Background. The development of tissue engineering provides a new method for the clinical treatment of bone defects, but the problems of slow formation and slow vascularization of tissue engineered bone have always existed. Studies have shown that the combined culture system of vascular endothelial cells and adipose stem cells is superior to single cell in repairing bone defects. With the excellent proliferation ability, secretion of synthetic collagen and a variety of regulatory factors and fibroblasts can differentiate into osteoblasts and have the potential to be excellent seed cells involved in tissue engineering bone construction. Objective. To investigate the effects of combined culture of fibroblasts, vascular endothelial cells, and adipose stem cells on proliferation and osteogenic differentiation of adipose stem cells. Methods. The cells were divided into 4 groups: adipose stem cell group, adipose stem cell+vascular endothelial cell coculture group, adipose stem cell+fibroblast coculture group, and adipose stem cell+vascular endothelial cell+fibroblast coculture group. The morphological changes of the cells were observed under an inverted microscope. After 1, 3, 5, 7, and 9 days of coculture, the proliferation of adipose stem cells in each group was detected by a CCK-8 method and the growth curve was plotted. Adipose stem cells in each group were stained with alizarin red and alkaline phosphatase at days 7, 14, 21, and 28. At the third week of coculture, Western blot was used to detect the expression level of bone morphogenetic protein 2 of adipose stem cells in each group. Results and Conclusions. (1) After 14 days of culture, some cells in the adipose stem cell+vascular endothelial cell+fibroblast coculture group fused into clumps and distributed in nests, while the adipose stem cells in the adipose stem cell group had a single cell morphology and no cell clusters were observed. (2) The cell growth curves were basically the same in each group, and the absorbance value increased gradually. The absorbance value of the adipocyte+vascular endothelial cell+fibroblast coculture group was the highest, followed by the adipocyte+fibroblast coculture group and then the adipocyte+fibroblast coculture group. (3) Alizarin red staining showed negative reaction in each group on the 7th day, and a small number of red positive cells gradually appeared in each group as time went on. On the 28th day, red positive cells were found in all groups, and most of them were in the coculture group of adipose stem cells+vascular endothelial cells+fibroblasts, showing red focal. The coculture group of adipose stem cells+vascular endothelial cells and adipose stem cells+fibroblasts was less, and the adipose stem cell group was the least. On day 28 of alkaline phosphatase staining, cells in each group had red positive particles, and the adipose stem cell+vascular endothelial cell+fibroblast coculture group and adipose stem cell+fibroblast coculture group had the most, followed by the adipose stem cell+vascular endothelial cell coculture group and then the adipose stem cell group. (4) Bone morphogenetic protein 2 was expressed in all groups, especially in adipose stem cell+fibroblast coculture group and adipose stem cell+vascular endothelial cell+ fibroblast coculture group. (5) Fibroblast could promote adipose stem cell osteogenic differentiation better than vascular endothelial cells, but the proliferation effect was not as good as vascular endothelial cells. The coculture system of fibroblast combined with vascular endothelial cells and adipose stem cells promoted the proliferation of adipose stem cells and the rapid and efficient differentiation of adipose stem cells into osteoblasts.
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.
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