Crustacean bifunctional proteins in mineralized exoskeleton – a model for biomimetic injectable bone substitutes

矿化外骨骼中的甲壳类双功能蛋白——仿生可注射骨替代品的模型

• 批准号: 538923079
• 负责人: Dr.-Ing. Benjamin Kruppke
• 金额: --
• 依托单位: Department of Life Sciences
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/538923079?language=en
• 关键词: Crustacean; bifunctional; proteins; mineralized; exoskeleton

With this resubmission of our project application, we would like to incorporate the suggestions that appeared at the initial review. We have completely revised the proposal and have highlighted in color the most significant improvements that primarily refer to the reviewers' advice. In particular, we have clarified the production of the recombinant proteins and their selection process. The use of proteins in chitin gels and their processing and distinction from porous sponge-like scaffolds were further clarified, as well as the perspective of the material for a future application in the context of bone regeneration. As suggested we have placed the latter point into perspective adding more emphasis on the basic research of mineralization influenced by crustacean proteins. 0This leads to an extensive revision and update of the state of the art and focus of the work plan. Complementary preliminary experiments performed at TUD and BGU are briefly explained and referred to. Thus, we believe, the risk is appropriate to the foreseeable gain in knowledge, as discussed in the newly added risk assessment section. We are still fascinated by the high mineralization potential of crustaceans during molting, which is superior to similar processes in mammals. We aim to make this potential accessible relying on the synergy of our materials science and biological expertise. The preliminary experiments, knowledge of in vitro and in vivo mineralization processes and our existing pipeline of protein candidates are enforcing the approach. To understand the mineralization processes occurring in crustaceans and under in vitro conditions, we will first perform binding analyses of the proteins to chitin-based gels. Subsequently, we will investigate the mineralization-mediating properties of the crustacean bifunctional proteins under defined conditions in a specially designed mineralization chamber. The osteoinductive potential of the resulting biomimetic material will be investigated using osteoblast cell cultures. The long-standing cooperation between BGU and TUD was established by Prof. Amir Sagi, Dr. Thomas Hanke, and Dr. Benjamin Kruppke based on a joint BMBF project in which we immobilized hemolymphatic proteins from crayfish on porous scaffolds. The added value of the interdisciplinary consortium was already evident here. In the future, bifunctional crayfish proteins found as part of the crayfish mineralized exoskeleton could be useful components of a chitin-containing biomimetic injectable bone substitute with rapid mineralization properties to support bone regeneration.

通过重新提交我们的项目申请，我们希望纳入初步审查时出现的建议。我们已经完全修改了提案，并以颜色突出显示了主要参考审查者建议的最重要的改进。特别是，我们已经阐明了重组蛋白的生产及其选择过程。进一步阐明了蛋白质在甲壳素凝胶中的使用及其加工和与多孔海绵状支架的区别，以及该材料在骨再生背景下未来应用的前景。因此，我们把后一点放在了透视图上，并更加重视甲壳动物蛋白质对成矿作用影响的基础研究。这导致对工作计划的最新技术和重点进行广泛的修订和更新。在TUD和BGU进行的补充初步实验进行了简要的解释和参考。因此，我们认为，风险与可预见的知识增长是适当的，正如新添加的风险评估部分所讨论的那样。我们仍然着迷于甲壳类动物在蜕皮过程中的高矿化潜力，这比哺乳动物的类似过程要优越上级。我们的目标是依靠我们的材料科学和生物专业知识的协同作用来实现这一潜力。初步实验，体外和体内矿化过程的知识以及我们现有的蛋白候选物管道正在实施该方法。为了了解发生在甲壳类动物和体外条件下的矿化过程，我们将首先进行结合分析的蛋白质几丁质为基础的凝胶。随后，我们将在一个专门设计的矿化室中，在限定的条件下研究甲壳动物双功能蛋白的矿化介导特性。将使用成骨细胞培养物研究所得仿生材料的骨诱导潜力。BGU和TUD之间的长期合作是由Amir Sagi教授、托马斯汉克博士和本杰明·克虏伯克博士基于BMBF联合项目建立的，在该项目中，我们将小龙虾的血淋巴蛋白固定在多孔支架上。跨学科联合会的附加值在这里已经很明显。在未来，作为小龙虾矿化外骨骼的一部分发现的双功能小龙虾蛋白可能是含有几丁质的仿生可注射骨替代物的有用组分，具有快速矿化特性，以支持骨再生。