THE EVALUATION OF OSTEOGEMC POTENTIAL OF CULTURED PERIOSTEUM-DERIVED CELLS

培养骨膜来源细胞的成骨潜力评估

• 批准号: 15592109
• 负责人: UENO Takaaki
• 金额: $ 1.98万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15592109/
• 关键词: Periosteal graft; Cultured periosteum derived cell; Bone morphogenetic protein-4; Bone repair; エレクトロポレーション; 遺伝子導入; BMP-4遺伝子; ラット骨膜細胞; 培養細胞; 細胞移植; 骨形成能

Periosteum has osteogenic potential and considerable attention has been focused on its use as a grafting material for the repair of bone and joint defects. Recently, the osteogenic potential of cultured periosteal cells has been reported. We present findings of bone formation induced from cultured periosteum-derived cells of rats tibiae. Periosteum was placed in culture medium with 10% Fetal Bovine Serum for 14 days. After reaching confluence, periosteal cells were resuspended with 025% trypsin/EDTA and then re-cultured three dimensionally on a collagen sponge for 7 days. Periosteal cell/collagen complex was grafted into rat submandibular muscle and A solution of plasmid DNA containing mouse BMP-4 (pMiw-BMP-4, kindly provided by Dr.Brigid I.M. Hogan and Dr.Kishimoto, Tohoku Univ.) was injected into grfated tissue and electric pulses were applied through paired-needle electrodes. As a control plasmid, Normal saline solution was injected. Immunosuppressant (FK506, 1.0 mg/kg/day) was administered intramuscularly. At 14, 21 and 35 days post grafting, grafted tissue was extirpated and compared histologically and radiographically with control groups. In the experimental group, periosteal cells had proliferated and differentiated into osteogenic cells by 14 days post grafting. Cell proliferation marker PCNA positive cells significantly increased compared with control groups. At 21 days post grafting, new bone formation was evident. At this stage, VEGF and BMP-4 positive cells significantly increased compared with control groups. By 35 days post grafting, bone growth was observed and new calcification was detected radiographically in the defect. New bone formation in experimental group developed and total calcified area was larger than control group. Cultured periosteum-derived cells treated with BMP-4 gene showed higher osteogenic potential in the early stage of periosteal cell graft model of rat.

骨膜具有成骨潜能，作为骨和关节缺损修复的植骨材料已受到广泛关注。近年来，培养的骨膜细胞具有成骨潜能。我们报道了用培养的大鼠胫骨骨膜来源细胞诱导骨形成的结果。将骨膜置于含10%胎牛血清的培养基中培养14天。汇合后，用025%胰蛋白酶/EDTA重悬骨膜细胞，然后在胶原海绵上三维培养7天。将骨膜细胞/胶原复合物移植到大鼠下颌下肌中，将含有小鼠BMP-4的质粒DNA溶液（pMiw-BMP-4，由东北大学brigid I.M. Hogan博士和岸本博士提供）注射到移植组织中，并通过对针电极施加电脉冲。注射生理盐水溶液作为对照质粒。肌肉注射免疫抑制剂FK506, 1.0 mg/kg/天。在移植后14、21和35天，将移植组织切除，并与对照组进行组织学和影像学比较。实验组骨膜细胞在移植后14天增殖分化为成骨细胞。细胞增殖标志物PCNA阳性细胞较对照组显著增加。移植21天后，新骨形成明显。与对照组相比，VEGF和BMP-4阳性细胞明显增多。移植后35天，观察骨生长，影像学检查发现缺损处出现新的钙化。实验组新生骨形成较发达，总钙化面积大于对照组。BMP-4基因处理培养的骨膜源性细胞在大鼠骨膜细胞移植模型早期表现出较高的成骨潜能。

项目成果

Immunohistochemical observation of cellular origin and proliferation in endochondral bone formation from rafted periosteum.

筏式骨膜软骨内骨形成中细胞起源和增殖的免疫组织化学观察。

• 发表时间: 2003
• 期刊: Journal of CranioMaxillofacial Surgery 31
• 作者: Takaaki Ueno;Toshimasa Kagawa;Miwa Kanou;Takashi Toshio Sugahara
• 通讯作者: Takashi Toshio Sugahara

Osteogenic Potential of Primed Periosteal Graft in the Rat Calvarial Model

大鼠颅骨模型中引发骨膜移植物的成骨潜力

• 发表时间: 2004
• 期刊: Annals Plastic Surgery 46
• 作者: Takaaki Ueno;Toshimasa Kagawa;Miwa Kanou;Toshio Sugahara
• 通讯作者: Toshio Sugahara

Immunohistochemical analysis of Sox9 in the periosteum of tibia nad calvaria after surgical release of perosteum.

手术释放骨膜后，对胫骨和颅骨骨膜中的 Sox9 进行免疫组织化学分析。

• 期刊: Acta Histochemica (in press)
• 作者: Fujii T;Ueno T;Kagawa T et al.
• 通讯作者: Kagawa T et al.

妊婦の歯科治療とカウンセリング妊婦歯科治療における留意点〜口腔外科の立場から〜

孕妇牙科治疗和咨询 孕妇牙科治疗的注意事项 ~ 从口腔外科的角度来看 ~

• 发表时间: 2004
• 作者: 分担執筆 植野高章

Ueno T, Kagawa T, Kanou M: "Immunolocalization of Vascular Endotherial Growth Factor (VEGF) during Heterotopic Bone Formation Induced from Grafted Periosteum"Annals of Plastic Surgery. 50(in press). (2004)

Ueno T、Kakawa T、Kanou M：“移植骨膜诱导异位骨形成过程中血管内皮生长因子 (VEGF) 的免疫定位”整形外科年鉴。