Characterization of the Subchondral Bone and Pain Behavior Changes in a Novel Bipedal Standing Mouse Model of Facet Joint Osteoarthritis.

BACKGROUND: The subchondral bone parallels with the progression of osteoarthritis (OA). However, the biomechanical properties and histopathological changes of subchondral bone changes in the lumbar facet joint (LFJ) after long-term axial loading on the spine have not been explored. In this study, we aimed to investigate the subchondral bone histopathological changes that occur in the LFJ and pain behaviors in a novel bipedal standing mouse model. METHODS: Sixteen 8-week-old male C57BL/6 mice were randomly assigned into bipedal standing and control groups. A finite element stimulate model based on the micro-CT data was generated to simulate the von Mises stress distribution on the LFJ during different positions. The spine pain behaviors tests were analysis. In addition, the change in the subchondral bone of the LFJ was assessed by histological and immunohistochemistry staining. RESULTS: The computerized simulation of the von Mises stress distribution in the superior articular process of LFJ at the spine level 5 in the lying position increased and reached a maximum value at the bipedal standing posture. The spine pain behavior test revealed that the threshold of pressure tolerance decreased significantly in bipedal groups relative to control groups, whereas the mechanical hyperalgesia of the hind paw increased significantly in bipedal groups relative to control groups. The axial load accelerates LFJ degeneration with increased histological scores in bipedal groups. The expression of type II collagen and aggrecan (ACAN) was significantly decreased in the bipedal groups compared with the control groups, whereas the expression of MMP13 was increased. Compared with the control groups, the osteoclast activity was activated with higher TRAP-positive staining and associated with increased CD-31-positive vessels and GCRP-positive nerve ending expression in the subchondral bone of LFJ. CONCLUSION: Collectively, long-term axial loading induces the development of spine hyperalgesia in mice associate with increased osteoclast activity and aberrant angiogenesis and nerve invasion into the subchondral bone of LFJ that stimulates the natural pathological change in human LFJ OA. These results indicate that aberrant bone remodeling associate with aberrant nerve innervation in the subchondral bone has a potential as a therapeutic target in LFJ OA pain.

Curcumin ameliorates monosodium urate-induced gouty arthritis through Nod-like receptor 3 inflammasome mediation via inhibiting nuclear factor-kappa B signaling.

BACKGROUND: Monosodium urate (MSU) crystals-induced inflammation is a key initiator in gouty arthritis. Curcumin is an active ingredient possessing anti-inflammatory efficacy. But the underlying mechanism is not fully understood and its effect on gouty arthritis remains elusive. METHODS: We evaluated the effects of curcumin on cell viability in primary rat abdominal macrophages with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). Then supernatants of MSU crystals-stimulated cells were collected and subjected to enzyme-linked immunosorbent assay for checking the modulation of curcumin on interleukin (IL)-1ß and tumor necrosis factor (TNF)-α. Meanwhile, cells were analyzed by using Western blot analysis and quantitative polymerase chain reaction (QPCR) to investigate the effects of curcumin on Nod-like receptor 3 (NLRP3) inflammasome/nuclear factor-kappa B (NF-κB) signaling. We also investigated the in vivo efficacy of curcumin with MSU-induced gouty arthritis rat models. RESULTS: Curcumin could reduce MSU crystals-induced IL-1ß and TNF-α in vitro. Western blot analysis and QPCR results revealed that curcumin regulated the production of these cytokines by suppressing the expression of inflammasome key components, including NLRP3, caspase-1. Further studies showed that the suppressive efficacy of curcumin on inflammasome was mediated by inhibiting MSU-induced NF-κB signaling activation. Intraperitoneal administration of curcumin could ameliorate symptoms of MSU-induced gouty arthritis, including the joint circumference, infiltration of neutrophils in knee joints, and production of IL-1ß, TNF-α, and elastase. Western blot analysis revealed that the levels of NLRP3, procaspase-1, caspase-1, pro-IL-1ß, and IL-1ß were downregulated by curcumin in vivo. CONCLUSIONS: These results indicated that curcumin could effectively ameliorate MSU crystal-induced gouty arthritis through NLRP3 inflammasome mediation via inhibiting NF-κB signaling both in vitro and in vivo, suggesting a promising active ingredient for the prevention and treatment of gouty arthritis.

Tetramethylpyrazine Facilitates Functional Recovery after Spinal Cord Injury by Inhibiting MMP2, MMP9, and Vascular Endothelial Cell Apoptosis.

BACKGROUND: Spinal cord injury (SCI) is a major public health issue that leads to neurological dysfunctions and morbidities in patients. Tetramethylpyrazine (TMP) plays a neuroprotective role in SCI; however, the underlying mechanism has not been fully elucidated. OBJECTIVE: In the present study, we aimed to investigate the mechanisms and therapeutic effects of TMP on SCI. METHODS: A contusion SCI model was established that used a modified Allen's method. In the TMP group, TMP (200 mg/kg) was injected daily for 5 days post-injury, while in the Negative Control (NC) group, an equal volume of normal saline was injected. Hindlimb motor function was evaluated using the Basso, Beattie, Bresnahan (BBB) scale. The effects of TMP on protein levels of the matrix metalloproteinases 2 (MMP2) and 9 (MMP9), Bax and cleaved caspase-3 were determined by western blotting. Apoptotic changes in vascular endothelial cells were evaluated using immunofluorescence and TUNEL staining. Alterations in 3D vessel morphology after treatment with TMP were assessed by synchrotron radiation micro-CT (SRµCT). RESULTS: TMP treatment significantly improved recovery in hindlimb motor function and attenuated vascular endothelial cell apoptosis in rats with SCI. Additionally, TMP treatment markedly decreased the protein levels of MMP2 and MMP9, pro-apoptotic bax and cleaved caspase-3 while promoting angiogenesis, as evidenced by vessel visualization using SRµCT. CONCLUSION: These results indicate that TMP attenuated SCI-induced neurological impairments by the down-regulation of the expression of MMP2 and MMP9 proteins, the inhibition of vascular endothelial cell apoptosis, and the promotion of angiogenesis.

[Tetramethylpyrazine accelerated spinal cord repair through regulation of caspase-3 and neurofilament protein expression: an acute spinal cord injury model in rats].

OBJECTIVE: To investigate the effect of tetramethylpyrazine on spinal cord repair through hypothesized pathways including regulation of caspase-3, neurofilament protein NF-L, NF-H, and NF-M expression. METHODS: Allenos weight drop method was used to establish acute spinal cord injury (SCI) rat model at T10 section and was conducted in 80 adult Sprague Dawley rats that were subsequently divided into pseudo-therapy group (sodium chloride group, n=40) and tetramethylpyrazine group (treatment group, n=40). Eight normal rats were included into the pseudo-surgery normal group (n=1 for each time point). The function of spinal cord was evaluated with animal behavioral scores by measuring modified Rivilin loxotic plate degree and counting BBB score at 1, 3, 7, 14, and 21 d postoperatively. The injured spinal cord tissue samples were harvested at 1 h, 3 h, 6 h, 1 d, 3 d, 7 d, 14 d, and 21 d postoperatively (n=5 for each time point) for the preparation of continuous histological sections that were analyzed the expression of caspase-3,NF-L,NF-H, and NF-M by immunohistochemistry method. RESULTS: At postoperative Day 7, 14, and 21, animal behavioral scores revealed higher modified Rivlin loxotic plate degree and BBB scores in the treatment group when compared with those of the control group (P<0.05). The cells in the injured spinal cord tissue expressed significantly less caspase-3 and significantly more NF (NF-L, NF-H and NF-M) in the treatment group than those in the control group at Day 3, 7, and 14 (P<0.05). There was positive correlation between modified Rivlin loxotic plate degree and NF expression, BBB score and NF expression; and negative correlations between BBB score and caspase-3 expression, caspase-3 expression and NF expression. CONCLUSION: Tetramethylpyrazine improves spinal cord healing through regulation of caspase-3 and neurofilament protein expression.

[Effect of low-intensity pulsed ultrasound on the enchondral bone formation in posterolateral lumbar fusion in rabbits].

OBJECTIVE: To observe the effect of daily low-intensity pulsed ultrasound (LIPUs) therapy on improving the enchondral bone formation in lumbar fusion in rabbit models, and to explore its possible mechanism. METHODS: Posterolateral noninstrumented bilateral fusions were performed at the L5 approximately L6 levels in 20 New Zealand rabbits. The autograft iliac bone was implanted on the left side, and the hydroxyapatite bioceramic artificial bone on the right. The rabbits were divided into a treatment group and a control group randomly. One week after the surgery, LIUPs was administered for 20 minutes per day for 4 weeks over the fusion site in the treatment group and false treatment was used in the control group. Post-anterior X-ray photographs were taken to determine the conditions of fusion area, and then, rabbits were killed and the fusion tissues were obtained. Chondrocytes were detected by histological and cytological methods. RESULTS: Compared with the control group, the fusion rate of the treatment group was significantly up-regulated (P<0.05). There was plenty bone trabecula in the fusion area in the treatment group, the number of chondrocytes was also higher than that of the control group (P<0.05), and there was no statistical difference in the number of chondrocytes between the iliac and artificial bone tissues after the treatment(P>0.05). CONCLUSION: Low-intensity pulsed ultrasound therapy improves the endochondral bone formation in the lumbar spine fusion in rabbit models.

[Study on exploring for oil and gas using reflectance spectra of surface soils].

Reflectance spectra in the visible and near-infrared wavelength region provide a rapid and inexpensive means for determining the mineralogy of samples and obtaining information on chemical composition. Hydrocarbon microseepage theory sets up a cause-and-effect relation between oil and gas reservoirs and some special surface alterations. Therefore the authors can explore for oil and gas by determining the reflectance spectra of surface alterations. This determination can be fulfilled by means of field work and hyperspectral remote sensing. In the present paper, firstly a macroscopical feature of reflectance spectra of typical observation points in gas fields is presented. Then a method is proposed in order to provide surface distribution information (e.g., classification) of alterations based on the reflectance spectra determined from the field, and obtain anomaly zones of the special alterations. This method has been applied to the analysis of the reflectance spectra determined in the field of Qinghai X X area, and the classification results tally with the existent gas fields in this area. A robustness analysis of the method shows that good results can be obtained when different combinations of parameters, such as samples, study band regions and thresholds, have been chosen in the process of classification. The valid classification samples and algorithms can be provided for the oil and gas exploration in progress in this area using NASA experimental hyperion hyperspectral satellite.

[Study on exploring for gas based on analysis of spectral absorption features].

Reflectance spectra in the visible and near-infrared wavelength region provide a rapid and inexpensive means for determining the mineralogy of samples and obtaining information on chemical composition. Hydrocarbon microseepage theory establishes a cause-and-effect relation between oil and gas reservoirs and some special surface anomalies. Therefore the authors can explore for oil and gas by determining the reflectance spectra of surface anomalies. This determination can be fulfilled by means of field work and hyperspectral remote sensing. In the present paper, based on the analysis of reflectance spectra determined in the field of Qinghai X X area, firstly, a macroscopic feature of the reflectance spectra of typical observation points in the gas fields is presented. Secondly, absorption-band parameters of spectra such as the position, depth, width, and asymmetry are extracted. Based on the spectral absorption features of the spectra of 144 samples collected from the field, a spectral library for the Qinghai X X area is built to make the detection of the mineral alterations more rapid and reliable. Thirdly, two methods are improved and proposed to detect hydrocarbon microseepage using hydrocarbon absorption bands of reflectance spectra determined from the field. Finally, a linear unmixing model is studied based on the spectra of 144 samples so as to semi-quantitatively determine the abundance fractions of main minerals in the authors' studied area.