CN110770164B - 磷酸八钙成型体的制造方法 - Google Patents
磷酸八钙成型体的制造方法 Download PDFInfo
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- CN110770164B CN110770164B CN201880039921.5A CN201880039921A CN110770164B CN 110770164 B CN110770164 B CN 110770164B CN 201880039921 A CN201880039921 A CN 201880039921A CN 110770164 B CN110770164 B CN 110770164B
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- YIGWVOWKHUSYER-UHFFFAOYSA-F tetracalcium;hydrogen phosphate;diphosphate Chemical compound [Ca+2].[Ca+2].[Ca+2].[Ca+2].OP([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O YIGWVOWKHUSYER-UHFFFAOYSA-F 0.000 title claims abstract description 244
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Abstract
本发明提供体积为2.0mm3以上的包含磷酸八钙的成型体的制造方法等,其特征在于,将在组成中含有Ca及PO4中的至少一者、在H2O中的溶解度高于磷酸八钙、体积大于2.0mm3的前体陶瓷组合物浸渍于下述溶液中,使所述前体陶瓷组合物反应,将所述前体陶瓷组合物的至少一部分转化为磷酸八钙,所述溶液包含作为磷酸八钙的组成的Ca、PO4、H2O的组成之中未被包含在所述前体陶瓷组合物中的组成。
Description
技术领域
本发明涉及可用于骨再生材料、有害分子吸附材料、催化剂担载材料、药剂担载材料等的磷酸八钙成型体的制造方法。
背景技术
磷酸八钙(OCP)是在溶液的pH为4~7的条件下优先结晶的磷酸钙的亚稳相,并且是磷灰石的前体。根据迄今为止的研究,已知该OCP可用作骨再生材料、有机分子吸附材料、及催化剂担载材料(参见非专利文献1~3)。另外,通过向该材料赋予多孔结构,可期待促进作为介由材料表面而发生的反应的骨再生反应、有机分子吸附反应、催化剂效果。
OCP无法烧结,无法利用在烧结过程中的成型体的固化工艺来制造复杂形状的成型体。就作为已知方法的LeGeros的滴加法、铃木等的三流管法而言,虽然能够制造OCP粉末、颗粒,但可形成的OCP的大小最大为1.0mm3以下(参见专利文献1、非专利文献4及5)。
该以往的OCP的成型体是利用晶体生长工艺制造的,因此颗粒大小存在极限,在用作骨再生材料的情况下,需要将大量颗粒埋入骨缺损部(参见非专利文献6)。另外,此时形成的、颗粒间的空隙的大小、形状未必呈现最适合用途的形状。此外,也存在由于血流、气流等主要因素而导致流出的风险。
为了解决这样的问题,需要使成型体的大小为2.0mm3以上。
现有技术文献
专利文献
专利文献1:日本特开平5-70113号公报
非专利文献
非专利文献1:J Biomed Mater Res 59:29-34,2002
非专利文献2:J Ceram Soc Jpn 115:425-428,2007
非专利文献3:Cell Mater 5:45-54,1995
非专利文献4:Calcif Tissue Int 37:194-197,1983
非专利文献5:Acta Biomater 6:3379-3387,2010
非专利文献6:J Dent Res 78:1682-1687,1999
发明内容
发明所要解决的课题
本发明的课题在于提供可适用于骨再生材料、有害分子吸附材料、催化剂担载材料、药剂担载材料等的大型的OCP成型体、及其制造方法。
用于解决课题的手段
本申请的发明人指出,迄今为止,使用硫酸钙二水合物(CSD)等溶解度高于含有碳酸的磷灰石、且包含Ca、PO4、CO3之中至少一者的陶瓷成型体作为前体,并浸渍于包含Ca、PO4、CO3之中陶瓷所未包含的全部离子的溶液中,从而引起液相介导溶解析出反应,由此形成作为热力学最稳定相的含有碳酸的磷灰石。已知经由该液相介导溶解析出反应而形成的含有碳酸的磷灰石(碳酸磷灰石、CO3Ap)成型体大致维持作为前体的陶瓷成型体的形状。
然而,OCP不是热力学上最稳定的相,可通过与液相的接触而被组成转化为磷灰石等。
本申请的发明人发现,通过组成转化反应,即使对于并非热力学最稳定相的OCP而言,也能够在特异性条件下进行制造,从而完成了本发明。即,发现了即使是亚稳相,只要溶解度小于浸渍于溶液中的陶瓷,则也能够经由组成转化反应而形成。另外,只要亚稳相的晶体进行缠结等从而在化学上保持成型体的结构,则能够得到包含亚稳相的成型体。这种情况下,由于稳定相晚于亚稳相而形成,因此需要在形成稳定相而取代亚稳相之前终止反应。
此处的稳定相是指在某温度、压力、化学组成、pH条件下,以充分的时间持续反应时而形成、且不再组成转化成其他相的相。是热力学上能级最低的相。在仅呈稳定相的情况下,当然不会发生进一步的反应,在溶液中的反应的情况下,不发生溶液的pH变化。
具体而言,本申请的发明人进行了各种研究,结果,通过将包含Ca、PO4中的至少一者、且在H2O中的溶解度高于OCP的溶解度(0.090g/L)的体积大于2.0mm3的前体陶瓷组合物浸渍于包含未被包含在前体陶瓷组合物的组成中的Ca、PO4、或H2O中的全部成分的溶液,并在反应完成时间点之前终止反应,从而成功制造了维持前体陶瓷组合物的基本形态、通过无机成分的化学键合、或者无机成分的晶体彼此的缠结或融合而固化的、体积为2.0mm3以上的包含10质量%以上的OCP的成型体,从而完成了本发明。
即,本发明如下所述。
[1]体积为2.0mm3以上的包含磷酸八钙的成型体的制造方法,其特征在于,将在组成中包含Ca及PO4中的至少一者、在H2O中的溶解度高于磷酸八钙、体积大于2.0mm3的前体陶瓷组合物浸渍于下述溶液中,使所述前体陶瓷组合物反应,将所述前体陶瓷组合物的至少一部分转化为磷酸八钙,所述溶液包含作为磷酸八钙的组成的Ca、PO4、H2O的组成之中未被包含在所述前体陶瓷组合物中的组成。
[2]如[1]所述的成型体的制造方法,其特征在于,前体陶瓷组合物在保持其基本形态的状态下被转化为包含磷酸八钙的成型体。
[3]如[1]或[2]所述的成型体的制造方法,其特征在于,制得的包含磷酸八钙的成型体是含有10质量%以上的磷酸八钙的成型体。
[4]如[1]~[3]中任一项所述的成型体的制造方法,其特征在于,在前体陶瓷组合物被组成转化为稳定相的物质的反应完成时间点之前,将成型体从溶液中取出。
[5]如[4]所述的成型体的制造方法,其特征在于,反应完成时间点的溶液的pH为4以上,在高于反应完成时间点的pH的pH条件下终止反应。
[6]如[5]所述的成型体的制造方法,其特征在于,反应完成时间点时的磷酸钙成分为磷灰石。
[7]如[4]~[6]中任一项所述的成型体的制造方法,其特征在于,在成型体的Ca/PO4的比率低于反应完成时间点的成型体的Ca/PO4的比率的时间点终止反应。
[8]如[4]~[7]中任一项所述的成型体的制造方法,其特征在于,在溶液的Ca/PO4的比率高于反应完成时间点的溶液的Ca/PO4的比率的时间点终止反应。
[9]如[4]所述的成型体的制造方法,其特征在于,反应完成时间点的pH低于4,在低于反应完成时间点的pH的pH条件下终止反应。
[10]如[9]所述的成型体的制造方法,其特征在于,反应完成时间点时的磷酸钙成分为无水磷酸氢钙或磷酸氢钙二水合物。
[11]如[4]、[9]或[10]所述的成型体的制造方法,其特征在于,在成型体的Ca/PO4的比率高于反应完成时间点的成型体的Ca/PO4的比率的时间点终止反应。
[12]如[4]、[9]、[10]或[11]所述的成型体的制造方法,其特征在于,在溶液的Ca/PO4的比率低于反应完成时间点的溶液的Ca/PO4的比率的时间点终止反应。
[13]如[1]~[12]中任一项所述的成型体的制造方法,其特征在于,通过使前体陶瓷组合物、及溶液中的至少一者含有在组成中包含含有与钙化学键合的官能团的物质的物质,从而将在组成中包含含有与钙化学键合的官能团的物质的物质担载于磷酸八钙晶体。
[14]如[13]所述的成型体的制造方法,其特征在于,将其他物质担载于磷酸八钙晶体中。
[15]成型体,其特征在于,其是通过无机成分的化学键合、或者无机成分的晶体彼此的缠结或融合而固化的成型体,所述成型体含有10质量%以上的磷酸八钙,且体积为2.0mm3以上。
[16]如[15]所述的成型体,其特征在于,所述成型体的97.5质量%以上由无机成分构成。
[17]如[15]或[16]所述的成型体,其特征在于,在其内部具有气孔径为0~2000μm的范围、且具有任意形状的多孔结构,气孔率为0~99%。
[18]如[15]~[17]中任一项所述的成型体,其特征在于,在磷酸八钙晶体上担载有在组成中具有包含与钙化学键合的官能团的物质的物质。
[19]如[18]所述的成型体,其特征在于,在磷酸八钙晶体中还担载有其他物质。
发明效果
利用本发明的制造方法,能够制造可适合用于骨再生材料、有害分子吸附材料、催化剂担载材料、药剂担载材料等的大型的OCP成型体。
附图说明
[图1-1](A)为实施例1中制造的前体陶瓷组合物即半水石膏(CSH)-磷酸二氢钠二水合物(NaDP)组合物的照片,(B)为XRD图谱(pattern)。
[图1-2]为实施例1中制造的成型体的照片,(A)为于37℃浸渍10天得到的成型体,(B)为于60℃浸渍1天得到的成型体,(C)为于80℃浸渍1天得到的成型体。
[图1-3]为实施例1中制造的成型体的XRD图谱。
[图1-4]为实施例1中制造的成型体(80℃、1天)的径向拉伸强度(DTS)。
[图1-5](A)~(D)为实施例1中制造的成型体(80℃、1天)的SEM照片,(A)及(B)表示表面,(C)及(D)表示截面。(E)~(H)为作为前体陶瓷组合物的CSH-NaDP组合物的SEM照片,(E)及(F)表示表面,(G)及(H)表示截面。
[图2-1](A)为实施例2中制造的前体陶瓷组合物即CSH-NaDP连通多孔结构组合物的照片,(B)为显微CT图像。
[图2-2](A)为实施例2中制造的成型体(80℃、1天)的照片,(B)为显微CT图像。
[图2-3]为实施例2中制造的成型体(80℃、1天)的XRD图谱。
[图2-4](A)及(B)为实施例2中制造的OCP连通多孔结构成型体的SEM照片,(C)及(D)为作为前体陶瓷组合物的CSH-NaDP连通多孔结构组合物的SEM照片。
[图2-5]为埋入兔股骨头后经过2星期后的HE染色组织图像,(A)为实施例1中制造的OCP成型体,(B)为实施例2中制造的OCP连通多孔结构成型体,(C)为作为对照的羟基磷灰石烧结体。
[图2-6]为埋入兔股骨头后经过4星期后的HE染色组织图像,(A)为实施例1中制造的OCP成型体,(B)为实施例2中制造的OCP连通多孔结构成型体,(C)为作为对照的羟基磷灰石(HAp)烧结体。
[图3-1](A)为实施例3中制造的磷酸氢钙二水合物(DCPD)组合物的照片,(B)为XRD图谱。
[图3-2]为实施例3中制造的OCP成型体的照片,(A)~(E)为于4℃~80℃浸渍1天得到的成型体,(F)~(J)为于4℃~80℃浸渍2天得到的成型体,(K)~(N)为于4℃~80℃浸渍3天得到的成型体,(O)~(R)为于4℃~80℃浸渍7天得到的成型体。
[图3-3]为实施例3中制造的OCP成型体(4℃~80℃、1天)的XRD图谱。
[图3-4]为实施例3中制造的OCP成型体(4℃、室温、7天)的XRD图谱。
[图3-5]为实施例3中制造的OCP成型体(37℃、3天、7天)的XRD图谱。
[图3-6]为实施例3中制造的OCP成型体(70℃、2天、3天、7天)的XRD图谱。
[图3-7]为实施例3中制造的OCP成型体(80℃、2天)的XRD图谱。
[图3-8]为实施例3中制造的OCP成型体(70℃、1天~7天)的DTS强度。
[图4-1]示出了实施例4中制造的OCP成型体的照片,(A)为浸渍于含有柠檬酸的磷酸氢二钠水溶液中后的OCP成型体,(B)为浸渍于含有琥珀酸的磷酸氢二钠水溶液中后的OCP成型体,(C)为浸渍于含有酒石酸的磷酸氢二钠水溶液中后的OCP成型体。
[图4-2]为实施例4中制造的OCP成型体的XRD图谱。
[图4-3]示出了实施例4中制造的OCP成型体(柠檬酸、琥珀酸)的FT-IR光谱。
[图4-4]为实施例4(7)中制造的含有柠檬酸的成型体的DTS强度。
[图4-5]为实施例4(7)中制造的含有琥珀酸的成型体的DTS强度。
[图5-1]为实施例5中制造的OCP成型体的照片。
[图5-2]为实施例5中制造的OCP成型体的XRD图谱。
[图5-3]为实施例5中制造的OCP成型体的照片。
[图5-4]为实施例5中制造的OCP成型体的XRD图谱。
[图6-1]为实施例6中制造的OCP成型体的照片。
[图6-2]为实施例6中制造的OCP成型体的XRD图谱。
[图7-1]为实施例7中制造的OCP成型体的照片,(A)为浸渍于聚乙二醇中得到的OCP成型体,(B)为浸渍于聚丙烯酸钠中得到的OCP成型体。
[图7-2]为实施例7中制造的OCP成型体的XRD图谱。
具体实施方式
本发明的体积为2.0mm3以上的包含磷酸八钙的成型体(以下,有时简称为成型体)的制造方法的特征在于,将前体陶瓷组合物浸渍于溶液中,使所述前体陶瓷组合物反应,将所述前体陶瓷组合物的至少一部分转化为磷酸八钙,所述前体陶瓷组合物在组成中包含Ca及PO4中的至少一者且在H2O中的溶解度高于磷酸八钙,所述前体陶瓷组合物的体积大于2.0mm3,所述溶液含有作为磷酸八钙的组成的Ca、PO4、H2O的组成之中未被包含在所述前体陶瓷组合物中的组成。
<前体陶瓷组合物>
本发明中的前体陶瓷组合物如上所述,在组成中包含Ca及PO4中的至少一者,在H2O中的溶解度高于磷酸八钙,体积为大于2.0mm3的大小。需要说明的是,PO4是指呈H3PO4、H2PO4 -、HPO4 2-、PO4 3-的形态的分子、或离子的总称,当然根据溶液的状态而存在混杂的情况。另外,PO4的浓度是指这些分子、离子的总和。
该前体陶瓷组合物的成型方法没有特别限定。例如,如石膏等那样,可以利用具有自固化性的前体陶瓷的固化反应而成型,可以利用烧结、干燥所引起的盐的析出反应而成型,也可以通过加压成型而制成压粉体。
从制造2.0mm3以上的OCP成型体的观点考虑,前体陶瓷组合物的体积大于2.0mm3。前体陶瓷组合物的大小基本在OCP成型体中得到保留,因此配合制造的目的适当调节大小即可。例如,可以为5.0mm3以上,也可以为10.0mm3以上。
作为本发明中的前体陶瓷组合物,可使用例如半水石膏(CSH)、二水石膏(CSD)、无水石膏(CSA)、磷酸氢钙二水合物(DCPD)、无水磷酸氢钙(DCPA)、α型磷酸三钙(α-TCP)、β型磷酸钙(β-TCP)、碳酸钙、磷酸二氢钙一水合物(MCPM)、磷酸二氢钙(MCPA)、无定形磷酸钙(ACP)、氢氧化钙等。上述可以单独使用,也可以同时使用2种以上。
前体陶瓷组合物在H2O中的溶解度高于OCP在H2O中的溶解度(0.090g/L)即可,优选高2倍以上。通过使溶解性为高水平,能够进一步促进前体陶瓷组合物的组成转化。
<浸渍溶液>
此处的溶液,是指溶质成分和溶剂成分在宏观状态下稳定地单一、且均匀地分散的液相。即使在微观上存在胶体粒子、分子簇、溶剂合分子、缔合簇等,但只要就整体而言具有流动性且是均匀的,则涵盖在溶液内。
即使在溶液中存在没有均匀地分散的物质、宏观上呈悬浮状态,在本发明的反应进行的情况下,仍然可以将其视为溶液。即,此处的溶液,是指电解质溶液、非电解质溶液、电解质悬浮液、非电解质悬浮液的总称。特别地,将用作溶剂的液体之中、水为50质量%以上的溶液称为水溶液。
就浸渍溶液而言,只要含有作为OCP的组成的Ca、PO4、H2O的组成之中、未被包含于上述陶瓷组合物的组成中的全部组成即可,没有特别限定。例如,作为含有PO4的溶液,可以举出磷酸、磷酸三钠、磷酸氢二钠、磷酸二氢钠、磷酸三钾、磷酸二氢钾、磷酸氢二钾、磷酸三铵、磷酸氢二铵、磷酸二氢铵等的溶液,作为含有Ca的溶液,可以举出氯化钙、硝酸钙、乙酸钙、乳酸钙、氢氧化钙、碳酸氢钙等的溶液。
在前体陶瓷组合物包含OCP的构成成分之中Ca及PO4这两者、无需从溶液得到Ca及PO4的情况下,也可以使用其他溶液。即,可以使用蒸馏水、碳酸钠、碳酸氢钠、硫酸钠、硫酸氢钠、氯化钠、氯化铵、硝酸铵、氢氧化钠、氢氧化钾等的溶液。另外,关于溶液的溶剂中的、H2O的比率没有特别限定。
在前体陶瓷组合物包含OCP的构成成分之中Ca及PO4中的至少一者、还含有H2O、无需从溶液得到H2O的情况下,可以将前体陶瓷组合物浸渍于含有Ca、PO4之中未被包含在前体陶瓷组合物中的成分、并且不含有H2O的溶剂中来制造OCP成型体。此外,在前体陶瓷组合物包含作为OCP的构成成分的、Ca、PO4及H2O中的全部成分的情况下,可以浸渍于不包含Ca、PO4及H2O中全部成分的溶液中来制造OCP成型体。需要说明的是,作为前体陶瓷组合物的H2O的含有形态,没有特别限定。可以举出例如结晶水、孔隙水、吸附水等。
作为浸渍的溶剂,除了水以外,可以举出以甲醇、乙醇、1-丙醇、1-丁醇、1-戊醇、1-己醇、1-庚醇、1-辛醇、1-壬醇、1-癸醇等伯醇、2-丙醇(异丙醇)、2-丁醇、2-戊醇、2-己醇、环己醇等仲醇、叔丁醇、2-甲基-2-丁醇、2-甲基-2-戊醇、2-甲基-2-己醇、3-甲基-3-戊醇、3-甲基-3-辛醇等叔醇为代表的一元醇;乙二醇、二乙二醇等二元醇;甘油等三元醇;苯酚等芳香环醇;聚乙二醇(PEG)、聚丙二醇(PPG)等聚醚;聚丙烯酸、聚氨基甲酸等聚羧酸;乙酸、戊酸、己酸、月桂酸、棕榈酸、硬脂酸、油酸、亚油酸等脂肪酸;戊烷、丁烷、己烷、庚烷、辛烷等链烷;二甲醚、甲基乙基醚、乙醚等醚;苯、甲苯、苦味酸、TNT等芳香族化合物;萘、薁、蒽等多环芳香族烃;氯甲烷、二氯甲烷、氯仿、四氯化碳等有机卤化物;乙酸乙酯、丁酸甲酯、水杨酸甲酯、甲酸乙酯、丁酸乙酯、己酸乙酯、乙酸辛酯、邻苯二甲酸二丁酯、碳酸亚乙酯、环硫乙烷这样的酯类;环戊烷、环己烷、萘烷等环烷;双环烷;丙酮、甲乙酮、二乙基甲酮等酮;甲醛、乙醛、丙醛、丁醛、戊醛、己醛、香草醛等醛;氨基甲烷、氨基乙烷、乙二胺、三乙胺、苯胺等胺化合物;葡萄糖、果糖、苏糖醇等糖类;甲硫醇、乙硫醇、丙硫醇、苯硫酚等硫醇类;二甲基硫醚、二苯基硫醚、芦笋酸、胱胺、胱氨酸等二硫化物等。上述可以单独使用,也可以混合多种进行使用。
<反应条件>
前体陶瓷组合物浸渍于溶液中的时间以OCP形成、反应在形成稳定相之前终止的方式进行调节。通常为10分钟~30天,优选为2小时~14天,进一步优选为2小时~7天。
在溶液中含有PO4及Ca离子的情况下,形成的磷酸钙的稳定相依赖于溶液的pH。此处的磷酸钙是指,在除了无机化合物中的水以外的组成之中,在组成中包含50质量%以上的Ca及PO4,且Ca在组成中的阳离子的比率中为最大,且PO4在组成中的阴离子的比率中为最大的无机化合物。
在溶液的pH为4以上的情况下,磷灰石呈稳定相。
此处的磷灰石是指被分类为磷酸钙的物质,并且是具有以下的化学组成、且在pH为4以上的溶液中较之不属于磷灰石的磷酸钙而言为热力学最稳定相的总称。即,磷灰石是指Ca10-xQy(PO4)6-zRwJv表示的物质。此处,Q为Na、Mg、Fe、K、Sr、Rb、Zn、Ga、Al、Mn、Cu、Mo、Ag、Au、Se、Te等中任一者的阳离子或空隙,R为HPO4、SO4、CO3、BO3、WO4、VO4、SiO4等中任一者的阴离子或空隙,J为OH、Cl、F、Br、I等中任一者的阴离子,X<5、Y<5、Z<3、W<3,且Ca及PO4的总和为50原子%以上。
具体而言,可以举出羟基磷灰石(Ca10(PO4)6(OH)2、HAp)、缺钙磷灰石(Ca9(HPO4)4(PO4)2(OH))、氟磷灰石(Ca10(PO4)6F2)、氯磷灰石(Ca10(PO4)6Cl2)、CO3Ap等物质。这些物质的Ca/PO4摩尔比率通常为1.50~2.50,在计量HAp的情况下为1.67。
已知在磷灰石从溶液中形成的情况下,有时经由亚稳相而形成。在溶液的pH为4以上的情况下,作为稳定相的磷灰石以外的相在磷灰石形成后,最终组成转化为磷灰石而消失。
OCP为磷灰石的亚稳相,因此若长时间浸渍于溶液、持续反应,则在pH为4以上的溶液中最终会形成磷灰石。但是,通过在经由OCP从前体形成的反应途径、来形成磷灰石的反应中,在磷灰石形成前终止反应,从而能够得到OCP。
即,在OCP从作为前体的陶瓷成型体形成后、在向磷灰石的组成转化完成之前终止反应,从而能够得到包含OCP的成型体。
为了终止反应,在成型体的情况下,需要使成型体从溶液中分离的操作。即,将成型体从所浸渍的溶液中取出,施以除去附着于成型体的溶液的操作,从而能够终止反应。本发明中,以将成型体从浸渍溶液中取出的时间点作为反应的终止时间。
就除去附着于成型体的溶液的操作而言,没有特别限定。通常,用蒸馏水、乙醇等溶剂多次洗涤成型体,将附着于成型体的溶液与这些溶剂置换后,用滤纸等除去剩余的溶剂,使其干燥。
OCP组成转化为HAp的情况下的化学反应式如下。
5Ca8H2(PO4)6·5H2O(OCP)
→4Ca10(PO4)6(OH)2(HAp)+6H3PO4+17H2O…(1)
由于在上述式(1)的反应中生成的H3PO4呈酸性,因此从OCP形成磷灰石时,周围的溶液的pH降低。即,较之OCP形成、残留的时间点的pH而言,进行向磷灰石的组成转化、成为磷灰石单一相的时间点的溶液的pH降低。利用这一点,从而可以将其作为用于在OCP组成转化为磷灰石之前终止反应的指标。
上述式(1)的反应中,溶液的pH降低,而其降低量依赖于溶液与成型体的质量比、及溶液中含有的离子所带来的缓冲作用。但是,在任一种情况下,在OCP消失、成为仅有磷灰石的反应完成时间点,pH不再变化。
本说明书中,在X射线粉末衍射法中,将磷酸钙形成磷灰石单一相、且pH变化消失的时间点定义为不再进一步发生反应的时间点,即反应完成时间点,将该时间点的pH定义为反应完成时间点的pH。
反应完成时间点的pH没有特别限定,优选为4以上,更优选为4~10,进一步优选为4~7。
关于反应终止时的溶液的pH,在反应完成时间点的pH为4以上的情况下,只要比其更高即可,没有特别限定。优选比反应完成时的pH高0.1以上,更优选高0.2以上。
另外,OCP的化学组成为Ca8H2(PO4)6·5H2O,因此化学计量的Ca/PO4比率为1.33。因此,在磷灰石从OCP形成的情况下,鉴于化学计量,需要将剩余的PO4释放至周围的溶液中,或从周围的溶液中吸收不足的量的Ca。
即,在磷灰石从OCP形成的情况下,溶液及成型体的Ca/PO4比率发生变化。利用这一点,从而可以将其作为用于在OCP组成转化为磷灰石之前终止反应的指标。另外,此时的溶液及成型体的Ca/PO4比率的变化依赖于成型体与溶液的质量的比率、溶液所含有的Ca、或PO4的浓度。但是,在任一种情况下,在OCP完全地组成转化为磷灰石、成为仅有稳定相的反应完成时间点,成型体和溶液的Ca/PO4不再变化。
关于反应完成时间点的成型体的Ca/PO4比率没有特别限定。在磷灰石为单相的情况下,通常为1.5~2.0。
关于反应终止时的成型体的Ca/PO4比率,在磷酸钙以磷灰石的形式构成反应完成时间点的成型体的情况下,只要高于该情况下的Ca/PO4比率即可,没有特别限定。
关于反应终止时的溶液的Ca/PO4比率,在磷酸钙以磷灰石的形式构成反应完成时间点的成型体的情况下,只要低于该反应完成时间点的溶液的比率即可,没有特别限定。
此处,作为磷灰石为稳定相的体系的一个实施方式,优选将酸性的前体陶瓷组合物浸渍于pH为4~14的溶液、优选pH为大于7且为12以下的溶液中,保持至pH成为4~7为止从而制造OCP成型体。该方式中,可以认为,从作为原料的前体陶瓷组合物经由ACP的生成而生成OCP,结果,ACP(在高pH区段其生成得以促进)在高pH溶液中高效地被生成,并且,OCP(在低pH区段其生成得以促进)由于前体陶瓷组合物自身的低pH而得以高效地被生成,由此从前体高效地组成转化为OCP成型体。
另一方面,在溶液的pH低于4的情况下,DCPA或DCPD成为稳定相。DCPA及DCPD的Ca/PO4理论上为1.00。DCPD为小于200℃时的稳定相,DCPA为200℃以上时的稳定相(Ame.Mine.2011,96,368-373,2011)。
在溶液的pH低于4的情况下,作为稳定相的DCPD或DCPA以外的相在DCPD或DCPA形成后,最终组成转化为DCPD或DCPA而消失。
OCP为DCPD或DCPA的亚稳相,因此若长时间浸渍于溶液、持续反应,则在pH小于4的溶液中最终会形成DCPD或DCPA。但是,在经由OCP从前体形成的反应途径、来形成DCPD或DCPA的反应中,通过在DCPD或DCPA形成前终止反应,从而能够得到OCP。
OCP组成转化为DCPD的情况下的化学反应式如下。
Ca8H2(PO4)6·5H2O(OCP)+11H2O
→6CaHPO4·2H2O(DCPD)+4OH-+2Ca2+…(2)
另外,OCP组成转化为DCPA的情况下的化学反应式如下。
Ca8H2(PO4)6·5H2O(OCP)+4H2O
→6CaHPO4(DCPA)+4OH-+2Ca2+…(3)
上述式(2)及(3)的反应中,DCPD或DCPA从OCP形成时,周围的溶液的pH上升。即,较之OCP形成、残留的时间点的pH而言,进行向DCPD或DCPA的组成转化、成为DCPD或DCPA的单一相的时间点的溶液的pH升高。利用这一点,从而可以将其作为用于在OCP组成转化为DCPD或DCPA之前终止反应的指标。
上述式(2)及(3)的反应中,溶液的pH上升,而其上升量依赖于溶液与成型体的质量比、及溶液中含有的离子所带来的缓冲作用。但是,在任一种情况下,在OCP消失、成为仅有DCPD及DCPA的反应完成时间点,pH不再变化。
本说明书中,在X射线粉末衍射法中,将磷酸钙成为仅DCPD或DCPA、且不再有pH变化的时间点定义为不再进一步发生反应的时间点,即反应完成时间点,将该时间点的pH定义为反应完成时间点的pH。
关于反应终止时的溶液的pH,在反应完成时间点的pH低于4的情况下,只要比其更低即可,没有特别限定。优选相较于反应完成时的pH而言低0.1以下,进一步优选相较于反应完成时的pH而言低0.2以下。
另外,OCP的化学组成为Ca8H2(PO4)6·5H2O,因此化学计量的Ca/PO4比率为1.33。因此,在DCPD从OCP形成的情况下,鉴于化学计量,需要将剩余的Ca释放至周围的溶液中,或从周围的溶液中吸收不足的量的PO4。
即,在DCPD及DCPA从OCP形成的情况下,溶液及成型体的Ca/PO4比率发生变化。利用这一点,从而可以将其作为用于在OCP组成转化为磷灰石之前终止反应的指标。另外,此时的溶液及成型体的Ca/PO4比率的变化依赖于成型体与溶液的质量的比率、溶液所含有的Ca、或PO4的浓度。但是,在任一种情况下,在OCP完全地组成转化为DCPD及DCPA、成为仅有稳定相的反应完成时间点,成型体和溶液的Ca/PO4不再变化。
关于反应完成时间点的成型体的Ca/PO4比率没有特别限定。在DCPD或DCPA为单相的情况下,通常为0.8~1.2。
关于反应终止时的成型体的Ca/PO4比率,在磷酸钙以DCPD或DCPA的形式构成反应完成时间点的成型体的情况下,只要低于该情况下的Ca/PO4比率即可,没有特别限定。
关于反应终止时的溶液的Ca/PO4比率,在磷酸钙以DCPD或DCPA的形式构成反应完成时间点的成型体的情况下,只要高于该情况下的Ca/PO4比率即可,没有特别限定。
具体而言,本发明中,可以预先在与所期望的制造条件中的温度和压力相同的条件下,探测反应中的pH的变化、Ca/PO4比率的变化,对直至成为稳定相为止的时间进行测定,基于该预备的时间条件来确定浸渍时间。
关于将前体陶瓷组合物浸渍于溶液中的温度,没有特别限定。通常为-80℃~270℃,优选为0℃~99℃,更优选为4℃~99℃。
<OCP成型体>
就本发明中的OCP成型体而言,优选OCP为组成中的10质量%以上,更优选OCP为组成中的50质量%以上,进一步优选OCP为组成中的70质量%以上,特别优选OCP为组成中的90质量%以上。特别地,在用作骨再生材料的情况下,除了上述的条件以外,还优选成型体中的HAp的比例为10质量%以下,更优选为5质量%以下,进一步优选为1质量%以下,特别优选无法检测。
此处的成型体,是指作为无机成分的构成成分的、以OCP为代表的无机物的晶体之间不存在胶原蛋白等物质、而是彼此通过化学键合、或者晶体彼此的缠结或融合而固化以保持形态的成型体,并且是至少在99.5%乙醇、或水中浸渍24小时也不会崩解而能够保持形态的成型体。因此,不包括对OCP粉末进行压粉并成型而得到的OCP压粉体。
此处的化学键合,是指在构成无机成分的晶体之间,存在1个以上被分类为共价键、离子键、氢键、金属键中任一者的化学键,利用该键将晶体彼此固定,确定晶体彼此的位置关系,维持成型体的基本形态。
此处的晶体彼此的缠结,是指在微观上构成成型体的多个晶体介由彼此的晶面而互相接触的结构,并且,利用该机构将多个晶体固定,维持成型体的基本形态。
此处的晶体彼此的融合,是指构成成型体的多个晶体在晶面处存在与其他晶体没有间隙地接触的部分,在该部分无法观察到晶界。
本发明中制造的成型体的形状保留前体陶瓷组合物的形状。因此,通过使用可以实现精密加工的前体陶瓷组合物,从而能够向成型体赋予连通多孔结构、蜂窝状结构等复杂的形状。
作为本发明中的OCP成型体的形状,没有特别限定。如上所述,OCP成型体基本保留前体陶瓷的形状,因此其形状也基本取决于前体陶瓷组合物的形状。具体而言,作为OCP成型体的形状,可以举出例如碟状(disk)、块状(block)、片状(sheet)。
作为碟状的大小没有特别限定,直径为例如1~20mm,优选为5~10mm。厚度为例如0.5~5mm,优选为1~3mm。
作为块状的大小没有特别限定,例如,为长1~15mm、宽1~50mm、高0.5~100mm,优选为长8~12mm、宽10~30mm、高10~50mm。
作为片状的大小没有特别限定,例如,为长1~20mm、宽1~20mm、厚0.01~0.5mm,优选为长5~15mm、宽5~15mm、厚0.05~0.3mm。
本发明中的OCP成型体的气孔径及内部结构没有特别限定。例如,气孔径通常为0~2000μm,优选为50~300μm。内部结构可以是单孔结构,也可以是具有任意形状的连通多孔结构。关于长径比,没有特别限定。关于气孔率(空隙率),为0~99%,优选为40~80%。
多孔结构的制造方法没有特别限定。可以通过晶体生长使前体陶瓷组合物的颗粒彼此结合而形成多孔结构,可以添加易溶性的盐作为空隙形成剂,可以通过挤出成型来制造多孔结构,也可以添加包含有在烧结时会被烧除的有机物的气孔形成剂来制造多孔结构。
可以将在组成中具有羧基、硅烷醇基、磷酸基、磺基、羟基、巯基等与钙化学键合的官能团的分子担载于本发明的OCP成型体。在欲将具有与钙化学键合的官能团的分子担载于OCP成型体的情况下,预先使浸渍溶液或前体陶瓷组合物中含有在组成中含有与钙化学键合的官能团的分子即可。需要说明的是,此处,将在组成中含有与钙化学键合的官能团的OCP定义为无机物。
此处,具有羧基的分子是指在组成中具有一个以上以-COOH表示的官能团的分子。
作为在OCP成型体中担载的具有羧基的分子,可以使用被分类为单羧酸、二羧酸、三羧酸、羧酸硫醇、卤代羧酸、氨基酸、芳香族酸、羟基酸、糖酸、硝基羧酸、聚羧酸等的物质、它们的衍生物、及使它们聚合而得到的物质。即,可以举出乙酸、丙酸、丁酸、甲酸、戊酸、琥珀酸、柠檬酸、巯基十一烷酸、硫代乙醇酸、芦笋酸(asparagusic acid)、α-硫辛酸、β-硫辛酸、二氢硫辛酸、氯乙酸、丙二酸、乌头酸、苹果酸、草酸、酒石酸、丙二酸、戊二酸、己二酸、富马酸、马来酸、草酰乙酸、α-氧代戊二酸、草酰琥珀酸、丙酮酸、异柠檬酸、α-丙氨酸、β-丙氨酸、精氨酸、天冬酰胺、天冬氨酸、半胱氨酸、谷氨酰胺、谷氨酸、甘氨酸、组氨酸、异亮氨酸、亮氨酸、赖氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、丝氨酸、苏氨酸、色氨酸、酪氨酸、缬氨酸、半胱氨酸、羟基脯氨酸、o-磷酸丝氨酸、锁链素、胭脂碱、章鱼碱、甘露碱、酵母氨酸、N-甲基甘氨酸、二甲基甘氨酸、三甲基甘氨酸、瓜氨酸、谷胱甘肽、肌酸、γ-氨基丁酸、茶氨酸、乳酸、亚叶酸、叶酸、泛酸、苯甲酸、水杨酸、邻苯二甲酸、间苯二甲酸、对苯二甲酸、烟酸、吡啶甲酸、没食子酸、苯六甲酸、肉桂酸、茉莉酮酸、十一烯酸、乙酰丙酸、艾杜糖醛酸、葡糖醛酸、半乳糖醛酸、甘油酸、葡糖酸、胞壁酸、唾液酸、甘露糖醛酸、乙醇酸、乙醛酸、乙二胺四乙酸(EDTA)、硝基乙酸、硝基氢化肉桂酸、硝基苯甲酸、聚丙烯酸、聚柠檬酸、聚衣康酸以及它们的盐等。上述可以单独进行担载,也可以复合使用。
此处,所谓具有硅烷醇基的分子,是指在组成中具有以-SiO4R3表示的官能团的分子。R为H或烷基。
作为使OCP成型体担载的具有硅烷醇基的分子,可以举出γ-甲基丙烯酰氧基丙基三甲氧基硅烷(γ-MPTS)、原硅酸四乙酯(TEOS)、硅酸钠、原硅酸、偏硅酸、二硅酸以及它们的盐等。上述可以单独进行担载,也可以复合使用。
此处,所谓具有磷酸基的分子,是指在组成中具有以-PO4R2表示的官能团。R为H或烷基。
作为使OCP成型体担载的具有磷酸基的分子,可以举出腺苷三磷酸(ATP)、腺苷二磷酸(ADP)、核苷酸、葡萄糖-6-磷酸、黄素单核苷酸、多聚磷酸、10-甲基丙烯酰氧基癸基二氢磷酸酯(MDP)、植酸、以及它们的盐等。上述可以单独进行担载,也可以复合使用。
此处,具有磺基的分子是指在组成中具有以-SO3R表示的官能团。R为H或烷基。
作为使OCP成型体担载的具有磺基的分子,可以举出苯磺酸、牛磺酸、直链烷基苯磺酸钠、二甲苯硅烷醇、溴酚蓝、甲基橙、4,4’-二异硫氰基-2,2’-芪二磺酸(DIDS)、偶氮玉红、苋菜红、靛蓝胭脂红、水溶蓝、甲酚红、考马斯亮蓝、刚果红、对氨基苯磺酸、酒石黄、百里酚蓝、对甲苯磺酰叠氮、胭脂红、8-羟基-1,3,6-芘三磺酸三钠(pyranine)、亚甲基蓝、羟乙基哌嗪乙磺酸(HEPES)、环己基氨基磺酸钠、糖精、牛磺胆酸、羟乙基磺酸、磺基丙氨酸、10-樟脑磺酸、4-羟基-5-氨基萘-2,7-二磺酸、甲磺酸、乙磺酸、以及它们的盐等。上述可以单独进行担载,也可以复合使用。
此处,所谓具有羟基的分子,是指在组成中具有以-OH表示的官能团。
作为使OCP成型体担载的具有羟基的分子,可以举出被分类为醇的化合物、甲基丙烯酸2-羟基乙酯(HEMA)、羟基胺、异羟肟酸、苯酚、被分类为羟醛的化合物、被分类为糖的化合物、被分类为甘醇(glycol)的化合物、肌醇、被分类为糖醇的化合物、泛酰巯基乙胺、以及它们的盐等。上述可以单独进行担载,也可以复合使用。
此处,所谓具有巯基的分子,是指在组成中具有以-SH表示的官能团。R为H或烷基。
作为使OCP成型体担载的具有巯基的分子,可以举出卡托普利、甲硫醇、乙硫醇、半胱氨酸、谷胱甘肽、苯硫酚、乙酰半胱氨酸、1,2-乙二硫醇、半胱胺、二硫赤藓糖醇、二硫苏糖醇、二巯基丙醇、硫代乙醇酸、硫普罗宁、2-萘硫醇、布西拉明、呋喃-2-基甲硫醇、D-青霉胺、放线硫醇、美司钠、3-甲基-2-丁烯-1-硫醇、3-巯基丙酮酸、以及它们的盐等。上述可以单独进行担载,也可以复合使用。
该盐是指化合物的盐、尤其是在使其与蒸馏水等溶剂接触时可良好地溶解、与上述化合物同样地发挥功能的盐。该盐不仅包括上述化合物的无水盐,当然也包括含水盐。作为该盐,可以举出例如钠盐、钾盐、锂盐、铷盐、铯盐等碱金属盐、镁盐、锶盐等碱土金属盐(不包括Ca盐)、铝盐、锌盐、铁盐、镍盐、钴盐、铜盐等过渡金属盐、铵盐等无机盐、三(羟基甲基)氨基甲烷盐、苯基甘氨酸烷基酯盐、二环己胺盐、N,N’-二苄基乙二胺盐、乙二胺盐、氨基葡萄糖盐、胍盐、二乙胺盐、三乙胺盐、N-甲基葡萄糖胺盐、叔辛胺盐、二苄胺盐、吗啉盐、普鲁卡因盐、二乙醇胺盐、N-苄基-N-苯乙胺盐、哌嗪盐、氯普鲁卡因盐、四甲基铵盐等有机胺盐、苯酚盐等。
浸渍溶液所含有的、在组成中包含与钙化学键合的官能团的物质的浓度没有特别限定。通常为0~2mol/L,优选为0.01~0.5mol/L。
在除了在组成中包含与钙化学键合的官能团的物质以外,还想担载其他物质(化合物)的情况下,需要预先向前体陶瓷组合物及浸渍溶液中的至少一者中添加化合物。需要说明的是,不仅担载有在组成中包含与钙化学键合的官能团的物质、还担载有其他化合物的OCP在此处也定义为无机物。
作为担载的化合物的种类,可以举出已知的抗菌剂、抗生素、抗癌剂、防腐剂、耐酸性提高剂等。
作为担载的化合物,可以举出银、硒、铂、金、钯、铱、锇、铼、镓、锗、碲、钛、铋等元素以及它们的离子、络合物、簇合物、纳米粒子、青霉素G·普鲁卡因(青霉素G-普鲁卡因盐)、苄星青霉素(苄星青霉素盐)、梭链孢酸、镰孢真菌素、磷霉素、莫匹罗星、溴莫普林(brodimoprim)、地红霉素、苄基青霉素、苯氧基甲基青霉素、甲氧西林、氨苄青霉素、氯唑西林、羧苄青霉素、匹氨西林(pivampicillin)、阿莫西林、酞氨西林(talampicillin)、巴氨西林、替卡西林、阿洛西林、美洛西林、匹美西林、哌拉西林、阿莫西林-克拉维酸、阿帕西林、替莫西林、替卡西林-克拉维酸、氨苄青霉素-舒巴坦、舒他西林、哌拉西林-他唑巴坦等青霉素系抗生素、链霉素等链霉素系抗生素、氯霉素、甲砜霉素等氯霉素系抗菌剂、万古霉素、替考拉宁等糖肽系抗生素、氯四环素、金霉素、氯霉素、土霉素、去甲基金霉素(demethylchlortetracycline)、去甲金霉素(ledermycin)、赖甲环素、多西环素、地美环素、米诺环素等四环素系抗生素、庆大霉素、新霉素、奇霉素、妥布霉素、阿米卡星、小诺霉素、异帕米星、阿贝卡星等氨基糖苷系抗生素、头孢呋辛、头孢克洛、头孢噻肟、头孢磺啶、头孢哌酮、头孢唑啉、头孢拉定、头孢羟氨苄、头孢孟多、头孢替安、头孢氨苄、头孢尼西、头孢匹胺、头孢哌酮-舒巴坦、头孢地嗪、头孢布坦(ceftibufen)、头孢泊肟、头孢地尼、头孢他美、头孢匹罗、头孢丙烯、头孢曲松、头孢甲肟、头孢他啶、头孢呋辛、头孢吡肟等头孢菌素系抗生素、黏菌素等多肽系抗生素、罗红霉素、阿奇霉素、麦迪霉素、红霉素、螺旋霉素、克拉霉素等大环内酯系抗生素、维吉尼霉素、普那霉素等链阳霉素系抗生素、氯碳头孢等碳头孢烯系抗生素、磺胺甲噻二唑、磺胺乙酰、磺胺甲嘧啶、磺胺二甲嘧啶、磺胺嘧啶等磺胺剂、左氧氟沙星、氟罗沙星、那氟沙星、诺氟沙星、氧氟沙星、环丙沙星、恩诺沙星、洛美沙星、芦氟沙星、西他沙星等喹诺酮系抗菌剂、红霉素等酮内酯系抗生素、帕尼培南-倍他米隆等碳青霉烯(carbapenem)系抗生素、克林霉素等林可霉素系抗生素、拉氧头孢、氟氧头孢等氧头孢烯系抗生素、亚胺培南-西拉司丁等碳青霉烯系抗生素、头孢西丁、头孢美唑、头孢替坦等头霉素系抗生素、氨曲南等单环内酰胺系抗生素、乙酰氨基酚、乙酰水杨酸、乙水杨胺、水杨酰胺、双氯芬酸、苯乙酸、吲哚美辛、洛索洛芬、布洛芬、酮基布洛芬、萘普生、吡罗昔康、米索前列醇、美洛昔康、氯诺昔康等非甾体性抗炎药等。
作为担载的抗癌剂,可以举出环三磷腈-铂络合物复合体、顺铂、奈达铂、奥沙利铂、卡铂这样的铂络合物、5-氟尿嘧啶(5-FU)、替加氟、替加氟·尿嘧啶、甲氨蝶呤这样的代谢拮抗剂、OK-432等抗癌溶血性链球菌制剂、云芝多糖、香菇多糖、西佐糖(sizofiran)、西佐喃(sonifiran)等抗癌多糖体、盐酸多柔比星、丝裂霉素C、放线菌素D、盐酸博来霉素、硫酸博来霉素、盐酸柔红霉素、新制癌菌素、盐酸阿柔比星(aclarubicin hydrochloride)、盐酸表阿霉素等抗癌抗生素、天冬酰胺酶这样的酶、长春碱这样的有丝分裂抑制剂、依托泊苷(etoposide)这样的拓扑异构酶抑制剂、干扰素这样的生物反应修饰剂、胞嘧啶阿拉伯糖苷、氧基脲(oxyurea)、N-{5-[N-(3,4-二氢-2-甲基-4-氧代喹唑啉-6-基甲基)-N-甲基氨基]-2-噻吩甲酰}-L-谷氨酸
(N-{5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl}-L-glutamic acid)这样的抗代谢剂、阿霉素、博来霉素这样的嵌入抗生素(intercalating antibiotics)、他莫昔芬这样的抗雌激素剂、长春新碱这样的植物生物碱、丝裂霉素C、放线菌素D、盐酸博来霉素、硫酸博来霉素、盐酸柔红霉素、盐酸多柔比星、新制癌菌素、盐酸阿柔比星、阿克拉霉素(Aclacinon)、盐酸表阿霉素这样的抗癌抗生素等。
除了上述的抗菌剂、抗生素、抗癌剂、耐酸剂以外,还可以根据用途而担载不属于上述的化合物。即,可以举出黄素单核苷酸、核黄素、腺嘌呤、鸟嘌呤、胸腺嘧啶、胞嘧啶、尿嘧啶、咖啡因、烟碱、阿托品、氮芥、碘解磷定(pralidoxime methiodide)、脱氧核糖、抗坏血酸、硫胺素、半乳糖胺、N-乙酰氨基半乳糖、艾杜糖、α-乙酰内酯、γ-丁内酯、甘油、乙二醇、碘仿、氯仿、溴仿、亚叶酸等。
担载的化合物的原子数没有特别限定。优选为200以下。
作为骨再生材料,优选的一个实施方式为仅由OCP和已作为骨置换型再生材料为人所知的材料构成的复合体,是实质上不含有其他成分的方式。
作为已作为骨置换型再生材料为人所知的材料,可以举出β-TCP、α-TCP、CSD、CSH、CSA、DCPD、碳酸钙、CO3Ap、ACP等。
就已作为骨置换型再生材料为人所知的材料与OCP的比率而言,只要构成为使得所含OCP的比例为成型体整体的10质量%以上即可,没有特别限定。可以考虑埋入部位及全身状态,根据用途而变更组成、比率。
另外,就本发明中的OCP成型体而言,优选97.5质量%以上由无机成分构成,更优选99.0质量%以上由无机成分构成。
以下,针对本发明的实施方式示出具体例,但本发明不限于这些具体例。
实施例
[实施例1]
(从以烧石膏作为主要成分的组合物制造OCP成型体)
(1)前体陶瓷组合物的制造
将均为自和光纯药工业购买的CSH(CaSO4·1/2H2O)3g、及NaDP(NaH2PO4·2H2O)2g与蒸馏水0.8mL一同在室温条件下、用刮刀以120rpm的混炼速度使用研钵混炼2分钟。将混炼体埋入φ6mm×3mm的组装模中进行成型。成型后于室温下静置12小时,充分地固化、干燥,制成作为前体陶瓷组合物之一的CSH-NaDP组合物。
该CSH-NaDP组合物是在H2O中的溶解度高于OCP的溶解度的陶瓷的混合物,并且,浸渍于蒸馏水时的周围的溶液的pH为4.5。CSH在蒸馏水(H2O)中的溶解度为2.60g/L,NaDP在蒸馏水(H2O)中的溶解度为949.0g/L。
图1-1(A)中示出了制得的前体陶瓷组合物即CSH-NaDP组合物的照片,(B)中示出了XRD图谱。
(2)利用组成转化反应的OCP成型体的制造
将上述(1)中制造的CSH-NaDP组合物于37℃、60℃、80℃浸渍于1mol/L磷酸氢二钠水溶液15mL中,使其反应12小时至10天。具体而言,使其于37℃反应12小时、1天、10天,于60℃反应12小时、1天、10天,于80℃反应12小时、1天、10天。反应后的成型体用蒸馏水洗涤3次以上,然后于37℃的恒温槽内静置6小时使其干燥。反应前的1mol/L磷酸氢二钠水溶液于室温的pH为9.5。于80℃反应1天后的水溶液的、室温的pH为6.8。该体系中,稳定相为磷灰石。
此处,1mol/L磷酸氢二钠水溶液是指下述溶液:使无水磷酸氢二钠141.96g、或磷酸氢二钠二水合物156.01g完全溶解于蒸馏水中,形成的溶液的总量为1L。只要磷酸氢二钠、或磷酸氢二钠二水合物与蒸馏水的比率相同,则也可以使所使用的磷酸氢二钠、磷酸氢二钠二水合物、及蒸馏水的量变化而进行制造。
(3)OCP成型体的表征
本实施例中制造的OCP成型体利用粉末X射线衍射(XRD)和傅里叶变换红外光谱法(FT-IR)进行了表征。关于XRD图谱,使用X射线衍射装置(D08 ADVANCE,Bruker AXS株式会社),射线源为CuKα射线、40kV、40mA,从3.0°至70.0°以0.02°的间隔使用步进扫描进行记录。使用OCP的JCPDS卡片编号、26-1056和HAp的JCPDS号9-432鉴定晶相。
(4)OCP成型体的机械特性的测定
本实施例中制造的OCP成型体的形状及微细结构使用以5kV的加速电压运作的Hitachi High-Technologies Corporation制扫描电子显微镜(SEM)S-3400N进行了研究。OCP成型体的空隙率如下算出:使用以67kV及160μA的加速电压运作的TOYO Corporation制显微CT拍摄系统Skyscan1076拍摄内部结构后,使用公共免费软件ImageJ分析而算出。关于OCP成型体的径向拉伸强度(DTS),使用岛津制作所制万能试验机AGS-J,以0.5mm/分钟的十字头移动速度进行测定。
(5)针对OCP成型体的形成而言的温度依赖性
图1-2中示出了将CSH-NaDP组合物于不同的温度浸渍于1mol/L磷酸氢二钠水溶液中而得到的成型体的照片。各成型体的大小为φ6mm×3mm。图1-3中示出了浸渍于1mol/L磷酸氢二钠水溶液中而得到的成型体的XRD图谱。
如图1-2及图1-3中所示,就于37℃浸渍于1mol/L磷酸氢二钠水溶液中的CSH-NaDP组合物而言,通过使其浸渍10天,形成了以OCP作为主要成分的成型体。就于60℃浸渍于1mol/L磷酸氢二钠水溶液中的CSH-NaDP组合物而言,通过使其浸渍1天,形成了以OCP作为主要成分的成型体。就于80℃浸渍于1mol/L磷酸氢二钠水溶液中的CSH-NaDP组合物而言,通过使其浸渍1天,形成了包含OCP单相的成型体。
(6)OCP成型体的机械特性
如图1-4所示,将CSH-NaDP组合物于80℃浸渍于1mol/L磷酸氢二钠水溶液中1天而制造的成型体的DTS强度为0.95±0.16MPa。
(7)OCP成型体的微细结构
如图1-5的SEM照片所示,观察到将CSH-NaDP组合物于80℃浸渍于1mol/L磷酸氢二钠水溶液中1天而制造的成型体呈长2~5μm、宽0.2~1μm、厚0.01~0.2μm的板状晶体致密地缠结的结构。由于在浸渍前的CSH-NaDP组合物中未观察到该结构,因此可认为其是OCP晶体。
[实施例2]
(从以烧石膏作为主要成分的具有连通多孔结构的组合物制造OCP连通多孔结构成型体)
(1)从前体陶瓷颗粒制造前体陶瓷连通多孔结构组合物
使用研钵和研杵将用与实施例1相同的方法制造的CSH-NaDP致密成型体(CSH-NaDP组合物)粉碎,进行颗粒化。使用分级筛对其进行筛分,使得粒径为200~300μm。将筛分后的颗粒装入φ6mm×3mm的组装模中。向其中滴加使CSH及NaDP饱和的70%乙醇0.2mL,使颗粒表面局部溶解。然后,于室温静置12小时以上使其干燥。由此,使颗粒彼此结合,制造了CSH-NaDP连通多孔结构组合物。
图2-1(A)中示出了制得的前体陶瓷组合物即CSH-NaDP连通多孔结构组合物的照片,(B)中示出了显微CT图像。
(2)利用组成转化反应的OCP连通多孔结构成型体的制造
将上述(1)中制造的CSH-NaDP连通多孔结构组合物于80℃浸渍于1mol/L磷酸氢二钠水溶液15mL中,使其反应1天(24小时)。
(3)OCP连通多孔结构成型体的表征
用与实施例1(3)同样的方法进行了表征。
(4)OCP连通多孔结构成型体的机械特性的测定
用与实施例1(4)同样的方法进行了测定。
(5)OCP连通多孔结构成型体的特性
图2-2(A)中示出了于80℃浸渍于1mol/L磷酸氢二钠水溶液中1天(24小时)而制造的成型体的照片,(B)中示出了显微CT图像。另外,图2-3中示出了XRD图谱。此外,图2-4(A)及(B)中示出了浸渍后的CSH-NaDP连通多孔结构组合物(OCP连通多孔结构成型体)的SEM照片,(C)及(D)中示出了浸渍前的CSH-NaDP连通多孔结构组合物的SEM照片。
如图2-2所示,在浸渍后也观察到维持了基本形态及多孔结构的情况。如图2-3所示,试样的XRD图谱表明浸渍后的连通多孔结构组合物呈OCP单一相。如图2-4所示,通过试样的SEM观察,在维持颗粒彼此连结的形状的同时,板状晶体形成为颗粒状。
[基于动物试验的本发明的成型体的有用性的确认]
在实验之前取得九州大学动物实验委员会的许可,然后进行了实验。(许可号A28-270-0)使用了体重为3.0~3.5kg的雄性日本大耳白家兔(Japanese white rabbit)(JapanSLC,Inc.)。臀部肌肉内注射氯胺酮(30mg/kg)和甲苯噻嗪(50mg/kg),镇静后,确保耳静脉,以氯胺酮(10mg/kg)和甲苯噻嗪(3mg/kg)的静脉麻醉进行维持。用消毒液对膝部进行消毒后,切开皮肤及骨膜,展开股骨头远位部。使用剥离子剥离骨膜后,在大腿骨内侧使用环钻杆(trephine bar)形成φ6.25mm×3mm的骨缺损。向其中埋入从CSH-NaDP组合物制造的OCP成型体(实施例1)、及从CSH-NaDP连通多孔结构组合物制造的OCP连通多孔结构成型体(实施例2)。另外,作为对照,同样地埋入羟基磷灰石烧结体。埋入试样的外径均为φ6mm×3mm。埋入后,缝合骨膜及皮肤,向手术野周边部注射2%利多卡因。
埋入后经过2星期、4星期后,通过过度麻醉进行安乐死,确认心跳停止、瞳孔反应消失后,将埋入物连同周围组织切除,用4%多聚甲醛(Paraformaldehyde)固定。固定后,将试样在10%EDTA水溶液、或急速脱钙水溶液(KC-X)中脱钙处理2周左右,然后水洗,使用阶段性的一系列乙醇脱水,石蜡包埋。取出股骨头的骨缺损部的中心,使用切片机沿试样的矢状切面制作约5μm的切片。对制作的切片进行苏木精-伊红(HE)染色,使用Keyence公司制的显微镜照相机(BZ-X710)以通常光进行拍摄。
基于利用HE染色的病理组织图像进行了研究。
图2-5中示出了埋入兔股骨头后经过2星期后的HE染色组织图像。(A)示出了实施例1中制造的OCP成型体,(B)示出了实施例2中制造的OCP连通多孔结构成型体,(C)示出了作为对照的羟基磷灰石烧结体。
如图2-5(A)及(B)所示,在埋入兔股骨头的骨缺损部的OCP成型体及OCP连通多孔结构成型体的周围观察到新生骨形成。OCP连通多孔结构成型体中,还观察到新生骨侵入多孔结构内部的一部分中的情况。此外,在OCP成型体、OCP连通多孔结构成型体中,均观察到骨将埋入试样的一部分进行置换的情况。在OCP成型体中,成型体周围的一部分被置换为骨,而在OCP连通多孔结构成型体中,观察到骨侵入至内部、将OCP连通多孔结构成型体的一部分进行置换的情况。
图2-6中示出了埋入兔股骨头后经过4星期后的HE染色组织图像。(A)为实施例1中制造的OCP成型体,(B)为实施例2中制造的OCP连通多孔结构成型体,(C)为作为对照的羟基磷灰石烧结体。
如图2-6所示,埋入4星期后的组织切片中,观察到较之2星期后而言,骨显著地置换OCP成型体及OCP连通多孔结构成型体的情况。OCP成型体中,成型体的3成左右被从外侧置换为新生骨。OCP连通多孔结构成型体中,除了上述的效果以外,还观察到骨侵入至OCP连通多孔结构成型体的中心部、即使在内部也活跃地发生骨置换的情况。
[实施例3]
(从DCPD组合物制造OCP成型体)
(1)前体DCPD组合物的制造
将从太平化学产业购买的α-TCP(Ca3PO4、α-TCP-B)0.1g埋入φ6mm×3mm的组装模中。向其中滴加2mol/L磷酸水溶液(H3PO4)200μL,静置3分钟,使其固化,制成作为前体陶瓷组合物之一的α-TCP/H3PO4组合物(前体DCPD组合物)。图3-1(A)中示出了制得的前体陶瓷组合物即DCPD组合物的照片,(B)中示出了XRD图谱。固化后的组合物显示出与DCPD对应的峰,将其定义为作为前体陶瓷组合物的DCPD组合物。
该DCPD组合物在H2O中的溶解度高于OCP的溶解度,另外,浸渍于蒸馏水时的周围的溶液的pH为6.7。DCPD在H2O中的溶解度为0.32g/L。
(2)利用组成转化反应的OCP成型体的制造
将上述(1)中制造的前体DCPD组合物于4℃、室温、37℃、70℃、80℃的恒温槽内浸渍于1mol/L磷酸氢二钠水溶液15mL,使其反应1天,制造OCP成型体。反应前的1mol/L磷酸氢二钠水溶液的pH为9.5。反应后的pH分别为9.0、8.7、8.3、7.7、7.4。该体系中,稳定相为磷灰石。
(3)OCP成型体的表征
OCP成型体的表征及机械特性的测定与实施例1的(3)、(4)同样地实施。
(4)针对OCP成型体的制造而言的温度依赖性
图3-2中示出了浸渍后的DCPD成型体(OCP成型体)的照片。
图3-3中示出了于4℃~80℃浸渍于1mol/L磷酸氢二钠水溶液中1天后的DCPD成型体(OCP成型体)的XRD图谱。
如图3-2所示,在浸渍于1mol/L磷酸氢二钠水溶液1天的情况下,在任意温度条件下均未观察到崩解的情况。
另外,如图3-3所示,在任意温度条件下浸渍后的成型体中,均观察到XRD图谱于4.7°附近处显示特征峰、OCP得以形成的情况。在4℃、室温、37℃的情况下,成型体均残留DCPD,成为DCPD和OCP的二相性的成型体。在70℃的情况下,成型体仅检测到OCP,成为OCP成型体。在80℃的情况下,成型体除了OCP以外还含有HAp。
(5)从前体陶瓷组合物制造OCP成型体与浸渍时间的关系
在相同的温度和溶液条件下,将浸渍时间设为2天、3天、7天,利用XRD进行了表征。其结果示于图3-4~图3-7。
首先,如图3-2所示,在全部的温度、时间条件下,前体DCPD成型体的基本形态得到了维持。如图3-4所示,在4℃及室温的情况下,即使浸渍7天,仍然观察到前体陶瓷组合物中的DCPD残留的情况。如图3-5所示,在37℃的情况下,针对3天的情况而言,DCPD残留,但在浸渍了7天的情况下,仅以痕迹程度残留。如图3-6所示,在70℃的情况下,浸渍2天、3天的情况下,得到了包含OCP单相的成型体。另外,浸渍7天后,得到了含有痕迹程度的HAp的OCP成型体。另一方面,如图3-7所示,可知在80℃的情况下,浸渍2天后,得到了基本由Hap形成的HAp-OCP的二相性的成型体,因此并未进行以上的研究。
(6)浸渍时间与机械强度的相关性
作为OCP成型体的机械强度的指标,使用与实施例1(4)同样的方法测定了径向拉伸强度(DTS)。使用下述体系进行了研究:在上述(5)的针对温度依赖性的研究中得到包含OCP单相的成型体的条件、即浸渍温度为70℃、浸渍时间为1天、2天、3天、7天的条件下浸渍于1mol/L磷酸氢二钠水溶液中。
如图3-8所示,浸渍于1mol/L磷酸氢二钠水溶液中1天的成型体的DTS强度为约2.12±0.23MPa。浸渍2天的成型体的DTS强度为约5.88±1.71MPa,较之浸渍1天的成型体的DTS强度而言显著增大。浸渍3天的成型体的DTS强度为约3.55±1.05MPa,浸渍7天的成型体的DTS强度为约3.71±1.47MPa,较之浸渍2天的成型体的DTS强度而言显著减少。
[实施例4]
(担载有在组成中包含羧基的分子的OCP成型体的制造)
(1)担载有在组成中包含羧基的分子的OCP成型体的制造
将使用与实施例3(1)同样的方法制造的前体DCPD组合物于70℃浸渍于含有0.2mol/L柠檬酸、琥珀酸、或酒石酸的1mol/L磷酸氢二钠水溶液中2天。
图4-1中示出了浸渍后的DCPD组合物(OCP成型体)的照片。图中,(A)为浸渍于含有柠檬酸的磷酸氢二钠水溶液后的成型体,(B)为浸渍于含有琥珀酸的磷酸氢二钠水溶液后的成型体,(C)为浸渍于含有酒石酸的磷酸氢二钠水溶液后的成型体。如图4-1所示,在任一种情况下,成型体的基本形态均在浸渍后也得到了维持。
(2)于含有在组成中包含羧基的分子的溶液中处理后的成型体的表征
利用通过与实施例1(3)同样的步骤测得的XRD图谱进行了表征。图4-2中示出了得到的XRD图谱。如图4-2所示,在分别含有0.2mol/L柠檬酸或琥珀酸的水溶液中进行处理而制造的OCP成型体的XRD图谱均显示成型体包含OCP单相。在含有0.2mol/L酒石酸的水溶液中进行处理而制造的OCP成型体的XRD图谱显示,成型体以OCP作为主要成分,并在其中含有DCPD。
(3)在组成中包含羧基的分子的担载量和担载形态
在能担载于上述OCP成型体的羧酸种类的研究中,针对得到了良好结果的柠檬酸、琥珀酸,对担载量及担载形态进行了研究。为了测定羧酸的浓度,在氩气气流中加热成型体,使用Yanaco Group制元素分析仪(MT-6),对成型体中的碳浓度进行碳-氢-氮分析(CHN分析)。由得到的成型体的碳含量、和尝试了担载的羧酸的分子量,测定针对OCP成型体的羧酸担载量。对于羧酸的担载形态,还并用了红外光谱法(FT-IR)。将试样固定于ATR棱镜上,在4,000-400cm-1的范围内以2cm-1分辨率利用FT-IR光谱仪(FT/IR-6300,日本分光株式会社)而得到。图4-3示出了得到的FT-IR光谱。
(4)OCP光谱和在组成中包含羧基的分子的担载形态的研究
作为OCP光谱数据的标准,使用了“Spectrochim Acta 23A:1781-1792,1967”的记载。
(5)在组成中包含羧基的分子对于OCP晶体结构的影响
如图4-3所示,在担载有0.2mol/L的柠檬酸的水溶液中制造的OCP成型体的IR光谱观察到1635cm-1附近的OCP晶体结构中的HPO4-OH层结构所导致的光谱带的形状显著地变化的情况。另外,在1300-1~400cm-1附近处观察到尝试了担载的柠檬酸及琥珀酸所导致的吸收带。由此表明,在HPO4-OH层结构中含有柠檬酸及琥珀酸。
(6)在组成中包含羧基的分子的担载量
根据元素分析的结果可知,在OCP成型体中含有大约2.6%的柠檬酸、1.1%的琥珀酸。
(7)担载有在组成中包含羧基的分子的OCP成型体的机械特性的测定
将DCPD成型体于70℃浸渍于分别以0.01mol/L、0.05mol/L、0.1mol/L、0.15mol/L、0.2mol/L的浓度含有柠檬酸及琥珀酸的1mol/L磷酸氢二钠水溶液中2天。另外,作为对照,同样地浸渍于不含有柠檬酸及琥珀酸的1mol/L磷酸氢二钠水溶液中。为了估计浸渍后的成型体的机械特性,使用与实施例1(4)同样的方法测定了DTS强度。
图4-4中示出了含有柠檬酸的OCP成型体的DTS强度,图4-5中示出了含有琥珀酸的OCP成型体的DTS强度。
[实施例5]
(在不含有PO4的溶液中的OCP成型体的制造)
将使用与实施例3(1)同样的方法制造的前体DCPD组合物在密封状态下于70℃浸渍于15mL的蒸馏水中1天。
图5-1中示出了浸渍后的DCPD成型体(OCP成型体)的照片,图5-2中示出了XRD图谱。
如图5-1所示,浸渍后的成型体维持了前体DCPD组合物的基本形态。另外,如图5-2所示,成为了含有OCP的成型体。
另外,将使用与实施例3(1)同样的方法制造的前体DCPD组合物在密封状态下于70℃浸渍于15mL的1mol/L硫酸钠水溶液中2天。
图5-3中示出了浸渍后的DCPD成型体(OCP成型体)的照片,图5-4中示出了XRD图谱。
如图5-3所示,浸渍后的成型体维持了前体DCPD组合物的基本形态。另外,如图5-4所示,成为了含有OCP的成型体。
[实施例6]
(在非电解质非水溶性溶液中的OCP成型体的制造)
将使用与实施例3(1)同样的方法制造的前体DCPD组合物在密封状态下于70℃浸渍于15mL的甲苯中2天。
图6-1中示出了浸渍后的DCPD成型体(OCP成型体)的照片,图6-2中示出了XRD图谱。
如图6-1所示,浸渍后的成型体维持了前体DCPD组合物的基本形态。另外,如图6-2所示,成为了含有OCP的成型体。
[实施例7]
(在非电解质水溶液溶液中的OCP成型体的制造)
将使用与实施例3(1)同样的方法制造的前体DCPD组合物在密封状态下于70℃浸渍于15mL的聚乙二醇(PEG400,和光纯药工业)、或聚丙烯酸钠(聚丙烯酸钠2,700~7,500,和光纯药工业)中2天。
图7-1(A)中示出了浸渍于聚乙二醇后的DCPD成型体(OCP成型体)的照片,(B)中示出了浸渍于聚丙烯酸钠后的DCPD成型体(OCP成型体)的照片。另外,图7-2中示出了XRD图谱。
如图7-1所示,浸渍后的OCP成型体维持了DCPD成型体的基本形态。另外,如图7-2所示,成为了含有OCP的成型体。
Claims (14)
1.体积为2.0mm3以上的包含70质量%以上的磷酸八钙的成型体的制造方法,其特征在于,将在组成中包含Ca及PO4中的至少一者、在H2O中的溶解度高于磷酸八钙、体积大于2.0mm3的前体陶瓷组合物浸渍于下述溶液中,使所述前体陶瓷组合物反应,将所述前体陶瓷组合物的至少一部分转化为磷酸八钙,所述溶液包含作为磷酸八钙的组成的Ca、PO4、H2O的组成之中未被包含在所述前体陶瓷组合物中的组成,并且所述溶液的pH大于7且为14以下。
2.如权利要求1所述的成型体的制造方法,其特征在于,前体陶瓷组合物在保持其基本形态的状态下被转化为包含磷酸八钙的成型体。
3.如权利要求1或2所述的成型体的制造方法,其特征在于,制得的包含磷酸八钙的成型体是含有90质量%以上的磷酸八钙的成型体。
4.如权利要求1所述的成型体的制造方法,其特征在于,在前体陶瓷组合物被组成转化为稳定相的物质的反应完成时间点之前,将成型体从溶液中取出。
5.如权利要求4所述的成型体的制造方法,其特征在于,反应完成时间点的溶液的pH为4以上,在高于反应完成时间点的pH的pH条件下终止反应。
6.如权利要求5所述的成型体的制造方法,其特征在于,反应完成时间点时的磷酸钙成分为磷灰石。
7.如权利要求4所述的成型体的制造方法,其特征在于,在成型体的Ca/PO4的比率低于反应完成时间点的成型体的Ca/PO4的比率的时间点终止反应。
8.如权利要求4所述的成型体的制造方法,其特征在于,在溶液的Ca/PO4的比率高于反应完成时间点的溶液的Ca/PO4的比率的时间点终止反应。
9.体积为2.0mm3以上的包含磷酸八钙的成型体的制造方法,其特征在于,将在组成中包含Ca及PO4中的至少一者、在H2O中的溶解度高于磷酸八钙、体积大于2.0mm3的前体陶瓷组合物浸渍于下述溶液中,使所述前体陶瓷组合物反应并在所述前体陶瓷组合物被组成转化为稳定相的物质的反应完成时间点之前从溶液中取出,从而将所述前体陶瓷组合物的至少一部分转化为磷酸八钙,所述溶液包含作为磷酸八钙的组成的Ca、PO4、H2O的组成之中未被包含在所述前体陶瓷组合物中的组成,
其中,反应完成时间点的pH低于4,在低于反应完成时间点的pH的pH条件下终止反应。
10.如权利要求9所述的成型体的制造方法,其特征在于,反应完成时间点时的磷酸钙成分为无水磷酸氢钙或磷酸氢钙二水合物。
11.如权利要求4、9或10所述的成型体的制造方法,其特征在于,在成型体的Ca/PO4的比率高于反应完成时间点的成型体的Ca/PO4的比率的时间点终止反应。
12.如权利要求4、9或10所述的成型体的制造方法,其特征在于,在溶液的Ca/PO4的比率低于反应完成时间点的溶液的Ca/PO4的比率的时间点终止反应。
13.如权利要求1所述的成型体的制造方法,其特征在于,通过使前体陶瓷组合物、及溶液中的至少一者含有在组成中包含与钙化学键合的官能团的物质,从而将在组成中包含与钙化学键合的官能团的物质担载于磷酸八钙晶体。
14.如权利要求13所述的成型体的制造方法,其特征在于,将其他物质担载于磷酸八钙晶体中。
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