アブストラクト（32巻1号：神奈川歯学）

神奈川歯学

Japanese

Title　： ヒト歯根膜由来線維芽細胞のコラーゲンゲル内培養の研究 - タンパク質生合成におけるゲル収縮の影響について -
Subtitle　： 原著
Authors　： 相原治美, 斎藤滋
Authors（kana）　：
Organization　： 神奈川歯科大学口腔生化学教室
Journal　： 神奈川歯学
Volume　： 32
Number　： 1
Page　： 1-11
Year/Month　： 1997 / 6
Article　： 原著
Publisher　： 神奈川歯科大学学会
Abstract　： 「緒言」 生体組織は, 細胞と細胞間物質より成り立つにもかかわらず, 今までの培養法は細胞間物質を欠如した状態で行われていた. 主な細胞間物質であるコラーゲンは, 生体を物理的に支持し保護するだけでなく, 細胞本来の形質や機能にも影響を与えている. コラーゲンゲル内培養法は, Bellらによって創傷治癒過程の真皮modelとして紹介されたのが初めである. この3次元培養系で線維芽細胞を培養するとコラーゲンゲルが収縮し, コラーゲン線維の再編を引き起こし, 実際の生体真皮に類似した構造を形成するとともに, 生体真皮で確認される線維芽細胞と同様に細胞増殖の抑制が起こることが知られている. コラーゲンゲル内で生じる線維芽細胞の張力の程度は, 瘢痕収縮や歯の萌出時の力に類似するといわれる. 線維芽細胞による瘢痕収縮は, 創傷治癒において創部を縮小させ治癒を早める目的に叶った機構である. しかし, この機構が慢性炎症においては組織の改変を引き起こすこともある.
Practice　： 歯科学
Keywords　： HPLF, collagen gel, gel contraction, ECM

English

Title　： Human Periodontal Ligament Fibroblasts within Collagen Gel Culture - Effect of Gel Contraction on Protein Biosynthesis -
Subtitle　： Original article
Authors　： Harumi AIHARA, Shigeru Saito
Authors（kana）　：
Organization　： Department of Oral Biochemistry, Kanagawa Dental College
Journal　： Kanagawa Shigaku
Volume　： 32
Number　： 1
Page　： 1-11
Year/Month　： 1997 / 6
Article　： Original article
Publisher　： Kanagawa Odontological Society
Abstract　： [Abstract] The cells of periodontal ligament were embedded in three-dimensional extracellular matrices (ECM) consisting of type I collagen and other non-collagenous components. I used an in vitro stabilized collagen lattice (gel) model to examine the cellular responses to fibroblast contraction which may moderate tensional forces. Most experiments on mechanoregulation have measured how cells react when they are subjected to increased mechanical stress. Equally important but less well studied is the cellular reaction to decreased stress. The present study was undertaken to determine the protein biosynthesis in collagen gel contraction by human periodontal ligament fibroblasts (HPLF). HPLF, cultured from explants, were added to the neutralized collagen solutions at a cell density of 2.5×105 cells/ml and cultured for seven days in D-MEM supplemented with 5% growth factor in serum (GFS) and 0.2 mM L-ascorbic acid-2-phosphate. To obtain mechanically stressed condition (No-cut), the matrices were allowed to remain anchored to the culture wells ; to obtain mechanically relaxed gels (Cut), the matrices were dislodged from the substratum and allowed to float in the medium during contraction. HPLF cultured for 7 days in the collagen gel were released from the wells. At day 8 culture period, HPLF anchored (No-cut), contracting (During cut) and floated (Cut) were labeled with 35S-methionine (met) for 24 hours. HPLF, in collagen gel, altered cell growth and the incorporation of 35S-met into proteins by decreased contraction. 35S-met labeled proteins in media were resolved by SDS-PAGE, and radioactive bands were observed by an autoradiography. The contraction was dependent, even in early stages, on the addition of exogenous factors such as the conditioned medium of HPLF that stimulated HPLF contractile activity. In the medium, there were some noteworthy differences between stressed and relaxed conditions. The 170 kDa, 110 kDa, 70 kDa, 45 kDa, 40 kDa and 30 kDa components labeled with 35S-met were increased by a stressed HPLF. On the other hand, the 90 kDa and 20 kDa were increased by a relaxed HPLF. These results demonstrate that a relaxed condition, which may modulate the reorganization of HPLF, might decrease the synthesis of extracellular macromolecular matrices. Therefore, I believe that the processes of gel contraction may mimic biomechanical responses in the periodontal ligaments.
Practice　： Dentistry
Keywords　： HPLF, collagen gel, gel contraction, ECM