Suppressive oligodeoxynucleotide-induced dendritic cells rein the aggravation of osteoarthritis in mice.

OBJECTIVE: To explore the effect of suppressive oligodeoxynucleotide-induced dendritic cells (S-DCs) in the osteoarthritis (OA) therapy. METHODS: S-DCs were prepared from splenic CD11c + cells by in vitro culture with suppressive oligodeoxynucleotide. The function and phenotypes of S-DCs were measured by ELISA and flow cytometry. The innate immune signaling pathways were detected by western blotting in the non-treated DCs and S-DCs upon stimulation. In vivo, we employed an iodoacetate-induced OA mice model. S-DCs were transferred by intravenous route. The weight bearing of mice was evaluated and pro-inflammatory factors in OA joint were measured by real-time PCR. Treg cell ratio and CD4 + IL10+ cells in spleen were detected by flow cytometry at day 5 post OA induction. RESULTS: The S-DCs showed less inflammatory phenotypes upon stimulation. The expression of pro-inflammatory cytokines and mature makers in the S-DCs were blunt, due to the impaired innate immune signal transduction. In an iodoacetate-induced OA model, transfer of S-DCs significantly controlled the process of OA. Restricted inflammatory responses were observed in the joint of S-DC recipients. Moreover, after S-DC transfer, Tregs and CD4 + IL10+ cells were mounted in the spleen. CONCLUSION: Transfer of suppressive oligodeoxynucleotides-induced autologous DCs may represent a potential agent to control the aggravation of OA in patients.

Asiatic acid improves high-fat-diet-induced osteoporosis in mice via regulating SIRT1/FOXO1 signaling and inhibiting oxidative stress.

Asiatic acid can attenuate osteoporosis through suppressing adipogenic differentiation and osteoclastic differentiation. Oxidative stress enhances osteoclastic differentiation but represses osteogenic differentiation to promote osteoporosis. However, the role and mechanism of asiatic acid in osteoporosis have not been reported. Firstly, mice were fed with high-fat-diet (HFD) with or without asiatic acid for 16 weeks. Data from an automatic biochemical analyzer showed that HFD induced down-regulation of high-density lipoprotein (HDL) and an increase of serum levels of triglyceride (TG), total cholesterol (TC) and low-density lipoprotein (LDL). However, asiatic acid administration attenuated the decrease of HDL and increase of serum TG, TC and LDL in osteoporotic mice. Secondly, HFD induced high levels of malondialdehyde (MDA) and reactive oxygen species (ROS), low levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in osteoporotic mice. However, the levels of MDA, ROS, SOD and GSH-Px in osteoporotic mice were reversed by asiatic acid administration. (this section is unclear and requires revision) Asiatic acid administration reduced expression of c-telopeptide of type 1 collagen (CTX-1), enhanced alkaline phosphatase (ALP) and procollagen type 1 N-terminal propeptide (P1NP) in HFD-induced osteoporotic mice. (this section is unclear and requires revision) Thirdly, asiatic acid promoted calcium deposition in bone marrow cells and osteogenic differentiation in osteoporotic mice, but decreased ALP in bone marrow cells. Lastly, asiatic acid enhanced SIRT1 and nuclear FOXO1 (Nu-FOXO1) expression, while it reduced Acetyl FOXO1 (Ac-FOXO1) in osteoporotic mice. In conclusion, asiatic acid might inhibit oxidative stress and promote osteogenic differentiation through activating SIRT1/FOXO1 to attenuate HFD-induced osteoporosis in mice.

Platelet rich plasma enhanced neuro-regeneration of human dental pulp stem cells in vitro and in rat spinal cord.

Background: Human dental pulp stem cells (hDPSCs) exhibit excellent differentiation potential and are capable of differentiating into several different cellular phenotypes, including neurons. Platelet-rich plasma (PRP) contains numerous growth factors that can stimulate stem cell differentiation. In this study, we investigated the potential stimulatory effects of PRP on neurogenic differentiation and anti-apoptosis of hDPSCs in injured spinal cords. Methods: The unipotential differentiation capacity of hDPSCs was analyzed by cell surface antigen identification and cell cycle analysis. A spinal cord injury rat model composed of 40 Sprague-Dawley (SD) rats was used to facilitate an in vivo study. Rats were divided into four groups: a double-treatment group (receiving both neurogenic-induced hDPSCs and PRP), two single-treatment groups (receiving neurogenic-induced hDPSCs or PRP) and a sham group (receiving normal saline). The Basso, Beattie, Bresnahan Locomotor Rating Scale was subsequently used to evaluate the motor function of the spinal cord. Cell viability and differentiation of hDPSCs in the damaged spinal cords were analyzed and apoptosis of neural cells was evaluated using the terminal uridine nucleotide end labeling (TUNEL) assay. Results: Growth pattern, cell surface marker and cell cycle analyses revealed that hDPSCs have a high degree of multi-directional differentiation potential and can be induced into neurons in vitro. In the rat spinal cord injury model, double-treatment with hDPSC/PRP or single treatment with hDPSCs or PRP significantly improved motor function compared with the sham group (P<0.05). Apoptosis of neural cells was observed to be significantly higher in the sham group compared to any of the treatment groups. Double-treatment with hDPSCs and PRP resulted in the lowest apoptotic rate among the groups analyzed. Conclusions: hDPSCs exhibit differentiation potential and are capable of transforming into neural cells both in vitro and in vivo. Significantly increased inhibition of neuronal apoptosis and improved motor function recovery of the spinal cord were observed following double-treatment with hDPSCs and PRP compared with the single-treatment groups.

Electrophysiological study on differentiation of rat bone marrow stromal stem cells into neuron-like cells in vitro by edaravone.

OBJECTIVE: To explore the electrophysiological properties of differentiation of rat bone marrow-derived stromal stem cells (rBMSCs) to neuron-like cells in vitro by edaravone, a new type of free radical scavenger. METHODS: Stromal stem cells were separated from rat bone marrow with Ficoll-Paque reagent and expanded in different culture medium in vitro. rBMSCs were induced by edaravone containing serum-free L-DMEM. Morphologic observation and Western blot analysis including the expression of Nav1.6, Kv1.2, Kv1.3, Cav1.2 were performed, and whole patch-clamp technique was used. RESULTS: Cyton contraction and long processes were shown in differentiated stromal stem cells. Nav1.6, Kv1.2, Kv1.3 and Cav1.2 were expressed in both differentiated and undifferentiated cells. However, the expression of channel proteins in differentiated cells was up-regulated. Consistently, their resting potential and outward currents were also enhanced in the differentiated cells, which was especially significant in the outward rectifier potassium current. CONCLUSION: In vitro, neuron-like cells derived from rBMSCs, induced by edaravone, possess electrophysiological properties of neurons.

Changes in bone mineral density and bone metabolic indexes in ankylosing spondylitis mouse model complicated with osteoporosis.

Changes in bone mineral density and bone metabolic indexes in a model of ankylosing spondylitis (AS) mice complicated with osteoporosis (OP) were investigated. BLAB/c mice were used as the subjects. AS was induced using proteoglycan, and OP was induced using tail suspension method. The mice were randomly divided into four groups: AS group, OP group, AS + OP group and negative control group. Changes in bone mineral density, bone strength, serum calcium (Ca), phosphorus, alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRACP), in mice of each group were detected and compared. There were statistically significant differences in bone mineral density and bone strength among groups. Compared with the negative control group, bone mineral density and bone strength in the AS, the OP and the AS + OP groups were significantly decreased, and the lowest bone mineral density and bone strength were found in the AS + OP group (P<0.05). There were no significant differences in bone mineral density and bone strength between the AS group and the OP group. Significant differences in serum Ca, ALP and TRACP but not in serum phosphorus were found among groups. Compared with the control group, serum levels of Ca and TRACP in the AS, the OP and the AS + OP groups were significantly increased, while levels of ALP were obviously decreased (P<0.05). Bone destruction in AS mice complicated with osteoarthritis was more serious than that in mice with simple AS.

Clindamycin Administration Increases the Incidence of Collagen-Induced Arthritis in Mice Through the Prolonged Impact of Gut Immunity.

The profound influence of gut flora on host immune system and its link with autoimmune disorders have been established. However, the role of certain antibiotic in progression of autoimmune disorder is still confusing. Here, we employed a collagen-induced arthritis (CIA) model to explore the role of clindamycin administration in different scenarios. In the first scenario, mice treated with antibiotics for 4 weeks were performed with the induction of CIA immediately. The results showed that clindamycin administration promoted the incidence and severity of CIA, while the recipients of vancomycin showed completed tolerance. We also found that increased gut-associated Th1 and Th17 cells might be related to the subsequent expansion of collagen-specific immune response. In the second scenario, mice treated with antibiotics for 4 weeks were performed with CIA induction 4 weeks later. Notably, clindamycin administration showed a prolonged impact on the incidence and severity of CIA, as well as the gut immunity as compared to vancomycin administration. In addition, antibody depletion of integrin α4ß7 systemically resulted in an impaired CIA response, underlining the influence of gut immunity. In the mice that received clindamycin, the abundance of anaerobic bacteria was significantly decreased and showed little recovery at 4 weeks later. Our observations highlighted the different characteristics of antibiotic administration on the development of autoimmune disorders and indicated its link with gut immunity.

[COMPARISON OF EFFECTIVENESS AND CHANGE OF SAGITTAL SPINO-PELVIC PARAMETERS BETWEEN MINIMALLY INVASIVE TRANSFORAMINAL AND CONVENTIONAL OPEN POSTERIOR LUMBAR INTERBODY FUSIONS IN TREATMENT OF LOW-DEGREE ISTHMIC LUMBAR SPONDYLOLISTHESIS].

OBJECTIVE: To compare the effectiveness and changes of sagittal spino-pelvic parameters between minimally invasive transforaminal lumbar interbody fusion and conventional open posterior lumbar interbody fusion in treatment of the low-degree isthmic lumbar spondylolisthesis. METHODS: Between May 2012 and May 2013, 86 patients with single segmental isthmic lumbar spondylolisthesis (Meyerding degree I or II) were treated by minimally invasive transforaminal lumbar interbody fusion (minimally invasive group) in 39 cases, and by open posterior lumbar interbody fusion in 47 cases (open group). There was no significant difference in gender, age, disease duration, degree of lumbar spondylolisthesis, preoperative visual analogue scale (VAS) score, and Oswestry disability index (ODI) between 2 groups (P>0.05). The following sagittal spino-pelvic parameters were compared between 2 groups before and after operation: the percentage of slipping (PS), intervertebral height, angle of slip (AS), thoracolumbar junction (TLJ), thoracic kyphosis (TK), lumbar lordosis (LL), sagittal vertical axis (SVA), spino-sacral angle (SSA), sacral slope (SS), pelvic tilt (PT), and pelvic incidence (PI). Pearson correlation analysis of the changes between pre- and post-operation was done. RESULTS: Primary healing of incision was obtained in all patients of 2 groups. The postoperative hospital stay of minimally invasive group [(5.1 ± 1.6) days] was significantly shorter than that of open group [(7.2 ± 2.1) days] (t = 2.593, P = 0.017). The patients were followed up 11-20 months (mean, 15 months). The reduction rate was 68.53% ± 20.52% in minimally invasive group, and was 64.21% ± 30.21% in open group, showing no significant difference (t = 0.725, P = 0.093). The back and leg pain VAS scores, and ODI at 3 months after operation were significantly reduced when compared with preoperative ones (P < 0.05), but no significant difference was found between 2 groups (P > 0.05). The postoperative other sagittal spino-pelvic parameters were significantly improved (P < 0.05) except PI (P > 0.05), but there was no significant difference between 2 groups (P > 0.05). The correlation analysis showed that ODI value was related to the SVA, SSA, PT, and LL (P < 0.05). CONCLUSION: Both minimally invasive transforaminal lumbar interbody fusion and conventional open posterior lumbar interbody fusion can significantly improve the sagittal spino-pelvic parameters in the treatment of low-degree isthmic lumbar spondylolisthesis. The reconstruction of SVA, SSA, PT, and LL are related to the quality of life.

Differentiation of human bone marrow mesenchymal stem cells into neuron-like cells in vitro.

STUDY DESIGN: Responses of human mesenchymal stem cells from bone marrow (hBMSCs) were analyzed under chemical conditions, and then characterization of ion channels was evaluated by whole-cell patch clamp. OBJECTIVE: To explore the possibility of differentiation of human bone marrow-derived mesenchymal stem cells into neuron-like cells in vitro under different conditions. SUMMARY OF BACKGROUND DATA: The generation of mesenchymal stem cells into neuron-like cells has been studied. However, few of these studies characterized functional properties of the differentiated hBMSCs. METHODS: hBMSCs (Passage 2) were expanded and cultured in vitro. After Passage 5 was subcultured, the cells were induced by cytokines and antioxidants. Morphologic observation, immunocytochemistry, Western blot analysis, and patch-clamp techniques were performed to evaluate properties of treated and control groups. RESULTS: The differentiated neuronal cells from hBMSCs not only expressed neuron phenotype and membrane channel protein including Nav1.6, Kv1.2, Kv1.3, and Cav1.2 but also exhibited functional ion currents. Both hBMSCs and differentiated cells expressed Nav1.6, Kv1.2, Kv1.3, and Cav1.2 and voltage-activated potassium currents, including delayed rectifier, noise-like and transient outward currents. However, expression of channel proteins, such as sodium channel Nav1.6 and potassium channels Kv1.2 and Kv1.3, were upregulated. Consistently, their potassium currents were also enhanced in the differentiated cells. CONCLUSION: hBMSCs possess of great potential to differentiate into functional neurons, indicating that hBMSCs may be an ideal cell source in managing a variety of clinical diseases such as spinal cord injury.