ObjectiveTo observe the in vivo three-dimensional (3-D) transient motion characteristics of the subaxial cervical spine in healthy adults. MethodsSeventeen healthy volunteers without cervical spine related diseases were recruited for this study, including 8 males and 9 females with a mean age of 26 years (range, 23-41 years). The vertebral segment motion of each subject was reconstructed with CT, and Rhinoceros 4.0 solid modeling software were used for 3-D reconstruction model of the subaxial cervical spine. In vivo cervical vertebral motion in flexionextension, left and right bending, left and right rotation was observed with dual fluoroscopic imaging system (DFIS). Coordinate systems were established at the vertebral center of C3-7 to obtain the intervertebral range of motion (ROM) and displacement at C3, 4, C4, 5, C5, 6, and C6, 7. The X-axis pointed to the left along the coronal plane, the Y-axis pointed to the back along the sagittal plane, and the Z-axis perpendicular to the X-Y plane pointed to the head. The ROM along X, Y, and Z axises were represented by rotation in flexion-extension (α), in left-right bending (β), and in left-right twisting (γ) respectively, and the displacement in left-right direction (x), in anterior-posterior direction (y), and in proximaldistal direction (z), respectively. ResultsIn flexion and extension, the displacement in anterior-posterior direction of C6, 7 was significantly less that of other segments (P<0.05), but the displacements in left-right direction and in proximaldistal direction showed no significant difference between segments (P>0.05); the ROM values in flexion-extension of C4, 5 and C5, 6 were significantly larger than those of C3, 4 and C6, 7 (P<0.05), and the ROM value in left-right twisting of C4, 5 was significantly larger than those of C5, 6 and C6, 7 (P<0.05), but the ROM value in left-right bending showed no significant difference between segments (P>0.05). In left and right bending, there was no significant difference in the displacement between other segments (P>0.05) except that the displacement in anterior-posterior direction of C3, 4 was significantly larger than that of C4, 5 (P<0.05), and that the displacement in proximal-distal direction of C6, 7 was significantly less than that of C3, 4 and C4, 5 (P<0.05); no significant difference was shown in the ROM value between segments (P>0.05), except that the ROM value in left-right twisting of C3, 4 was significantly larger than that of C5, 6 and C6, 7 (P<0.05). In left and right rotation, the ROM value in left-right twisting of C3, 4 was significantly larger than that of C4, 5 and C6, 7 (P<0.05), and the displacement and ROM value showed no significant differece between other segments (P>0.05). ConclusionThe intervertebral motions of the cervical spine show different characters at different levels. And the 6-degree-of-freedom data of the cervical vertebrae are obtained, these data may provide new information for the in vivo kinematics of the cervical spine.
Objective To evaluate the influence of PKH26 labeling on the biological function of the goat nucleus pulposus cells and the biological function of seeded cells in nude mice by in vivo imaging techonology. Methods Primary nucleus pulposus cells were isolated by enzymatic digestion from the nucleus pulposus tissue of the 1-year-old goat disc. The nucleus pulposus cells at passage 1 were labeled with PKH26 and the fluorescent intensity was observed under the fluorescence microscopy. The labeled cells were stained with toluidine blue and collagen type II immunocytochemistry. The cells viability and proliferation characteristics were assessed by trypan blue staining and MTT assay, respectively. Real-time fluorescent quantitative PCR was used to detect the gene expressions of collagen types I and II, and aggrecan. The fluorescent intensity and scope of the nucleus pulposus cells-scaffold composite in vivo for 6 weeks after implanting into 5 6-week-old male nude mice were measured by in vivo imaging technology. Results Primary nucleus pulposus cells were ovoid in cell shape, showing cluster growth, and the cells at passage 1 showed chondrocyte-like morphology under the inverted phase contrast microscope. The results of toluidine blue and collagen type II immunocytochemistry staining for nucleus pulposus cells at passage 1 were positive. The fluorescent intensity was even after labeling, and the cell viability was more than 95% before and after PKH26 labeling. There was no significant difference in cell growth curve between before and after labeling (P gt; 0.05). The real-time fluorescent quantitative PCR showed that there was no significant difference in gene expressions of collagen types I and II, and aggrecan between before and after labeling (P gt; 0.05). Strong fluorescence in nucleus pulposus cells-scaffold composite was detected and by in vivo imaging technology. Conclusion The PKH26 labeling has no effect on the activity, proliferation, and cell phenotype gene expression of the nucleus pulposus cells. A combination of PKH26 labeling and in vivo imaging technology can track the biological behavior of the cells in vivo.
Objective To investigate the in vivo degradable properties of new calcium phosphate cement (CPC) containing poly lactic-co-glycolic acid (PLGA) so as to lay a foundation for the future clinical application. Methods A novel CPC containing PLGA (CPC/PLGA) was prepared according to a ratio of 45% dicalcium phosphate anhydrous ∶ 45% partially crystallized calcium phosphates ∶ 10% PLGA. Thirty-two adult New Zealand rabbits (weighing 2.2-3.0 kg, male or female in half) were divided into the experimental group (n=17) and the control group (n=15). The bone defect models of the bilateral femoral condyles (4.5 mm in diameter and 1.5 cm in depth) were made by drilling hole. Defect at the right side was repaired with CPC/ PLGA in the experimental group and with CPC in the control group, while defect at the left side was not treated as blank control. The general condition of rabbits was observed after operation; the histological observation and bone histomorphometric analysis were performed at 2, 4, 8, 16, and 24 weeks; and scanning electronic microscope (SEM) observation was performed at 8 and 16 weeks after operation. Results All rabbits survived to the end of experiment. The histological observation showed: CPC/PLGA degraded gradually, and the new-born bone trabecula ingrew; bone trabeculae became rough and b; and CPC/PLGA almost biodegraded at 24 weeks in the experimental group. The CPC degradation was much slower in the control group than in the experimental group. The total bone tissue percentage was 44.9% ± 23.7% in the experimental group, and 25.7% ± 10.9% in the control group, showing significant difference between 2 groups (t=3.302, P=0.001); and the bone tissue percentage showed significant difference between 2 groups at 8, 16, and 24 weeks (P lt; 0.05). The results of SEM observation showed that the pore size was 100-300 μm at 8 weeks after operation, new-born bone trabecula grew into the pores and combined bly with residual cement in the experimental group. Conclusion Novel CPC/PLGA has good in vivo degradable properties, and it can be an ideal bone substitute in future clinical application.
Objective To monitor the importance of establishing lung cancer models for immunological treatment through in vivo imaging system (IVIS). Methods In this study, a new optical bioluminescence IVIS was used to confirm the tumour formation and luminescence in male BALB/c nude mice by injecting A549-luc cells. First, A549-luc cells which expressed luciferase stably were transferred into nude mice by tail vein injection in order to establish a stable and reliable model of lung cancer. Then, D-fluorescein potassium salt was intraperitoneally injected every other week. The tumor formation and growth were dynamically observed on day 7th, 14th and 21st by IVIS Spectrum and pathological exam with hematoxylin-eosin staining. Results Animal model of lung cancer was successfully established, and the development of lung cancer was effectively monitored by IVIS real-time fluorescence value which was consistent with pathological exam, and tumor volume was correlated with fluorescence intensity (r=0.7996, P<0.01). Conclusions IVIS has multiple benefits, including high sensitivity and specificity, simple operation, and no radiation. IVIS Spectrum can measure the fluorescence of tumor formed by injection of A549-luc cells in nude mice metastasis of lung cancer in a non-invasive, real-time and dynamic mode, which is worthy of promotion for using in clinical research.
ObjectiveThis study construct a pulmonary fibrosis model in vivo to study anti-pulmonary fibrosis effect of ampelopsis.MethodsWe constructed a pulmonary fibrosis model by bleomycin in BALB/c mice. The mice were divided by weight random number table into a blank control group, a model control group, a dexamethasone treatment group (intervened with dexamethasone in a dose of 2.5 mg/kg), and three ampelopsis treatment groups intervened with ampelopsis in dose of 200, 100, and 50 mg/kg, respectively. Bleomycin solution (3 mg/kg) was intratracheally injected respectively on 1st and 14th day, except the blank group. Twenty-eight days later, the relevant indicators were collected, including respiratory function (airway resistance, dynamic lung compliance, maximal ventilator volume), level of hydroxyproline and histopathological changes in the lungs.ResultsAfter 28 days, the model control group mice had severe respiratory resistance, dynamic lung compliance and maximal ventilator volume were decreased. The high dose ampelopsis treatment could enhance respiratory function (P<0.05). Lung coefficient was lower in the treatment groups than that in the model control group (P<0.05). The hydroxyproline of the treatment groups was less than that of the model control group (P<0.05). Histopathological examination showed that the degree of fibrosis increased in the model control group (P<0.05), but decreased in the treatment groups (P<0.05).ConclusionAmpelopsis can resist bleomycin-induced pulmonary fibrosis in mice, relieve the symptoms of respiratory failure, reduce the formation of collagen, and produce anti-pulmonary fibrosis effect.
ObjectiveTo evaluate the most efficient method for transfection of human umbilical cord mesenchymal stem cells (HUMCSs) in vivo. MethodsHUCMSCs were isolated from human umbilical cord and cultured, which were labelled by PKH26 and lentivirus-GFP, then were observed by using a fluorescence microscope. Sixty SD rats were randomly divided into PKH26 transfection group and lentivirus-GFP transfection group. The right hepatic lobe of rat was resected, then the transfected stem cells were injected into portal vein. The rats were sacrificed on day 3, 8, and 13 after transfection. The liver specimens were observed by using a fluorescence microscope. Flow cytometry was used to evaluate the percentage of transfected stem cells and the apoptotic stem cells. ResultsThe third generation of HUCMSCs labelled by PKH26 and lentivirus-GFP were spindle shaped. PKH26 red dye was evenly distributed in the cell membrane of HUCMSCs and could be clearly labelled. The HUCMSCs labelled by lentivirus-GFP were green fluorescence under the fluorescence microscope, and it was clear and stable. The HUCMSCs were clear and could be clearly distinguished on day 3 after transfection by two methods in vivo. As the time went by, red was faded and blurred, then was gradually disappeared on day 13 after transfection in the HUCMSCs stansfected by PKH26; but the color in the HUCMSCs stansfected by lentivirus-GFP were clear at all the time points. The transfection rate of the lentivirus-GFP was significantly higher that that of the PKH26 (P < 0.05), the rate of apoptotic stem cells had no significant differences at all the time points between these two groups (P > 0.05). ConclusionLentivirus-GFP transfection is a higher efficient method for stem cell labelling in vivo, it could be used to observe transplantation cells for a long time in future.
In vivo transplantation of tracheal grafts utilizes natural environment in vivo to improve cell adhesion, growth and scaffold properties, which can not only promote graft revascularization, but also induce immune tolerance and increase postoperative survival rate. Decellularized trachea with stem cells covering the outside layer and airway epithelial cells covering the inside layer can achieve complete mucosa re-epithelialization, cartilage cell growth and revascularization, using own body as a natural bioreactor to boost the maturity of tissue engineered trachea. Then transplantation at a normotopic in situ positioning is performed. This transplantation strategy provides a promising approach for the treatment of long-segment tracheal defects. This review focuses on the significance and research progress of constructing tissue engineered trachea in vivo.
ObjectiveTo review the researches of in vivo kinematics in lumbar degenerative spondylol isthesis (DS). MethodsRelated literature concerning the in vivo kinematics in patients with lumbar DS was extensively reviewed and comprehensively analyzed in 4 terms of the instabil ity of lumbar DS, vertebral motion pattern, the morphological changes of spinal canal, and intraoperative biomechanical measurement. ResultsWhether there is lumbar segmental instabil ity in lumbar DS patients is still controversial, which should be based on degenerative stage of lumbar spine and grade of slip. The hypomobility of the lumbar spinous processes and the facet joint is seen in DS. The diameter, cross-sectional area, and volume of spinal canal in lumbar DS patients are significantly smaller than those of the normal control. Because of its invasive procedure and medical ethics, the use of the intraoperative measurement device is limited. These reported researches of in vivo kinematics in DS are almost on the sagittal plane. However, few data have been reported on the 6-degree-of-freedom (6DOF) kinematics of the diseased levels under physiological loading conditions. ConclusionThe 6DOF kinematics data can accurately reflect the segmental motion characteristics in lumbar DS patients, recent studies have been reported, further studies are still needed.
ObjectiveTo review the application and research progress of in vivo bioreactor as vascularization strategies in bone tissue engineering. MethodsThe original articles about in vivo bioreactor that can enhance vascularization of tissue engineered bone were extensively reviewed and analyzed. ResultsThe in vivo bioreactor can be created by periosteum, muscle, muscularis membrane, and fascia flap as well as biomaterials. Using in vivo bioreactor can effectively promote the establishment of a microcirculation in the tissue engineered bones, especially for large bone defects. However, main correlative researches, currently, are focused on animal experiments, more clinical trials will be carried out in the future. ConclusionWith the rapid development of related technologies of bone tissue engineering, the use of in vivo bioreactor will to a large extent solve the bottleneck limitations and has the potential values for clinical application.
ObjectiveTo explore effect and mechanism of the carcinoma associated fibroblasts (CAFs) of breast cancer on growth and metastasis of breast cancer induced in nude mice by inoculation of CAFs and breast cancer cells. MethodsBreast cancer cell line of MDA-MB-231 (abbreviated as MDA), CAFs, and normal breast tissue fibroblasts (NFs) of the same breast cancer patient were collected, and mixed with normal saline (NS) or SDF-1 ligand blockers of four nitrogen heterocyclic fourteen alka (AMD3100, abbreviated as AMD) for inoculation of nude mice in vivo. According to the different combination, 36 nude mice were randomly divided into 6 groups:MDA+NS group, NFs+NS group, MDA+NFs+NS group, MDA+NFs+AMD group, MDA+CAFs+AMD group, and MDA+CAFs+NS group. Forty six days after the inoculation and feeding, volume of tumor, metastasis of lymph node, lung or liver were observed. In addition, level of plasma SDF-1 was tested by using ELISA method, and expressions of SDF-1 mRNA and protein in tumor specimens were detected by using real-time PCR and Western blot method respectively. ResultsExcept for NFs+ NS group, implanted tumor could be seen in nude mice of other 5 groups. In MDA+CAFs+NS group, the volume of tumor[(9.092±2.662) cm3], level of plasma SDF-1[(75.25±16.23) ng/L], and expression levels of SDF-1 mRNA (the median level was 14.714) and protein (the median level was 0.673). of tumor tissue were significantly greater or higher than those of the other 5 groups (P < 0.050). In addition, lymph node metastasis were found in 4 mice in MDA+CAFs+NS group, and 2 in MDA+NS group. The tumor metastasis of lung and liver was not found in all nude mice. ConclusionsCAFs can promote growth and lymph node metastasis of breast cancer, whose mechanism is related with SDF-1 secreted by CAFs and SDF-1/CXC chemokine receptor 4 (CXCR4), signal pathway.