Evaluation of the concentration of growth associated protein-43 and glial cell-derived neurotrophic factor in degenerated intervertebral discs of the lumbosacral region of the spine.

Important neurotrophic factors that are potentially involved in degenerative intervertebral disc (IVD) disease of the spine's lumbosacral (L/S) region include glial cell-derived neurotrophic factor (GDNF) and growth associated protein 43 (GAP-43). The aim of this study was to determine and compare the concentrations of GAP-43 and GDNF in degenerated and healthy IVDs and to quantify and compare the GAP-43-positive and GDNF-positive nerve fibers. The study group consisted of 113 Caucasian patients with symptomatic lumbosacral discopathy (confirmed by a specialist surgeon), an indication for surgical treatment. The control group included 81 people who underwent postmortem examination. GAP-43 and GDNF concentrations were significantly higher in IVD samples from the study group compared with the control group, and the highest concentrations were observed in the degenerated IVDs that were graded 4 on the Pfirrmann scale. In the case of GAP-43, it was found that as the degree of IVD degeneration increased, the number of GAP-43-positive nerve fibers decreased. In the case of GDNF, the greatest number of fibers per mm2 of surface area was found in the IVD samples graded 3 on the Pfirrmann scale, and the number was found to be lower in samples graded 4 and 5. Hence, GAP-43 and GDNF are promising targets for analgesic treatment of degenerative IVD disease of the lumbosacral region of the spine.

Usefulness of Detecting Brain-Derived Neurotrophic Factor in Intervertebral Disc Degeneration of the Lumbosacral Spine.

BACKGROUND In determining the etiology of pain of discogenic origin, attention is paid to the role of neurotrophic factors, such as brain-derived neurotrophic factor (BDNF). Considering the potential role of BDNF in the etiology of pain during intervertebral disc degeneration (IVDD), this study aimed to assess changes in the number of BDNF-positive nerve fibers and levels of BDNF in IVDD of the lumbosacral spine in comparison to intervertebral discs (IVDs) of the control group (cadavers). MATERIAL AND METHODS The study group comprised 113 patients with IVDD of the lumbosacral spine. The control group consisted of 81 people (cadavers). We performed hematoxylin-eosin staining to assess IVD structures (degeneration), immunohistochemistry to determine the number of BDNF-positive nerve fibers, and an enzyme-linked immunosorbent assay and western blot to quantify BDNF levels in IVDs. RESULTS Levels of BDNF in the study group were significantly higher than in the control group (17.91±19.58 pg/mg; P<0.05). Furthermore, BDNF levels were significantly higher in the annulus fibrosus compared to the nucleus pulposus of the intervertebral disc (5.50±6.40 pg/mg; P<0.05). Neither the number of BDNF-positive nerves (P=0.359) nor BDNF concentration (P=0.706) were significantly correlated with the degree of perceived pain. The number of BDNF-positive fibers per 1 mm2 was not found to differ significantly according to the radiological degree of degeneration of the lumbosacral spine based on the Pfirrmann scale (P=0.735). CONCLUSIONS The level of BDNF expression may be indicative of IVD degeneration, although it does not predict the degree of this degeneration.

Pathomechanism of the IVDs Degeneration and the Role of Neurotrophic Factors and Concentration of Selected Elements in Genesis of Low Back Pain.

Degenerative disc disease of the lumbosacral spine is a very common medical problem. An episode of sciatica occurs at least once in the life of 60-90% of the human population. A phenomenon that is closely related to the process of lowering the pH of the extracellular matrix degenerating the intervertebral disc (IVD) is the precipitation of calcium salts, especially pyrophosphate dehydrate and hydroxyapatite. In such an altered environment of the IVD, we can observe an increased influx of monocytes, macrophages, T-lymphocytes, as well as non-immunocompetent cells, which are a source of cytokines, e.g., tumor necrosis alpha (TNF-α), interleukin- (IL-1ß, IL-8). The above-mentioned mediators of an inflammatory condition contribute to an increase in the expression of Brain-Derived Neurotrophic Factor (BDNF) and Glial cell Derived Neurotrophic Factor (GDNF) in mast cells and chondrocytes, as well as to the descending transport of these mediators along the nerve endings. In the process of degeneration of the IVD as a result of repeated and even slight injuries, there is damage to the connections of the endplate of the vertebral bodies with the IVD, which results in an impairment of the penetration of nutritional substances and water into the disc. As a consequence, there is an overexpression of the brain-derived neurotrophic factor GDNF, as well as neuromodulin (GAP-43) in the mast cells and chondrocytes of the IVDs, while descending transport of these mediators along the nerve fibers is also observed.

Changes in Elements and Relationships among Elements in Intervertebral Disc Degeneration.

Intervertebral disc degeneration (IVDD) is a complex and progressive process of disc aging. One of the most important causes of changes in the internal environment, leading to IVDD, can be changes in the concentration of individual metal elements. This study aimed to analyze the concentrations of copper, iron, manganese, lead, zinc, sodium, potassium, phosphorus, and calcium in the degenerated intervertebral discs of the lumbosacral spine, compared to healthy intervertebral discs. The study group (S) consisted of 113 Caucasian patients, qualified by a specialist surgeon for IVDD of the lumbosacral spine. The control group (C) consisted of 81 individuals. The biological material was obtained from Caucasian human cadavers during post-mortem examination. The concentrations of individual elements were assessed using inductively coupled plasma−optical emission spectroscopy (ICP-OES). Statistically significant differences in the concentrations of microelements, depending on the degree of pain intensity, were noted for only potassium (p < 0.05). Statistically significant differences in the concentrations of the assessed microelements, depending on the degree of radiological advancement of the lesions, were noted for copper and iron (p < 0.05). In the degenerated intervertebral discs, the strongest relationships were noted between the concentrations of zinc and lead (r = 0.67; p < 0.05), zinc and phosphorus (r = 0.74; p < 0.05), and zinc and calcium (r = 0.77; p < 0.05). It has been indicated that, above all, the concentrations of copper and iron depend on the advancement of radiological changes, according to the Pfirrmann scale; however, no influence on the pain intensity, depending on the concentration of the assessed elements, was found.

Changes in the Expression Pattern of DUSP1-7 and miRNA Regulating their Expression in the Keratinocytes Treated with LPS and Adalimumab.

BACKGROUND: Increased levels of phosphorylated ERK and p38 MAPK proteins have been observed in psoriatic skin biopsies compared to controls, which may be associated with an impaired expression pattern of dual activity protein phosphatase (DUSP). OBJECTIVE: The purpose of this study was to assessment changes in expression profile of mRNA DUSP 1-7 and miRNA regulating their expression in human keratinocyte cells (HaCaT) had exposed to the liposaccharide A (LPS). METHODS: HaCaT was exposed to 1 µg/ml LPS and next adalimumab by 2,8,24h compared to untreated cells. The microarray method was used to analyze expression pattern of mRNAs, miRNAs, and ELISA to evaluate changes in the level of the proteins. RTqPCR was used to validate the microarray data. Transcriptome Analysis Console and Statistica Software 13 PL were used in statistical analysis (p<0.05). RESULTS: The highest changes in expression was observed for DUSP2 (FC +11.12) and DUSP5 (FC +5.53) in HaCaT culture after 2 hours exposition on adalimumab. It was observed that miR- 1275 (FC -2.39) and miR-34a (FC +6.52) might regulate level of DUSP2, and miR-27a (FC +3.55), miR-27b (FC +2.87) are involved in DUSP5 expression. CONCLUSION: The results obtained suggest that DUSP2 and DUSP5 may be considered as complementary molecular markers in the diagnosis and monitoring of the effectiveness of psoriasis therapy. It was confirmed that hsa-miR-34a, hsa-miR-1275, hsa-miR-3188, hsa-miR-382, hsa-miR- 27a, hsa-miR-27b, hsa-miR-16 have the highest influence on the expression pattern of DUSP1-7.