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首页> 美国卫生研究院文献>Journal of Functional Biomaterials >Is Macroporosity Absolutely Required for Preliminary in Vitro Bone Biomaterial Study? A Comparison between Porous Materials and Flat Materials
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Is Macroporosity Absolutely Required for Preliminary in Vitro Bone Biomaterial Study? A Comparison between Porous Materials and Flat Materials

机译：初步体外生物材料研究是否绝对需要大孔？多孔材料与平板材料的比较

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Porous materials are highly preferred for bone tissue engineering due to space for blood vessel ingrowth, but this may introduce extra experimental variations because of the difficulty in precise control of porosity. In order to decide whether it is absolutely necessary to use porous materials in in vitro comparative osteogenesis study of materials with different chemistries, we carried out osteoinductivity study using C3H/10T1/2 cells, pluripotent mesenchymal stem cells (MSCs), on seven material types: hydroxyapatite (HA), α-tricalcium phosphate (α-TCP) and β-tricalcium phosphate (β-TCP) in both porous and dense forms and tissue culture plastic. For all materials under test, dense materials give higher alkaline phosphatase gene (Alp) expression compared with porous materials. In addition, the cell density effects on the 10T1/2 cells were assessed through alkaline phosphatase protein (ALP) enzymatic assay. The ALP expression was higher for higher initial cell plating density and this explains the greater osteoinductivity of dense materials compared with porous materials for in vitro study as porous materials would have higher surface area. On the other hand, the same trend of Alp mRNA level (HA > β-TCP > α-TCP) was observed for both porous and dense materials, validating the use of dense flat materials for comparative study of materials with different chemistries for more reliable comparison when well-defined porous materials are not available. The avoidance of porosity variation would probably facilitate more reproducible results. This study does not suggest porosity is not required for experiments related to bone regeneration application, but emphasizes that there is often a tradeoff between higher clinical relevance, and less variation in a less complex set up, which facilitates a statistically significant conclusion. Technically, we also show that the base of normalization for ALP activity may influence the conclusion and there may be ALP activity from serum, necessitating the inclusion of “no cell” control in ALP activity assay with materials. These explain the opposite conclusions drawn by different groups on the effect of porosity.
机译：由于用于血管向内生长的空间，多孔材料非常适合用于骨组织工程，但是由于难以精确控制孔隙率，因此这可能会带来额外的实验变化。为了确定在绝对不同化学性质的材料的体外比较成骨研究中是否绝对需要使用多孔材料，我们使用C3H / 10T1 / 2细胞，多能间充质干细胞（MSCs）对七种材料类型进行了骨诱导研究：多孔和致密形式的羟基磷灰石（HA），磷酸α-三钙（α-TCP）和β-磷酸三钙（β-TCP）和组织培养塑料。对于所有被测材料，与多孔材料相比，致密材料具有更高的碱性磷酸酶基因（Alp）表达。此外，通过碱性磷酸酶蛋白（ALP）酶法评估了细胞密度对10T1 / 2细胞的影响。对于较高的初始细胞铺板密度，ALP表达较高，这解释了与用于体外研究的多孔材料相比，致密材料的骨诱导性更高，因为多孔材料具有更高的表面积。另一方面，多孔和致密材料的Alp mRNA含量趋势相同（HA>β-TCP>α-TCP），验证了使用致密平板材料对具有不同化学性质的材料进行比较研究的可靠性，从而更可靠如果没有明确定义的多孔材料，则进行比较。避免孔隙率变化可能会促进结果的重现性。这项研究并不表明与骨再生应用相关的实验不需要孔隙度，而是强调在较高的临床相关性和较简单的结构中较少的变异之间通常需要权衡取舍，这有利于统计上有意义的结论。从技术上讲，我们还表明标准化ALP活性的基础可能会影响结论，并且血清中可能存在ALP活性，因此有必要在材料的ALP活性测定中包括“无细胞”对照。这些解释了不同小组对孔隙率影响得出的相反结论。

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