Objective To further study the influence of the co-cultivation of vascular endothel ial cells (VECs) and adi pose-derived stromal cells (ADSCs) on cell osteogenic differentiation in vitro and provide experimental evidences of the probabil ity of the co-cultivation of VECs and ADSCs as the seed cells of tissue engineering. Methods The VECs derived fromcord blood and ADSCs were prepared by full-term pregnancy SD rats and 18-week-old SD rats, to carry on the morphological observation and immunohistochemical staining identification. The third generation of ADSCs and the VECs induced by conditioned medium for 6 weeks were cultured and were divided into groups A, B, and C as the experimental group according to cell ratios of 3 ∶ 1, 1 ∶ 1, and 1 ∶ 3, respectively. ADSCs or VECs was cultured alone in groups D and E as control groups. ALP and al izarin red staining were done respectively on the 7th day and 14th day; ALP and osteocalcin (OC) were detected respectively on the 4th day, 7th day, and 14th day. Results The VECs derived from cord blood showed mixed growth of short spindle and polygonal cells after 6 weeks of induction, the immunofluorescent staining result of von Willebrand factor was positive. ADSCs showed adherent mononuclear cells and spindle-shaped growth without dupl ication; the immunofluorescent staining result of CD90 was positive and no positive cells were seen in the control group. On the 7th day of cell culture, ALP staining showed that the results were negative in groups A, D, and E, and some positive cells were seen in groups B and C; on the 14th day, the results were still negative in groups D and E, and positive cells fused to sheet form in groups A, B, and C. von Kossa staining showed that the results were negative in all groups on the 7th day; few positve cells were seen in groups A, B, and C, and no positive cells were seen in groups D and E on the 14th day. The ALP contents increased gradually in all groups,which was highest in group B at every time point, showing significant difference (P lt; 0.01) between group B and other groups, between groups A, C and groups D, E. The OC value increased gradually in every group, which was highest in group B on the 7th and 14th days, showing significant difference between group B and other groups (P lt; 0.01), between group C and group D (P lt; 0.01) on the 4th and the 14th days, between groups A, C and group E (P lt; 0.05) on the 14th day. Conclusion ADSCs have potential of osteogenic differentiation by VECs in the system of co-culturing VECs and ADSCs in vitro, the influence on osteogenic differentiation is the best in a ratio of 1 ∶ 1.
ObjectiveTo investigate the heterotopic osteogenesis of tissue engineered bone using the co-culture system of vascular endothelial cells (VECs) and adipose-derived stem cells (ADSCs) as seed cells.MethodsThe partially deproteinized biological bone (PDPBB) was prepared by fibronectin combined with partially deproteinized bone (PDPB). The ADSCs of 18-week-old Sprague Dawley (SD) rats and VECs of cord blood of full-term pregnant SD rats were isolated and cultured. Three kinds of tissue engineered bone were constructed in vitro: PDPBB+VECs (group A), PDPBB+ADSCs (group B), PDPBB+co-cultured cells (VECs∶ADSCs was 1∶1, group C), and PDPBB was used as control group (group D). Scanning electron microscopy was performed at 10 days after cell transplantation to observe cell adhesion on scaffolds. Forty-eight 18-week-old SD rats were randomly divided into groups A, B, C, and D, with 12 rats in each group. Four kinds of scaffolds, A, B, C, and D, were implanted into the femoral muscle bags of rats in corresponding groups. The animals were killed at 2, 4, 8, and 12 weeks after operation for gross observation, HE staining and Masson staining histological observation, and the amount of bone collagen was measured quantitatively by Masson staining section.ResultsScanning electron microscopy showed that the pores were interconnected in PDPB materials, and a large number of lamellar protein crystals on the surface of PDPBB modified by fibronection were loosely attached to the surface of the scaffold. After 10 days of co-culture PDPBB and cells, a large number of cells attached to PDPBB and piled up with each other to form cell clusters in group C. Polygonal cells and spindle cells were mixed and distributed, and some cells grew along bone trabeculae to form cell layers. Gross observation showed that the granulation tissue began to grow into the material pore at 2 weeks after operation. In group C, a large number of white cartilage-like substances were gradually produced on the surface of the material after 4 weeks, and the surface of the material was uneven. At 12 weeks, the amount of blood vessels on the surface of group A increased, and the material showed consolidation; there was a little white cartilage-like material on the surface of group B, but the pore size of the material did not decrease significantly; in group D, the pore size of the material did not decrease significantly. Histological observation showed that there was no significant difference in the amount of bone collagen between groups at 2 weeks after operation (F=2.551, P=0.088); at 4, 8, and 12 weeks after operation, the amount of bone collagen in group C was significantly higher than that in other 3 groups, and that in group B was higher than that in group D (P<0.05); there was no significant difference between group A and groups B, D (P>0.05).ConclusionThe ability of heterotopic osteogenesis of tissue engineered bone constructed by co-culture VECs and ADSCs was the strongest.
ObjectiveTo investigate the effectiveness of the posterior-posterior triangulation technique for arthroscopic posterior cruciate ligament (PCL) reconstruction by comparing with the anteroposterior approach.MethodsRetrospective analysis was performed on 40 patients who underwent arthroscopic PCL reconstruction between February 2016 and February 2020. The PCLs were reconstructed via anteroposterior approach in 20 patients (anteroposterior approach group) and posterior-posterior triangulation technique in 20 patients (posterior-posterior triangulation technique group). There was no significant difference in gender, age, cause of injury, injury side, disease duration, preoperative International Knee Documentary Committee (IKDC) score, and Lysholm score between the two groups (P>0.05). The operation time, surgical complications, and postoperative posterior drawer test, Lysholm score, and IKDC score were recorded and compared between the two groups.ResultsThe operation time was (65.25±10.05) minutes in the anteroposterior approach group and (56.15±8.15) minutes in the posterior-posterior triangulation technique group, and the difference was significant (t=3.145, P=0.003). All incisions healed by first intention, and there was no complication such as vascular and nerve injuries or infection. Patients were followed up (27.05±11.95) months in the anteroposterior approach group and (21.40±7.82) months in the posterior-posterior triangulation technique group, with no significant difference (t=1.770, P=0.085). At last follow-up, the posterior drawer tests were positive in 4 cases (3 cases of stageⅠand 1 case of stage Ⅱ) of the anteroposterior approach group and in 1 case (stageⅠ) of the posterior-posterior triangulation technique group, showing no significant difference between the two groups (P=0.342). At last follow-up, Lysholm score and IKDC score in both groups were significantly higher than those before operation (P<0.05). The above functional scores in the posterior-posterior triangulation technique group were significantly higher than those in the anteroposterior approach group (P<0.05). Imaging reexamination showed that the position, shape, and tension of the grafts were well in both groups, and the grafts were covered with the synovium in the posterior-posterior triangulation technique group, the meniscofemoral ligaments were well preserved. There was no re-rupture of the reconstructed ligament during follow-up.ConclusionCompared to the anteroposterior approach, the posterior-posterior triangulation technique provides a clearer view under arthroscopy, no blind spot, sufficient operating space, and relative safety. Moreover, it is easier to retain the remnant and the meniscofemoral ligaments, and can obtain good short-term effectiveness.
Objective To review the studies about the tibial-graft fixation methods on anterior cruciate ligament (ACL) reconstruction, in order to provide clinical reference. MethodsThe literature about the tibial-graft fixation methods on ACL reconstruction at home and abroad was extensively reviewed, and the factors that affect the selection of fixation methods were summarized. Results The knee flexion angle, graft tension, and graft fixation device are mainly considered when the tibial-graft was fixed on ACL reconstruction. At present, the graft is mainly fixed at 0°/30° of knee flexion. The study shows that the knee joint is more stable after fixed at 30°, while the incidence of knee extension limitation decrease after fixed at 0°. In terms of graft tension, a good effectiveness can be obtained when the tension level is close to 90 N or the knee flexion is 30° to recover the affected knee over-restrained 2 mm relative to the healthy knee. In terms of the graft device, the interference screw is still the most commonly used method of tibial-graft fixation, with the development of all-inside ACL reconstruction in recent years, the cortical button fixation may become the mainstream. Conclusion Arthroscopic reconstruction is the main treatment of ACL rupture at present. However, there is no optimal fixation method for the tibial-graft, the advantages and disadvantages of each fixation methods need to be further studied.