Cell life-or-death events in osteoporosis: All roads lead to mitochondrial dynamics.

Mitochondria exhibit heterogeneous shapes and networks within and among cell types and tissues, also in normal or osteoporotic bone tissues with complex cell types. This dynamic characteristic is determined by the high plasticity provided by mitochondrial dynamics and is stemmed from responding to the survival and functional requirements of various bone cells in a specific microenvironments. In contrast, mitochondrial dysfunction, induced by dysregulation of mitochondrial dynamics, may act as a trigger of cell death signals, including common apoptosis and other forms of programmed cell death (PCD). These PCD processes consisting of tightly structured cascade gene expression events, can further influence the bone remodeling by facilitating the death of various bone cells. Mitochondrial dynamics, therefore, drive the bone cells to stand at the crossroads of life and death by integrating external signals and altering metabolism, shape, and signal-response properties of mitochondria. This implies that targeting mitochondrial dynamics displays significant potential in treatment of osteoporosis. Considerable effort has been made in osteoporosis to emphasize the parallel roles of mitochondria in regulating energy metabolism, calcium signal transduction, oxidative stress, inflammation, and cell death. However, the emerging field of mitochondrial dynamics-related PCD is not well understood. Herein, to bridge the gap, we outline the latest knowledge on mitochondrial dynamics regulating bone cell life or death during normal bone remodeling and osteoporosis.

Cell death regulation: A new way for natural products to treat osteoporosis.

Osteoporosis is a common metabolic bone disease that results from the imbalance of homeostasis within the bone. Intra-bone homeostasis is dependent on a precise dynamic balance between bone resorption by osteoclasts and bone formation by mesenchymal lineage osteoblasts, which comprises a series of complex and highly standardized steps. Programmed cell death (PCD) (e.g., apoptosis, autophagy, ferroptosis, pyroptosis, and necroptosis) is a cell death process that involves a cascade of gene expression events with tight structures. These events play a certain role in regulating bone metabolism by determining the fate of bone cells. Moreover, existing research has suggested that natural products derived from a wide variety of dietary components and medicinal plants modulate the PCDs based on different mechanisms, which show great potential for the prevention and treatment of osteoporosis, thus revealing the emergence of more acceptable complementary and alternative drugs with lower costs, fewer side effects and more long-term application. Accordingly, this review summarizes the common types of PCDs in the field of osteoporosis. Moreover, from the perspective of targeting PCDs, this review also discussed the roles of currently reported natural products in the treatment of osteoporosis and the involved mechanisms. Based on this, this review provides more insights into new molecular mechanisms of osteoporosis and provides a reference for developing more natural anti-osteoporosis drugs in the future.

Association of spinopelvic index with proximal junctional failure developing in adult spinal deformity after surgical treatment: an observational study.

BACKGROUND: Those pelvic parameters of sacral slope (SS) and pelvic tilt (PT) correlated significantly to lumbar spine and hip joints respectively. We proposed the match between SS and PT, namely spinopelvic index (SPI), in order to investigate whether the SPI correlated to proximal junctional failure (PJF) in adult spinal deformity (ASD) after correction surgery. METHODS: Ninety-nine ASD patients who had undergone long-fusion (≥ 5 vertebras) surgeries were reviewed retrospectively in two medical institutions from January 2018 to December 2019. Those SPI were calculated with the equation: SPI = SS/PT, and analyzed using the receiver operating characteristic curve (ROC) analysis. All participants were subdivided into the observational and control group. Comparisons of demographics, surgical and radiographic data between the two groups were performed. A Kaplan-Meier curve and log-rank test was used to analyze the differences in PJF-free survival time, and the 95% confidence intervals (CI) were recorded respectively. RESULTS: Nineteen patients suffering from PJF had much smaller postoperative SPI (P = 0.015), but much larger TK postoperatively (P < 0.001). ROC analysis determined the best cutoff value of 0.82 for SPI (sensitivity = 88.5%, specificity = 57.9%; AUC = 0.719, 95%CI: 0.612-0.864; P = 0.003). There were 19 and 80 cases in the observational (SPI ≤ 0.82) and control group (SPI > 0.82) respectively. The incidence of PJF in the observational group was much higher (11/19 VS 8/80, P < 0.001); further logistic regression analysis showed that SPI ≤ 0.82 was associated with increased odds of PJF (odds ratio: 12.375; 95%CI: 3.851-39.771). PJF-free survival time in the observational group decreased significantly (P < 0.001, log-rank test), moreover, multivariate analysis demonstrated that a value of SPI ≤ 0.82 (HR 6.626, 95%CI: 1.981-12.165) was significantly associated with PJF. CONCLUSIONS: For ASD patients underwent long-fusion surgeries, the SPI should be over 0.82. The incidence of PJF may increase by 12-fold in such individuals with the immediate SPI ≤ 0.82 postoperatively.

Factors associated with intervertebral cage subsidence in posterior lumbar fusion.

BACKGROUND: The interbody fusion apparatus is a key component of the operation and plays a key role in the postoperative efficacy. Cage subsidence is one of the common complications after lumbar fusion and internal fixation. Clinical studies on the risk factors of cage subsidence are incomplete and inaccurate, especially paravertebral muscle atrophy and intervertebral bone fusion time. METHODS: Among the patients who underwent PLIF surgery in our hospital from January 2016 to January 2019, 30 patients with cage subsidence and 30 patients without cage subsidence were randomly selected to be included in this study. The differences between the two groups were compared, and the relevant factors of cage subsidence were explored by single factor comparison and multiple logistic regression analysis. RESULTS: Bone mineral density (T) of the subsidence group [(- 1.84 ± 1.81) g/cm2 vs (- 0.87 ± 1.63) g/cm2, P = 0.018] was significantly lower than that of the normal group. There were 4 patients with end plate injury in the subsidence group (P = 0.038). Preoperative end plate Modic changes [I/II/III, (7/2/2) vs (2/5/8), P = 0.043] were significantly different between the two groups. In the subsidence group, preoperative rCSA of psoas major muscle [(1.43 ± 0.40) vs (1.64 ± 0.41), P = 0.043], CSA of paravertebral muscle [(4530.25 ± 776.55) mm2 vs (4964.75 ± 888.48) mm2, P = 0.047], paravertebral muscle rCSA [(3.03 ± 0.72) vs (3.84 ± 0.73), P < 0.001] and paravertebral muscle rFCSA [(2.29 ± 0.60) vs (2.89 ± 0.66), P < 0.001] were significantly lower than those in normal group. In the subsidence group, the vertebral body area [(1547.81 ± 309.89) mm2 vs (1326.48 ± 297.21) mm2, P = 0.004], the height of the immediately corrected vertebral space [(2.86 ± 1.10) mm vs (1.65 ± 1.02) mm, P = 0.020], immediately SL corrective Angle [(5.81 + 4.71)° vs (3.24 + 3.57) °, P = 0.009), postoperative PI-LL [(11.69 + 6.99)° vs (6.66 + 9.62) °, P = 0.029] and intervertebral fusion time [(5.38 ± 1.85) months vs (4.30 ± 1.49) months, P = 0.023] were significantly higher than those in the normal group. Multivariate logistic regression analysis showed that the time of intervertebral fusion (OR = 1.158, P = 0.045), the height of immediate intervertebral space correction (OR = 1.438, P = 0.038), and the Angle of immediate SL correction (OR = 1.101, P = 0.019) were the risk factors for cage subsidence. Bone mineral density (OR = 0.544, P = 0.016) and preoperative paravertebral muscle rFCSA (OR = 0.525, P = 0.048) were protective factors. CONCLUSION: Intervertebral fusion time, correctable height of intervertebral space, excessive Angle of immediate SL correction, bone mineral density and preoperative paravertebral muscle rFCSA are risk factors for cage subsidence after PLIF.

Wnt-3a improves functional recovery after spinal cord injury by regulating the inflammatory and apoptotic response in rats via wnt/ß-catenin signaling pathway.

The specific molecular mechanism of neuroprotective effects of wnt-3a on spinal cord injury (SCI) has not been elucidated. In our study, we evaluated the recovery of motor function after SCI by BBB, observed neuronal apoptosis by western blot and TUNEL, observed the changes of neuronal inflammation by western blot and immunofluorescence staining, and observed the changes of motoneurons and spinal cord area in the anterior horn of the spinal cord via Nissl and HE staining. We found that wnt-3a could significantly promote the recovery of motor function, reduce the loss of motor neurons in the anterior horn of the spinal cord, promote the recovery of injured spinal cord tissue, inhibit neuronal apoptosis and inflammatory response, and ultimately promote neuronal function after SCI. However, when XAV939 inhibits the wnt/ß-catenin signaling pathway, the neuroprotective effects of wnt-3a are also significantly inhibited. The above results together indicated that wnt-3a exerts its neuroprotective effect on after SCI via activating the wnt/ß-catenin signaling pathway.

Will coronal alignment postoperatively be deteriorating in adult spinal deformity after long-fusion surgery?

BACKGROUND: To investigate whether the coronal alignment (CA) will deteriorate, and identify the risk factors for coronal malalignment (CM) developing in adult spinal deformity (ASD) after long-fusion surgery. METHODS: A multi-center retrospective study was performed, which included a total of 161 ASD patients who had undergone the surgical procedure of long-fusion (≥ 5 vertebras) with instrumentations in three medical centers. All of the participants were retrospectively reviewed, and subsequently assigned into the consistency group (C7 plumb line (C7PL) shifting towards the convex side of the main curve), and the opposition group (C7PL shifting towards the concave side). CM was considered if the coronal balance distance (CBD) being over 30 mm. A Kaplan-Meier curve and log-rank test were used to analyze the differences in CM-free survival during follow-up. Multivariate analysis via a Cox proportional hazards test was used to analyze the risk factors. RESULTS: Patients showing CM equaled 35 (21.7%) at the pre-operation, and that increased significantly up to 51 (31.7%) at the final follow-up (P = 0.04). In the consistency group, the incidence of CM at the final follow-up was much higher than that preoperatively (35:16, P = 0.002). CM-free survival time decreased significantly in patients with larger CBD correction, pelvic fixation and more instrumented segments, respectively, during follow-up (P < 0.05, log-rank test). Age ≥ 60 years, the consistency CA, pelvic fixation, CBD-correction ≥ 30 mm and fixed-vertebra ≥ 8 were risk factors for CM happening after surgery using multivariate regression analysis (P < 0.05). CONCLUSIONS: The coronal alignments in ASD patients underwent long-fusion surgeries may deteriorate during follow-up, for which the risk factors include the consistency CA, age ≥ 60, fixed-vertebra ≥ 8, CBD-correction ≥ 30 mm and pelvic fixation.

Construction of a nomogram to predict the probability of new vertebral compression fractures after vertebral augmentation of osteoporotic vertebral compression fractures: a retrospective study.

Objective: This study aimed to develop and validate a new nomogram model that can predict new vertebral fractures after surgery for osteoporotic compression fractures to optimize surgical plans and reduce the incidence of new vertebral compression fractures. Methods: 420 patients with osteoporotic vertebral compression fractures were randomly sampled using a computer at a fixed ratio; 80% of the patients were assigned to the training set, while the remaining 20% were assigned to the validation set. The least absolute shrinkage and selection operator (LASSO) regression method was applied to screen the factors influencing refracture and construct a predictive model using multivariate logistic regression analysis. Results: The results of the multivariate logistic regression analysis showed a significant correlation between bone cement leakage, poor cement dispersion, the presence of fractures in the endplate, and refractures. The receiver operating characteristic curve (ROC) results showed that the area under the ROC curve (AUC) of the training set was 0.974 and the AUC of the validation set was 0.965, which proves that this prediction model has a good predictive ability. The brier score for the training set and validation set are 0.043 and 0.070, respectively, indicating that the model has high accuracy. Moreover, the calibration curve showed a good fit with minimal deviation, demonstrating the model's high discriminant ability and excellent fit. The decision curve indicated that the nomogram had positive predictive ability, indicating its potential as a practical clinical tool. Conclusion: Cement leakage, poor cement dispersion, and presence of fractures in the endplate are selected through LASSO and multivariate logistic regressions and included in the model development to establish a nomogram. This simple prediction model can support medical decision-making and maybe feasible for clinical practice.

Bushen Huoxue Formula Inhibits IL-1ß-Induced Apoptosis and Extracellular Matrix Degradation in the Nucleus Pulposus Cells and Improves Intervertebral Disc Degeneration in Rats.

Background: The method of action of Bushen Formula (BSHXF) in the treatment of intervertebral disc degeneration (IVDD) was uncovered in this work using in vivo and in vitro tests. To clarify the mechanism of action of BSHXF, we validated the rat intervertebral disc degeneration model and the nucleus pulposus cell degeneration model. Methods: In an in vivo model of IVDD the study explores the impact of BSHXF on mitochondrial function, pro-inflammatory cytokines, pro-apoptotic factors, and matrix metalloproteinases. Additionally, it evaluates the induced degeneration of nucleus pulposus (NP) cells in an in vitro model stimulated by interleukin-1 ß (IL-1ß). The study measures the effects of BSHXF on both the inflammatory response and mitochondrial function. Results: The MRI results showed that BSHXF reduced intervertebral disc volume reduction and degradation of NP tissue. HE, SO-FG and immunofluorescence further confirmed the protective effect of BSHXF on degenerative intervertebral discs. BSHXF reduced the inflammatory levels of IL-6 IL-1ß and TNF-α in degenerative intervertebral disc tissue. Meanwhile, JC-1, mPTP and ROS detection revealed that BSHXF can restore mitochondrial function by regulating the expression of antioxidant proteins, playing a protective role in NP cells. Finally, the WB results showed that BSHXF can alleviate IL-1ß mediate the degeneration of NP cells. BSHXF can alleviate NP cell apoptosis by inhibiting the expression of bax, cleaved caspase-3, caspase-3, and cyt-c, and increasing the expression of Bcl-2. Conclusion: This study reveals that BSHXF inhibits the development of inflammatory factors, which may play a significant role in intervertebral disc degeneration. This implies that BSHXF is a suitable herbal medication for future research into inflammatory cytokine treatment.

Magnetic resonance imaging classification in a percutaneous needle injury rat model of intervertebral disc degeneration.

BACKGROUND: During the development of disease-modifying intervertebral disc degeneration (IDD) drugs, the rat model of IDD is frequently used for disease progression assessment. The aim of this study was to describe a magnetic resonance (MRI) scoring system for the assessment of different disc conditions in puncture-induced IDD, allowing standardization and comparison of results obtained by different investigators. METHODS: A total of 36 Sprague-Dawley rats were utilized in the present study. The animals were divided into two groups: a sham group and an IDD group caused by puncture. The rats in the IDD group were subsequently divided into six categories based on time frames, with five rats in each category. The sham group was divided into two sub-groups (n = 3) for 28 and 56 days, respectively. T2-weighted images of rats consecutively studied with MRI of the coccygeal discs were classified according to the time course using the corresponding histological data. Additional scoring of the micro-CT was employed to identify the progression of bone destruction of the rat model of IDD. RESULTS: A comparison of the MRI results between the sham group and the IDD group revealed a significant reduction in NP height, area, T2WI value, and DHI in the latter group (P < 0.05). The micro-CT results demonstrated that following acupuncture, there was a notable decline in the BV, Tb.N, and height of the coccygeal vertebra, while the BS/BV and Tb.Sp exhibited a significant increase (P < 0.05). The histological results were analogous to the MRI results, indicating a progressive exacerbation of IDD and a corresponding increase in NP score (P < 0.05). The results of the MRI were found to be consistent with those of the micro-CT and histological analyses (P < 0.05). The results of the study demonstrate a robust correlation between MRI analysis and histological findings. Live animals are employed for MRI analysis to improve experiment comparability. The reliability of the MRI scoring system ensures assessment of disease progression in live animals, while promoting cost savings and animal welfare by avoiding the sacrifice of animals at different times. CONCLUSIONS: The described scoring paradigm has quantitatively been found to differentiate IDD disease progression in an in vivo rat model. Hence, we suggest employing it to evaluate the rat IDD model and assess the effects of treatments in this model.

Association of ADAMTS-5 gene polymorphisms with the susceptibility to knee osteoarthritis in a Chinese Han population.

BACKGROUND: Osteoarthritis (OA) is the most prevalent type of arthritis and the main reason for progressive disability in middle-aged and older people. Studies of candidate genes may provide a novel insight and treatment strategy for knee osteoarthritis (KOA). The aim of this study was to investigate the relationship between KOA susceptibility and single-nucleotide polymorphism (SNP) of the ADAMTS-5 gene. MATERIALS AND METHODS: The case group included 188 patients from Luoyang Orthopedic Hospital with clinically and radiographically diagnosed primary KOA, and the control group included 100 age-matched individuals without KOA. Fifteen ADAMTS-5 SNPs were assayed using MALDI-TOF MS. Allelic and haplotypic frequencies were compared between the groups. The relationship between genotype distribution and risk of KOA was analyzed by multivariate logistic regression. RESULTS: The frequency of A allele in rs2249350 site in the KOA group was significantly lower (odds ratio [OR]: 0.761; 95% confidence interval [95% CI]: 0.612-0.947; P = 0.016), while that of C allele was higher than that in the control group (OR: 1.176; 95% CI: 1.025-1.351; P = 0.016). AA genotype and gene model, especially recessive gene model at rs2249350 locus, negatively correlated with KOA risk after adjustment for sex, body mass index, age, and occupation (AA vs. CC: OR: 0.288; 95% CI: 0.124-0.669; P = 0.004; AA vs. CA + CC: OR: 0.348; 95% CI: 0.162-0.749; P = 0.007). Meanwhile, one protective haplotype, GA (rs229054, rs2249350) (OR: 0.763; 95% CI: 0.614-0.949; P = 0.017), and one high-risk haplotype, GC (rs229054, rs2249350) (OR: 1.259; 95% CI: 1.032-1.537; P = 0.019), were found in this study. CONCLUSION: Despite a limited sample size, our study suggests that the rs2249350 polymorphism in the ADAMTS-5 gene is one of the genetic factors influencing the risk of KOA. The A allele and AA genotype of rs2249350 may protect from KOA, whereas C allele and CC genotype increase the risk of KOA. In addition, the GA haplotype (rs229054, rs2249350) might be associated with a decreased risk of KOA, whereas the GC haplotype (rs229054, rs2249350) may be a risk factor for KOA. Additional larger-sized studies in more ethnically diverse populations are needed to confirm these findings.

Pyroptosis: A spoiler of peaceful coexistence between cells in degenerative bone and joint diseases.

BACKGROUND: As people age, degenerative bone and joint diseases (DBJDs) become more prevalent. When middle-aged and elderly people are diagnosed with one or more disorders such as osteoporosis (OP), osteoarthritis (OA), and intervertebral disc degeneration (IVDD), it often signals the onset of prolonged pain and reduced functionality. Chronic inflammation has been identified as the underlying cause of various degenerative diseases, including DBJDs. Recently, excessive activation of pyroptosis, a form of programed cell death (PCD) mediated by inflammasomes, has emerged as a primary driver of harmful chronic inflammation. Consequently, pyroptosis has become a potential target for preventing and treating DBJDs. AIM OF REVIEW: This review explored the physiological and pathological roles of the pyroptosis pathway in bone and joint development and its relation to DBJDs. Meanwhile, it elaborated the molecular mechanisms of pyroptosis within individual cell types in the bone marrow and joints, as well as the interplay among different cell types in the context of DBJDs. Furthermore, this review presented the latest compelling evidence supporting the idea of regulating the pyroptosis pathway for DBJDs treatment, and discussed the potential, limitations, and challenges of various therapeutic strategies involving pyroptosis regulation. KEY SCIENTIFIC CONCEPTS OF REVIEW: In summary, an interesting identity for the unregulated pyroptosis pathway in the context of DBJDs was proposed in this review, which was undertaken as a spoiler of peaceful coexistence between cells in a degenerative environment. Over the extended course of DBJDs, pyroptosis pathway perpetuated its activity through crosstalk among pyroptosis cascades in different cell types, thus exacerbating the inflammatory environment throughout the entire bone marrow and joint degeneration environment. Correspondingly, pyroptosis regulation therapy emerged as a promising option for clinical treatment of DBJDs.

A repeated strike loading organ culture model for studying compression-associated chronic disc degeneration.

Mechanical stress has been viewed as one of the key risk factors in accelerating the intervertebral disc degeneration process. The goal of the present study was to employ a repeated strike loading bovine caudal disc system to elucidate the pathophysiological impacts of cumulative mechanical stress on the disc. The discs in the model groups were subjected to two different mechanical stresses: one strike loading or repeated strike loading. The following indices were analyzed: histological morphology, glycosaminoglycan release, disc height, cell viability, apoptosis-related protein expression, and catabolism-related gene expression. Both mechanical stress modes induced degenerative changes in the discs by day 11, such as clefts and delamination of the annulus fibrosus; they increased glycosaminoglycan release. Cell viability was significantly decreased and catabolic gene expression was significantly up-regulated in the degenerative loading group and repeated strike loading group by day 9. These alterations remained evident in the annulus fibrosus tissue of the repeated strike loading group on day 11. Our data suggests that the repeated strike loading model adopted in this study could lead to degenerative changes in the disc organ model. Annulus fibrosus cells displayed a more noticeable response to mechanical stress damage and a slower recovery process, suggesting that the annulus fibrosus serves as a pivotal factor in disc degeneration due to mechanical stress injuries. The study also indicates that due to the gradual self-repair of intervertebral disc cells after injury, it is necessary to apply repeated strike loading on the disc at specific intervals when researching the repair of chronic disc injuries.

Naringin promotes osteoblast differentiation and effectively reverses ovariectomy-associated osteoporosis.

BACKGROUND: Osteoporosis is a common pathological condition that influences 20 % of women over 50 years of age. This condition decreases bone strength and increases the risk of bone fracture. Naringin is a major flavonoid found in grapefruit and an active compound extracted from a Chinese herbal medicine (Rhizoma Drynariae). Studies have shown that naringin possesses many pharmacological effects. The current study evaluated the influence of naringin on osteoblastic cell differentiation and proliferation, and assessed its therapeutic effects on a rat osteoporosis model. METHOD: The proliferation, differentiation, and function of rat bone marrow stromal cells (BMSCs) were determined following treatment with various concentrations of naringin. Ovariectomy (OVX)-induced osteoporotic rats were orally administered naringin daily at low, medium, and high dosages, while a control group received PBS for 2 months. Femoral X-ray images and microCT scans were used for bone mineral density (BMD) and BV/TV (bone volume/total volume) analyses, and histological assessments of left tibiae were employed to check for changes in trabecular thickness (Tb.Th) and trabecular space (Tb.Sp) in the groups. RESULTS: Naringin was effective at enhancing the proliferation and osteogenic differentiation of BMSCs, and a concentration of 10 µg/ml prompted the highest levels of osteocalcin expression among the in vitro study groups. There appeared to be a delayed response pattern of BMSCs to the naringin treatment. Naringin also effectively reversed OVX-induced bone loss via increasing BMD, bone volume, and trabecular thickness. The medium dose (300 mg/kg) appeared to be the optimal dosage for delivering satisfactory therapeutic effects. CONCLUSION: Naringin promotes the proliferation and differentiation of BMSCs, and increases osteocalcin expression. Naringin also effectively reverses ovariectomy-induced osteoporosis in rats. The study suggests that naringin administration may represent an effective treatment for osteoporosis.

3D,2D-QSAR study and docking of novel quinazolines as potential target drugs for osteosarcoma.

Background: Quinazolines are an important class of benzopyrimidine heterocyclic compounds with a promising antitumor activity that can be used for the design and development of osteosarcoma target compounds. Objective: To predict the compound activity of quinazoline compounds by constructing 2D- and 3D-QSAR models, and to design new compounds according to the main influencing factors of compound activity in the two models. Methods: First, heuristic method and GEP (gene expression programming) algorithm were used to construct linear and non-linear 2D-QSAR models. Then a 3D-QSAR model was constructed using CoMSIA method in SYBYL software package. Finally, new compounds were designed according to molecular descriptors of 2D-QSAR model and contour maps of 3D-QSAR model. Several compounds with optimal activity were used for docking experiments with osteosarcoma related targets (FGFR4). Results: The non-linear model constructed by GEP algorithm was more stable and predictive than the linear model constructed by heuristic method. A 3D-QSAR model with high Q2 (0.63) and R 2 (0.987) values and low error values (0.05) was obtained in this study. The success of the model fully passed the external validation formula, proving that the model is very stable and has strong predictive power. 200 quinazoline derivatives were designed according to molecular descriptors and contour maps, and docking experiments were carried out for the most active compounds. Compound 19g.10 has the best compound activity with good target binding capability. Conclusion: To sum up, the two novel QSAR models constructed were very reliable. The combination of descriptors in 2D-QSAR with COMSIA contour maps provides new design ideas for future compound design in osteosarcoma.

Achyranthoside D attenuates chondrocyte loss and inflammation in osteoarthritis via targeted regulation of Wnt3a.

BACKGROUND: Achyranthes bidentata Blume (A. bidentata) is a common Chinese herb used to treat osteoarthritis (OA). Achyranthoside D (Ach-D) is a glucuronide saponin isolated from A. bidentata. PURPOSE: To assess the mechanisms of action of Ach-D and its effects on OA. METHODS: The effects of Ach-D were evaluated in rats underwent anterior cruciate ligament transection (ACLT) with medial meniscectomy (MMx) and in interleukin (IL)-1ß-induced chondrocytes. Histological changes in rat cartilage tissues were detected using Safranin O-Fast green and haematoxylin-eosin staining. Immunohistochemical staining, qRT-PCR, ELISA, immunoblotting, and immunofluorescence were conducted to examine cartilage degeneration-related and inflammation-related factor expression. CCK-8, LDH assay, and EdU staining were performed to detect chondrocyte death. RESULTS: Ach-D dose-dependently reduced the Osteoarthritis Research Society International (OARSI) scores, alleviated cartilage injury, and decreased the serum concentrations of CTX-II and COMP in ACLT-MMx models. Ach-D increased the expression levels of collagen II and aggrecan and decreased the levels of cartilage degeneration-related proteins, ADAMTS-5, MMP13, and MMP3, in rat cartilage tissues. Additionally, nod-like receptor protein 3 (NLRP3)-related inflammation was reduced by Ach-D, as shown by the significantly inhibited expression levels of NLRP3, ASC, GSDMD, IL-6, TNF-α, IL-1ß, and IL-18 in rat cartilage tissues. In primary rat chondrocytes, Ach-D protected against IL-1ß-induced viability loss and LDH release. Wnt3a is the target protein of Ach-D. Mechanistically, Ach-D alleviated OA by inhibiting Wnt signalling. CONCLUSION: ACH-D may reduce inflammation and cartilage degeneration by inhibiting the Wnt signalling pathway, thereby reducing OA.

How does the correction in lumbar lordosis affect the spinopelvic realignments in degenerative lumbar scoliosis underwent scoliosis surgery?

BACKGROUND: To evaluate the effects of correction in lumbar lordosis (LL) that have on full-body realignments in patients with degenerative lumbar scoliosis (DLS) who had undergone long sacroiliac fusion surgery. METHODS: A multi-center retrospective study including 88 DLS patients underwent the surgical procedure of long sacroiliac fusion with instrumentations was performed. Comparisons of radiographic and quality-of-life (QoL) data among that at the pre-operation, the 3rd month and the final follow-up were performed. The correlations between the LL correction and the changes in other spinopelvic parameters were explored using Pearson-correlation linear analysis and linear regression analysis. The correlation coefficient (r) and the adjusted r2 were calculated subsequently. RESULTS: All radiographic and QoL data improved significantly (P < 0.001) after the surgical treatments. The LL correction correlated (P < 0.001) with the changes in the sacral slope (SS, r = 0.698), pelvic tilt (PT, r = -0.635), sagittal vertical axis (SVA, r = -0.591), T1 pelvic angle (TPA, r = -0.782), and the mismatch of pelvic incidence minus lumbar lordosis (PI-LL, r = -0.936), respectively. Moreover, LL increased by 1° for each of the following spinopelvic parameter changes (P < 0.001): 2.62° for SS (r2 = 0.488), -4.01° for PT (r2 = 0.404), -4.86° for TPA (r2 = 0.612), -2.08° for the PI-LL (r2 = 0.876) and -15.74 mm for SVA (r2 = 0.349). Changes in the thoracic kyphosis (r = 0.259) and pelvic femur angle (r = 0.12) were independent of the LL correction, respectively. CONCLUSIONS: LL correction correlated significantly to the changes in spinopelvic parameters; however, those independent variables including the thoracic spine and hip variables probably be remodeled themselves to maintain the full-body balance in DLS patients underwent the correction surgery.

IRF8 and its related molecules as potential diagnostic biomarkers or therapeutic candidates and immune cell infiltration characteristics in steroid-induced osteonecrosis of the femoral head.

PURPOSE: Steroid-induced osteonecrosis of the femoral head (SONFH) was a refractory orthopedic hip joint disease in the young and middle-aged people, but the pathogenesis of SONFH remained unclear. We aimed to identify the potential genes and screen potential therapeutic compounds for SONFH. METHODS: The microarray was obtained for blood tissue from the GEO database, and then it identifies differentially expressed genes (DEGs). The DEGs were analyzed to obtain the differences in immune cell infiltration. The gene functional enrichment analysis of SONFH was analyzed. The PPI of DEGs was identified through the STRING database, and the cluster modules and hub genes were ascertained using MCODE and CytoHubba, and the ROC curve of hub genes was analyzed, and the tissues distribution of hub genes was understood by the HPA, Bgee and BioGPS databases. The hub genes and target miRNAs and corresponding upstream lncRNAs were predicted by TargetScan, miRDB and ENCORI database. Subsequently, we used CMap, DGIdb and L1000FWD databases to identify several potential therapeutic molecular compounds for SONFH. Finally, the AutoDockTools Vina, PyMOL and Discovery Studio were employed for molecular docking analyses between compounds and hub genes. RESULTS: The microarray dataset GSE123568 was obtained related to SONFH. There were 372 DEGs including 197 upregulated genes and 175 downregulated genes by adjusted P value < 0.01 and |log2FC|> 1. Several significant GSEA enrichment analysis and biological processes and KEGG pathway associated with SONFH were identified, which were significantly related to cytoskeleton organization, nucleobase-containing compound catabolic process, NOD-like receptor signaling pathway, MAPK signaling pathway, FoxO signaling pathway, neutrophil-mediated immunity, neutrophil degranulation and neutrophil activation involved in immune response. Activated T cells CD4 memory, B cells naïve, B cells memory, T cells CD8 and T cells gamma delta might be involved in the occurrence and development of SONFH. Three cluster modules were identified in the PPI network, and eleven hub genes including FPR2, LILRB2, MNDA, CCR1, IRF8, TYROBP, TLR1, HCK, TLR8, TLR2 and CCR2 were identified by Cytohubba, which were differed in bone marrow, adipose tissue and blood, and which had good diagnostic performance in SONFH. We identified IRF8 and 10 target miRNAs that was utilized including Targetsan, miRDB and ENCORI databases and 8 corresponding upstream lncRNAs that was revealed by ENCORI database. IRF8 was detected with consistent expression by qRT-PCR. Based on the CMap, DGIdb and L1000FWD databases, the 11 small molecular compounds that were most strongly therapeutic correlated with SONFH were estradiol, genistein, domperidone, lovastatin, myricetin, fenbufen, rosiglitazone, sirolimus, phenformin, vorinostat and vinblastine. All of 11 small molecules had good binding affinity with the IRF8 in molecular docking. CONCLUSION: The occurrence of SONFH was associated with a "multi-target" and "multi-pathway" pattern, especially related to immunity, and IRF8 and its noncoding RNA were closely related to the development of SONFH. The CMap, DGIdb and L1000FWD databases could be effectively used in a systematic manner to predict potential drugs for the prevention and treatment of SONFH. However, additional clinical and experimental research is warranted.

Bushen Huoxue Formula Modulates Autophagic Flux and Inhibits Apoptosis to Protect Nucleus Pulposus Cells by Restoring the AMPK/SIRT1 Pathway.

Background: Low back pain (LBP) has the characteristics of chronic and persistence, which is a heavy social burden. Intervertebral disc degeneration (IVDD) is a major cause of LBP. The typical features of IVDD are extracellular matrix (ECM) degradation and nucleus pulposus cell (NP) apoptosis. Bushen Huoxue Formula (BSHXF) has good clinical effects on LBP. However, the mechanism of BSHXF affecting ECM and NP cells is still unclear. Aim of the Study. In this study, the impact of BSHXF on autophagy and apoptosis of NP cells was studied through the AMPK/SIRT1 pathway. Material and Methods. NP cells were extracted through the digestion of collagenase and trypsin, and the components of BSHXF were identified. Cell Counting Kit-8 was applied to detect the impact of BSHXF on NP cells. Mitochondrial function was detected using MitoTracker assay, ATP kit, and SOD kit. Autophagy and apoptosis were detected by RT-qPCR, western blotting, and flow cytometry. Results: BSHXF promoted NP cell survival in a concentration-dependent manner, and the elimination of rat serum did not increase cell proliferation; TNF-α accelerated ECM degradation, ROS accumulation, and NP cell apoptosis and decreased autophagic flux. BSHXF restored mitochondrial function and autophagic flux. In addition, AMPK/SIRT1 pathway activation was associated with IVDD. Conclusions: BSHXF regulates autophagy and enhances autophagic flux to suppress excessive ROS production and restore mitochondrial function in an AMPK/SIRT1-dependent manner. However, the protection of BSHXF on TNF-α-treated cells was eliminated by 3-MA. Furthermore, the protective impact of BSHXF on ECM degradation and apoptosis induced by TNF-α was restrained by an AMPK inhibitor. Therefore, maintaining the proper autophagy illustrates treatment strategy for IVDD.

The common regulatory pathway of COVID-19 and multiple inflammatory diseases and the molecular mechanism of cepharanthine in the treatment of COVID-19.

Background: Similar pathogenesis makes Corona Virus Disease 2019 (COVID-19) associated with rheumatoid arthritis (RA), ankylosing spondylitis (AS) and gouty arthritis (GA), and it is possible to introduce common drugs for the treatment of RA, AS and GA into the treatment of COVID-19. That is, "homotherapy for heteropathy", especially cytokine inhibitors. But little is known about the specific link between the diseases. In addition, "new use of old drugs" is an important short-term strategy for the treatment of COVID-19. Cepharanthine (CEP), a monomer component of traditional Chinese medicine (TCM), is mainly used in the treatment of leukopenia and has recently been proved to have a good therapeutic effect on COVID-19, but its specific molecular mechanism has not been clearly explained. The purpose of this work is to explore the common targets and signaling pathways among COVID-19, RA, AS, and GA by means of network pharmacology (NP), and to infer the potential mechanism of CEP in the treatment of COVID-19. Methods: Firstly, SwissTargetPrediction was used to predict the targets of CEP, and the pathogenic targets of COVID-19, RA, AS and GA were searched in GeneCards, OMIM, TTD, PharmGKB database and literature, respectively. Then, the protein interaction network of CEP and COVID-19 cross targets and the common targets of COVID-19, RA, AS and GA was constructed. Cytosscape 3.7.2 software was used to construct CEP-common targets-signaling pathways-COVID-19 network, module function analysis, gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG). Finally, the molecular docking of hub targets and CEP was carried out by AutoDock software. Results: The results showed that the common targets of the four diseases were tumor necrosis factor (TNF), interleukin (IL)-6 and IL-1ß, and involved Coronavirus disease, IL-17 signaling pathway and TNF signaling pathway. CEP has a good binding force with AKT Serine/Threonine Kinase 1 (AKT1), phosphatidylinositol 3-kinase (PIK3) CA, PIK3CD and Angiotensin-converting enzyme 2 (ACE2), and plays a role in the treatment of COVID-19 by regulating PI3K-Akt signaling pathway, Relaxin signaling pathway, VEGF signaling pathway and HIF-1 signaling pathway. Conclusion: Therefore, this study not only confirmed the potential mechanism of CEP in the treatment of COVID-19 at the molecular level, but also found that TNF and IL-17 inhibitors, which are commonly used in the treatment of RA, AS and GA, may also affect the treatment of COVID-19, which provides new clues and theoretical basis for the rapid discovery of effective therapeutic drugs for COVID-19.

Surgical treatment of thoracolumbar fracture in ankylosing spondylitis: A comparison of percutaneous and open techniques.

BACKGROUND CONTEXT: Posterior percutaneous long-segment internal fixation and open fixation with long-segment screws have been used to treat thoracolumbar fractures in ankylosing spondylitis patients. PURPOSE: To observe the clinical effect of posterior percutaneous long-segment internal fixation in 26 ankylosing spondylitis (AS) patients with thoracolumbar fractures. STUDY DESIGN: Retrospective cohort study. PATIENT SAMPLE: Forty-seven AS patients who were diagnosed with thoracolumbar fractures and treated from December 2014 to December 2018. OUTCOME MEASURES: Visual analog scale score, Cobb angle, American Spinal Injury Association Grade, SF-Qualiveen score, pedicle screw misplacement rate, operative duration, blood loss, complications, bed rest duration and modified MacNab score. METHODS: All patients were divided into the percutaneous group (PG) and the open group. Twenty-six patients were treated with percutaneous long-segment internal fixation, and the remaining 21 underwent open fixation with long-segment screws. The minimum follow-up period was 12 months. RESULTS: The operations were successful in both groups. A patient in the PG showed class C wound healing, while the others showed class A healing, and some patients experienced perioperative complications. All patients were followed up for 12-48 months (mean, 33.81 months), and all patients showed clinical osseous fracture healing. Significant differences were found in operative duration, intraoperative blood loss and postoperative bed rest duration between the two groups (P < 0.05). No significant difference was found in improvement of the visual analog scale score, Cobb angle of spinal kyphosis or neurological function after the operation (P > 0.05). CONCLUSIONS: As a minimally invasive procedure, posterior percutaneous long-segment internal fixation requires less time, results in less blood loss and causes less trauma. This procedure can also improve patients' pain, neurological function and kyphotic deformity and achieve effects similar to those of traditional methods. With this curative clinical effect, this procedure can be used as an ideal surgical treatment for thoracolumbar fractures in AS patients, especially for elderly patients with underlying diseases and high surgical risk.