フィブロネクチン固定化ポリ乳酸─ポリグリコール酸共重合体フィルムの作製と生物学的評価

DOI
  • 村上 洋
    日本大学松戸歯学部口腔インプラント学講座
  • 二木 由峰
    日本大学松戸歯学部歯科臨床検査医学講座 中国・四国支部（にき歯科医院）
  • 若井 広明
    日本大学松戸歯学部歯科臨床検査医学講座 関東・甲信越支部（若井歯科医院）
  • 中西 一
    日本大学松戸歯学部歯科臨床検査医学講座 中国・四国支部（まこと歯科医院）
  • 井山 禎之
    日本大学松戸歯学部歯科臨床検査医学講座 中国・四国支部（にき歯科医院）
  • 古市 嘉秀
    日本大学松戸歯学部歯科臨床検査医学講座 中部支部（グリーン歯科医院）
  • 布施 恵
    日本大学松戸歯学部歯科臨床検査医学講座

書誌事項

タイトル別名
  • Preparation and Biological Evaluation of Fibronectin Immobilized Polylactide-co-glycolide (PLGA) Copolymer Film

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<p>Polylactic acid (PLA) and polyglycolic acid (PGA) are widely used as guided tissue regeneration and/or guided bone regeneration membranes in the dental field. However, these membranes do not have any biological function such as acceleration of cell attachment, cell spreading, proliferation and differentiation. In the present study, we aimed to immobilize fibronectin, cell adhesive protein, to polylactide-co-glycolide (PLGA) film. The biological evaluation of fibronectin immobilized PLGA was investigated by human umbilical vein endothelial cell (HUVEC) culture.</p><p>Three types of PLGA (PLA : PGA = 90 : 10 (PLGA10), PLA : PGA = 80 : 20 (PLGA20), PLA : PGA = 70 : 30 (PLGA30)) films were used. The PLGA films were immersed in 0.5 M NaOH solution at 37℃ for 3 h for surface hydrolysis. The hydrolyzed PLGA film was immersed in MES buffer containing water-soluble carbodiimide. Afterwards, the film was immersed in 0.05% fibronectin solution in MES buffer solution for 48 h at 4℃. The fibronectin immobilized PLGA films, namely FN-PLGA10, FNPLGA20, and FN-PLGA30, were obtained. The contact angle of the FN-PLGA surface with respect to double distilled water was measured. Cell assay was performed by using human umbilical vein endothelial cells (HUVECs). The seeded HUVECs were allowed to incubate for 90 min, 3 and 7 days under standard culture conditions. The number of attached cells was evaluated and the morphologies of attached sells were observed by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM).</p><p>The contact angle of each FN-PLGA surface decreased compared with PLGA (p＜0.05). The presence of fibronectin on each FN-PLGA was confirmed by attenuated total reflection Fourier transform infrared spectroscopy (FTIR-ATR). The number of attached cells on FNPLGA10 was significantly greater than that on PLGA10 after 90 min, 3 and 7 days (p＜0.05). For FN-PLGA20, the number of attached cells was significantly higher than that on PLGA20 after 90 min and 7 days (p＜0.05). FN-PLGA30 showed a significantly greater number of attached cells than FN-PLGA30 after 90 min (p＜0.05). The number of attached cells on FN-PLGA10 was significantly greater than that on FN-PLGA20 and FN-PLGA30 after 3 and 7 days (p＜0.05). HUVECs attached to each FN-PLGA exhibited spread morphologies and the formation of pseudopodia. For each PLGA, rounded morphologies of cells and less formation of pseudopodia were observed. More actin filaments and stress fibers on FNPLGA10 were observed compared with PLGA10 by CLSM observation.</p><p>In conclusion, it was revealed that fibronectin immobilization of PLGA film was effective for biological activity. In addition, the ratio of PLA and PGA in PLGA influenced the attachment and proliferation of cells.</p>

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