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Objective:To establish a scoring system based on lumbar magnetic resonance imaging (MRI) images to evaluate bone mineral density and evaluate its correlation with T score of dual energy X-ray absorptiometry (DEXA).Methods:The clinical data of 82 patients with lumbar degenerative diseases who were admitted to the Clinical Medical College of Yangzhou University from January 2019 to August 2020 were analyzed retrospectively. According to the lower value of T value of femoral neck and total hip bone mineral density detected by DEXA, they were divided into normal bone mass group ( n=40) and abnormal bone mass group ( n=42). The vertebral body bone mass (VBQ) score of the patient was calculated by dividing the average signal intensity of L 1-4 vertebral body by the signal intensity of L 3 level cerebrospinal fluid on T 1 weighted image of MRI. The receiver operating characteristic (ROC) curve was drawn to evaluate the ability of VBQ score to distinguish between normal bone mass and abnormal bone mass and the accuracy of predicting the occurrence of abnormal bone mass. Further, the correlation between VBQ score and T value was determined by regression analysis. Results:The lowest T value measured by DEXA in the abnormal bone mass group were significantly lower than those in the normal bone mass group, and the VBQ score was significantly higher than that in the normal bone mass group(all P<0.001). The area under curve (AUC) of VBQ score for predicting abnormal bone mass was 0.93, the cut-off value was 2.98, with sensitivity 81.6%, and specificity 88.6%. The VBQ score was corrected with the lowest T value measured by DEXA ( r=-0.77). Conclusions:VBQ score could effectively distinguish normal bone mass from abnormal bone mass and was negatively correlated with the lowest T value of DEXA.
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Osteoporotic thoracolumbar fracture in the elderly will seriously reduce their quality of life and life expectancy. For osteoporotic thoracolumbar fracture in the elderly, spinal reconstruction is necessary, which should comprehensively consider factors such as the physical condition, fracture type, clinical characteristics and osteoporosis degree. While there lacks relevant clinical norms or guidelines on selection of spinal reconstruction strategies. In order to standardize the concept of spinal reconstruction for osteoporotic thoracolumbar fracture in the elderly, based on the principles of scientificity, practicality and progressiveness, the authors formulated the Clinical guideline for spinal reconstruction of osteoporotic thoracolumbar fracture in elderly patients ( version 2022), in which suggestions based on evidence of evidence-based medicine were put forward upon 10 important issues related to the fracture classification, non-operative treatment strategies and surgical treatment strategies in spinal reconstruction after osteoporosis thoracolumbar fracture in the elderly, hoping to provide a reference for clinical treatment.
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Objective:To analyze the curative effect of discectomy under Quadrant expandable channel associated with annulus repair for the treatment of adolescent lumbar disc herniation (ALDH).Methods:10 ALDH patients received discectomy under Quadrant expandable channel associated with annulus repair (annulus repair group) and 12 patients received discectomy (control group) in Clinical Medical College of Yangzhou University from January 2013 to December 2017 were retrospectively collected. The length of skin incision, amount of intraoperative bleeding, operation time and duration of hospitalization were compared. The degree of pain was assessed by visual analogue scale (VAS) before operation, 24 h, 1 week and 1 year after operation, and the lumbar function was assessed by Oswestry disability index (ODI) before operation, 1 week and 1 year after operation. At the last follow-up, the curative effect was evaluated by MacNab Scale, and the recurrence of lumbar disc herniation during the follow-up was recorded.Results:There were no significant differences in the amount of intraoperative bleeding, operation time and duration of hospitalization between the two groups ( P>0.05). The skin incision length of the annulus repair group was less than that of control group ( P<0.05). The postoperative VAS score and ODI score at each follow up time point in both groups were significantly improved when compared with the preoperative ones ( P<0.05). There were no statistically significant difference between the two groups in the VAS score and ODI score 1 week postoperative and 1 year postoperative ( P>0.05). According to the MacNab criteria, there was no statistically significant difference between the two groups in the excellent and good rate (annulus repair group: 9/10, control group: 10/12; P>0.05). There was no recurrence case in the annulus repair group, but two cases of recurrence (one recovered by conservative treatment, the other needed second operation) in the control group ( P<0.05). Conclusions:Discectomy under Quadrant expandable channel associated with annulus repair can achieve early satisfied outcome, reduce surgery related trauma, pain in the early period postoperative and recurrence rate after operation in the treatment of ALDH.
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Objective:Through TTC staining, immunohistochemical analysis, RT-PCR and hind limb motor function evaluation and other experimental methods, to explore the regulatory mechanism of metformin on anti-apoptosis in rats with spinal cord injury (SCI).Methods:Establish a rat spinal cord injury model. Through Basso-Beattie -Bresnahan locomotor rating scale (BBB) and cant test to evaluate the recovery of hindlimb motor function in rats. The changes of necrotic area of spinal cord tissue were compared by TTC staining. Extraction of rat spinal cord tissue, by Dot blot analysis and immunohistochemical detection of the hydroxyl of DNA methylation level. By qPCR, Western Blot detection TET2mRNA and protein expression level, and the changes in the scope of spinal cord injury were detected by inhibiting the expression of TET2. The interaction between TET2 and Foxo3a was detected by immunoblotting and immunoprecipitation. Through RT-PCR assay Foxo3a downstream related changes in the level of gene expression.Results:Compared with the SCI+NS group, the necrotic area of the spinal cord tissue was reduced after metformin treatment, and the BBB score and the incline test score were higher ( P<0.05). At the same time, we found that the levels of TET2mRNA and protein increased significantly after SCI at 24 h, and the 5-hmC level of DNA increased. The levels of TET2mRNA and protein and 5-hmC increased further after the use of metformin. After using SC-1, compared with the SCI+MET group, the level of 5-hmC decreased and the area of infarction increased. After SCI, the mRNA levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the mRNA levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). After SCI, the 5-hmC levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the 5-hmC levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). Conclusion:Metformin can promote the interaction between TET2 and Foxo3a, increase the 5-hmC level of the overall DNA, and inhibit the activation of related apoptosis genes, thereby improving tissue damage and nerve function recovery after spinal cord injury.
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Objective:To investigate the effects of allicin on the proliferation, migration and angiogenesis of rat vascular endothelial cells (RVES), and to explore the influencing mechanism of allicin on epidural fibrosis.Methods:According to the results of preliminary experiments, RVECs were divided into control group (0 mg/L), low concentration group (25 mg/L), medium concentration group (50 mg/L) and high concentration group (100 mg/L). The morphology, viability, migration rate, cell cycle, apoptosis rate and cell lumen formation ability were measured using fluorescence microscope, AnnexinV-FITC double staining, PI/RN-asestaining, scratch assay and Transwell experiments test. Western Blot was used to measure the protein expression level of JAK2, STAT3, p-STAT3, PCNA, Bax and Bcl-2 protein. Using random number method, 36 adult male SD rats were divided into sham operation group, saline group and allicin group, with 12 rats in each group. Hematoxylin-eosin (HE) staining, Masson staining and immunohistochemical staining were used to analysis the epidural fibrosis in each group.Results:With the increase of concentration of allicin, cell viability, cell migration and lumen formation ability significantly lower than that of control group ( P<0.05). With the increase of allicin concentration, the percentage of cells in the G1 and S phases gradually decreased ( P<0.05), the percentage of cells in the G2 phase and the apoptosis rate gradually increased ( P<0.05), and the cells were blocked in the G2/M phase. With the increase of allicin concentration, the protein expression levels of JAK2, STAT3, p-STAT3, PCNA and Bcl-2 were gradually down-regulated ( P<0.05), while the protein expression level of Bax was gradually up-regulated ( P<0.05), the ratio of p-STAT3/STAT3 was decreased ( P<0.05), and the ratio of Bax/Bcl-2 was increased ( P<0.05). There was no death, infection or abnormal gait in all the experimental animals. Dense scar tissue could be observed in the extradural area of the sham operation group and the epidural area of the control group, but there was obvious space between the epidural scar and the dura mater in the allicin group, and the density of collagen, the number of blood vessels, and the protein density of p-STAT3 were significantly lower than those in the control group ( P<0.05). Conclusion:Allicin inhibits angiogenesis and the severity of epidural scar after laminectomy, and the mechanism may be through inhibiting the JAK2/STAT3 signaling pathway of vascular endothelial cells.
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Objective To compare the effect of different size needle gauges to the degenerative response in rat caudal discs.Methods A total of 40 Sprague Dawley (SD) rats,level 5/6,7/8 and 9/10 interverbral discs of rat caudal spine were punctured with 18 or 21 or 25-gauge needles respectively.Radiographs and magnetic resonance imaging (MRI) were obtained at 1,2,4 and 6 weeks postsurgery.At each time point,ten rats from each group were sacrificed for histological analysis.Real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to examine mRNA expression level.Results Significant differences were identified in the disc height index (DHI ％) and MRI grade between 18 G and normal group,MRI grade,histological score between 21 G and normal group at 2,4,and 6 weeks postsurgery.Significant differences were also identified in the histological score and mRNA expression levels between 18 G and normal group,alcian blue stain and hypoxia inducible factor-1α (HIF-1 α) mRNA expression level between 21G and normal group at all time point postsurgery.Significant differences existed in DHI％,type Ⅱ collagen and aggrecan mRNA expression levels between 21 G and normal group,all type mRNA expression levels between 25 G and normal group at 4,6 weeks.There were significant differences in MRI grade and histological score between 25 G and normal group at 6 weeks.Significant differences existed in almost all parameters compared between 18 G and 25 G at all time point.There were significant differences in DHI％,MRI grade,histological score and HIF-1α mRNA expression levels between 18 G and 21 G at 4,6 weeks.There were significant differences in type Ⅱ collagen and aggrecan mRNA expression levels between 18 G and 21 G at all time point.Significant differences exist in DHI％ and HIF-1α mRNA expression level between 21 G and 25 G at 6 weeks.Compared with the 25 G group,the DHI％ and Pfirrmann scores and the pathological score of each time at 2,4 and 6 weeks after operation in 18 G group have significant difference (P ＜ 0.05).Conclusions The middle size needle (21G) is better to induce disc degeneration.The 2-week time point may be the better time frame to further experimental treatments.
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Objective To compare the clinical results of percutaneous endoscopic interlaminar discectomy (PEID) and percutaneous endoscopic transforaminal discectomy (PETD) in L5/S1 disc herniation.Methods A total of 102 patients with L5/S1 disc herniation in our hospital from September 2014 to June 2016 were enrolled in this study.Fifty-two patients underwent percutaneous endoscopic interlaminar discectomy (PEID group) and 50 patients underwent percutaneous endoscopic transforaminal discectomy (PETD group).The surgical effectiveness was assessed according to Visual Analog Scale (VAS),Oswestry Disability Index (ODI),and modified MacNab criteria.The frequencies of intraoperative radiation exposure,operation time and complication rates were compared between the groups.Results All the patients completed follow up with a mean of 15.0 months (range,10-20 months).In the PEID group,the mean operation time was 44-72 (58.3 ± 12.0)minutes and the intraoperative frequencies of radiation exposure were 3-6(3.8 ±2.1)seconds.For the PETD group,the mean operation time was 52-96(82.4 ± 16.0) minutes and the intraoperative radiation time was 13-34 (24.1 ± 10.1) seconds.There were significant differences in operation time and radiation frequency between the two groups (P ＜ 0.05).The postoperative VAS and ODI were obviously improved in both groups when compared with preoperation (P ＜ 0.05),but there were no significant differences between the two groups (P ＞ 0.05).There was no statistically significant difference considering the satisfactory rates according to the MacNab criteria between PEID group (96.1％) and PETD group (96.0％).Conclusions The treatment of L5/S1 disc herniation through lamina or intervertebral foramen approach is both effective.PEID can significantly reduce the frequencies of intraoperative radiation exposure and operation time.
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Objective To explore the early clinical effects of lumbar discectomy associated with annulus repair in the treatment of lumbar disc herniation.Methods A prospective study was conducted to review 56 patients with lumbar disc herniation who accepted discectomy surgery in Subei People's Hospital of Jiangsu Province from January 2014 to September 2015,including 28 cases of discectomy associated with annulus repair (repair group) and 28 cases of discectomy (control group).Oswestry disability index and visual analog scale scores were recorded.Simultaneously,incision length,operative time,blood loss,hospitalization time,surgical complications,and postoperative recurrence of lumbar disc herniation were recorded.Results All patients completed the follow-up for 12 to 18 months (14.5 ± 1.3).There was no difference between the repair and control groups in the incision length,blood loss and hospitalization time (P ＞ 0.05).The operative time of the repair group was longer than that of the control group,but the difference was not statistically significant (P ＞ 0.05).The Oswestry disability index and visual analog scale scores for lumbar and lower limb pain significantly decreased in both groups after surgery (P ＜ 0.05).The visual analog scale scores at 24 hours and 3 days after surgery in the repair group were less than that in the control group (P ＜ 0.05).The satisfactory rate of treatment in the repair group was slightly higher than that in the control group,but the difference was not statistically significant (P ＞ 0.05).There was no recurrence in the repair group,but 2 recurrence cases in the control group (P ＞ 0.05).Conclusions These findings indicate that discectomy associated with annulus repair is a safe and reliable method to obtain remarkable early clinical results and can reduce the recurrent rate in the treatment of lumbar disc herniation.
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BACKGROUND:Statins can promote the mRNA expression of bone morphogenetic protein 2, aggrecan and type II col agen in intervertebral disc cel s, and they also can reverse the phenotype of dedifferentiated nucleus pulposus cel s to slow disc degeneration process. OBJECTIVE:To review the research progress of simvastatin modulating the biological characteristics and mobilizing endogenous stem cel s for the repair of intervertebral disc degeneration. METHODS:The first author retrieved the PubMed and CNKI databases for relevant articles published before January 2016 using the key words of“disc degeneration factor, Simvastatin AND stem cel s, endogenous stem cel s AND disc degeneration”in English and Chinese, respectively. Initial y, 102 relevant articles were retrieved, but only 48 articles were included in result analysis fol owing elimination of duplicate studies.RESULTS AND CONCLUSION:By summarizing a large number of studies on the treatment of intervertebral disc degeneration worldwide, we found that simvastatin may modulate the biological characteristics and function of nucleus pulposus mesenchymal stem cel s via promoting the expression of hypoxia-inducible factor 1αfor the endogenous stem cel-based therapy of intervertebral disc degeneration.
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BACKGROUND: Percutaneous vertebroplasty has been gradual y used to treat Kümmel disease because of less trauma and quick pain relief, but there is stil a high rate of bone cement leakage. OBJECTIVE: To investigate the clinical efficacy of percutaneous vertebroplasty with high-viscosity bone cement plus hyperextension position reset for treatment of Kümmel ’s disease. METHODS: The clinical data of 17 patients with Kümmel ’s disease were retrospectively analyzed, including 5 males and 12 females, aged 55-83 years, and al underwent percutaneous vertebroplasty with high-viscosity bone cement plus hyperextension position reset. The visual analog scale, Oswestry disability index score, vertebral body height and vertebral kyphosis angle were determined. The bone cement leakage, pulmonary embolism, adjacent vertebral fractures and other complications were recorded. RESULTS AND CONCLUSION: At the 12th Oswestry disability index scores and vertebral kyphosis angle of patients were significantly lower than those before treatment (P < 0.05), the vertebral body height was significantly higher than that before month of follow-up, the visual analog scale scores, treatment (P < 0.05). After treatment, there were three cases of bone cement leakage, which had no special discomfort and neurological symptoms, and one case of new fractures. These results demonstrate that hyperextension position reset combined with percutaneous vertebroplasty with high-viscosity bone cement in treatment of Kümmel ’s disease can effectively relieve back pain, improve function of the lower back, partial y restore vertebral height and reduce kyphosis angle.
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BACKGROUND:Preliminary experiments have reported the influence of serum and nerve growth factor on olfactory ensheathing cells proliferation in vitro, but there are less studies concerning choice of serum concentration and growth time for in vitro culture of olfactory ensheathing cells. OBJECTIVE:To find out the influence of different blood serum concentration and growth time on olfactory ensheathing cells from the olfactory mucosa of adult rats based on the growth curve of olfactory ensheathing cells. METHODS:The olfactory ensheathing cells from the olfactory mucosa of adult rats were separated, culture and identified in vitro. Sulforhodamine B and microplate reader were employed to measure absorbance values and plot growth curve of olfactory ensheathing cells. RESULTS AND CONCLUSION:When cultured for the same time in blood serum of different concentrations, absorbance values, especial y in the groups 10%, 20%, 30%, 40%, tended to increase with time except the 0%group. When cultured in the same serum for different time, absorbance values increased within the first 9 days, then promoted rapidly in the groups 10%, 20%, 30%, 40%at 13 days, entered the plateau phase at 19 days, and decreased at 23 days;meanwhile, in the other groups (50%, 60%, 70%, 80%, 90%) the absorbance values peaked at the 13th day and then decreased gradual y. These findings indicate that different serum concentrations and different growth time in vitro affect cellgrowth and survival of olfactory ensheathing cells significantly, which should be ful y considered when cells are cultured in an in vitro condition.
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Objective To study the neuroprotective effect of sevoflurane on controlled hypotension in patients with coronary heart disease undergoing craniocerebral surgery. Methods Twenty-six patients with coronary heart disease undergoing craniocerebral surgery were randomly divided into two groups,receiving either inhaled 2%-5%sevoflurane plus intravenous sodium nitroprusside (treatment group,n=13) or intravenous sodium nitroprusside 3-8 μg·kg-1 alone (contorl group,n=13) for blood pressure control. The hemodynamic changes were recorded during the operation. Patient satisfaction with surgeons and the duration hospital stay after surgery were recorded. The levels of cardiac troponin I (TNI),creatine kinase-MB (CK-MBM),neuron specific enolase ( NSE) and S100 calcium binding proteinβ( S-100βprotein) in serum were detected at one day pre-operation,the end of the operation,and one day post-operation. Results The duration of hospital stay after surgery was significantly shortened in treatment group [(20. 3±3. 8) versus (23. 9±4. 6) d,P<0. 05) compared with control group. The average heart rate significantly decreased, patient satisfaction significantly increased, and serum levels of NSE and S-100β protein one day post-operation significantly decreased as compared to control group (all P<0. 05). Compared with the day before operation,serum levels of NSE and S-100βprotein in the two groups increased significantly at the end of surgery (P<0. 05),and the levels of TNI,CK-MBM,NSE and S-100βsignificantly elevated one day post-operation (P<0. 05). Compared with the end of operation,serum levels of NSE and S-100βin contorl group incarcerated remarkably (P<0. 05) Conclusion Sevoflurane plays an important neuroprotective role,as evidenced by improving patients' satisfaction,reducing hospital stay after surgery,and maintaining the balance of myocardial oxygen delivery and consumption in craniocerebral surgery patients with coronary heart disease during controlled hypotension. However,it can not prevent postoperative myocardial injury in these patients.
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BACKGROUND:Vertebroplasty and kyphoplasty have been widely applied in the treatment of osteoporotic thoracolumbar compression fracture. However, cement leakage is a major problem in the application of this technology, especial y for the vertebral posterior wal ruptured patients. OBJECTIVE:To investigate the therapeutic efficacy of high viscosity bone cement and vertebroplasty in the treatment of osteoporotic thoracolumbar compression fracture. METHODS:A retrospective study was conducted in 20 cases receiving high viscosity bone cement and vertebroplasty surgery for osteoporotic thoracolumbar compression fracture. Clinical outcomes were evaluated mainly with use of Visual Analog Scale for lower back pain. Function of lower back pain was assessed using Oswestry Disability Index questionnaire. Quality of life was evaluated using 36-Item Short Form Health Survey and Frankel score was applied to evaluate neurological function. The anterior vertebral height of the fractured vertebrae was assessed with X-ray. The bone cement leakage, pulmonary embolism, incidence of nearby vertebral fractures and other complications were evaluated during fol ow-up. RESULTS AND CONCLUSION:Al patients were fol owed up for 12-18 months. The anterior vertebral height of the fractured vertebrae, the lower back pain and function, and quality of life were improved significantly after treatment (P<0.05). Al patients got the same neurological symptoms before surgery. The bone cement dispersion was good after treatment, detected by X-ray and CT scan, only two cases appeared with bone cement leakage, but no clinical symptoms were found. There was no cement toxicity or al ergic complications, pulmonary embolism, infection, nerve injury or new fractures. The high viscosity bone cement used in the treatment of osteoporotic thoracolumbar vertebral compression fractures can significantly relieve thoracic back pain, improve lower back function and quality of life, and greatly reduce the risk of bone cement leakage.
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Objective To study the effects of controlled hypotension with sevoflurane on hymo-dynamics and cerebral metabolic rate of oxygen in neurosurgery.Methods Thirty-seven ASA Ⅰ or Ⅱpatients undergoing elective intracranial surgery were enrolled.The patients were treated with general anesthesia and assigned to two experimental groups,sevoflurane group (group S,n=20)and sodium nitroprusside group (group N,n=1 7).HR,BP,SpO2 and ECG were continuously monitored during operation.Blood samples were taken from radial artery and internal jugular bulb to determine cerebral arteriovenous oxygen differences (Da-jvO2 ) and cerebral metabolic rate of oxygen (CMRO2 ). Results The HR in group S were decreased during hypotension,5 min,10 min,20 min,and 30 min compared with group N (P <0.05).Da-jvO2 and CMRO2 both in two groups were decreased during hypotension(P < 0.05 ),but there was no difference between the two groups.RPP was lower in group S than in group N during hypotension.Conclusion Controlled hypotension with sevoflurane does not increase HR and improves cerebral blood flow-metabolism coupling.
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BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent. OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor. METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified. RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.
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BACKGROUND:Scaffolds made of chitosan and its derivatives play an important role in cellmigration and axonal regeneration. Chitosan and its derivatives have good histocompatibility, which is easy to make stem cells grow on the surface, thereby having a more broad application prospect in the nerve tissue engineering. OBJECTIVE:To fabricate a thermosensitive hydrogel scaffold using chitosan/hydroxypropyltrimethyl ammonium chloride chitosan/sodium glycerophosphate (CS/HACC/GP), which is suitable for cellgrowth, and then, to observe the growth and proliferation of bone marrow mesenchymal stem cells on the scaffold. METHODS:Chitosan was modified using quaternary ammonium salt and confirmed by Fourier transform infrared spectroscopy. The chitosan and quaternary ammonium salt of chitosan was mixed at a ratio of 8:1 to successful y prepare stable CS/HACC/GP thermosensitive hydrogel scaffold. Then, the gel ing was observed, and biosafety test was conducted. RESULTS AND CONCLUSION:Fourier transform infrared spectroscopy showed the characteristic peak of quaternary ammonium groups. Cytotoxicity test showed that rat bone marrow mesenchymal stem cells cultured in hydrogel extracts had no toxicity. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test showed that hydrogel extracts exerted no significant effect on the increase in body weight, and the biological safety of the scaffold was good. Under the scanning electron microscopy, bone marrow-derived mesenchymal stem cells grew and proliferated normal y in the scaffold. The results confirmed that the CS/HACC/GP thermosensitive hydrogel scaffold was successful y prepared in the experiment, which is suitable for the growth and proliferation of bone marrow mesenchymal stem cells.
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BACKGROUND:In recent years, chitosan-based thermosensitive hydrogel, as scaffold materials, have received more and more attentions in the field of tissue repair because of good biocompatibility, biodegradability and drug-sustained release. OBJECTIVE:To explore the directed differentiation and growth of rat bone marrow mesenchymal stem cells on the quaternary chitosan thermosensitive hydrogel scaffold and to look for more ideal tissue engineering materials for the treatment of nervous system damage. METHODS:The thermosensitive hygrogel scaffold was prepared using hydroxypropyltrimethyl ammonium chloride chitosan (HACC) andβ-glycerophosphate (β-GP). The spatial structure of scaffold was observed by scanning electronic microscope. Effect of leaching liquor from the HACC/β-GP scaffold on the viability of bone marrow mesenchymal stem cells was detected by (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The albumin from bovine serum was combined with the scaffold, and the slow-release effect of the scaffold was detected by ultraviolet absorption spectrometry. Bone marrow mesenchymal stem cells were incubated onto the compound scaffold at 3 passages. The adhesion, growth and differentiation of bone marrow mesenchymal stem cells on the compound scaffold were observed by the scanning electron microscope. Neuron-specific enolase was detected by immunofluorescence. RESULTS AND CONCLUSION:The porosity and thermal sensitivity of HACC/β-GP scaffold and slow-release effect of glial cellline-derived neurotrophic factor were apparent. The results of MTT showed that the compound scaffold cannot take apparent negative effects to the proliferation of bone marrow mesenchymal stem cells. After inoculation, bone marrow mesenchymal stem cells permeated the porous structure of the scaffold and adhered to the scaffold. Under the role of glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells showed neuron-like cellmorphology and cells co-cultured with the compound scaffold expressed the marker of neurons, neuron-specific enolase. Under the role of slow-release glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells can grow wel in vitro and differentiate into neuron-like cells on the HACC/β-GP scaffold.
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BACKGROUND:L-type calcium channels, as a kind of voltage-dependent calcium channel, is the main way of extracellular calcium ions into the cell, and play an important role in maintaining cellmorphology and physiological activities, characterized by a large single-channel conductance, slow channel attenuation, and longer duration of channel opening. Previous studies showed that basic fibroblast growth factor can promote the proliferation of chondrocytes cultured in vitro. OBJECTIVE:To explore the effect of the L-type calcium channels on regulating chondrocyte proliferation and differentiation in response to basic fibroblast growth factor with patch-clamp. METHODS:The chondrocytes were harvested from the joints of 3-day-old New Zealand rabbits. The second passage of chondrocytes was divided into experimental group and control group. Chondrocytes were incubated in media containing 10μg/L basic fibroblast growth factor and media alone separately. The opening of L-type calcium channels under the action of basic fibroblast growth factor was detected by patch-clamp. The intracellular calcium concentration was detected with laser confocal microscopy in the chondrocytes after 2 weeks of culture with basic fibroblast growth factor. Chondrocyte proliferation was analyzed by cellTiter kit after 8 days of culture. Type Ⅱ col agen was assessed quantitatively by immunohistochemistrical staining after 10 days of culture. RESULTS AND CONCLUSION:Basic fibroblast growth factor has an inhibitory effect on the opening of the L-type calcium channels, resulting in a decrease in intracellular free calcium concentration (P<0.01). cellnumber was higher after culture with basic fibroblast growth factor than that cultured under conventional condition (P<0.01), and staining area of type II col agen significantly increased (P<0.05). Results verified that basic fibroblast growth factor can maintain intracellular Ca2+concentration at a low level by inhibiting the opening of L-type calcium channels, which can promote the proliferation and differentiation of chondrocytes.
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Objective To compare the clinical outcomes of dilator-kyphoplasty (DKP) and balloonkyphoplasty (BKP) in treatment of osteoporotic vertebral compression fractures. Methods From May 2007 to March 2009, 23 cases with 26 vertebrae were treated with DKP, while 29 cases with 29 vertebrae were treated with BKP procedures. The operating time, bleeding volume and injecting volume of cement were recorded during operation. The distribution of cement, the restoration of vertebral height and Cobb angle were observed. The patients' visual analogue scales (VAS) score and Oswestry disability index (ODI) score were evaluated after operation. Results There were no differences in operative time, bleeding volume of every vertebrae and cement injected volume between these two groups (P＞0.05). The vertebral height, Cobb angle, VAS and ODI scores were significantly improved than those of pre-operation in these two groups (P＜0.05). The height of the anterior vertebrae and Cobb angle in DKP groups were restored significantly than those in BKP groups (P＜ 0.05). There were 1 case (1 vertebra, 3.8%) underwent cement leakage in DKP groups and 5 cases (5 vertebrae, 17.2%) in BKP groups. Conclusion DKP and BKP were effective in the treatment of osteoporotic vertebral compression fractures. The height of the anterior vertebrae and Cobb angle in DKP groups were restored significantly than those in BKP groups.
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AbstractBACKGROUND: Fas and P53 are important regulator and control gene which can promote apoptosis. They belong to the receptor family part of tumor necrotic factor/nerve growth factor. Their expression products have effects on apoptosis signal transmission, and can regulate and control cell apoptosis in cerebral ischemia-reperfusion injury. And puerarin can alleviate the level of cell apoptosis.OBJECTIVE: To observe the effect of puerarin on Fas and P53, the apoptosis-related gene of nerve cell in hippocampns CA1 region of rats af ter cerebral resuscitation.DESIGN: Randomized controlled trial. SETTING: Department of Emergency, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: The experiment was carried out at Emergency Department, Tongji Hospital, Tongji Medical College, Huazhong University ofScience and Technology from September 2001 to Februray 2002. Totally 45 of 3 months old Wistar rats of clean grade were selected, and randomly divided into 3 groups: sham operation group, model control group and puerarin treatment group with 15 rats in each group.METHODS: Acute global brain ischemia-reperfusion models were established in rats of puerarin treatment group and model control group. In rats of sham operation group, stigmata of both flanks of the first cervical vertebrae were isolated, but bilateral vertebral arteries were not electric coagulated, and biolateral common carotid arteries were only isolatedwithout clamping close. Rats in puerarin treatment group were given puerarin injection 100 mg/kg, 1 hour before ischemia, and model control group were given normal saline in equivalence while rats in sham operation group were not given medicine. Death of rats in each group was performed separately in the 3rd, 6th, 12th, 24th and 48th hours after cerebral ischemia-reperfusion with 3 rats per group in each time. Hippocampus tissues of rats were isolated, and tissue slices were preparated. And the changes of- the protein expression levels and the number of apoptosis cells of rats in each group at different time point after cerebral ischemia-reperfusion were detected in immuno-histochemical method and end labelling in situ method. MAIN OUTCOME MEASURES: ① The number of positive cells inprotein expression of Fas and P53 in hippocampus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion was studied. ② Comparison of the number of apoptosis cells in hippocampus CA1 region of rats between groups at different time point after cerebral ischemia-reperfusion were studied, too.RESULTS: All the 45 rats enrolled in research were entered the stage of result analysis: ① The number of positive cells in protein expression of Fas in hippocampus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion: Obvious gene expression of Fas was not found in sham operation group. In contrast with model control group, obvious decrease was found at all time points after cerebral ischemi a-reperfusion in puerarin treatment group, and in the 6th, 12th, 24th and 48th hour the differences were significant [(15.0±4.3), (13.5±4.9); (40.7±3.4), (27.2±3.1); (37.0±4.8), (22.0±2.1); (24.7±4.1), (18.9±5.3)/mm; P < 0.05,P < 0.01]. ② The number of positive cells in protein expression of P53 in hippocanpus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion: Obvious gene expression of P53 was not found in sham operation group. In contrast with model control group, obvious decrease was found in the 24th and 48th hour after cerebral ischemiareperfusion in puerarin treatment group [(25.3±4.4), (12.8±2.7); (24.3±3.6), (10.9±3.0)/mm; P < 0.01]. ③ Comparison of the number of apoptosis cells in hippocampus CA1 region of rats between groups at different time point after cerebral ischemia-reperfusion :In contrast with model control group,obvious decrease was found in the 12th, 24th and 48th hour after cerebral is chemia-reperfusion in puerarin treatment group [(34.0±3.7), (21.0±3.7); (41.0±4.2), (33.0±4.8); (71.0±5.5), (41.0±3.4)/mm; P < 0.01].CONCLUSION: In rats which were given puerarin treatment, the expression of Fas decrease obviously in 6 to 48 hours after cerebral ischemiareperfusion, and the expression of P53 decreased obviously in the 24th to 48th hour after cerebral ischemia-reperfusion, and a descent tendency could be found in the number of apoptosis cells. These can further prove the cerebral protective effect of puerarin, and indicate that the inhibition of puerarin to cell apotosis after cerebral resuscitation is related to its effect on the decrease in protein expression of apoptosis-promoting gene, Fas and P53.Puerarin has a protective effect on cerebral ischemia-reperfusion injury of rats. In comparison with model control group, the expression of Fas in puerarin treatment group has an obvious decrease inthe 6th to 48th hour after cerebral ischemia-reperfusion, the expression of P53 has an obvious decrease in the 24th to 48th hour after cerebral ischemia-reperfusion, and the number of apoptosis cells decrease obviously, too, which further improves the cerebral protective effect of puerarin and indicates that the inhibition of puerarin to cell apoptosis after cerebral resuscitation is related to its effect on the decrease in protein expression of apoptosis-promoting gene Fas and P53.