Wang Z.,Xian Honghui Hospital Of Xian Jiaotong University |
Zhang J.,Xian Honghui Hospital Of Xian Jiaotong University |
Wang D.-F.,Xian Honghui Hospital Of Xian Jiaotong University |
Shao Y.-X.,Xian Honghui Hospital Of Xian Jiaotong University |
And 3 more authors.
Chinese Journal of Tissue Engineering Research | Year: 2015
BACKGROUND: Many studies have confirmed that recombinant human bone morphogenetic protein 2 plays a very important role in bone formation and fracture healing, but recombinant human bone morphogenetic protein 2 alone implanted is prone to diffusion and degradation, which is unable to play a persistent role in new bone formation. OBJECTIVE: To explore the effect of recombinant human bone morphogenetic protein 2 composite bone in the rabbit lumbar fusion. METHODS: Thirty New Zealand white rabbits were selected to make posterior lumbar intertransverse fusion models, and then were randomly divided into three groups, in which, L5-6 intertransverse implantation of autologous iliac bone, allogeneic bone and recombinant human bone morphogenetic protein 2 composite bone (recombinant human bone morphogenetic protein 2 and allogeneic bone comptex) was done respectively. At 6 weeks after implantation, gross observation, X-ray examination and histological observation were performed. RESULTS AND CONCLUSION: Fusion rate and percentage of new bone area were higher in the composite bone group than the autologous iliac bone and allogeneic bone groups (P < 0.05); the tensile strength was lower in the allogeneic bone group than the other two groups (P < 0.05), but there was no difference between these two groups except the allogeneic bone group. X-ray films showed callus formation in the implanted region of the three groups. In the autologous iliac bone group, a large amount of cartilage tissues formed along with a small amount of bone trabeculae and a certain amount of woven bones. In the allogeneic bone group, the implant was covered with a large amount of fibrous tissues, bone island was seen and there was also a small amount of bone trabeculae and cartilage tissues. In the composite bone group, a great amount of bone trabeculae and cartilage tissues were visible to form woven bone and cortical bone. These findings indicate that the recombinant human bone morphogenetic protein 2 composite bone can obtain good effect in the rabbit lumbar fusion. © 2015, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.
Lu M.,Second Affiliated Hospital Of Xian Jiaotong University |
Dong J.,Second Affiliated Hospital Of Xian Jiaotong University |
Lu T.,Second Affiliated Hospital Of Xian Jiaotong University |
Lv H.,First Affiliated Hospital Of Xian Jiaotong University |
And 7 more authors.
International Journal of Molecular Sciences | Year: 2015
Transplantation of olfactory ensheathing cells (OEC) is a promising therapy in spinal cord injury (SCI) treatment. However, the therapeutic efficacy of this method is unstable due to unknown reasons. Considering the alterations in the culture environment that occur during OEC preparation for transplantation, we hypothesize that these changes may cause variations in the curative effects of this method. In this study, we compared OEC cultured in medium containing different types and concentrations of serum. After purification and passage, the OEC were cultured for 7 days in different media containing 5%, 10%, 15% or 20% fetal bovine serum (FBS) or rat serum (RS), or the cells were cultured in FBS-containing medium first, followed by medium containing RS. In another group, the OEC were first cultured in 10% FBS for 3 days and then cultured with rat spinal cord explants with 10% RS for another 4 days. An MTT assay and P75 neurotrophin receptorimmunofluorescence staining were used to examine cell viability and OEC numbers, respectively. The concentration of neurotrophin-3 (NT-3), which is secreted by OEC into the culture supernatant, was detected using the enzyme-linked immunosorbent assay (ELISA). RT-PCR was applied to investigate the NT-3 gene expression in OEC according to different groups. Compared with FBS, RS reduced OEC proliferation in relation to OEC counts (χ2 = 166.279, df = 1, p < 0.01), the optical density (OD) value in the MTT assay (χ2 = 34.730, df = 1, p < 0.01), and NT-3 concentration in the supernatant (χ2 = 242.997, df = 1, p < 0.01). OEC cultured with spinal cord explants secreted less NT-3 than OEC cultured alone (F = 9.611, df = 5.139, p < 0.01). Meanwhile, the order of application of different sera was not influential. There was statistically significant difference in NT-3 gene expression among different groups when the serum concentration was 15% (χ2 = 64.347, df = 1, p < 0.01). In conclusion, different serum conditions may be responsible for the variations in OEC proliferation and function. © 2014 by the authors; licensee MDPI, Basel, Switzerland.
Zhao Q.-P.,Xian Honghui Hospital Of Xian Jiaotong University
Chinese Journal of Tissue Engineering Research | Year: 2015
BACKGROUND: Bone marrow mesenchymal stem cells are non-hematopoietic stem cells from the bone marrow and can differentiate into chondrocytes, osteocytesand adipocytes under different induction conditions, which are the most promising seed cells for tissue engineering. OBJECTIVE: To evaluate the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cel-under directional induction. METHODS: Bone marrow samples from Japanese rabbits were extracted aseptically to isolate and purify bone marrow mesenchymal stem cells using density gradient centrifugation and cell adherent method. Then, the cells were cultured in osteogenic and adipogenic induction media as experimental group. Another cells cultured in DMEM medium acted as control group. Sudan IV staining, Von Kossa staining and alkaline phosphatase activity detection were performed at different time after culture to compare the osteogenic and adipogenic rates. RESULTS AND CONCLUSION: After 7 days of adipogenic induction, small lipid droplets were visible and the cells arranged disorderly; at 21 days of induction, see lipid droplets with high refractivity were sen in the cytoplasm. Sudan IV staining showed that there were a large amount of red lipid droplets in the cloning center of bone marrow mesenchymal stem cells, but only 5% bone marrow mesenchymal stem cells differentiated into adipocytes in the control group. At 7 days of osteogenic induction, induced cells were confluent in a flagstone shape, and there were many nodules; at 14 days of culture, the center of brown mineralized nodules were visible; at 21 days of culture, small pieces of mineralized nodules formed. The osteogenic and adipogenic rates were 40% and 20% in the experimental group, respectively, which were higher than those in the control group (5%, 5%). These findings indicate that under certain conditions, bone marrow mesenchymal stem cells can partially differentiate into fat cells part, and another part diiferentiate into osteocytes, suggesting there is a certain link between them, that is, more adipocytes and fewer osteoblasts, or vice versa. © 2015, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.