Wang Lulu, Sun Yan, Zhang Ruihao, Pan Kehou, Li Yuhang, Wang Ruibing, Zhang Lin, Zhou Chengxu, Li Jian, Li Yun, Zhu Baohua, Han Jichang
Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao, 266003, China.
College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo, 315200, China.
Biotechnol Biofuels Bioprod. 2023 Sep 14;16(1):136. doi: 10.1186/s13068-023-02389-x.
The silicified cell wall of diatoms, also known as frustule, shows huge potential as an outstanding bio-nanomaterial for hemostatic applications due to its high hemostatic efficiency, good biocompatibility, and ready availability. As the architectural features of the frustule determine its hemostatic performance, it is of great interest to develop an effective method to modify the frustule morphology into desired patterns to further improve hemostatic efficiency.
In this study, the gene encoding Silicalemma Associated Protein 2 (a silicalemma-spanning protein) of Cyclotella cryptica (CcSAP2) was identified as a key gene in frustule morphogenesis. Thus, it was overexpressed and knocked down, respectively. The frustule of the overexpress lines showed no obvious alteration in morphology compared to the wild type (WT), while the size, specific surface area (BET), pore volume, and pore diameter of the knockdown strains changed greatly. Particularly, the knockdown frustules achieved a more pronounced coagulation effect and in vivo hemostatic performance than the WT strains. Such observations suggested that silicalemma proteins are ideal genetic encoding targets for manipulating frustule morphology associated hemostatic properties. Furthermore, the Mantel test was adopted to identify the key morphologies associated with C. cryptica bleeding control. Finally, based on our results and recent advances, the mechanism of frustule morphogenesis was discussed.
This study explores a new strategy for enhancing the hemostatic efficiency of the frustule based on genetic morphology modification and may provide insights into a better understanding of the frustule morphogenesis mechanism.
硅藻的硅化细胞壁,也称为壳面,由于其高止血效率、良好的生物相容性和易于获取,作为一种出色的生物纳米材料在止血应用中显示出巨大潜力。由于壳面的结构特征决定其止血性能,因此开发一种有效的方法将壳面形态修饰成所需模式以进一步提高止血效率具有重要意义。
在本研究中,隐环藻(Cyclotella cryptica)编码硅质外层相关蛋白2(一种跨硅质外层的蛋白)的基因(CcSAP2)被确定为壳面形态发生的关键基因。因此,分别对其进行了过表达和敲除。与野生型(WT)相比,过表达系的壳面形态没有明显变化,而敲除菌株的尺寸、比表面积(BET)、孔体积和孔径发生了很大变化。特别是,敲除的壳面比WT菌株具有更显著的凝血效果和体内止血性能。这些观察结果表明,硅质外层蛋白是操纵与壳面形态相关的止血特性的理想基因编码靶点。此外,采用Mantel检验来确定与隐环藻出血控制相关的关键形态。最后,基于我们的结果和最新进展,讨论了壳面形态发生的机制。
本研究探索了一种基于基因形态修饰提高壳面止血效率的新策略,并可能为更好地理解壳面形态发生机制提供见解。