IL-10 and TGF-ß1 may weaken the efficacy of preoperative anti-tuberculosis therapy in older patients with spinal tuberculosis.

Spinal tuberculosis is a common extrapulmonary type that is often secondary to pulmonary or systemic infections. Mycobacterium tuberculosis infection often leads to the balance of immune control and bacterial persistence. In this study, 64 patients were enrolled and the clinicopathological and immunological characteristics of different age groups were analyzed. Anatomically, spinal tuberculosis in each group mostly occurred in the thoracic and lumbar vertebrae. Imaging before preoperative anti-tuberculosis therapy showed that the proportion of abscesses in the older group was significantly lower than that in the younger and middle-aged groups. However, pathological examination of surgical specimens showed that the proportion of abscesses in the older group was significantly higher than that in the other groups, and there was no difference in the granulomatous inflammation, caseous necrosis, inflammatory necrosis, acute inflammation, exudation, granulation tissue formation, and fibrous tissue hyperplasia. B cell number was significantly lower in the middle-aged and older groups compared to the younger group, while the number of T cells, CD4+ T cells, CD8+ T cells, macrophages, lymphocytes, plasma cells, and NK cells did not differ. Meaningfully, we found that the proportion of IL-10 high expression and TGF-ß1 positive in the older group was significantly higher than that in the younger group. TNF-α, CD66b, IFN-γ, and IL-6 expressions were not different among the three groups. In conclusion, there are some differences in imaging, pathological, and immune features of spinal tuberculosis in different age groups. The high expression of IL-10 and TGF-ß1 in older patients may weaken their anti-tuberculosis immunity and treatment effectiveness.

Mycobacterium tuberculosis Induced Osteoblast Dysregulation Involved in Bone Destruction in Spinal Tuberculosis.

Disturbance of bone homeostasis caused by Mycobacterium tuberculosis (Mtb) is a key clinical manifestation in spinal tuberculosis (TB). However, the complete mechanism of this process has not been established, and an effective treatment target does not exist. Increasing evidence shows that abnormal osteoclastogenesis triggered by an imbalance of the receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) axis may play a key role in the disturbance of bone homeostasis. Previous studies reported that RANKL is strongly activated in patients with spinal TB; however, the OPG levels in these patients were not investigated in previous studies. In this study, we investigated the OPG levels in patients with spinal TB and the dysregulation of osteoblasts caused by Mtb infection. Inhibition of the Mce4a gene of Mtb by an antisense locked nucleic acid (LNA) gapmer (Mce4a-ASO) was also investigated. Analysis of the serum OPG levels in clinical samples showed that the OPG levels were significantly decreased in patients with spinal TB compared to those in the group of non-TB patients. The internalization of Mtb in osteoblasts, the known major source of OPG, was investigated using the green fluorescent protein (GFP)-labeled Mycobacterium strain H37Ra (H37RaGFP). The cell-associated fluorescence measurements showed that Mtb can efficiently enter osteoblast cells. In addition, Mtb infection caused a dose-dependent increase of the CD40 mRNA expression and cytokine (interleukin 6, IL-6) secretion in osteoblast cells. Ligation of CD40 by soluble CD154 reversed the increased secretion of IL-6. This means that the induced CD40 is functional. Considering that the interaction between CD154-expressing T lymphocytes and bone-forming osteoblast cells plays a pivotal role in bone homeostasis, the CD40 molecule might be a strong candidate for mediating the target for treatment of bone destruction in spinal TB. Additionally, we also found that Mce4a-ASO could dose-dependently inhibit the Mce4a gene of Mtb and reverse the decreased secretion of IL-6 and the impaired secretion of OPG caused by Mtb infection of osteoblast cells. Taken together, the current finding provides breakthrough ideas for the development of therapeutic agents for spinal TB.

Single Posterior Approach versus Combined Anterior and Posterior Approach in the Treatment of Spinal Tuberculosis: A Meta-Analysis.

BACKGROUND: Spinal tuberculosis is the most common form of tuberculosis affecting bone and often needs surgical treatment. Single anterior, single posterior, and combined anterior and posterior approaches are the 3 most commonly used approaches in surgical treatment. Clinically, the choice of optimal surgical approach remains controversial. The purpose of this meta-analysis was to evaluate clinical efficacy of single posterior approach versus combined anterior and posterior approach. METHODS: Studies comparing surgical treatment of spinal tuberculosis by single posterior approach versus combined anterior and posterior approach were identified in a literature search conducted from study inception to July 2020. Selection of studies, extraction of data, and evaluation of bias risk of studies were performed independently by 2 authors, and meta-analysis was conducted using RevMan 5.3 software. RESULTS: The meta-analysis included 15 studies and 793 spinal tuberculosis cases. Single posterior approach was used in 397 patients, and combined anterior and posterior approach was used in 396 patients. There were no statistical differences in visual analog scale score (P = 0.51), correction of Cobb angle (P = 0.14), neurological improvement (P = 0.71), erythrocyte sedimentation rate (P = 0.32), C-reactive protein after operation (P = 0.81), and loss of correction at final follow-up (P = 0.44) between approaches. Single posterior approach was associated with less intraoperative hemorrhage (P < 0.00001), shorter operative time (P < 0.00001), shorter length of hospital stay (P < 0.00001), and fewer complications (P < 0.00001). Combined anterior and posterior approach was associated with shorter fusion time (P = 0.04). CONCLUSIONS: Both approaches can achieve satisfactory clinical outcomes. Posterior-only approach can safely and effectively achieve lesion débridement, decompression, and stability reconstruction and maintenance with advantages of less invasive surgery, less bleeding, shorter surgery time and hospital stay, and fewer complications and seems to be superior to combined posterior-anterior approach.

Clinical Significance of M1/M2 Macrophages and Related Cytokines in Patients with Spinal Tuberculosis.

BACKGROUND: Macrophages are important immune cells involved in Mycobacterium tuberculosis (M.tb) infection. To further investigate the degree of disease development in patients with spinal tuberculosis (TB), we conducted research on macrophage polarization. METHODS: Thirty-six patients with spinal TB and twenty-five healthy controls were enrolled in this study. The specific morphology of tuberculous granuloma in spinal tissue was observed by hematoxylin-eosin (H&E) staining. The presence and distribution of bacilli were observed by Ziehl-Neelsen (ZN) staining. Macrophage-specific molecule CD68 was detected by immunohistochemistry (IHC). M1 macrophages play a proinflammatory role, including the specific molecule nitric oxide synthase (iNOS) and the related cytokine tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). M2 macrophages exert anti-inflammatory effects, including the specific molecule CD163 and related cytokine interleukin-10 (IL-10). The above markers were all detected by quantitative real-time PCR (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and IHC. RESULTS: Typical tuberculous granuloma was observed in the HE staining of patients with spinal TB. ZN staining showed positive expression of Ag85B around the caseous necrosis tissue and Langerhans multinucleated giant cells. At the same time, IHC results indicated that CD68, iNOS, CD163, IL-10, TNF-α, and IFN-γ were expressed around the tuberculous granuloma, and their levels were obviously higher in close tissue than in the distant tissue. RT-PCR and ELISA results indicated that IL-10, TNF-α, and IFN-γ levels of TB patients were also higher than those of the healthy controls. CONCLUSION: The report here highlights that two types of macrophage polarization (M1 and M2) are present in the tissues and peripheral blood of patients with spinal TB. Macrophages also play proinflammatory and anti-inflammatory roles. Macrophage polarization is involved in spinal TB infection.

Relationship Between IL-10 Gene Polymorphism and Spinal Tuberculosis.

BACKGROUND To investigate the relation between interleukin-10 (IL-10) gene rs1800871 (A/G) polymorphism and spinal tuberculosis. MATERIAL AND METHODS A total of 129 patients with spinal tuberculosis (spinal tuberculosis group) and 106 healthy subjects receiving physical examination (control group) were enrolled in this study. The general data of these subjects were collected, and the C-reactive protein, erythrocyte sedimentation rate (ESR) and baseline hematologic function were examined. The rs1800871 (A/G) polymorphism in IL-10 gene was detected by TaqMan-MGB probe method. RESULTS The C-reactive protein, ESR, white blood cell count, absolute neutrophil count and relative neutrophil count in spinal tuberculosis group were higher than those in control group, while the absolute lymphocyte count and relative lymphocyte count were lower than those in control group (p<0.05). Compared with AA genotype, GG and AG+GG genotypes showed statistically significant difference in distribution frequency (p<0.05), but no significant difference was detected between AG genotype and AA genotype (p>0.05). In spinal tuberculosis group, the frequency of G allele was higher than that of A allele (p<0.01). The C-reactive protein, ESR, white blood cell count and relative neutrophil count in GG genotype were increased compared with those in AG+GG genotype (p<0.05). CONCLUSIONS The rs1800871 (A/G) polymorphism in IL-10 gene is related to the susceptibility to spinal tuberculosis. Moreover, carrying G allele increases the risk of spinal tuberculosis.

Correlation analysis of ADAMTS-4, VCAM-1, and TAK1 expression in cartilage tissue from spine tuberculosis.

A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) can effectively degrade articular cartilage matrix proteoglycan and damage the intervertebral disc of spinal tuberculosis patients, resulting in deterioration of the physical properties of articular cartilage. Transforming growth factor ß activated kinase 1 (TAK1) is similar to vascular cell adhesion molecule 1 (VCAM-1) and closely related to a variety of pathophysiological processes. This study intended to explore the expression of ADAMTS-4, VCAM-1, and TAK1 in cartilage tissue obtained from spinal tuberculosis patients and their inter-relationships, aiming to provide new treatment approaches for spinal tuberculosis. Patients with spinal tuberculosis (N = 60) from the department of orthopedics and patients with traumatic spinal fracture (N = 60, controls) were recruited for the study. ADAMTS-4, VCAM-1, and TAK1 expression was detected by immunohistochemistry. SPSS 19.0 software was used for data processing and analysis. The score values of ADAMTS-4, TAK1, and VCAM-1 were 1.45 ± 0.10, 1.33 ± 0.09, and 1.54 ± 0.11, respectively, which were significantly higher than those in normal controls (P < 0.05). ADAMTS-4 showed positive correlation with VCAM-1 and TAK1. ADAMTS-4, TAK1, and VCAM-1 expressions increased in spinal tuberculosis patients. They could provide clinical reference for spinal tuberculosis diagnosis and new treatment strategies can be devised by focusing on their positive correlation.

The in vitro and in vivo effects of microRNA-133a on intervertebral disc destruction by targeting MMP9 in spinal tuberculosis.

AIMS: We aim to investigate the role of microRNA-133a (miR-133a) in intervertebral disc destruction by targeting MMP9 in spinal tuberculosis (TB). MAIN METHODS: Rabbit models with spinal TB were established and assigned to the blank, miR-133a mimic, miR-133a inhibitor and negative control (NC) groups. Primary notochordal cells were extracted and separately transfected with miR-133a mimics, miR-133a inhibitor, miR-nonsense sequence control (NC), si-NC and si-MMP9. QRT-PCR and Western blot assay were used to detect the expression of MMP-9, Collagen I, Collagen II and Collagen-X. Gelatin Zymography was performed to detect MMP9 activity. Immunohistochemistry was used to detect the expression of Collagen I, Collagen II and Collagen-X proteins. Osteoclast morphology and the number of osteoclast cells were observed after Tartrate resistant acid phosphatase staining. KEY FINDINGS: MMP9, Collagen-X and Collagen I expression and MMP9 activity were higher while the expression of Collagen II was lower in the miR-133a mimic group than the miR-NC group. MMP9, Collagen-X Collagen I and MMP9 activities were lower and Collagen II expression was higher in the miR-133a inhibitor group than the miR-NC group. Compared with the si-NC group, the si-MMP9 group showed increased Collagen II expression but decreased expression of MMP9, Collagen-X and Collagen I and MMP9 activity. A reduced amount of osteoclast cells exhibited in the miR-133a mimic group while an increased number was seen in the miR-133a inhibitor group compared to the blank group. SIGNIFICANCE: MiR-133a could inhibit Collagen degradation by down-regulating MMP-9 expression to attenuate the destructive effects of spinal TB on intervertebral disc.

Tuberculosis in early medieval Switzerland--osteological and molecular evidence.

Lesions consistent with skeletal tuberculosis were found in 13 individuals from an early medieval skeletal sample from Courroux (Switzerland). One case of Pott's disease as well as lytic lesions in vertebrae and joints, rib lesions and endocranial new bone formation were identified. Three individuals with lesions and one without were tested for the presence of Myobacterium tuberculosis complex (MTBC) ancient DNA (aDNA), and in two cases, evidence for MTBC aDNA was detected. Our results suggest the presence of tuberculosis in the analysed material, which is in accordance with other osteological and biomolecular research that reported a high prevalence of tuberculosis in medieval skeletons.

Expression of TNF-α, IFN-γ, TGF-ß, and IL-4 in the spinal tuberculous focus and its impact on the disease.

To investigate the expression of TNF-α, IFN-γ, TGF-ß, and IL-4 in the spinal tuberculous focus and its relationship with the lesions type, severity, and bone destruction. The pathological samples of patients with spinal tuberculosis (TB) were divided into hyperplasia group and necrosis group according to their intra-operative and post-operative pathological findings. Normal bone tissues were taken as the control group. Pathology and expression of TNF-α, IFN-γ, TGF-ß, and IL-4 in different tissues were compared among these three groups using immunohistochemical staining, quantitative image analysis, and measurement of bone tissue. 286 granulomas observed in the 14 samples in the hyperplasia group, which included 84 necrotizing and 202 non-necrotizing granulomas. As for the 20 samples in the necrosis group, there were 356 necrotizing and 186 non-necrotizing granulomas among all the 542 granulomas. The proportion of necrotizing granulomas in the necrosis group was significantly higher than that of the hyperplasia group. By inter-group comparison, expression of TNF-α, IFN-γ of granulomas in the hyperplasia group was significantly higher than that of the necrosis group, while the expression of TGF-ß, IL-4 of granulomas in the necrosis group was significantly higher than that of the hyperplasia group. Also, expression of IFN-γ of non-necrotizing granulomas was significantly higher than that of necrotizing granulomas in the hyperplasia group, and expression of TGF-ß in necrotizing granulomas was significantly higher than that of non-necrotizing granulomas in the necrosis group. The lesions were mainly bone resorption in the hyperplasia group, whereas mostly necrotic bones accompanied by local fibrosis in the necrosis group. Expression levels of TNF-α, IFN-γ in the hyperplasia group have a positive correlation to bone loss, whereas expression levels of TGF-ß, IL-4 in the necrosis group have a positive correlation to the bone formation. The high expressions of TNF-α, IFN-γ in the spinal tuberculous focus were associated with protective immune cells. TGF-ß and IL-4 were related to allergic lesions, fibrosis and osteogenesis. Expression imbalance of TNF-α, IFN-γ, TGF-ß, and IL-4 might aggravate the allergy of TB.

Transcript levels of major interleukins in relation to the clinicopathological profile of patients with tuberculous intervertebral discs and healthy controls.

OBJECTIVES: The purpose of the present study was to simultaneously examine the transcript levels of a large number of interleukins (ILs; IL-9, IL-10, IL-12, IL-13, IL-16, IL-17, IL-18, IL-26, and IL-27) and investigate their correlation with the clinicopathological profiles of patients with tuberculous intervertebral discs. METHODS: Clinical data were collected from 150 patients participating in the study from January 2013 to December 2013. mRNA expression levels in 70 tuberculous, 70 herniated, and 10 control intervertebral disc specimens were determined by real-time polymerase chain reaction. RESULTS: IL-10, IL-16, IL-17, IL-18, and IL-27 displayed stronger expression in tuberculous spinal disc tissue than in normal intervertebral disc tissue (P<0.05). Our results illustrated multiple correlations among IL-10, IL-16, IL-17, IL-18, and IL-27 mRNA expression in tuberculous samples. Smoking habits were found to have a positive correlation with IL-17 transcript levels and a negative correlation with IL-10 transcript levels (P<0.05). Pain intensity, symptom duration, C-reactive protein levels, and the erythrocyte sedimentation rate exhibited multiple correlations with the transcript levels of several ILs (P<0.05). CONCLUSIONS: The experimental data imply a double-sided effect on the activity of ILs in tuberculous spinal intervertebral discs, suggesting that they may be involved in intervertebral discs destruction. Our findings also suggest that smoking may affect the intervertebral discs destruction process of spinal tuberculosis. However, further studies are necessary to elucidate the exact role of ILs in the intervertebral discs destruction process of spinal tuberculosis.

Transcript levels of major MMPs and ADAMTS-4 in relation to the clinicopathological profile of patients with tuberculous intervertebral discs and healthy controls.

OBJECTIVES: The purpose of the present study was to simultaneously examine the transcript levels of a large number of MMPs (-3, -8, -9, -13 and -14) and ADAMTS-4 and to investigate their correlation with the clinicopathological profile of patients suffering from tuberculous intervertebral discs. DESIGN AND METHODS: Clinical data were collected from 130 patients participating in the study from March 2011 to April 2012. mRNA expression levels were determined by means of the real-time polymerase chain reaction in 60 tuberculous (TB), 60 herniated, and 10 control intervertebral disc (ID) specimens. RESULTS: MMP-8, -9, -13, and -14 that showed a stronger expression in spinal TB disc tissue compared to normal ID tissue (P<0.05). Our results showed multiple positive correlations among MMP-8, -9, and -13 mRNA in TB samples. Smoking habits were found to significantly up-regulate the transcript levels of MMP-3 and -13 (P<0.05). Pain intensity, duration of symptoms, CRP, and ESR significantly affected the transcript levels of several MMPs (P<0.05). CONCLUSIONS: The MMPs may drive extracellular matrix destruction in spinal tuberculosis intervertebral discs. The experimental data imply a synergistic effect on the activity of these MMPs in spinal tuberculosis intervertebral discs. Furthermore, the experimental data suggest that smoking plays an unfavourable role in the prognosis of spinal tuberculosis intervertebral discs. Moreover, pain intensity, duration of symptoms, CRP, and ESR may affect the process of extracellular matrix destruction by increasing the expression of MMPs in spinal tuberculosis intervertebral disc samples.

[Comparison of three antituberculous drugs in serum and cold abscesses of patients with spinal tuberculosis].

OBJECTIVE: Pharmacokinetics of INH, RFP and OFLX in serum and cold abscesses of patients with spinal tuberculosis was analyzed to provide reference to choosing clinical therapeutic regimen. METHOD: The aspiration specimens of abscesses and venous blood were collected from 8 patients with spinal tuberculosis at 0.5, 0.75, 1.0, 1.5, 2.0, 4.0, 6.0, 9.0, 12.0, 16.0 and 24.0 h after administration of antituberculous drugs. The specimens were assessed by high performance liquid chromatography (HPLC). The data were processed with software 3P87. RESULT: The Cmax of INH, RFP and OFLX in serum were 10.87 +/- 7.09 micrograms/ml, 9.98 +/- 3.53 micrograms/ml, and 5.29 +/- 0.72 micrograms/ml, while the Cmax of INH, RFP and OFLX in cold abscesses were 2.84 +/- 1.63 micrograms/ml, 0.57 +/- 0.26 microgram/ml and 3.19 +/- 1.29 micrograms/ml respectively. CONCLUSION: After administration, the Cmax of INH and OFLX in the cold abscesses of patients with spinal tuberculosis, reached the level beyond their MIC and appeared and disappeared more slowly than that in the serum; RFP was not easy to permeate into the cold abscesses, the Cmax of RFP in the cold abscesses just reached its MIC.

Mycobacterium tuberculosis chaperonin 10 stimulates bone resorption: a potential contributory factor in Pott's disease.

Pott's disease (spinal tuberculosis), a condition characterized by massive resorption of the spinal vertebrae, is one of the most striking pathologies resulting from local infection with Mycobacterium tuberculosis (Mt; Boachie-Adjei, O., and R.G. Squillante. 1996. Orthop. Clin. North Am. 27:95-103). The pathogenesis of Pott's disease is not established. Here we report for the first time that a protein, identified by a monoclonal antibody to be the Mt heat shock protein (Baird, P.N., L.M. Hall, and A.R.M. Coates. 1989. J. Gen. Microbiol. 135:931-939) chaperonin (cpn) 10, is responsible for the osteolytic activity of this bacterium. Recombinant Mt cpn10 is a potent stimulator of bone resorption in bone explant cultures and induces osteoclast recruitment, while inhibiting the proliferation of an osteoblast bone-forming cell line. Furthermore, we have found that synthetic peptides corresponding to sequences within the flexible loop and sequence 65-70 of Mt cpn10 may comprise a single conformational unit which encompasses its potent bone-resorbing activity. Our findings suggest that Mt cpn10 may be a valuable pharmacological target for the clinical therapy of vertebral tuberculosis and possibly other bone diseases.

The penetration of drugs into the lesions of spinal tuberculosis.

The concentration of antituberculous drugs in blood and pus from tuberculous spinal lesions was measured initially, and again after 3 to 5 months of treatment in 4 cases. The pre- and post-chemotherapy drug concentrations were almost the same. This indicated that healing does not interfere with the penetration of the drugs into the lesion.

[Changes in the levels and composition of glycolipids in various sections of the spinal cord of patients with spinal tuberculosis]. / Izmenenie soderzhaniia i sostava glikolipidov v raznykh otdelakh spinnogo mozga u bol'nykh tuberkulezom pozvonochnika.

The post mortem content and composition of cerebrosides, sulfocerebrosides and gangliosides found in different sections of the spinal marrow of the healthy persons and 5 patients with spinal tuberculosis were analysed. It was shown that the changes in the content of gangliosides were varying in the lesion focus of different patients. In all patients with chronic tuberculous spondylitis, a significant drop in the content of cerebrosides (myelin markers) and sulfocerebrosides in the lesion focus, as compared with controls, was detected. There were some changes found in the content of cerebrosides and gangliosides in the spinal marrow lesion focus, which are also typical of the disease associated with demyelination of the nervous system. The findings suggest that identification of blood anti-cerebroside antibodies in patients with spinal tuberculosis allows one to make an early diagnosis of abnormalities in the cellular structures of nervous tissue in this form of the disease and to predict its course.

[Changes in the content and composition of phospholipids in various portions of the spinal cord in patients with tuberculosis of the spine]. / Izmenenie soderzhaniia i sostava fosfolipidov v raznykh otdelakh spinnogo mozga u bol'nykh tuberkulezom pozvonochnika.

Content and composition of phospholipids in various sections of the spinal marrow were investigated in patients with spinal tuberculosis. In the patients with tuberculous spondylitis, changes in the content of phospholipids in the spinal marrow were observed which were especially pronounced in the injured areas. The changes varied in different patients: in some patients, the content of the lipids was changed insignificantly while in the others it was markedly lowered (5-6-fold). Investigation of the phospholipid composition revealed in some patients decreased relative contents of the phosphatidyl ethanolamine plasmogenic form characteristic of myelin. It was suggested that the spinal marrow cells had undergone destruction in the injured areas.