Yunji Wang1, Jie Li1, Jianpin Zhou1, Ye Qiu2, Jinlin Song3. 1. College of Stomatology, Chongqing Medical University, Chongqing, China; Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing, China; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, China. 2. College of Stomatology, Chongqing Medical University, Chongqing, China; Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing, China; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, China. Electronic address: 500694@hospital.cqmu.edu.cn. 3. College of Stomatology, Chongqing Medical University, Chongqing, China; Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing, China; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, China. Electronic address: songjinlin@hospital.cqmu.edu.cn.
Abstract
BACKGROUND: Alveolar bone loss is one of the most common consequence for periodontitis, which is a major obstacle in periodontal regeneration. Bone marrow stromal cells (BMSCs) have shown significant promise in the treatment of various disease, which also contribute to the natural bone repair process. Low-intensity pulsed ultrasound (LIPUS) is a therapeutic ultrasound used in our previous studies to promotes alveolar bone regeneration. In addition, LIPUS was found to be a promising method to enhance mesenchymal stromal cell-based therapies. In the current study, we have investigated the effects of LIPUS combined with BMSCs therapies on BMSCs homing and its potential to promote alveolar bone regeneration. METHODS: BMSCs were isolated from rat and characterized by multilineages differentiation assay. Then these cells were labeled with luciferase and green fluorescent protein (GFP) by lentivirus in vitro. Periodontal bone defect was made on the mesial area of the maxillary first molar in rats. A total of 1 × 106 Luc-GFP labeled BMSCs were injected into rat tail vein. Bioluminescence imaging was utilized to track BMSCs in vivo. The rats were sacrificed eight weeks after surgery and the samples were harvested. Micro-computed tomography (Micro-CT) was performed to evaluate alveolar bone regeneration. Paraffin sections were made and subject to hematoxylin-eosin staining, masson staining and immunohistochemistry staining. RESULTS: BMSCs display a fibroblast-like morphology and can differentiate into adipocytes or osteoblasts under appropriate condition. The transfected BMSCs are strongly positive for GFP express. Bioluminescence imaging showed that most of BMSCs were trapped in the lung. A small portion BMSCs were homed to the alveolar bone defect area in BMSCs group, while more cells were observed in BMSCs/LIPUS group compare to other groups on day 3 and 7. Micro-CT results showed that BMSCs/LIPUS group resulted in more new bone formation than other groups. Immunohistochemical results showed higher expression of COL-I and osteopontin in BMSCs/LIPUS group compared with the other groups. CONCLUSIONS: These results suggested that LIPUS can enhance BMSCs-based periodontal alveolar bone regeneration. This study provides new insights into how LIPUS might provide therapeutic benefits by promoting BMSCs homing.
BACKGROUND: Alveolar bone loss is one of the most common consequence for periodontitis, which is a major obstacle in periodontal regeneration. Bone marrow stromal cells (BMSCs) have shown significant promise in the treatment of various disease, which also contribute to the natural bone repair process. Low-intensity pulsed ultrasound (LIPUS) is a therapeutic ultrasound used in our previous studies to promotes alveolar bone regeneration. In addition, LIPUS was found to be a promising method to enhance mesenchymal stromal cell-based therapies. In the current study, we have investigated the effects of LIPUS combined with BMSCs therapies on BMSCs homing and its potential to promote alveolar bone regeneration. METHODS: BMSCs were isolated from rat and characterized by multilineages differentiation assay. Then these cells were labeled with luciferase and green fluorescent protein (GFP) by lentivirus in vitro. Periodontal bone defect was made on the mesial area of the maxillary first molar in rats. A total of 1 × 106 Luc-GFP labeled BMSCs were injected into rat tail vein. Bioluminescence imaging was utilized to track BMSCs in vivo. The rats were sacrificed eight weeks after surgery and the samples were harvested. Micro-computed tomography (Micro-CT) was performed to evaluate alveolar bone regeneration. Paraffin sections were made and subject to hematoxylin-eosin staining, masson staining and immunohistochemistry staining. RESULTS: BMSCs display a fibroblast-like morphology and can differentiate into adipocytes or osteoblasts under appropriate condition. The transfected BMSCs are strongly positive for GFP express. Bioluminescence imaging showed that most of BMSCs were trapped in the lung. A small portion BMSCs were homed to the alveolar bone defect area in BMSCs group, while more cells were observed in BMSCs/LIPUS group compare to other groups on day 3 and 7. Micro-CT results showed that BMSCs/LIPUS group resulted in more new bone formation than other groups. Immunohistochemical results showed higher expression of COL-I and osteopontin in BMSCs/LIPUS group compared with the other groups. CONCLUSIONS: These results suggested that LIPUS can enhance BMSCs-based periodontal alveolar bone regeneration. This study provides new insights into how LIPUS might provide therapeutic benefits by promoting BMSCs homing.