Abstract
BackgroundIntervertebral disc degeneration (IDD) is a natural progression of age-related processes. Associated with IDD, degenerative disc disease (DDD) is a pathologic condition implicated as a major cause of chronic lower back pain, which can have a severe impact on the quality of life of patients. As degeneration progression is associated with elevated levels of inflammatory cytokines, enhanced aggrecan and collagen degradation, and changes in the disc cell phenotype. The purpose of this study was to investigate the biological and cytological characteristics of rabbit nucleus pulposus mesenchymal stem cells (NPMSCs)—a key factor in IDD—and to determine the effect of the growth and differentiation factor-5 (GDF5) on the differentiation of rabbit NPMSCs transduced with a lentivirus vector.MethodsAn in vitro culture model of rabbit NPMSCs was established and NPMSCs were identified by flow cytometry (FCM) and quantitative real-time PCR (qRT-PCR). Subsequently, NPMSCs were randomly divided into three groups: a transfection group (the lentiviral vector carrying GDF5 gene used to transfect NPMSCs); a control virus group (the NPMSCs transfected with an ordinary lentiviral vector); and a normal group (the NPMSCs alone). FCM, qRT-PCR, and western blot (WB) were used to detect the changes in NPMSCs.ResultsThe GDF5-transfected NPMSCs displayed an elongated shape, with decreased cell density, and significantly increased GDF5 positivity rate in the transfected group compared to the other two groups (P < 0.01). The mRNA levels of Krt8, Krt18, and Krt19 in the transfected group were significantly higher in comparison with the other two groups (P < 0.01), and the WB results were consistent with that of qRT-PCR.ConclusionsGDF5 could induce the differentiation of NPMSCs. The lentiviral vector carrying the GDF5 gene could be integrated into the chromosome genome of NPMSCs and promoted differentiation of NPMSCs into nucleus pulposus cells. Our findings advance the development of feasible and effective therapies for IDD.
Highlights
Intervertebral disc degeneration (IDD) is a natural progression of age-related processes
Degenerative disc disease (DDD) is a pathological condition recognized as a major contributor to chronic lower back pain (LBP) with a severe impact on the quality of life of patients
Apoptosis of the nucleus pulposus cells (NPCs) and a decrease in cell viability are widely recognized as key factors for disc degeneration, leading to a decrease in extracellular matrix syntheses such as collagen type II and glycoprotein [1, 2]
Summary
Intervertebral disc degeneration (IDD) is a natural progression of age-related processes. Associated with IDD, degenerative disc disease (DDD) is a pathologic condition implicated as a major cause of chronic lower back pain, which can have a severe impact on the quality of life of patients. Degenerative disc disease (DDD) is a pathological condition recognized as a major contributor to chronic lower back pain (LBP) with a severe impact on the quality of life of patients. It is associated with intervertebral disc degeneration (IDD) which is a natural progression of. The intervertebral disc (IVD) is an important component of the spinal column enabling blending, flexion, and torsion of the spine It is composed of a proteoglycan-rich nucleus pulposus (NP), which is constrained by the surrounding annulus fibrosus and the cartilaginous endplates. As a promising treatment approach for DDD, therapies are aimed at the prevention or reversal of IDD
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