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象牙質における抗う蝕性イオンのマルチスケール解析

内藤, 克昭 大阪大学 DOI:10.18910/82158

2021.03.24

概要

現在，高齢者における露出した根面象牙質に発生するう蝕が社会的な問題となっている[1–4]．う蝕は，細菌由来の酸性代謝産物により象牙質の無機成分の崩壊を引き起こし，その結果発生するイオンが再石灰化に働くという，イオンの循環を示す複雑な病態である[5–9]．それゆえ，う蝕の詳細な病態解明には，イオンの動的状態を捉えることが必要である．

象牙質は，エナメル質あるいはセメント質に囲まれた無機有機複合体であり，有機質であるⅠ型コラーゲン線維と無機質であるハイドロキシアパタイトナノ結晶と水分から構成されている[10,11]．象牙質や骨などの生体アパタイト(BioAp)は，ハイドロキシアパタイト(HAp)だけでなく，リン酸オクタカルシウム（OCP）[12,13]や非晶質リン酸カルシウム（ACP）[14,15]，第２リン酸カルシウム２水塩[16,17]など多様なリン酸カルシウム相を含有する極めて複雑な結晶である[18,19]．さらに，象牙質のHApの内部には空孔に加えて，炭酸イオン（CO32-），マグネシウム（Mg），フッ化物（F），ナトリウム（Na），亜鉛（Zn）などの微量元素やイオンが含有されている[20–24]．空孔や微量元素は，HApの結晶性や脱灰，再石灰化など，生体における化学的動態に複雑に関与していると考えられるが[25]，これらの機能は，未だ一部分しか判明していない．

歯質の耐酸性に対するFの有効性は，invitroの研究だけでなく[26]，歯磨剤や水道水のフロリデーションを通じてFを歯質内へ導入することにより，疫学的にう蝕予防に成功している[27–31].しかし，先進国においては成人における根面う蝕の増加を認めるなど，う蝕の撲滅はできていない[32]．これらを背景に昨今，F単独よりも他元素と組み合わせることで，歯質のさらなる抗う蝕性の向上が期待され始めている[33–37]．Fと組み合わせる微量元素として，SrやZn,Sn,Tiなどが候補として挙げられ，本研究では，とくにZnに着目した．Znは12族元素であり，d軌道が閉殻されており，MgやSrを代表とするアルカリ土類金属と同様の性質を示す[38–40]．Caと同族であるMgやSrに関しては，象牙質に対しての作用が数多く報告されている[41,42]．一方，Znの抗う蝕性については，抗菌作用[43]，マトリックスメタロプロテアーゼ阻害作用[44]，脱灰抑制作用[45]などが報告されているが，原子スケールを含めた詳細なメカニズムの解析はほとんどされていなかった．

近年，げっ歯類のエナメル質においてAtom probe tomographyを用いた原子スケールでのイオン分布解析により，結晶粒界に存在するMg-ACPにFが置換することで抗う蝕作用を発揮することが報告された[35]．この成果は，原子スケールでのイオン動態を解明することで，これまでのう蝕予防および治療方法が大きく変わる可能性を示したものである．象牙質は，無機成分として多様な結晶と40wt%近い有機成分から成り，構造的に複雑であるため，これまでの汎用的な手法では原子スケールでのイオン動態を捉えることが困難であった[46]．象牙質の結晶構造へのイオン動態については，X線回折法（XRD）による長距離秩序の解析[47]や透過型電子顕微鏡（TEM）による観察が行われているが[48–50]，これらの手法では，周期的な構造変化を評価できるが，イオン置換に本質的に関与している局所構造を見落としている可能性がある．

そこで本研究では，大気陽子線励起X線/ガンマ線分析法（In-airμ-PIXE/PIGE）およびX線吸収微細構造測定（XAFS）を用いて，人為的に象牙質内に導入した抗う蝕性イオンの分布，結合状態，および機能を解析することを目的とした．Inairμ-PIXE/PIGE法は，陽子線の衝突により発生する蛍光X線およびガンマ線を計測することで，軽元素を含む多元素を大気中で高感度に検出できる[51–56]．また，歯質内の元素分布および濃度を非破壊的かつ経時的に分析することができる．XAFSは，X線を照射し，対象物の電子状態や，化学結合状態を探索する手法であり，ガラスのような非晶質な材料であっても各構成元素周囲の構造を同定することができ，微量な元素に対する感度も高い[57,58]．

これらのイオン動態の解析に加えて，従来の走査型電子顕微鏡（SEM）による形態観察，マイクロコンピュータ断層撮影法(μCT)による非破壊構造解析，XRDによる長距離秩序解析，X線光電子分光法(XPS)による化学結合状態分析を合わせることにより，マルチスケールでのイオンの動態解析を実現することが可能となる．さらにこのマルチスケール解析によって，う蝕における脱灰と再石灰化に伴う化学的な反応を材料から溶出するイオンで能動的に制御し，う蝕の予防および進行抑制を達成する方法を模索することができる．本研究では，旧来の画一的な治療方法である”drillandfill”から脱却し，イオン動態の見地から生物学的なアプローチに基づく削らないう蝕治療を実現することを目ざしている．

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