Kruppke Benjamin, Farack Jana, Weil Simy, Aflalo Eliahu David, Poláková Dagmar, Sagi Amir, Hanke Thomas
Max Bergmann Center of Biomaterials and Institute of Materials Science, Technische Universität Dresden, Dresden, Germany.
Department of Life Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
J Biomed Mater Res A. 2020 Mar;108(3):694-708. doi: 10.1002/jbm.a.36849. Epub 2019 Dec 8.
Crustacean chitin-hemocyanin-calcium mineral complexes were designed as bone biomimetics, with emphasis on their ability to bind or release calcium ions. Chitin scaffolds were prepared by dissolving chitin flakes in LiCl/dimethylacetamide, followed by gel formation and freeze-drying. Some of these scaffolds were modified by incorporation of CaCO . In some of the chitin-CaCO scaffolds, macroporosity was introduced by HCl treatment. Hemocyanin from the crayfish Cherax quadricarinatus was used to further modify the chitin scaffolds by dip coating. Cytocompatibility, cellular adherence and proliferation of human mesenchymal stem cells (hMSCs) were evaluated in terms of cell number as reflected in lactate dehydrogenase activity. The chitin, chitin-CaCO , and porous chitin-CaCO scaffolds were all found to facilitate cell attachment. Hemocyanin dip-coating of these scaffolds led to increased initial cell adhesion, enhanced proliferation, and osteogenic differentiation. Since the hemocyanin loading of the scaffolds was impaired by sterilization by gamma-irradiation (as required for biomedical applications), the hemocyanin loading was performed on previously sterilized scaffolds. All scaffolds facilitated osteogenic differentiation of osteoblasts, with the highest cell ALP-activity being found on hemocyanin-modified porous chitin-CaCO scaffolds. Thus, chitin-hemocyanin scaffolds enhanced the initial stages of bone cell development and could serve as promising biomaterials for bone regeneration.
甲壳类几丁质-血蓝蛋白-钙矿物复合物被设计为骨仿生材料，重点在于它们结合或释放钙离子的能力。几丁质支架通过将几丁质薄片溶解在LiCl/二甲基乙酰胺中制备，随后形成凝胶并冻干。其中一些支架通过掺入CaCO₃进行改性。在一些几丁质-CaCO₃支架中，通过HCl处理引入大孔隙率。来自四脊滑螯虾的血蓝蛋白用于通过浸涂进一步修饰几丁质支架。根据乳酸脱氢酶活性反映的细胞数量评估人间充质干细胞（hMSCs）的细胞相容性、细胞黏附和增殖。发现几丁质、几丁质-CaCO₃和多孔几丁质-CaCO₃支架均有助于细胞附着。这些支架的血蓝蛋白浸涂导致初始细胞黏附增加、增殖增强和成骨分化。由于支架的血蓝蛋白负载因γ射线辐照灭菌（生物医学应用所需）而受损，因此血蓝蛋白负载是在先前灭菌的支架上进行的。所有支架均促进成骨细胞的成骨分化，在血蓝蛋白改性的多孔几丁质-CaCO₃支架上发现最高的细胞碱性磷酸酶活性。因此，几丁质-血蓝蛋白支架增强了骨细胞发育的初始阶段，可作为有前途的骨再生生物材料。
