Investigating the mechanism of placental amnion-chorion membranes on periodontal regeneration

胎盘羊膜绒毛膜促进牙周再生的机制研究

• 批准号: 21K16974
• 负责人: 吉田 航
• 金额: $ 3万
• 依托单位: Tokyo Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K16974/
• 关键词: 歯周組織再生療法; ヒト胎盤由来羊膜・絨毛膜; 成長因子

2022年度の研究実績In vivoでは，10週齢のWistarラットの上顎第一臼歯近心に規格化欠損を形成後ルートプレーニングし，歯周組織欠損を作製した。HACM＋HPC応用群，HPC単独群，Unfilled群に分け，術後4週で形態学的 (マイクロCT)，組織学的(H-E染色)にて検討した。2021年度では術後2週での評価を行い，マイクロCT画像による歯槽骨形成量の解析では，HACM+HPC群で新生骨様構造を認めた。さらに，骨梁構造解析でHACM+HPC群はHPC，Unfilled群と比較し骨体積率で有意に高い値を示した。2022年度では術後4週での評価を行い，HACM+HPC群はHPC，Unfilled群と比較し骨体積率で有意に高い値を示した。術後4週のH-E染色においてHACM+HPC 群では，Unfilled，HPC群と比較し，新生骨様構造が多く観察され，術後2週と同様の傾向を示した。また,免疫組織化学染色(VEGF，α-SMA)を行い血管新生についても評価を行った。In vitroでは，HACMをPBS中に溶解したHACM extract (100-500 ug/ml)を作製し，ELISAにてVEGFおよびFGF-2の濃度を測定した。ラットより採取した歯根膜由来細胞の培養液中にHACM extractを滴下し，細胞増殖能をWST-1，細胞遊走能をWound healing assayにより評価した。2022年度も引き続き同様の評価を行い，VEGFおよびFGF-2の成長因子が含めれている100 ug/ml のHACM Extractは，Control群と比較して歯根膜由来細胞の増殖および遊走の促進を認めた。以上のことからHACMの含有成分が歯根膜由来細胞の増殖および遊走を促すことで，歯周組織治癒を促進させることが示唆された。

Results of the study in 2022: In vivo, 10 weeks after the formation of the first maxillary dental proximal normalization deficit, the first maxillary dental tissue deficit was controlled. HACM+HPC group, HPC single group, Unfilled group were analyzed morphologically (CT) and histologically (H-E staining) at 4 weeks postoperatively. In 2021, the evaluation was carried out at 2 weeks after operation, and the new bone structure of HACM+HPC group was identified. In addition, HACM+HPC group is higher than HPC group in structural analysis of bone beam, and the ratio of bone volume is higher than HPC group. In 2022, the HACM+HPC group was evaluated at 4 weeks after surgery, and the Unfilled group was evaluated at a significantly higher bone volume rate. H-E staining at 4 weeks postoperatively showed that HACM+HPC group was negative, Unfilled, HPC group was negative, new bone structure was more observed, and the same trend was observed at 2 weeks postoperatively. Immunohistochemical staining (VEGF, α-SMA) was used to evaluate angiogenesis. In vitro, HACM was dissolved in PBS and HACM extract (100-500 ug/ml) was prepared, and VEGF and FGF-2 concentrations were determined by ELISA. HACM extract was added to the culture medium of root membrane-derived cells, and the cell proliferation ability was evaluated by WST-1 and Wound healing assay. In 2022, the growth factors of VEGF and FGF-2 were found to be higher than 100 ug/ml in HACM Extract, and the Control group was compared with the growth and migration promotion of dental root membrane-derived cells. The above HACM components promote proliferation and migration of dental-derived cells, and promote healing of periapical tissues.

项目成果

To evaluate the effect of Human dehydrated amnion-chorion membranes on periodontal healing in vivo and in vitro.

评价人脱水羊膜绒毛膜在体内和体外对牙周愈合的影响。

• 发表时间: 2021
• 作者: Imamura K;Hamada Y;Yoshida W;Yoshikawa K;Saito A
• 通讯作者: Saito A

Investigating the Effects of Dehydrated Human Amnion-Chorion Membrane on Periodontal Healing.

• DOI: 10.3390/biom12060857
• 发表时间: 2022-06-20
• 期刊: Biomolecules
• 影响因子: 5.5

ヒト胎盤由来羊膜・絨毛膜メンブレン（HACM）の応用が歯周組織の創傷治癒に及ぼす影響

人胎盘源性羊膜绒毛膜（HACM）应用对牙周组织创面愈合的影响

• 发表时间: 2021
• 作者: Imamura K;Hamada Y;Yoshida W;Murakami T;Nakane-Koyachi S;Yoshikawa K;Saito A.;今村健太郎，中根咲，吉田航，吉川幸輝，齋藤淳
• 通讯作者: 今村健太郎，中根咲，吉田航，吉川幸輝，齋藤淳