Abstract
BackgroundThe stem cell-based therapies for intervertebral disc degeneration have been widely studied. However, the mechanisms of mesenchymal stem cells interacting with intervertebral disc cells, such as nucleus pulposus cells (NPCs), remain unknown. Exosomes as a vital paracrine mechanism in cell–cell communication have been highly focused on. The purpose of this study was to detect the role of exosomes derived from bone marrow mesenchymal stem cells (BM-MSCs) and NPCs in their interaction with corresponding cells.MethodsThe exosomes secreted by BM-MSCs and NPCs were purified by differential centrifugation and identified by transmission electron microscope and immunoblot analysis of exosomal marker proteins. Fluorescence confocal microscopy was used to examine the uptake of exosomes by recipient cells. The effects of NPC exosomes on the migration and differentiation of BM-MSCs were determined by transwell migration assays and quantitative RT-PCR analysis of NPC phenotypic genes. Western blot analysis was performed to examine proteins such as aggrecan, sox-9, collagen II and hif-1α in the induced BM-MSCs. Proliferation and the gene expression profile of NPCs induced by BM-MSC exosomes were measured by Cell Counting Kit-8 and qRT-PCR analysis, respectively.ResultsBoth the NPCs and BM-MSCs secreted exosomes, and these exosomes underwent uptake by the corresponding cells. NPC-derived exosomes promoted BM-MSC migration and induced BM-MSC differentiation to a nucleus pulposus-like phenotype. BM-MSC-derived exosomes promoted NPC proliferation and healthier extracellular matrix production in the degenerate NPCs.ConclusionOur study indicates that the exosomes act as an important vehicle in information exchange between BM-MSCs and NPCs. Given a variety of functions and multiple advantages, exosomes alone or loaded with specific genes and drugs would be an appropriate option in a cell-free therapy strategy for intervertebral disc degeneration.
Highlights
The stem cell-based therapies for intervertebral disc degeneration have been widely studied
Identification of exosomes derived from Bone marrow mesenchymal stem cell (BM-Mesenchymal stem cell (MSC)) and nucleus pulposus cell (NPC) Exosomes derived from BM-MSCs and NPCs were isolated with the methods described previously [17] and observed by transmission electron microscopy (TEM)
Reciprocal uptake of exosomes by BM-MSCs and NPCs To assay the internalization of exosomes derived from different cells, the PKH67-labeled exosomes were added into the recipient cells
Summary
The stem cell-based therapies for intervertebral disc degeneration have been widely studied. The mechanisms of mesenchymal stem cells interacting with intervertebral disc cells, such as nucleus pulposus cells (NPCs), remain unknown. The purpose of this study was to detect the role of exosomes derived from bone marrow mesenchymal stem cells (BM-MSCs) and NPCs in their interaction with corresponding cells. There are numerous potential pain generators in the lumbar spine, symptomatic disc degeneration is considered a significant contributor to LBP. The primary therapeutic methods for degenerative disc diseases (DDD) are conservative treatment, surgical discectomy, intervertebral disc displacement and, as a last resort, spinal fusion. In the past decades, many studies have been carried out to find a solution in the field of molecular biology and stem cell therapy
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