Back pain (BP) due to degenerative disc disease (DDD) is a severe, often disabling condition. The aim of this study was to determine the association between the expression level of proinflammatory cytokines (IL-1β, IL-6, and IL-17), angiogenesis markers (VEGF-A and CD31) in intervertebral disc (IVD) tissue and IVD degeneration in young people with discogenic BP. In patients who underwent discectomy for a disc herniation, a clinical examination, magnetic resonance imaging of the lumbar spine, histological and immunohistochemical analyses of these factors in IVD were performed in comparison with the parameters of healthy group samples (controls). Histology image analysis of IVD fragments of the DDD group detected zones of inflammatory infiltration, combined with vascularization, the presence of granulation tissue and clusters of chondrocytes in the tissue of nucleus pulposus (NP). Statistically significant increased expression of IL-1β, IL-6, IL-17, VEGF-A and CD31 was evident in the samples of the DDD group compared with the controls, that showed a strong correlation with the histological disc degeneration stage. Our results denote an immunoinflammatory potential of chondrocytes and demonstrates their altered morphogenetic properties, also NP cells may trigger the angiogenesis.

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BrookeSpiegler syndrome (Brooke-Spiegler syndrome; OMIM #605041) is a rare, autosomal dominant inherited monogenic disease caused by mutations in gene CYLD with its different penetrance. It is clinically manifested by the development of multiple neoplasms of skin appendages such as spiradenoma, cylindroma, spiradenocylindroma and trichoepithelioma. Several phenotypic variants with mutations in gene CYLD have been described in the scientific literature. They are classic BrookeSpiegler syndrome; multiple familial trichoepithelioma syndrome (multiple trichoepitheliomas without cylindromas, spiradenomas, etc.); familial cylindromatosis (multiple cylindromas on the scalp i.e., "turban tumor"); syndrome of multiple spiradenomas or spiradenocylindromas without other neoplasms of the skin appendages. For four diseases associated with CYLD mutations, the prevalence is 1/1 000 000. The scientific literature describes more than 200 cases of BrookeSpiegler syndrome, which is more common in women. Phenotypic manifestations of the CYLD mutation are variable, so there is no reliable statistics on the frequency of occurrence of clinical variants of BrookeSpiegler syndrome. The variety of neoplasms of the skin appendages, the commonality of their histogenesis, the similarity of clinical and histological patterns significantly complicate the diagnosis. Verification of the diagnosis of BrookeSpiegler syndrome is based on histological examination and sequencing of the CYLD gene.

Multipotent mesenchymal stromal cells (MSCs) can be used as a model for the development of gene and cell technologies and as a means of delivering nucleic acids to the body, including as part of tissue-engineering constructs. Small interfering RNA (siRNA) molecules acting by the RNA interference mechanism are a high-precision tool for genetic silencing of target mRNA transcripts. The search for low toxic and highly efficient transfection agents for delivery of siRNA or other nucleic acids to MSCs is an urgent task for the development of therapy based on these molecules. A comparative evaluation of five transfection agents showed that compounds based on cationic polymers were more efficient in delivering siRNA molecules than liposomes, while the cytotoxicity of all tested reagents was independent of their chemical structure. For two of the three transfection agents selected according to their efficiency and belonging to different classes, TurboFect and Lipofectamine® 3000, a moderate effect on cell viability was revealed. The results obtained allow us to recommend TurboFect and Lipofectamine® 3000 as highly efficient and relatively low-toxic agents for transfection of MSCs cultures. multipotent mesenchymal stromal cells, siRNA, transfection, lipofection, cationic lipids, cationic polymers, polyethyleneimine. The authors are grateful to staff of functional genomics laboratory of Research Centre for Medical Genetics Skoblova M.Yu. and Krivosheeva I.A., for technical and methodological assistance. The work was financially supported by the Ministry of Science and Higher Education of the Russian Federation within the state assignment for Research Centre for Medical Genetics.

Back pain (BP) due to degenerative spinal injury is a severe, often disabling disease and it tends to decrease age of onset. The cellular and molecular basis of the disc and adjacent structures degradation remains in the focus of pathophysiologists and pathomorphologists close attention. The aim of the study was to determine the expression level of proinflammatory cytokines (IL-1β, IL-6, IL-17) and angiogenesis markers (VEGF-A and CD31) in intervertebral disc (IVD) tissue, their association between each other and between disc degeneration in young people with discogenic BP. In patients who underwent discectomy for a disc herniation, a clinical examination, magnetic resonance imaging (MRI) of the lumbar spine, histological and immunohistochemical analyses of these factors in the disc tissue were performed in comparison with the parameters of healthy group samples (controls). Histology image analysis of IVD fragments of the degenerative disc disease (DDD) group detected zones of inflammatory infiltration, combined with vascularization, the presence of granulation tissue and clusters of chondrocytes in the tissue of nucleus pulposus (NP). Statistically significant expression of IL-1β, IL-6, IL-17 and VEGF-A (especially on the chondrocytes surface) and CD31 (in the disc matrix) was evident in the samples of the DDD group compared with the controls (p < 0.0001), that showed a strong correlation with the histological disc degeneration stage (r > 0.5; p < 0.0001). This denotes a high immunoinflammatory potential of chondrocytes and demonstrates their altered morphogenetic properties. The coincidence of the spatial expression of IL-1β and IL-17 in the perivascular zone endothelium and in the vascular lumen (p < 0.01) was found, which point at the inflammatory synergy of these cytokines. High expression of VEGF-A prevailed on the surface of chondrocytes in cell clusters compared to the matrix (p < 0.0001), indicating that the NP cells trigger the angiogenesis. The absence of CD31 in the cracks of NP with high expression in the disc matrix states the secondary nature of IVD defects due to degeneration, and not due to vascular ingrowth. A high number of patients with Modic changes according to MRI data shown the contribution of cytokines to the formation of reactive spondylitis and the clinical course of BP. These results obtained will help to development molecular/cellular targets and basic strategies for therapy of BP with DDD in the early stages in young people.