bluuebunny/crossref_metadata_2025_split huggingface dataset:
Isolation of Stem Cells of the Degenerated Lumbar Intervertebral Disc Using a Low Cost and Very Effective Technique
Authors: Falavigna A. , Peletti-Figueiró M. , Aguiar I. Silvera , Roesch-Ely M. , Machado D. Cantarelli , Pêgas Henriques J. A. 
Published: May 2014

Introduction The limitations of regenerative therapy of the tissue using stem cells are, among others, the length of time until a satisfactory amount of viable cells is obtained for the therapy and the high financial cost. The purpose of this study, was to report a low cost and very effective technique to obtain large amounts of adult mesenchymal stem cells from the degenerated intervertebral disc in a short time. Materials and Methods The methodology of the isolation in this study gave rise to an intellectual property claim deposited at INPI under number BR 10 2013 031330 0. The intervertebral disc in this study was obtained from patients with lumbar disc degenerative disease refractory to conservative treatment, who required surgery as therapy. Isolation consisted of incubating the degenerated intervertebral disc without going through enzymatic or mechanical dissociation in DMEM medium supplemented with 10% SFB and 1% P/S in a humidified oven at 37°C for 3 days. To prove that these cells were adult mesenchymal stem cells, the supernatant cells were collected with a Pasteur pipette and cultured conventionally in culture containers adding growth factors (0.02 ng/µL of EGF and FGF) for another 3 days. For this purpose, cellular adhesion tests, with fibroblastic characteristics, of the presence/absence of specific surface antigens through flow cytometry (CD 105+, CD73+, CD90+, CD45-, CD34-, CD14-, CD11b-, CD19-, HLA-DR-) and of cell-differentiation for adipogenic and osteogenic tissue were performed. Results Using this process, a confluence of ∼100% of adult mesenchymal stem cells was found in the culture containers 3 days after the patient's intervertebral disc was removed. The isolated cells presented percentages of antigens with specific surfaces in flow cytometry that were positive for mesenchymal stem cells, could differentiate into adipogenic and osteogenic tissue, and had an adhesive characteristic with fibroblastic morphology. The greater the degeneration, the smaller was the percentage of growth and isolation of cells, however its differentiation was satisfactory for adipocytes and osteocytes. Conclusion The benefits of the technique described are that it allows: (1) obtaining quickly and (2) a large amount of adult mesenchymal cells, using a methodology (3) with a low financial cost. Disclosure of Interest A. Falavigna: Conflict with CNPq, FAPERGS, AOSpine Latin America M. Peletti-Figueiró: None declared I. Aguiar: None declared M. Roesch-Ely: None declared D. Machado: None declared J. A. Henriques: None declared
SDC2 and SDC4 Working Together to Keep the Homeostasis of IVD? A Preliminary Study
Authors: Zheng Z. , Wang J. , Liu H. 
Published: May 2014

Introduction Our previous work have proved that Syndecan 4 (SDC4) plays an important role in TNFa and IL1b induced disc degeneration by promoting extracellular matrix degradation, Syndecan 2 (SDC2) and SDC4 both are members of SDC family, however, expression and role of SDC2 in intervertebral disc is still unclear. The role of this study is to investigate the expression pattern and regulation of SDC2 and its and role in intervertebral disc degeneration. Materials and Methods Nucleus pulposus (NP) cells and annulus fibrosis (AF) cells were cultured separately. mRNA were extracted and expression of SDC2 in NP and AF cells were examined by q-PCR method. Expression of SDC2 was examined by q-PCR in 4, 8, 24hs after NP cells were treated by TNFa (20mg/L), IL1-b(10mg/L) and TGF-b (20mg) retrospectively. NP cells were pretreated with p38, JNK and NFkB signaling pathway inhibitors before treatment with TNFa, then expression of SDC2 was examined by qPCR. Results SDC2 was expressed in both NP and AF cells. NP cells have a higher expression level than AF cells of SDC2 in mRNA level. TNFa and IL1b could inhibit SDC2 expression in NP cells robustly. What's more, SDC2 expression was induced after TGFb treatment in NP cells. Inhibition of SDC2 expression by TNFa could be reduced by p38 and NFkB pathway inhibitor. JNK inhibitor has no role in Inhibition of SDC2 expression by TNFa. Conclusion Inflammatory cytokines could inhibit SDC2 expression and induce SDC4 expression in NP cells, p38 and NFkB pathway participant in this process. The different regulation pattern of SDC2 and SDC4 might play important role in keeping intervertebral disc homeostasis. Disclosure of Interest None declared
Adjacent Segment Disease Developing Dependence on Preoperative Adjacent Disc Degenerative Changes
Authors: Masevnin S. , Ptashnikov D. , Michailov D. , Smekalenkov O. , Hao M. , Zaborovskii N. 
Published: May 2014

Introduction The causes of adjacent segment disease (ASD) after lumbar spinal fusion are controversial today. Although the role of initial degenerative changes in the adjacent discs is not fully studied many authors have noted the special importance of them. Object To determine the importance of the preoperative degenerative changes of adjacent segment in the development of symptomatic ASD after short lumbar fusion. Materials and Methods Retrospective study of 78 patients who underwent short 360° fusion lumbar surgery from 2006 to 2010 for the treatment of degenerative lumbar spine with follow-up at least 3 years. All patients had no preoperative radiographic instability signs of the adjacent levels with normal height of the adjacent discs. We compare 2 groups: first group include 38 patients with the pre existing initial degenerative changes of adjacent discs according MRI and scored by Pfirrmann modified classification with 2-5 grade. The second group include 40 patients with no pre existing changes of the adjacent discs according MRI. There were 67% females. Mean age of 58 years (range 23-78). Mean follow-up of 4 years (3-7 years). MRI evaluation of adjacent segment condition and bend lateral radiographs were performed on the preoperative and follow-up visits. Adjacent segment condition was evaluated by MRI and scored with Pfirrmann modified classification. Results In the I group symptomatic ASD was found in 3 cases (7,9%) during 1 year follow-up. There were no cases with increasing the stage of degenerative changes by Pfirrmann according MRI. 3 years f/up in I group showed increasing symptomatic ASD to the 17 cases (44,7%). 76% of them required revision surgery. 11 patients had increasing the degenerative changes to 6-7 stage by Pfirrmann with no clinical manifestation. In the II group at 1 year f/up 2 patients (5%) had symptomatic ASD. At 3 years f/up the number of patients with symptomatic ASD was increased to 6 (15%). 50% of them required revision surgery. Conclusion Our study showed that the great role in the development of the symptomatic ASD after short lumbar fusion is belong to pre existing initial degenerative changes of adjacent discs. Patients with preoperative disc degenerative changes have statistically significant increasing risks of developing symptomatic ASD. Disclosure of Interest None declared References Hilibrand AS, Robbins M. Adjacent segment degeneration and adjacent segment disease: the consequences of spinal fusion? Spine J 2004;4(6, Suppl)190S-194S Park P, Garton HJ, Gala VC, Hoff JT, McGillicuddy JE. Adjacent segment disease after lumbar or lumbosacral fusion: review of the literature. Spine 2004;29(17):1938–1944
TGFβ Suppresses Galectin-3 Expression through Smad3 Signaling by NP Cells and its Role in Degenerative Disc Disease
Authors: Tian Y. , Cao P. , Yuan W. , Li J. , Shapiro I. M , Risbud M. 
Published: May 2014

Introduction Objective To investigate TGFβ regulation of galectin-3 expression in nucleus pulposus (NP) cells and its role in degenerative disc disease. Materials and Methods qRT-PCR and Western blot were used to measure galectin-3 expression in NP. Transfections were used to determine role of Smad3 on TGFβ-mediated galectin-3 promoter activity. Lentiviral knockdown of Smad3 was performed to assess the role of Smad3 in galectin-3 expression. Results A decrease in galectin-3 expression and promoter activity was observed in NP cells after TGFβ treatment. The same trend was observed in degenerative disc samples. CA-ALK5 treatment resulted in a decrease in basal galectin-3 promoter activity. While DN-ALK5 treatment can block the abolished the suppressive effect of TGFβ on galectin-3 promoter activity. The suppressive effect of Smad3 on galectin-3 promoter was confirmed through gain and loss of function studies. When cells are treated with DN-Smad3 (dominant negative Smad3) or Smad7, cytokine-mediated suppression in galectin-3 promoter activity is completely abolished. To further validate the role of Smad3 and to determine if there is cell type specificity, we measured galectin-3 promoter activity in Smad3 null and wild type mouse fibroblasts. Only in wild type cells is the promoter activity cytokine suppressible. Lentiviral transduction with sh-Smad3 significantly blocked TGFβ dependent decrease in galectin-3 expression. Conclusion By controlling the activation of Smad3 signaling, TGFβ modulates the expression of galectin-3 in NP cells. Galectin-3 may play an important role in degenerative disc diseases. To our knowledge, this is first study that shows that in NP cells TGFβ control galectin-3 transcription in Smad3-dependent fashion. Disclosure of Interest None declared
Role of BMI in Matrix Delamination during Intervertebral Disc Degeneration
Authors: Friedmann A. , Schwan S. , Heilmann A. , Goehre F. , Meisel H. J. 
Published: May 2014

Introduction Intervertebral disc (IVD) degeneration is a natural process during aging, contributed by mechanical, genetic, and biochemical parameters. Interactions between these parameters cause an instance of matrix degeneration in IVD tissue on the macroscopic and microscopic scale. The process of disc degeneration by means of matrix destruction should be promoted by an increased body mass index (BMI) representing increased mechanical loadings. The objective is to determine and correlate tissue changes of IVD degeneration in different states under consideration of patients’ BMI value. Materials and Methods As part of a clinical study, human IVD tissues from 60 patients removed during standard sequestrectomy as neurosurgical treatment after disc herniations are investigated regarding their microstructure using scanning electron microscopy. Specimens’ microstructure is investigated with focus on the delamination of the tissue matrix, as well as cellular changes. With digital image analyzing software, degeneration characteristics are evaluated and correlated with clinical relevant parameters. Results Matrix delamination is identified by digital image analysis as microstructural destruction in disc degeneration closely related to patients BMI on the one hand but on the other hand not related to patients’ age or gender. Cellular variations are proved as decrease in cell density and increase in cell size while creeping reduction of the thickness of the extracellular matrix. Conclusion Due to the future development of regenerative therapies for IVD degeneration it is essential to understand the mechanism of degenerative processes in disc tissue. This study demonstrates that structural variations in the microscale are characteristically for disc degeneration and may be the link or trigger for prolapse of degenerated disc tissue. Disclosure of Interest None declared
FasL on Human Nucleus Pulposus Cells Prevents Angiogenesis in the Disc by Inducing Fas-Mediated Apoptosis of Vascular Endothelial Cells
Authors: Sun Z. , Luo Z. 
Published: May 2014

Introduction The intervertebral disc is the largest avascular organ in the human body. However, with the progress of intervertebral disc degeneration (IDD), the disc tends to be vascularized increasingly via angiogenesis. It is well established that both human nucleus pulposus (NP) cells and vascular endothelial cells express FasL and Fas. However, the issue remains open as to whether there are certain active mechanisms preventing angiogenesis in the disc via the FasL-Fas machinery. Materials and Methods The study groups consisted of NP from cadavers and patients as below: Normal cadaveric donors as control ( n = 8; average age 41.8 ± 3.6 [range 37-47]) and patients with IDD undergoing discectomy as degenerative NP samples ( n = 20; average age 43.4 ± 5.4 [range 35-52]). Here, we established a coculture system of human NP cells and vascular endothelial (HMEC-1) cells. In adition, upregulated FasL of NP cells were cocultured with HMEC-1 cells. Results We found that normal NP cells were more capable of inducing apoptosis in HMEC-1 cells (14.2 ± 3.4%) than degenerate NP cells (6.7 ± 1.9%), p < 0.05 By upregulating the FasL expression in degenerate NP cells, we found that FasL played an essential role in the mediation of HMEC-1 cell apoptosis with the activation of downstream FADD and caspase-3. Moreover, we found an increased Fas expression in HMEC-1 cells following cocultured with NP cells, which might be closely linked with FasL produced by NP cells and enhance their interaction. Conclusion This is the first study showing FasL-Fas network might plays an important role in the molecular mechanisms of angiogenesis avoidance of human disc. Consequently, our findings might shed light on the pathogenesis in human IDD and provide a novel target for the treatment strategies for IDD. Disclosure of Interest None declared
Experimental Research on Repair of the Degenerated Intervertebral Disc by Bone Marrow Stromal Stem Cell
Authors: Wang J. , Cui X. , Xiang Q. , Zhou Y. 
Published: May 2014

Introduction Objective To observe the effects of halting, reversing degeneration of the intervertebral disc repair or repairing the degenerated intervertebral disc after bone marrow, stromal stem cells were transplanted to the degenerated rat intervertebral disc. Materials and Methods Rats aged 2 weeks were selected and sacrificed by breaking the neck. The bone marrow stroma stem cells of the rats were separated, cultured, detected for their adherence rate, growth, and diving. The Wistar rats aged 8 weeks were divided into four groups at random with 15 rats for each group. Each group was further divided into three subgroups with five rats for each subgroup. Experimental animal models were established. The three subgroups of animals were sacrificed at appropriate time, respectively, observed for morphological changes of the intervertebral disc tissue under a light microscope, and immunohistochemically detected for expression of type I collagen in the fibrous rings. Results Primary culture and continuous culture indicate that the bone marrow stroma stem cells of the rats have an active proliferation and multiplication capacity. Hematoxylin and eosin staining indicates that group A is the normal group and the intervertebral disc shape is regular during various periods of time; in group B, the intervertebral space becomes narrow and the fibrous rings are arranged irregularly; in group C, the intervertebral space of the specimens is becoming narrow and the fibrous rings are arranged irregularly in an early stage; the histological changes of various periods of time are consistent with that of group B. An immunohistochemical qualitative analysis on the fibrous ring type I collagen indicates that the expression of fibrous ring type I collagen is more in group A and the expression of the fibrous ring type I collagen in group B is decreasing. In group C, the intervertebral disc structure is damaged and the expression of type I collagen in the fibrous rings decreases. In group D, the morphology of the fibrous rings and expression of type I collagen during various periods of time are similar to that of group B indicating that the alginate serves as a carrier for the stem cells and does not play a role in repairing the intervertebral disc. Conclusion The degeneration of the intervertebral disc may be delayed or the degenerated intervertebral disc repaired after the bone marrow stromal stem cells are transplanted to the degenerated rat intervertebral disc. Disclosure of Interest None declared
Collagen-Derived Peptide N-Acetylated Proline-Glycine-Proline in Intervetebral Disc Recruits Cartilage Endplate Stem Cells by Binding with CXC Receptor 2
Authors: Feng C. , Huang B. , Li H. , Zhou Y. 
Published: May 2014

Introduction Degenerative disc disease (DDD) is a prevalent musculoskeletal disorder and is the major cause of low back pain. But our comprehension about development of the intervertebral disc (IVD) and pathogenesis of disc degeneration is unclear, which is an obstacle for appropriate therapies. Materials and Methods Recently, some reports identified stem/progenitor cells within particular IVD niches and cartilage endplate. These cells together with mesenchymal stem cells are recruited to nucleus pulposus (NP) and annulus fibrosus (AF) to generate regenerative functions in a healthy IVD. The possibility of stimulating these stem cells holds promise for future therapy for DDD. But factors which recruit stem cells to NP and AF are unknown. There are increasing evidences that N-acetylated proline-glycine-proline (N-AC-PGP), which is a degradation fragment of collagen, plays an important role in inflammatory cell recruitment. But whether it presents in IVD and whether it is a chemokine of stem cell have remained elusive. Results In this study, we demonstrated the presence of N-AC-PGP in IVD and found the correlation between its content in IVD and degeneration grade of IVD. Then we described the generation pathway of N-AC-PGP in IVD. At last, we demonstrated that N-AC-PGP can recruit cartilage endplate stem cells (CESC) by binding with CXC receptors 2 on CESC. Conclusion These findings reveal a possible regeneration mechanism in IVD, which depends on the chemotaxis of N-AC-PGP for CESC and indicate a new therapy by using N-AC-PGP to stimulate stem cells in IVD. Disclosure of Interest None declared
Electrospun Scaffold Containing TGF-β1 Promotes Human Mesenchymal Stem Cell Differentiation toward a Nucleus Pulposus-Like Phenotype under Hypoxia
Authors: Zhou Y. , Cui X. , Wang J. 
Published: May 2014

Introduction Intervertebral disc degeneration is considered to have a close relation with nucleus pulposus calcification. Electrospunning provides promising strategies for fabricating nanofibrous scaffolds that resemble the extracellular matrix. Mesenchymal stem cells (MSCs) have great potential ability of proliferation and differentiation, which are suitable stem cell source for nucleus pulposus regeneration. Materials and Methods In this study, we fabricated two kinds of nanofibrous scaffolds. The first kind of scaffold was fabricated by blending TGF-β1, polyvinyl alcohol (PVA), and poly (lactic-co-glycolic acid) (PLGA) nanofibers and electrospinning (group I). The other kind of scaffold was fabricated using coaxial electrospinning, in which TGF-β1 and PVA formed the central core and PLGA formed the outer layer (group II). Human MSCs were seeded on both kinds of scaffolds and cultured in a hypoxia chamber (2% O2). Scaffold characterizations were examined using scanning electron microscopy, transmission electron microscopy, scaffold characterization, attenuated total reflectance-Fourier transform infrared spectroscopy. Cell proliferation and differentiation were evaluated after 3 weeks of cell culture. Results Results showed that both kinds of scaffolds shared similar diameter distributions and protein release. However, group I scaffolds were more hydrophilic than that of group II. Both kinds of scaffolds induced the MSCs to differentiate toward the nucleus pulposus-type phenotype in vitro. In addition, the expression of nucleus pulposus-associated genes (aggrecan, type II collagen, and Sox-9) in group I increased more than that in group II. Conclusion Our results indicated that electrospinning nanofibrous scaffolds containing TGF-β1 support the differentiation of MSCs toward the pulposus-type phenotype in a hypoxia chamber. Electrospinning scaffolds by blending TGF-β1, PVA, and PLGA nanofibers would be a more appropriate choice for nucleus pulposus regeneration. Disclosure of Interest None declared
Does Site of Symptoms Influence the Surgical Results for Discogenic Low Back Pain Treatment?
Authors: Ohtori S. , Orita S. , Yamauchi K. , Miyagi M. , Sakuma Y. , Kubota G. , Oikawa Y. , Inage K. , Sainoh T. , Sato J. , Fujimoto K. , Suzuki M. , Takahashi K. 
Published: May 2014

Introduction Surgical results for discogenic low back pain (DLBP) treatment remain controversial. The method of diagnosis of DLBP generally used is MRI and discography. However, this imaging sometime fails to diagnose DLBP. Symptoms from DLBP are varied and include back pain alone, referred leg pain, and referred inguinal pain. In the current study, we aimed to evaluate whether the site of symptoms influences surgical results for DLBP treatment. Materials and Methods The site of symptoms before surgery and the surgical result were retrospectively evaluated in 62 DLBP patients. Symptom sites before surgery were divided into (1) back pain alone, (2) back pain + referred inguinal pain, (3) back pain + referred thigh pain, and (4) back pain + referred lower leg pain. The patients showed disc degeneration only at one level (L4-5 or L5-S1) on MRI, pain provocation on discography, and underwent anterior interbody fusion. Visual analog scale (VAS) of back pain and the Roland-Morris disability questionnaire (RDQ) were evaluated before and 2 years after surgery. Results VAS and RDQ were not significantly different between the four groups before surgery, but were significantly improved in all four groups 2 years after surgery ( p < 0.05). The greatest improvement of back pain was found in the group with back pain + referred inguinal pain compared with the other three groups ( p < 0.05). Less improvement of back pain in the back pain alone group was seen compared with the back pain + referred thigh and back pain + referred lower leg pain groups ( p < 0.05). Conclusion If DLBP is strictly diagnosed using several imaging modes, surgical results were superior in patients with both back pain and either referred inguinal or leg pain compared with those having back pain alone. Disclosure of Interest None declared