Neck shoulder pain or lumbocrural pain caused by intervertebral disc degeneration (IDD) could seriously affect the qualities life of patients. Current treatments mainly focus on alleviating pain and the symptoms of nerve compression, which could not radically stop the process of intervertebral disc degeneration, but conversely lead to high recurrence rate. In recent years, scholars have turned to study the biological treatment for repair and rebuild the intervertebral disc by biological molecular therapy, gene therapy, cell therapy and tissue engineering to solve the problem of intervertebral disc degeneration, while most of the above methods are still in animal experiments or in vitro experiments and the clinical application is still a long way to go.
Animals ; Biological Therapy ; Genetic Therapy ; Humans ; Intervertebral Disc ; metabolism ; Intervertebral Disc Degeneration ; genetics ; metabolism ; therapy

Intervertebral disc degeneration is considered as a primary cause of clinical low back pain, however the molecular mechanism is not clear yet. Recently, researches on the molecular basis of intervertebral disc degeneration have become a hotspot. The special structure and biomechanics properties of the disc contribute to its propensity toward degeneration. Intervertebral disc degeneration is associated with the changes of the cytological behavior,including the increase in cell death and the degradation of extracellular matrix. However, the mechanism of cell death including cell apoptosis and autophagy in intervertebral disc degeneration remains unclear. Further study on the molecular mechanism of intervertebral disc degeneration is the foundation of improving and treating the intervertebral disc degeneration in the future. Although some progresses are made in the aspect of biological study, the biological environment of intervertebral disc itself is still a challenge for the development of biological treatment. This article is to review the latest advance on the biological characteristics of normal intervertebral disc and the cell death in the process of the intervertebral disc degeneration.
Animals ; Apoptosis ; Cell Death ; Extracellular Matrix ; metabolism ; Humans ; Intervertebral Disc ; cytology ; metabolism ; Intervertebral Disc Degeneration ; metabolism ; physiopathology

OBJECTIVE: To investigate the expression and clinical significance of receptor interacting protein serine-threonine kinases 1 (RIPK1) in the nucleus pulposus of patients with lumbar disc herniation (LDH). METHODS: Nucleus pulposus tissue specimens of 40 patients with LDH patients underwent surgical treatment from January 2016 to January 2018 as the case group, and nucleus pulposus tissue specimens of 30 patients with lumbar spine fracture underwent surgical treatment at the same time as the control group. The expression of RIPK1 mRNA and protein of receptor interaction were detected by polymerase chain reaction (PCR) and Western blot, respectively. The expression of RIPK1 protein in the nucleus pulposus were detected by immunohistochemical staining. The concentrations of RIPK1 and tumor necrosis factor-α (TNF-α) in nucleus pulposus were detected by ELISA method. The relationship between the concentrations of RIPK1, TNF-α in nucleus pulposus and the Pearce grade of LDH patients was analyzed by one-way ANOVA. The correlation between RIPK1 and TNF-α was analyzed by Pearson. RESULTS: RIPK1 was weakly positively expressed in nucleus pulposus of control group, and RIPK1 protein was positively or strongly positively expressed in case group. The expression of RIPK1 mRNA in nucleus pulposus of case group was higher than that of control group ( CONCLUSION The expression levels of RIPK1 mRNA and protein in the intervertebral disc tissues of LDH patients are higher than those of normal intervertebral disc tissues, and increased with the increase of Pearce grade, which may be an important factor involved in LDH inflammatory disease.
Humans ; Intervertebral Disc/metabolism* ; Intervertebral Disc Degeneration ; Intervertebral Disc Displacement/genetics* ; Nucleus Pulposus ; Receptor-Interacting Protein Serine-Threonine Kinases/genetics* ; Tumor Necrosis Factor-alpha/metabolism*

OBJECTIVESTo investigate the expression of chondromodulin-1 (ChM-I) in human adult degenerative intervertebral disc (IVD) cells and the relationship between ChM-I expression and disc degeneration.

METHODSThree degenerated disc specimens obtained from patients in the treatment of disc degenerative disease from March to April 2009 were used for cell culture. ChM-I expression in IVD cells was examined by RT-PCR and Western blot. The effect of basic fibroblast growth factor (bFGF) on the expression of ChM-I was assessed by real-time PCR and Western blot. From October 2008 to October 2009, 26 human IVD tissues were obtained from patients in the surgical treatment of disc degenerative disease at different stage of degeneration according to MRI. Six IVD tissues removed from patients with metastatic spinal tumor were used as normal control. The expression of ChM-I determined by immunohistochemical analysis was correlated with MRI degeneration grade.

RESULTSRT-PCR and Western blot examination showed that ChM-I was expressed in both adult degenerative anulus fibrosus and nucleus pulposus cells. The mRNA and protein expression of ChM-I were both down-regulated by administration of bFGF with dose-dependent way (P < 0.05). Immunohistochemical analysis showed the percent of ChM-I immunopositive cells in the control group was 0.12 ± 0.03, and the number increased significantly in the advanced degeneration group (P < 0.05).

CONCLUSIONSThe current results demonstrate that IVD cells express ChM-I. Administration of bFGF down-regulates the expression of ChM-I. The expression of ChM-I is correlated with the degree of IVD degeneration which means it may involve in the process of IVD degeneration.

Adult ; Cells, Cultured ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Intervertebral Disc ; metabolism ; Intervertebral Disc Degeneration ; metabolism ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; RNA, Messenger ; genetics

BACKGROUNDThis study aimed to investigate the effect of pcDNA3.1-vascular endothelial growth factor (VEGF)165 vector on vertebral cartilage endplate vascular buds and intervertebral discs.

METHODSRabbits were randomly assigned to the control and experimental groups with 10 in each. In the experimental group, we anesthetized the rabbits and exposed the front vertebral body. Using the mark of the longitudinal ossature of the front vertebral body of the lumbar vertebrae, we advanced a needle at the central point of the front fourth and fifth lumbar intervertebral discs and injected 20 µl pcDNA3.1-VEGF165. Similarly, in the control group, we injected 20 µl pcDNA3.1. At 4 and 8 weeks post-injection, we examined the changes of the vertebral cartilage endplate using X-ray radiograph, histology, and scanning electron microscopy.

RESULTSThe vertebral cartilage endplate calcification and degeneration in the experimental group were less than those in the control group at 8 weeks post-operation. The average number and diameter of vascular buds obviously increased in the experimental group at 4 and 8 weeks post-operation. The number of vascular buds and the diameter in the region of the inner annulus increased when compared to those in the area near the nucleus pulposus.

CONCLUSIONSThe pcDNA3.1-VEGF165 plasmid can increase the average number and diameter of vascular buds and decelerate intervertebral disc degeneration.

Animals ; Female ; Intervertebral Disc Degeneration ; genetics ; metabolism ; Intervertebral Disc Displacement ; genetics ; metabolism ; Male ; Microscopy, Electron, Scanning ; Plasmids ; genetics ; Rabbits ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

BACKGROUNDThe development of mechanically active culture systems helps increase the understanding of the role of mechanical stress in intervertebral disc (IVD) degeneration. Motion segment cultures allow for preservation of the native IVD structure, and adjacent vertebral bodies facilitate the application and control of mechanical loads. The purpose of this study was to establish loading and organ culture methods for rabbit IVD motion segments to study the effect of static load on the whole disc organ.

METHODSIVD motion segments were harvested from rabbit lumbar spines and cultured in no-loading 6-well plates (control conditions) or custom-made apparatuses under a constant, compressive load (3 kg, 0.5 MPa) for up to 14 days. Tissue integrity, matrix synthesis, and the matrix gene expression profile were assessed after 3, 7, and 14 days of culturing and compared with those of fresh tissues.

RESULTSThe results showed that ex vivo culturing of motion segments preserved tissue integrity under no-loading conditions for 14 days whereas the static load gradually destroyed the morphology after 3 days. Proteoglycan contents were decreased under both conditions, with a more obvious decrease under static load, and proteoglycan gene expression was also downregulated. However, under static load, immunohistochemical staining intensity and collagen Type II alpha 1 (COL2A1) gene expression were significantly enhanced (61.54 ± 5.91, P = 0.035) and upregulated (1.195 ± 0.040, P = 0.000), respectively, compared with those in the controls (P < 0.05). In contrast, under constant compression, these trends were reversed. Our initial results indicated that short-term static load stimulated the synthesis of collagen Type II alpha 1; however, sustained constant compression led to progressive degeneration and specifically to a decreased proteoglycan content.

CONCLUSIONSA loading and organ culture system for ex vivo rabbit IVD motion segments was developed. Using this system, we were able to study the effects of mechanical stimulation on the biology of IVDs, as well as the pathomechanics of IVD degeneration.

Animals ; Gene Expression Regulation ; Immunohistochemistry ; Intervertebral Disc ; metabolism ; physiology ; Intervertebral Disc Degeneration ; metabolism ; physiopathology ; Male ; Nucleus Pulposus ; metabolism ; physiology ; Organ Culture Techniques ; methods ; Rabbits ; Stress, Mechanical

OBJECTIVETo explore the role of transforming growth factor-β1 (TGF-β1) in intervertebral disc degeneration and its association with the pathological grading of disc degeneration.

METHODSNormal and degenerative intervertebral disc tissues were collected were classified into 5 grades of increasing degenerative changes. HE staining, immunohistochemistry, TUNEL staining and RT-PCR were used to detect the expression of TGF-β1 in the disc tissues.

RESULTSImmunohistochemistry and RT-PCR showed positive expressions of TGF-β1 and Bcl-2 in normal disc tissues, where Bax was expressed at have a trace level. In the degenerative disc tissues, TGF-β1 expression increased with the pathological grades; the expression levels of TGF-β1 showed significant differences between degenerative and normal tissues and between grade IV and grade I disc tissues (P<0.01).

CONCLUSIONTGF-β1 is an important factor participating in the disc degeneration and its expression level is closely related to the pathological grade of degenerative discs.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc ; pathology ; Intervertebral Disc Degeneration ; classification ; metabolism ; pathology ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; metabolism

Traditional Chinese medicine has unique advantages in the treatment of degenerative bone and joint diseases, and its widely used in clinical practice. In recent years, many scholars have conducted a large number of basic studies on the delay of intervertebral disc degeneration by herbal compound and monomeric components from different perspectives. In order to further elucidate its mechanism of action, this paper summarizes the in vivo and in vitro experimental studies conducted at the level of both herbal compound and single components, respectively, in order to provide references for the basic research on the treatment of lumbar intervertebral disc degeneration by Chinese medicine. A summary shows that commonly used herbal compound prescriptions include both classical prescriptions such as Duhuo Jisheng Decoction, as well as clinical experience prescriptions such as Yiqi Huoxue Recipe. Angelicae Sinensis Radix, Chuanxiong Rhizoma, Rehmanniae Radix Praeparata, Achyranthis Bidentatae Radix, and Eucommiae Cortex were used most frequently. Tonic for deficiency and blood stasis activators were used most frequently. The most utilized monomeric components include icariin, ginsenoside Re, salvianolic acid B and aucubin. The main molecular mechanisms by which herbal compound and monomeric components delay of lumbar intervertebral disc degeneration include improving the intervertebral disc microenvironment, promoting the synthesis of aggregated proteoglycans and type Ⅱ collagen in the intervertebral disc, reducing the degradation of the extracellular matrix, and inhibiting apoptosis in the nucleus pulposus cells, etc. The main signaling pathways involved include Wnt/β-catenin signaling pathway, MAPK-related signaling pathway, mTOR signaling pathway, Fas/FasL signaling pathway, PI3 K/Akt signaling pathway, NF-κB signaling pathway, JAK/STAT signaling pathway, and hedgehog signaling pathway, etc.
China ; Drugs, Chinese Herbal/therapeutic use* ; Hedgehog Proteins/metabolism* ; Humans ; Intervertebral Disc Degeneration/metabolism* ; Nucleus Pulposus/metabolism* ; Wnt Signaling Pathway

BACKGROUNDThe bacterial endotoxins test (BET) is a method used to detect or quantify endotoxins (lipo-polysaccharide, LPS) and is widely used in the quality control of parenteral medicines/vaccines and clinical dialysis fluid. It is also used in the diagnosis of endotoxemia and in detection of environment air quality control. Although BET has been adopted by most pharmacopoeias, result judgment algorithms (RJAs) of the test for interfering factors in the BET still differ between certain pharmacopoeias. We have evaluated RJAs of the test for interfering factors for the revision of BET described in the Chinese Pharmacopoeia 2010 (CHP2010).

METHODSOriginal data from 1 748 samples were judged by RJAs of the Chinese Pharmacopoeia 2010, the Japanese Pharmacopoeia 2011 (JP2011), the European Pharmacopoeia 7.0 (EP7.0), the United States Pharmacopoeia 36 (USP36), and the Indian Pharmacopoeia 2010 (IP2010), respectively. A SAS software package was used in the statistical analysis.

RESULTSThe results using CHP2010 and USP36, JP2011, EP7.0, and IP2010 had no significant difference (P = 0.7740). The results using CHP2010 of 1 748 samples showed that 132 samples (7.6%) required an additional step; nevertheless there was no such requirement when using the other pharmacopeias. The kappa value of two RJAs (CHP2010 and EP7.0) was 0.6900 (0.6297-0.7504) indicating that the CHP2010 and other pharmacopoeias have good consistency.

CONCLUSIONSThe results using CHP2010 and USP36, JP2011, EP7.0, and IP2010 have different characteristics. CHP2010 method shows a good performance in Specificity, mistake diagnostic rate, agreement rate, predictive value for suspicious rate, and predictive value for passed rate. The CHP2010 method only had disadvantages in sensitivity compared with other pharmacopeias. We suggest that the Chinese pharmacopoeia interference test be revised in accordance with the USP36, JP2011, EP7.0, and IP2010 judgment model.

Adult ; Algorithms ; Asian Continental Ancestry Group ; Diskectomy ; Endotoxins ; metabolism ; Female ; Humans ; Intervertebral Disc Degeneration ; surgery ; Intervertebral Disc Displacement ; surgery ; Low Back Pain ; surgery ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo observe the morphology and phenotypes of cells extracted from the endplate in the intervertebral discs and identify the factors affecting their biological characteristic.

METHODSThe intervertebral disc endplate were digested enzymatically, and the morphology of the obtained cells was examined under light microscope. Immunhistochemical analysis of collagen II and real-time PCR was carried out, and the morphologies, viability, cell growth, apoptosis and chondrocyte matrix production were compared between the cells isolated from the degenerative and normal vertebral endplates.

RESULTSThe cells in primary culture presented with spherical and oval morphology, and the cytoplasm was stained blue with toluidine blue. The morphologies of the cartilage endplate cells and the articular cells were almost identical. All the freshly isolated cells expressed collagen II. The degenerative vertebral endplate cells showed decreased expression of collagen II with increased apoptotic cells as compared with normal vertebral endplate cells.

CONCLUSIONThe intervertebral disc endplate cells, like articular cartilage cells, express cartilage-specific matrix proteins. Degenerative vertebral endplate cells show decreased cell vitality with increases cell apoptosis.

Adult ; Apoptosis ; physiology ; Cartilage ; metabolism ; pathology ; Cells, Cultured ; Chondrocytes ; metabolism ; pathology ; Collagen ; metabolism ; Female ; Growth Plate ; metabolism ; pathology ; Humans ; Intervertebral Disc ; metabolism ; pathology ; Intervertebral Disc Degeneration ; metabolism ; pathology ; Lumbar Vertebrae ; metabolism ; pathology ; Male ; Young Adult