Effects of Tissue-engineered Bone by Coculture of Adipose-derived Stem Cells and Vascular Endothelial Cells on Host Immune Status.

AIM: The study aimed to explore the effects of tissue-engineered bone constructed with partially deproteinized biologic bone (PDPBB) and coculture of adipose-derived stem cells (ADSCs) and vascular endothelial cells (VECs) on host immune status, providing a very useful clue for the future development of bone engineering. METHODS: Tissue-engineered bones constructed by PDPBB and ADSCs, VECs or coculture of them were implanted into the muscle bag of bilateral femurs of Sprague-Dawley rats. Partially deproteinized biologic bone alone and blank control were also implanted. After transplantation, the proliferation of implanted seed cells in tissue-engineered bones was labeled by bromodeoxyuridine staining. Moreover, the changes of T-lymphocyte subpopulations, including CD3 + CD4+ and CD3 + CD8+ in peripheral blood were then detected using flow cytometry to analyze the immune rejection of tissue-engineered bone implantation based on peripheral blood CD4/CD8 ratios. RESULTS: After transplantation, the proliferation of implanted seed cells was observed in tissue-engineered bones of different groups. At different time points after transplantation, the CD4+/CD8+ ratio in peripheral blood of PDPBB + ADSCs, PDPBB + coculture, and blank control groups did not exhibit significant change. Although the CD4+/CD8+ ratio in peripheral blood of PDPBB + VECs group was significantly higher than other group at 1 week after transplantation, that of PDPBB + VECs and PDPBB + coculture group was significantly decreased at 8 week after transplantation compared with that of blank control group. CONCLUSIONS: Our results indicated that there was no significant immune rejection after transplantation of tissue-engineered bone constructed with PDPBB and coculture of ADSCs and VECs as seed cells.

Combined treatment with Prunella vulgaris and Radix bupleuri activated the Bax/Bcl-2 and Caspase-3 signal pathways in papillary thyroid carcinoma cells.

To explore the effect of Prunella vulgaris (PV) combined with Radix bupleuri (RB) on apoptosis of papillary thyroid carcinoma cells. Our study was divided into four groups: the control group, the PV group, the RB group, and the PV combined with the RB group. The viability of cells from different treatment groups was assessed by the CCK-8 assay. Cell migration and invasion were assessed by healing wounding and the transwell assay, respectively. Cell apoptosis rate and cell cycle arrest were detected by a flow cytometry assay. The protein expression of Bcl-2, Bax, Caspase-3, CyclinA1, CyclinB1, and CDK1 was detected using a western blot assay. Our results indicated that, compared with the control group, PV combined with RB group could significantly alter the cell morphology, inhibit cell migration and invasion, decrease the number of cells in the G0/G1 phase and increase the number of cells in the G2/M phase, and promote the cell apoptosis. Moreover, PV combined with RB treatment also obviously increased the expression of Bax/Bcl2 and caspase-3 proteins and decreased the expression of Cyclin A1, Cyclin B1, and CDK1 proteins. Overall, our results indicated that PV combined with RB could activate the Bax/Bcl-2 and Caspase-3 signal pathways to induce cell apoptosis in papillary thyroid carcinoma cells; this also provides a new way to treat thyroid cancer.

Comparison of Rapid Rehabilitation after Anterior Cruciate Ligament Reconstruction with Tensioning Technique and Traditional Rehabilitation.

To investigate the efficacy of a fast rehabilitation program for the recovery of knee joint function after arthroscopic autologous hamstring tendon transplantation for reconstruction of the anterior cruciate ligament (ACL), from January 1, 2017, to March 31, 2019, a total of 65 patients with ACL injury were randomly divided into a study group and a control group. Both groups were treated with autologous hamstring tendon to reconstruct the anterior cruciate ligament, arthroscopic transplantation, and decompression techniques. The research group was treated with a fast rehabilitation program. The control group was treated with traditional rehabilitation program. Knee flexion angles were measured at 2, 4, and 8 weeks postoperatively. KT-1000 knee anterior stability was measured at 3, 6, and 12 months after operation. Knee function was assessed by subjective knee function assessment scale (IKDC) and Lysholm knee score. The knee curvature, KT-1000 measurement, IKDC score, and Lysholm score were compared between the two groups before and after treatment. KT-1000 measured value, IKDC score, and Lysholm score in 2 groups were significantly improved 3, 6, and 12 months compared with those before treatment, and the difference was statistically significant (P < 0.001). Comparison between the two groups: 2 weeks, 4 weeks, and 8 weeks after treatment, the knee curvature in the study group was better than that in the control group, and the difference was statistically significant (P < 0.001); there was no significant difference in the measured values of KT-1000 between the two groups 3, 6, and 12 months after treatment (P > 0.05); IKDC score and Lysholm score in the study group 3 and 6 months after treatment were significantly better than those in the control group, with statistical significance (P < 0.001); there was no significant difference in IKDC score and Lysholm score between the two groups 12 months after treatment (P >0.05). Autograft hamstring tendon transplantation and tense-reducing technique for anatomical reconstruction of anterior cruciate ligament under arthroscopy combined with rapid rehabilitation program can quickly, safely, and effectively restore the knee function of patients, greatly shortening the rehabilitation period of patients.

Construction of tissue engineering bone with the co­culture system of ADSCs and VECs on partially deproteinized biologic bone in vitro: A preliminary study.

Scaffold­based bone tissue engineering has therapeutic potential in the regeneration of osseous defects. The present study aimed to explore the adhesion and cell viability of a co­culture system composed of vascular endothelial cells PI­/Annexin V+ represents early apoptotic cells, and PI+/Annexin V+ represents late apoptotic cells (VECs) and adipose­derived stem cells (ADSCs) on partially deproteinized biologic bone (PDPBB) in vitro, and determine the optimum time period for maximum cell viability that could possibly be used for standardizing the scaffold transplant into the in vivo system. VECs and ADSCs were isolated from pregnant Sprague­Dawley rats and confirmed by immunostaining with von Willebrand factor and CD90, respectively. PDPBB was prepared using standardized protocols involving coating partially deproteinized bone with fibronectin. PDPBB was incubated in a mono­culture with VECs or ADSCs, or in a co­culture with both of these cells at a ratio of 1:1. An MTT assay was used to assess the adhesion and cell viability of VECs and ADSCs on PDPBB in the three different cultures. Scanning electron microscopy was used to observe the adhesion, cell viability and morphology of the different types of cells on PDPBB. It was observed that the absorbance of each group increased gradually and peaked on the 10th day; the highest absorbance was found for the co­cultured cells group. The difference of cell viability between each cell group was statistically significant. On the 10th day, in the co­cultured cells group, several cells adhered on the PDPBB material and a nest­like distribution morphology was observed. Therefore, the adhesion and cell viability of the co­cultured cells was higher compared with the mono­cultures of VECs or ADSCs. As cell viability was highest on the 10th day, this could be the optimal length of time for incubation and therefore could be used for in vivo experiments.

Dynamic alterations of spontaneous neural activity in patients with amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a multi-system disease featured by movement disorder. Studies on ALS using static neuroimaging indexes demonstrated inconsistent results. However, recent work indicated that the intrinsic brain activity was time-varying, and the abnormal temporal dynamics of brain activity in ALS remains unknown. Resting-state functional magnetic resonance imaging data were first obtained from 54 patients with ALS and 54 healthy controls (HCs). Then the dynamic regional homogeneity (d-ReHo) was calculated and compared between the two groups. Correlation analyses between altered d-ReHo and clinical scores were further performed. Compared with HCs, ALS patients showed higher d-ReHo in the left lingual gyrus while lower d-ReHo in the left rectus gyrus and left parahippocampal gyrus. Moreover, the d-ReHo in the left lingual gyrus exhibited correlation with disease progression rate in ALS at a trend level. Our findings suggested that altered dynamics in intrinsic brain activity might be a potential biomarker for diagnosing of ALS.

The use of a 3D-printed individualized navigation template to assist in the anatomical reconstruction surgery of the anterior cruciate ligament.

BACKGROUND: To explore the location accuracy and early clinical outcomes of using a 3D-printed individualized navigation template to assist in the reconstruction of the anterior cruciate ligament (ACL). METHODS: A single center randomized control study was conducted. Patients with ACL injury were treated with a conventional operation or an operation assisted by a 3D-printed individualized navigation template (the 3D group). The primary endpoint was the accuracy of the actual reconstruction compared with the planned position. RESULTS: There were 20 and 23 participants in the conventional group and the 3D group, respectively. There were no differences in the bone tunnel position between the actual postoperative position and the preoperative design in the 3D group (P>0.05). Compared with the 3D group, the positioning of the femoral tunnel was more inferior and shallower in the conventional group (P<0.05). The position of the tibia tunnel was closer to the anterior and medial edge of the tibial platform in the conventional group compared to the 3D group (P<0.05). The intraoperative positioning time was shorter in the 3D group than in the conventional group (3.3±1.0 vs. 5.9±1.8 minutes, P<0.001). The Lysholm and International Knee Documentation Committee scores did not differ between the two groups (P>0.05 for both), and all patients improved after surgery (P<0.001). CONCLUSIONS: The 3D-printed individualized navigation template showed good location accuracy and resulted in reduced intraoperative positioning time compared to the traditional method for ACL reconstruction.

Static and dynamic alterations in the amplitude of low-frequency fluctuation in patients with amyotrophic lateral sclerosis.

BACKGROUND: Static changes in local brain activity in patients suffering from amyotrophic lateral sclerosis (ALS) have been studied. However, the dynamic characteristics of local brain activity are poorly understood. Whether dynamic alterations could differentiate patients with ALS from healthy controls (HCs) remains unclear. METHODS: A total of 54 patients with ALS (mean age = 48.71 years, male/female = 36/18) and 54 (mean age = 48.30 years, male/female = 36/18) HCs underwent magnetic resonance imaging scans. To depict static alterations in cortical activity, amplitude of low-frequency fluctuations (ALFF) which measures the total power of regional activity was computed. Dynamic ALFF (d-ALFF) from all subjects was calculated using a sliding-window approach. Statistical differences in ALFF and d-ALFF between both groups were used as features to explore whether they could differentiate ALS from HC through support vector machine method. RESULTS: In contrast with HCs, patients with ALS displayed increased ALFF in the right inferior temporal gyrus and bilateral frontal gyrus and decreased ALFF in the left middle occipital gyrus and left precentral gyrus. Furthermore, patients with ALS demonstrated lower d-ALFF in widespread regions, including the right lingual gyrus, left superior temporal gyrus, bilateral precentral gyrus, and left paracentral lobule by comparison with HCs. In addition, the ALFF in the left superior orbitofrontal gyrus had a tendency of correlation with ALSFRS-R score and disease progression rate. The classification performance in distinguishing ALS was higher with both features of ALFF and d-ALFF than that with a single approach. CONCLUSIONS: Decreased dynamic brain activity in the precentral gyrus, paracentral gyrus, lingual gyrus, and temporal regions was found in the ALS group. The combined ALFF and d-ALFF could distinguish ALS from HCs with a higher accuracy than ALFF and d-ALFF alone. These findings may provide important evidence for understanding the neuropathology underlying ALS.

Effect of SDF-1 with biphasic ceramic-like bone graft on the repair of rabbit radial defect.

BACKGROUND: This study aimed to investigate the effects of stromal cell-derived factor-1 (SDF-1) on biphasic ceramic-like biologic bone (BCBB) in vivo on the repair of large segment bone defect in rabbits. METHODS: A large-segment radius defect model of the rabbits was constructed. In the experimental group, BCBB with SDF-1 sustained-release system were implanted into the bone defect site. Other three groups including normal control, autologous bone graft, and BCBB implantation without SDF-1 were set. After surgery, general observation, X-ray radiography and scoring, and tissue section staining were performed at 2, 4, 8, 12, and 24 weeks post-implantation. RESULTS: By general observation, X-ray radiography and grading and tissue section staining observation, we found that the BCBB carrying SDF-1 was better than those in the group of BCBB without SDF-1 (P < 0.05). BCBB scaffold had certain bone conduction capacity, and the BCBB scaffold carrying SDF-1 had improved bone conduction ability and possessed bone induction ability. In the case of carrying SDF-1, it can be used to repair large bone defects in a shorter time than simply using BCBB, which is equivalent to the effect of autologous bone. CONCLUSION: BCBB scaffold carrying SDF-1 can promote the repair effect on a large bone defect, which is equivalent to the effect of autologous bone.