PDGF-BB inhibits intervertebral disc cell apoptosis in vitro.
Degeneration of the intervertebral disc (IVD) results in deterioration of the spinal motion segment and can lead to debilitating back pain. Given the established mitotic and anti-apoptotic effects of recombinant human platelet-derived growth factor-BB (rhPDGF-BB) in a variety of cell types we postulated that rhPDGF-BB might delay disc cell degeneration through inhibition of apoptosis. To address this hypothesis, we treated human IVD cells isolated from five independent patients with rhPDGF-BB in monolayer and 3D pellet cultures. The anti-apoptotic potential, cell proliferative capacity, morphology/pellet differentiation, and gene expression of PDGF-treated IVD cells were evaluated via flow cytometry/immunohistochemistry, MTT assays, histology, and quantitative RT-PCR, respectively. We found that rhPDGF-BB treatment significantly inhibited cell apoptosis, increased cell proliferation and matrix production, and maintained mRNA expression of critical extracellular matrix genes. This study suggests two possible mechanisms for the anti-degenerative effects of rhPDGF-BB on human IVD cells. First, PDGF treatment strongly inhibited IVD cell apoptosis in 3D cultures. Second, rhPDGF-BB acts as an anabolic agent, promoting maintenance of IVD cell phenotype in 3D culture, based on the molecular and protein expression analysis. We speculate that rhPDGF-BB may be used as a biologic treatment to target early degenerative IVD disease in the future.
Departments of Orthopaedic Surgery, University of Connecticut Health Center, 263 Farmington Avenue, MARB Building, 4th Floor, Farmington, Connecticut, 06030.
Steven M
SM
Presciutti
David N
DN
Paglia
Teja
T
Karukonda
Do Yu
do Y
Soung
Rosa
R
Guzzo
Hicham
H
Drissi
Isaac L
IL
Moss
D000328:Adult; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D000077214:Becaplermin; D053148:Caspase 3 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007403:Intervertebral Disc / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D055959:Intervertebral Disc Degeneration / Q000473:pathology*; D008875:Middle Aged; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology
D017304:Annexin A5; D016326:Extracellular Matrix Proteins; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; D000077214:Becaplermin; D053148:Caspase 3
PDGF; apoptosis; growth factors; intervertebral disc degeneration; pellet culture; spinal therapeutics