ABSTRACT
<p><b>BACKGROUND</b>Several studies have revealed that adipose-derived mesenchymal stem cells (ADSCs) can be used as seed cells for the treatment of spinal cord injury (SCI). Chondroitinase ABC (ChABC) decomposes chondroitin sulfate proteoglycans in the glial scar that forms following SCI, allowing stem cells to penetrate through the scar and promote recovery of nerve function. This study aimed to establish ADSCs that stably express ChABC (ChABC-ADSCs) and evaluate the migratory capability of ChABC-ADSCs in vitro.</p><p><b>METHODS</b>ADSCs were obtained from Sprague-Dawley rats using secondary collagenase digestion. Their phenotypes were characterized using flow cytometry detection of cell surface antigens and their stem cell properties were confirmed by induction of differentiation. After successful culture, ADSCs were transfected with lentiviral vectors and ChABC-ADSCs were obtained. Proliferation curves of ChABC-ADSCs were determined using the Cell Counting Kit-8 method, ChABC expression was verified using Western blotting, and the migration of ChABC-ADSCs was analyzed using the transwell assay.</p><p><b>RESULTS</b>Secondary collagenase digestion increased the isolation efficiency of primary ADSCs. Following transfection using lentiviral vectors, the proliferation of ChABC-ADSCs was reduced in comparison with control ADSCs at 48 h (P < 0.05). And the level of ChABC expression in the ChABC-ADSC group was significantly higher than that of the ADSC group (P < 0.05). Moreover, ChABC-ADSC migration in matrigel was significantly enhanced in comparison with the control (P < 0.05).</p><p><b>CONCLUSIONS</b>Secondary collagenase digestion can be used to effectively isolate ADSCs. ChABC-ADSCs constructed using lentiviral vector transfection stably express ChABC, and ChABC expression significantly enhances the migratory capacity of ADSCs.</p>
Subject(s)
Animals , Male , Rats , Adipocytes , Cell Biology , Metabolism , Adipose Tissue , Cell Biology , Cell Differentiation , Physiology , Cell Movement , Physiology , Cell Proliferation , Physiology , Cells, Cultured , Chondrocytes , Cell Biology , Metabolism , Chondroitin ABC Lyase , Metabolism , Flow Cytometry , Mesenchymal Stem Cells , Cell Biology , Metabolism , Osteoblasts , Cell Biology , Metabolism , Rats, Sprague-DawleyABSTRACT
OBJECTIVE@#To observe the effect of daily low-intensity pulsed ultrasound (LIPUs) therapy on improving the enchondral bone formation in lumbar fusion in rabbit models, and to explore its possible mechanism.@*METHODS@#Posterolateral noninstrumented bilateral fusions were performed at the L5 approximately L6 levels in 20 New Zealand rabbits. The autograft iliac bone was implanted on the left side, and the hydroxyapatite bioceramic artificial bone on the right. The rabbits were divided into a treatment group and a control group randomly. One week after the surgery, LIUPs was administered for 20 minutes per day for 4 weeks over the fusion site in the treatment group and false treatment was used in the control group. Post-anterior X-ray photographs were taken to determine the conditions of fusion area, and then, rabbits were killed and the fusion tissues were obtained. Chondrocytes were detected by histological and cytological methods.@*RESULTS@#Compared with the control group, the fusion rate of the treatment group was significantly up-regulated (P0.05).@*CONCLUSION@#Low-intensity pulsed ultrasound therapy improves the endochondral bone formation in the lumbar spine fusion in rabbit models.