Gene therapy using sonoporation for maxillofacial distraction

使用声孔进行颌面牵引的基因治疗

• 批准号: 20592340
• 负责人: YOSHIOKA Izumi
• 金额: $ 2.91万
• 依托单位: Kyushu Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20592340/
• 关键词: 顎骨延長法; BMP; Smad; ヒアルロン酸; 骨芽細胞; 超音波遺伝子導入; マイクロバブル; ソノポレーション法

Bone morphogenetic protein-2 (BMP-2) is expected to be utilized to fill bone defects and promote healing of fractures. However, it is unable to generate an adequate clinical response for use in bone regeneration. Recently, it was reported that glycosaminoglycans, including heparin, heparan sulfate, keratan sulfate, dermatan sulfate, chondroitin-4-sulfate, chondroitin-6-sulfate, and hyaluronic acid (HA), regulate BMP-2 activity, though the mechanism by which HA regulates osteogenic activities has not been fully elucidated. The aim of this study was to investigate the effects of HA on osteoblast differentiation induced by BMP-2.Monolayer cultures of osteoblastic lineage MG63 cells were incubated with BMP-2 and HA for various time periods. To determine osteoblastic differentiation, alkaline phosphatase (ALP) activity in the cell lysates was quantified. Phosphorylation of Smad 1/5/8, p38, and ERK proteins was determined by Western blot analysis. To elucidate the nuclear translocation of phosphorylated Smad 1/5/8, stimulated cells were subjected to immunofluorescence microscopy. To further elucidate the role of HA in enhancement of BMP-2-induced Smad signaling, mRNA expressions of the BMP-2 receptor antagonists noggin and follistatin were detected using real-time RT-PCR.BMP-2-induced ALP activation, Smad 1/5/8 phosphorylation, and nuclear translocation were up-regulated when MG63 cells were cultured with both BMP-2 and HA. Western blot analysis revealed that phosphorylation of ERK protein was diminished by HA. Furthermore, the mRNA expressions of noggin and follistatin induced by BMP-2 were preferentially blocked by HA.These results indicate that HA enhanced BMP-2 induces osteoblastic differentiation in MG63 cells via down-regulation of BMP-2 antagonists and ERK phosphorylation.

骨形态发生蛋白-2（BMP-2）有望用于填充骨缺损并促进骨折愈合。然而，它无法产生足够的临床反应用于骨再生。最近，据报道，糖胺聚糖，包括肝素、硫酸乙酰肝素、硫酸角质素、硫酸皮肤素、4-硫酸软骨素、6-硫酸软骨素和透明质酸（HA），可调节 BMP-2 活性，但其机制HA 调节成骨活性尚未完全阐明。本研究的目的是探讨HA对BMP-2诱导的成骨细胞分化的影响。将成骨细胞谱系MG63细胞的单层培养物与BMP-2和HA一起孵育不同时间段。为了确定成骨细胞分化，对细胞裂解物中的碱性磷酸酶（ALP）活性进行了定量。通过蛋白质印迹分析确定 Smad 1/5/8、p38 和 ERK 蛋白的磷酸化。为了阐明磷酸化 Smad 1/5/8 的核转位，对受刺激的细胞进行免疫荧光显微镜检查。为了进一步阐明HA在增强BMP-2诱导的Smad信号传导中的作用，使用实时RT-PCR检测了BMP-2受体拮抗剂noggin和卵泡抑素的mRNA表达。BMP-2诱导的ALP激活，Smad 1当 MG63 细胞与 BMP-2 和 HA 一起培养时，/5/8 磷酸化和核易位上调。蛋白质印迹分析显示，HA 减弱了 ERK 蛋白的磷酸化。此外，BMP-2诱导的noggin和卵泡抑素的mRNA表达优先被HA阻断。这些结果表明HA增强BMP-2通过下调BMP-2拮抗剂和ERK磷酸化诱导MG63细胞中的成骨细胞分化。

项目成果

Correlation of mandibular bone quality with neurosensory disturbance after sagittal split ramus Osteotomy

升支矢状劈裂截骨术后下颌骨骨质量与感觉神经障碍的相关性

• 发表时间: 2010
• 作者: Izumi Yoshioka; 他
• 通讯作者: 他

Post-operative skeletal stability and accuracy of a new combined Le Fort I and-horseshoe osteotomy for superior repositioning of the maxilla

新型 Le Fort I 和马蹄形截骨术相结合的术后骨骼稳定性和准确性，可实现上颌骨的出色重新定位

• 发表时间: 2009
• 作者: I Yoshioka

顎関節症の疾患概念と治療概念

颞下颌关节紊乱病的疾病概念及治疗理念

• 发表时间: 2009
• 作者: 冨永和宏

A comparison of stability following mandibular setback between vertical ramus and sagittal split osteotomies

垂直升支截骨术与矢状劈裂截骨术下颌后缩后稳定性的比较

• 发表时间: 2008
• 作者: Izumi Yoshioka

A novel modification of a bone repositioning device and a new technique for reestablishing facial contours after mandibular resection surgery

骨复位装置的新颖改进和下颌切除术后重建面部轮廓的新技术

• 发表时间: 2009
• 作者: 冨永和宏