medium

HPRThypoxanthine-guanine phosphoribosyltransferase

ID1inhibitor of DNA binding

ITSinsulin-transferrin-selenium

IVDintervertebral disc

MMP13matrix metalloproteinase 13

MRImagnetic resonance imaging

NCnotochordal cells

NOGnoggin

NPnucleus pulposus

NPCnucleus pulposus cells

p-spenicillin-streptomycin

P2passage 2

qPCRreal-time quantitative polymerase chain reaction

rhBMP-7recombinant human bone morphogenetic protein 7

ROIregion of interest

RPS19ribosomal protein S19

Ssignal intensity

SDHAsuccinate dehydrogenase complex, subunit A, flavoprotein variant

T1-TFET1-turbo field echo

TEecho time

TGF-βtransforming growth factor-β

TIMP1tissue inhibitor of metalloproteinase 1

TRrepetition time

TSEturbo-spin echo

TSLspin-lock time

Electronic supplementary materialThe online version of this article doi:10.1186-s13075-015-0625-2 contains supplementary material, which is available to authorized users.

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Autor: Nicole Willems - Frances C Bach - Saskia G M Plomp - Mattie HP van Rijen - Jeannette Wolfswinkel - Guy CM Grinwis - Cle

Fuente: https://link.springer.com/



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