D-RADA16-RGD-Reinforced Nano-Hydroxyapatite/Polyamide 66 Ternary Biomaterial for Bone Formation
D-RADA16-RGD-Reinforced Nano-Hydroxyapatite/Polyamide 66 Ternary Biomaterial for Bone Formation
复制标题
D-RADA16-RGD增强纳米羟基磷灰石/聚酰胺66三元骨形成生物材料
DOI：
10.1007/s13770-018-0171-5
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发表时间：
2019-04-01
影响因子：
3.6
通讯作者：
Jiang, Dianming
中科院分区：
文献类型：
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作者：
Zhao, WeiKang;He, Bin;Jiang, Dianming
背景纳米羟基磷灰石/聚酰胺66（nHA/PA66）是一种广泛应用于骨缺损修复的复合材料。然而，这种材料的生物活性不足。相比之下，D-RADA16-RGD自组装肽（包含所有D-氨基酸的D-RADA16-RGD序列为Ac-RADARADARADARADARGDS-CONH2）在细胞培养和骨再生方面显示出令人钦佩的生物活性。在这里，我们描述了一种良好的生物材料（nHA/PA66/D-RADA16-RGD）的制造。方法采用蛋白酶K和圆二色光谱分别测试肽D-RADA16-RGD的稳定性和二级结构特性。使用扫描电子显微镜（SEM）和透射电子显微镜（TEM）来表征这些材料的表面。体外共聚焦激光扫描（CLS）、细胞计数试剂盒8测试（CCK-8）、茜素红S染色、细胞免疫荧光分析和Western blotting。并对它们的生物安全性和体内生物活性进行了评价。结果蛋白酶K和圆二色光谱表明nHA/PA66中的D-RADA16-RGD能够形成稳定的片状二级结构。 SEM和TEM显示D-RADA16-RGD材料宽度为7~33 nm，长度为130~600 nm，交织孔径为40~200 nm。 CLS 表明 nHA/PA66/D-RADA16-RGD 组的细胞与具有更多肌动蛋白丝的相邻细胞相关。 CCK-8分析表明nHA/PA66/D-RADA16-RGD具有良好的生物相容性。茜素红 S 染色和蛋白质印迹以及体内成骨的结果表明 nHA/PA66/D-RADA16-RGD 表现出更好的生物活性。结论本研究表明我们的 nHA/PA66/D-RADA16-RGD 复合材料表现出合理的机械性能、生物相容性和促进骨形成的生物活性。
BACKGROUNDNano-hydroxyapatite/polyamide 66 (nHA/PA66) is a composite used widely in the repair of bone defects. However, this material is insufficient bioactivity. In contrast, D-RADA16-RGD self-assembling peptide (D-RADA16-RGD sequence containing all D-amino acids is Ac-RADARADARADARADARGDS-CONH2) shows admirable bioactivity for both cell culture and bone regeneration. Here, we describe the fabrication of a favorable biomaterial material (nHA/PA66/D-RADA16-RGD).METHODSProteinase K and circular dichroism spectroscopy were employed to test the stability and secondary structural properties of peptide D-RADA16-RGD respectively. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to characterize the surface of these materials. Confocal laser scanning (CLS), cell counting kit-8 tests (CCK-8), alizarin red S staining, cell immunofluorescence analysis and Western blotting were involvedin vitro. Also biosafety and bioactivity of them have been evaluatedin vivo.RESULTSProteinase K and circular dichroism spectroscopy demonstrated that D-RADA16-RGD in nHA/PA66 was able to form stable-sheet secondary structure. SEM and TEM showed that the D-RADA16-RGD material was 7–33 nm in width and 130–600 nm in length, and the interwoven pore size ranged from 40 to 200 nm. CLS suggests that cells in nHA/PA66/D-RADA16-RGD group were linked to adjacent cells with more actin filaments. CCK-8 analysis showed that nHA/PA66/D-RADA16-RGD revealed good biocompatibility. The results of Alizarin-red S staining and Western blotting as well as vivo osteogenesis suggest nHA/PA66/D-RADA16-RGD exhibits better bioactivity.CONCLUSIONThis study demonstrates that our nHA/PA66/D-RADA16-RGD composite exhibits reasonable mechanical properties, biocompatibility and bioactivity with promotion of bone formation.