ObjectiveTo study the effect of Schwann cells (SCs) promoting the function of nitric oxide (NO) secretion of bone marrow mesenchymal stem cells (BMSCs) derived endothelial cells so as to lay the experimental foundation for research of the effect of nerves on vessels during the process of tissue engineering bone formation. MethodsSCs were collected from 1-day-old Sprague Dawley (SD) rats,and identified through S100 immunohistochemistry (IHC).BMSCs were collected from 2-week-old SD rats and induced into endothelial cells (IECs),which were identified through von Willebrand factor (vWF) and CD31 immunofluorescence (IF).Transwell system was used for co-culture of SCs and IECs without contact as the experimental group,and simple culture of IECs served as the control group.The NO concentration in the medium was measured at 1,3,5,and 7 days after culture; the mRNA expressions of nitric oxide synthetase 2 (NOS2) and NOS3 were detected by real-time fluorescence quantitative PCR (RT-qPCR) at 1,3,7,and 10 days. ResultsSCs and IECs were identified through morphology and immunology indexes of S100 IHC,vWF and CD31 IF.Significant differences were found in the NO concentration among different time points in 2 groups (P<0.05); the NO concentration of the experimental group was significantly higher than that of the control group at the other time points (P<0.05) except at 3 days.NOS2 mRNA expression of the experimental group was significantly higher than that of the control group (P<0.05); difference was significant in the NOS2 mRNA expression among different time points in 2 groups (P<0.05).NOS3 mRNA expression of the experimental group was significantly higher than that of the control group at the other time points (P<0.05) except at 10 days.No significant difference was found in NOS3 mRNA expression among different time points in the experimental group (F=6.673,P=0.062),but it showed significant differences in the control group (F=36.581,P=0.000). ConclusionSCs can promote NO secretion of BMSCs derived endothelial cells,which is due to promoting the activity of NOS.
In order to establish the best procedure to store the femur samples from the biomechanical viewpoint, we compared the effects of different storage methods on the mechanical properties of mouse femurs. We obtained femurs surgically from twenty C57BL/6J female mice, 12 weeks old, and randomly divided them into 5 groups, i.e. fresh control group, 4% paraformaldehyde fixation group, 4℃ storage group, -20℃ storage group and -80℃ storage group, respectively, with five mice in each group. For the three low-temperature storage groups, each group was stored for 1 week, 2 months, 6 months at their respective temperatures. After rewarming, three-point bending test was performed to test the load and deflection changes. The results showed that both the elastic modulus and deflection decreased significantly in the 4% paraformaldehyde group. The maximum load and elastic modulus of the samples in the 4℃ group after one week storage was significantly reduced; The mechanical properties were close to the fresh control group in the -20℃ group stored for 2 months but the maximum load was also reduced after 6 months. However, mechanical properties, such as elastic load, maximum load and elastic modulus, were not changed obviously in the -80℃ storage group. Accordingly, -80℃ cryopreservation had little influence on the mechanical properties of bone tissues, which proved that the temperature -80℃ is a suitable one for long-term preservation.
ObjectiveTo explore the effectiveness of excision and reconstruction of bone tumor by using operation guide plate made by variety of three-dimensional (3-D) printing techniques, and to compare the advantages and disadvantages of different 3-D printing techniques in the manufacture and application of operation guide plate. MethodsBetween September 2012 and January 2014, 31 patients with bone tumor underwent excision and reconstruction of bone tumor by using operation guide plate. There were 19 males and 12 females, aged 6-67 years (median, 23 years). The disease duration ranged from 15 days to 12 months (median, 2 months). There were 13 cases of malignant tumor and 18 cases of benign tumor. The tumor located in the femur (9 cases), the spine (7 cases), the tibia (6 cases), the pelvis (5 cases), the humerus (3 cases), and the fibula (1 case). Four kinds of 3-D printing technique were used in processing operation guide plate:fused deposition modeling (FDM) in 9 cases, stereo lithography appearance (SLA) in 14 cases, 3-D printing technique in 5 cases, and selective laser sintering (SLS) in 3 cases; the materials included ABS resin, photosensitive resin, plaster, and aluminum alloy, respectively. Before operation, all patients underwent thin layer CT scanning (0.625 mm) in addition to conventional imaging. The data were collected for tumor resection design, and operation guide plate was designed on the basis of excision plan. Preoperatively, the operation guide plates were made by 3-D printing equipment. After sterilization, the guide plates were used for excision and reconstruction of bone tumor. The time of plates processing cycle was recorded to analyse the efficiency of 4 kinds of 3-D printing techniques. The time for design and operation and intraoperative fluoroscopy frequency were recorded. Twenty-eight patients underwent similar operations during the same period as the control group. ResultsThe processing time of operation guide plate was (19.3±6.5) hours in FDM, (5.2±1.3) hours in SLA, (8.6±1.9) hours in 3-D printing technique, and (51.7±12.9) hours in SLS. The preoperative design and operation guide plate were successfully made, which was used for excision and reconstruction of bone tumor in 31 cases. Except 3 failures (operation guide plate fracture), the resection and reconstruction operations followed the preoperative design in the other 28 cases. The patients had longer design time, shorter operation time, and less fluoroscopy frequency than the patients of the control group, showing significant differences (P<0.05). The follow-up time was 1-12 months (mean, 3.7 months). Postoperative X-ray and CT showed complete tumor resection and stable reconstruction. Conclusion3-D printing operation guide plates are well adapted to the requirements of individual operation for bone tumor resection and reconstruction. The 4 kinds of 3-D printing techniques have their own advantages and should be chosen according to the need of operation.