RESUMO
Nucleus pulposus cells (NPCs) degeneration is an essential pathological basis of intervertebral disc diseases, and autologous cell transplantation is a means of regeneration of NPCs. This study aimed to evaluate the effects of autologous facet joint chondrocytes (CHs) with Sirtuin 1 (sirt1)-overexpression on NPCs degeneration. We used human NPCs and CHs isolated from the patients' tissue and transduced CHs with the plasmid vector to overexpress the sirt1 gene. Further, NPCs were seeded as monolayers and treated with IL-1ß to obtain the degeneration, and the sirt1-overexpressed CHs (sirt1-CHs) in the transwell insert were co-cultured in the same well. The NPCs' degenerated degree was determined by the levels of living cells, proliferation, p16, and collagen I/II, and aggrecan expression at the time point of 1, 3, or 5 days. Besides, the ROS accumulation, antioxidative enzymes, sirt1, and inflammatory factors gene expression were also tested. After IL-1ß treatment, when co-cultured with sirt1-CHs, NPCs accumulated more living cells, proliferation, collagen II, aggrecan, but less p16 and collagen I expression than cultured without sirt1-CHs. Additionally, SOD1, CAT, and TIMP4 mRNA were protected, and the production of TNF-α, IL-6, MMP3, and ROS were alleviated with the presence of sirt1-CHs. Thus, co-culture with sirt1-CHs delays NPCs' degeneration via the suppression of ROS accumulation and inflammatory response. Transplanting autologous CHs with sirt1-overexpressed into the NP tissue might be a novel treatment for intervertebral disc degeneration.
Assuntos
Núcleo Pulposo , Células Cultivadas , Condrócitos/metabolismo , Técnicas de Cocultura , Humanos , Núcleo Pulposo/metabolismo , Sirtuína 1/genética , Sirtuína 1/metabolismo , Sirtuína 1/farmacologiaRESUMO
BACKGROUND: Osteoarthritis (OA) is a chronic joint disease characterized by the degradation of articular cartilage. Polyphyllin I (PPI) has anti-inflammatory effects in many diseases. However, the mechanism of PPI in OA remains unclear.
Methods: HC-a cells treated with IL-1ß were identified by immunofluorescence staining and microscopic observation. The expression of collagen II and DAPI in HC-a cells was detected by immunofluorescence. The effects of gradient concentration of PPI on IL-1ß-induced cell viability, apoptosis, senescence, and inflammatory factor release were detected by MTT, flow cytometry, SA-ß-Gal assay and ELISA, respectively. Expressions of apoptosis-related genes, extracellular matrix (ECM)- related genes, and TWIST1 were determined by qRT-PCR and western blot as needed. The above-mentioned experiments were conducted again after TWIST1 overexpression in IL-1ß-induced chondrocytes.
Results: IL-1ß reduced the number of chondrocytes and the density of collagen II. PPI (0.25, 0.5, 1 µmol/L) had no effect on cell viability, but it dose-dependently elevated the inhibition of cell viability regulated by IL-1ß. The elevation of cell apoptosis, senescence and expression of IL-6 and TNF-α were suppressed by PPI in a dosedependent manner. Additionally, PPI reduced the expression of cleaved caspase-3, bax, MMP-3, and MMP-13 and promoted the expression of collagen II. TWIST1 expression was diminished by PPI. TWIST1 overexpression reversed the abovementioned effects of PPI on chondrocytes.
Conclusion: PPI suppressed apoptosis, senescence, inflammation, and ECM degradation of OA chondrocytes by downregulating the expression of TWIST1.
RESUMO
Excess inflammatory microglia activation deteriorates the pathological degree of spinal cord injury (SCI). We here employed microglia samples in vitro and murine model in vivo to trace the role of inhibition of Arhgef3 in inflammatory response post SCI. From the specimen analysis of lipopolysaccharide (LPS)-induced inflammatory microglia, we found that Arhgef3 expression was positively relative to microglia activation. In vitro, LPS caused the microglia inflammatory activation and induced upregulation of the Arhgef3 expression. Interestingly, presence of Arhgef3 could activate RhoA through promoting Rho GTPases, but silencing of Arhgef3 decreased RhoA activation and inhibited the microglia inflammation. Moreover, disruption of Arhgef3 inhibited the GTP-RhoA, resulted in a suppression of proinflammatory cytokines, and alleviated the LPS-elicited inflammatory genes expression. Moreover, artificially decreasing Arhgef3 expression remarkedly reduced ROS generation after LPS treatment. In vivo of a mouse mechanical contusion-induced SCI model, inhibition of Arhgef3 reduced the ratio of GTP-RhoA/Total-RhoA, and prevented SCI via mitigating the microglial inflammatory phenotype and decreased secondary neurological injury. Besides, inhibition of Arhgef3 prevented alleviated the degree of demyelination but did not affect neuronal regeneration. Meaningfully, absence of Arhgef3 improved mouse locomotor recovery post SCI. Taken together, Arhgef3 involves the microglial activation and inflammatory response following neural injury, and targeted disrupting of which may indicate a promising therapeutic direction in preventing SCI.
Assuntos
Microglia/metabolismo , Fatores de Troca de Nucleotídeo Guanina Rho/antagonistas & inibidores , Fatores de Troca de Nucleotídeo Guanina Rho/biossíntese , Traumatismos da Medula Espinal/metabolismo , Proteína rhoA de Ligação ao GTP/antagonistas & inibidores , Proteína rhoA de Ligação ao GTP/metabolismo , Animais , Expressão Gênica , Técnicas de Silenciamento de Genes/métodos , Locomoção/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fatores de Troca de Nucleotídeo Guanina Rho/genética , Traumatismos da Medula Espinal/genéticaRESUMO
OBJECTIVE: To explore the effectiveness of the calcaneal plate bridge reconstruction plate for acetabular fracture involving quadrilateral surface via modified Stoppa approach. METHODS: Between January 2015 and December 2017, 18 patients with acetabular fracture involving quadrilateral surface were treated with the calcaneal plate bridge reconstruction plate via the modified Stoppa approach. There were 12 males and 6 females. The age ranged from 28 to 63 years (mean, 39 years). The cause of injury was traffic accident in 13 cases and falling from height in 5 cases. According to the Letournel-Judet classification, there were 10 cases of anterior and posterior column fractures, 6 cases of T-shaped fractures, and 2 cases of anterior column and posterior semi-transevere fractures. The interval from injury to operation was 6 to 24 days (mean, 8.6 days). The reduction quality was assessed by postoperative X-ray film and CT according to the criteria proposed by Matta. The hip joint function was assessed by the modified Merled'Aubigné-Postel score. RESULTS: The operation time was 120-240 minutes (mean, 165 minutes) and the intraoperative blood loss was 600-1 400 mL (mean, 850 mL). All patients were followed up 18-30 months (mean, 24.5 months). There were 2 cases of the fat liquefaction of abdominal incisions, 3 cases of intraoperative injury of lateral femoral cutaneous nerve, 1 case of lower limb thrombosis, and 1 case of abdominal pain and hematuria due to intraoperative accidental bladder injury. According to the criteria proposed by Matta, the reduction quality rated as anatomic reduction in 12 cases, satisfactory reduction in 5 cases, and unsatisfied reduction in 1 case, and the satisfaction rate was 94.4%. All fractures healed with the healing time of 3-5 months (mean, 3.4 months). During follow-up, no internal fixator loosening, breakage, or fracture displacement occurred. At last follow-up, according to modified Merled'Aubigné-Postel score, hip joint functions rated as excellent in 11 cases, good in 4 cases, fair in 2 cases, and poor in 1 case. The excellent and good rate was 83.3%. CONCLUSION: Application of calcaneal plate bridge reconstruction plate via the modified Stoppa approach for the acetabular fracture involving the quadrilateral surface can obtain satisfactory effectiveness.
Assuntos
Calcâneo , Fraturas do Quadril , Acetábulo/cirurgia , Adulto , Placas Ósseas , Feminino , Fixação Interna de Fraturas , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Objective: To research the feasibility and effectiveness of percutaneous kyphoplasty (PKP) by improved injecting tube through unipedicular puncturing. Methods: Between January 2012 and Junuary 2016, 60 cases (68 vertebrae) of osteoporotic vertebral compression fractures (OVCF) were treated. PKP was performed through unipedicular puncturing with routine injecting tube in 30 cases (34 vertebrae, routine group), and with improved injecting tube in 30 cases (34 vertebrae, improved group). There was no significant difference in age, gender, disease duration, fracture level, preoperative visual analogue scale (VAS), or vertebral height between 2 groups ( P>0.05). The operation time, the volume of bone cement injected, preoperative and postoperative VAS, and preoperative and postoperative vertebral height, and postoperative distribution coefficient of bone cement were recorded and compared between 2 groups. Results: Good healing of puncture points was achieved in 2 groups after PKP, and no serious complication occurred. There was no significant difference in operation time and the volum of bone cement injected between 2 groups ( t=0.851, P=0.399; t=1.672, P=0.101). Bone cement leakage was observed in 2 cases of 2 groups respectively. The distribution coefficient of bone cement in routine group was significantly less than that in improved group ( t=13.049, P=0.000). All patients were followed up 12-36 months (mean, 19 months). The postoperative VAS and vertebral height were significantly improved when compared with preoperative ones in 2 groups ( P<0.05), but there was no significant difference in VAS between at 2 days after operation and at last follow-up, in vertebral height between at 2 days after operation and at 1 year after operation, and between 2 groups after operation ( P>0.05). X-ray films showed vertebral compression fractures in 6 cases of routine group and in 1 case of improved group during follow-up. Conclusion: PKP by improved injecting tube through unipedicular puncturing can improve the distribution of bone cement, restore the height and strength of vertebral body, and reduce the incidence of re-fracture.