Transplantation of mesenchymal stem cells in a canine disc degeneration model.

Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa, 259-1193, Japan.
Journal of Orthopaedic Research (Impact Factor: 2.88). 06/2008; 26(5):589-600. DOI: 10.1002/jor.20584
Source: PubMed

ABSTRACT Transplantation of mesenchymal stem cells (MSCs) is effective in decelerating disc degeneration in small animals; much remains unknown about this new therapy in larger animals or humans. Fas-ligand (FasL), which is only found in tissues with isolated immune privilege, is expressed in IVDs, particularly in the nucleus pulposus (NP). Maintaining the FasL level is important for IVD function. This study evaluated whether MSC transplantation has an effect on the suppression of disc degeneration and preservation of immune privilege in a canine model of disc degeneration. Mature beagles were separated into a normal control group (NC), a MSC group, and the disc degeneration (nucleotomy-only) group. In the MSC group, 4 weeks after nucleotomy, MSCs were transplanted into the degeneration-induced discs. The animals were followed for 12 weeks after the initial operation. Subsequently, radiological, histological, biochemical, immunohistochemical, and RT-PCR analyses were performed. MSC transplantation effectively led to the regeneration of degenerated discs. FACS and RT-PCR analyses of MSCs before transplantation demonstrated that the MSCs expressed FasL at the genetic level, not at the protein level. GFP-positive MSCs detected in the NP region 8 weeks after transplantation expressed FasL protein. The results of this study suggest that MSC transplantation may contribute to the maintenance of IVD immune privilege by the differentiation of transplanted MSCs into cells expressing FasL.

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    ABSTRACT: Intervertebral disc degeneration is associated with low back pain. Mesenchymal stem cells (MSCs) have been used to "regenerate" the disc. The aim of this study was to perform a systematic review of comparative controlled studies assessing the safety and efficacy of using MSCs in animal in disc regeneration. Literature databases were extensively searched. Trial design, MSC sources, injection method, disc assessment, outcome intervals, and complication events were assessed. Validity of each study was performed. Twenty-four animal studies were included with 20.8% of the studies reporting randomization of groups. The studies represented 862 discs that were injected with MSCs and 1,603 discs as controls. All three types of MSCs (i.e. bone marrow, synovial and adipose tissue) showed successful inhibition of disc degeneration. Bone marrow-derived MSCs demonstrated superior quality of repair compared with other non-MSC treatments. A 2.7% overall complication rate was noted, whereby complications were noted only in rabbits. Overall, evidence suggested that MSCs increased disc space height in the majority of animal models. This is the first systematic review to assess the safety and efficacy of MSC for the treatment of disc degeneration. Short-term MSC transplantation is safe and effective; however, additional, larger and higher-quality studies are needed to assess the long-term safety and efficacy. Inconsistency in methodological design and outcome parameters prevent any robust conclusions. Recommendations are further made to improve efficacy, reduce potential complications, and standardize techniques for future studies.
    Stem Cells and Development 07/2014; · 4.20 Impact Factor
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    ABSTRACT: Introduction: Link N is a naturally occurring peptide that can stimulate proteoglycan synthesis in intervertebral disc (IVD) cells. IVD repair can also potentially be enhanced by mesenchymal stem cell (MSC) supplementation to maximize extracellular matrix production. In a previous study we have shown that Link N can inhibit osteogenesis and increase the chondrogenesis of MSCs in vitro. The aim of the present study was to determine the potential of MSCs and Link N alone or together on the repair the degenerate discs. Materials and Methods: Bovine IVDs with trypsin-induced degeneration were treated with MSCs, Link N or a combination of MSCs and Link N. Trypsin-treated discs were also injected with PBS to serve as a degeneration control. The extracellular matrix (ECM) proteins and proteoglycans were extracted from the inner nucleus pulposus of the discs and sulfated glycosaminoglycans (GAGs) were analyzed by the dimethyl methylene blue (DMMB) dye-binding assay. The expression of type II collagen was analyzed by western blot. To track the MSCs after injection, MSCs were labeled with PKH67 and observed under confocal microscopy after the two week culture period. Results: The GAG content significantly increased compared to the degeneration control when degenerate discs were treated with MSCs, Link N or with a combination of both Link N and MSCs. Histological analysis revealed that the newly synthesized proteoglycan was able to diffuse throughout the ECM and restore tissue content even in areas remote from the cells. The quantity of extractable type II collagen was also increased when the degenerate discs were treated with MSCs and Link N, either alone or together. MSCs survived and integrated in the tissue and were found distributed throughout the nucleus pulposus after the two week culture period. Conclusion: MSCs and Link N can restore GAG content in degenerate discs, when administered separately or together. Treatment with MSCs and Link N can also increase the expression of type II collagen. The results support the concept that biological repair of disc degeneration is feasible, and that the administration of either MSCs or Link N has therapeutic potential in early stages of the disease.
    Tissue Engineering Part A 04/2014; · 4.64 Impact Factor
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    ABSTRACT: To establish a minimally invasive rat model of lumbar intervertebral disc degeneration (IDD) to better understand the pathophysiology of the human condition. The annulus fibrosus of lumbar level 4-5 (L4-5) and L5-6 discs were punctured by 27-gauge needles using the posterior approach under C-arm fluoroscopic guidance. Magnetic resonance imaging (MRI), histological examination by hematoxylin and eosin (H&E) staining, and reverse transcription polymerase chain reaction (RT-PCR) were performed at baseline and 2, 4, and 8 weeks after disc puncture surgery to determine the degree of degeneration. All sixty discs (thirty rats) were punctured successfully. Only two of thirty rats subjected to the procedure exhibited immediate neurological symptoms. The MRI results indicated a gradual increase in Pfirrmann grade from 4 to 8 weeks post-surgery (P<0.05), and H&E staining demonstrated a parallel increase in histological grade (P<0.05). Expression levels of aggrecan, type II collagen (Col2), and Sox9 mRNAs, which encode disc components, decreased gradually post-surgery. In contrast, mRNA expression of type I collagen (Col1), an indicator of fibrosis, increased (P<0.05). The procedure of annular puncture using a 27-gauge needle under C-arm fluoroscopic guidance had a high success rate. Histological, MRI, and RT-PCR results revealed that the rat model of disc degeneration is a progressive pathological process that is similar to human IDD.
    Experimental Animals 01/2014; 63(2):227-34. · 1.46 Impact Factor


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May 21, 2014