Preparation and in vitro biocompatibility of hydroxyapatite/polyphenylene sulfide composites

doi:10.3969/j.issn.2095-4344.2014.52.015

Abstract

Abstract:

BACKGROUND: It is difficult to combine nano-hydroxyapatite with polyphenylene sulfide by traditional method, thus which is unable to take advantage of nanomaterials. There is still a lack of the research of biocompatibility of nano-hydroxyapatite/polyphenylene sulfide composites.

OBJECTIVE: To prepare the nano-hydroxyapatite/polyphenylene sulfide composites and to evaluate the in vitro biocompatibility.

METHODS: Liquid composite technology was applied to prepare the nano-hydroxyapatit/polyphenylene sulfide composites; infrared spectrometer and scanning electron microscope were applied to analyze the composite constituent structure. Contact angle test, hemolysis test and cytotoxicity test in vitro were conducted with contact angle meter, cony blood and L929 mouse fibroblast cells.
RESULTS AND CONCLUSION: The nano-hydroxyapatite/polyphenylene sulfide composites were intermixed evenly and closely; the composites owned good hydrophilic performance (sheet: 42.5°, powder: 16.1°). The hemolysis ratio of nano-hydroxyapatite/polyphenylene sulfide composites was less than 5% (1.66%), which showed good blood compatibility. L929 cells grew well in the leaching liquor of the composites. The cell counting kit-8 test showed the cytotoxicity in vitro belongs to non-toxic category (level 0-1). All these results prove that the nano-hydroxyapatite/polyphenylene sulfide composites have been prepared successfully and show excellent histocompatibility in vitro.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: nanoparticles, hydroxyapatites, materials testing

CLC Number:

R318

Fan Ke-xia, Ma Yuan2, Yu Si-xun, Kuang Yong-qin, Yang Li-bin, Xia Xun, Zhao You-guang, Yang Wei-zhong, Gu Jian-wen. Preparation and in vitro biocompatibility of hydroxyapatite/polyphenylene sulfide composites [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(52): 8443-8449.

Figures/Tables 3

2.1 羟基磷灰石/聚苯硫醚复合材料成品图1A，B分别为羟基磷灰石/聚苯硫醚复合材料片材及粉体；图1C为利用羟基磷灰石/聚苯硫醚复合材料片材制备的浸提液。 2.2 红外光谱分析羟基磷灰石/聚苯硫醚复合材料的分子结构由羟基磷灰石/聚苯硫醚复合粉的红外光谱分析图(图2)可见：聚苯硫醚的特征吸收峰分别为3 500，1 573，1 470，1 180，810 cm-1，其中特征峰1 573 cm-1和1 470 cm-1为苯环的骨架振动吸收峰，特征峰1 180 cm-1为芳环上的C-S键振动吸收峰，特征峰810 cm-1是芳环对位取代质子的面外弯曲振动吸收峰；羟基磷灰石的特征峰：OH-为 3 571 cm-1，H2O 为3 431 cm-1和1 634 cm-1，PO43-为 1 095，1 036，604 cm-1。以上结果证实了复合粉中羟基磷灰石和聚苯硫醚两相的共同存在。 2.3 扫描电子显微镜观察羟基磷灰石/聚苯硫醚复合材料的微观形貌羟基磷灰石/聚苯硫醚复合材料的扫描电镜图显示，纳米羟基磷灰石与聚苯硫醚形成均匀复合，有机相与无机相结合紧密，无明显相界面(图3)。 2.4 接触角测试比较羟基磷灰石/聚苯硫醚复合材料与聚苯硫醚亲水性聚苯硫醚粉体的接触角为81.25°(图4A)，而羟基磷灰石/聚苯硫醚复合材料粉体接触角为16.1° (图4B)。聚苯硫醚片材的接触角为83.5°(图4C)，羟基磷灰石/聚苯硫醚复合材料片材接触角为42.5°(图4D)。对比可知，纳米羟基磷灰石的加入明显改善了单纯聚苯硫醚材料的亲水性能。 2.5 羟基磷灰石/聚苯硫醚复合材料与聚苯硫醚的溶血率分析对比溶血实验结果显示：聚苯硫醚溶血率为6.43%，高于5%，显示聚苯硫醚具有溶血作用，血液相容性不佳；通过本方案复合纳米羟基磷灰石后，羟基磷灰石/聚苯硫醚复合材料溶血率为1.66%，小于5%，显示其血液相容性良好(表1)。 2.6 CCK-8法检测羟基磷灰石/聚苯硫醚复合材料细胞毒性由细胞生长曲线可知：实验组和阴性对照组的A值随着时间延长而同步增长，表明两组细胞增殖能力及趋势基本一致(图5)。按公式1计算每日细胞相对增殖率，见表2。统计学分析组间的细胞毒性差异(F=261.263)，实验组与阴性对照组差异无显著性意义(P=0.446)。根据6级毒性评分标准，可知羟基磷灰石/聚苯硫醚复合材料细胞毒性级别为0至1级反应，属合格范畴。 2.7 倒置显微镜观察L929细胞在材料浸提液中细胞形态变化 24 h后，实验组细胞完全贴壁，形态良好，呈梭形，胞质均匀、胞核较大，并可见圆形分裂细胞，分布均匀，细胞形态与阴性对照组无明显差别；阳性对照组细胞大多呈悬浮状，很少贴壁。48 h后，实验组及阴性对照组细胞数目明显增多，而阳性对照组细胞则大部分停止生长，成圆形，仅少数细胞贴壁，死亡细胞数明显增多。72 h后，实验组与阴性对照组细胞进一步增殖，排列密集，细胞呈梭形、三角形或不规则形；阳性对照组细胞数量明显减少，正常形态消失，核固缩伴溶解(图6)。依据细胞形态分析标准，判断羟基磷灰石/聚苯硫醚复合材料体外细胞毒性程度属无毒范畴。"

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