Desai Krupansh, Sankaran Shrikrishnan, Del Campo Aránzazu, Trujillo Sara
INM - Leibniz Institute for New Materials, Campus D2 2, Saarbrücken, 66123, Germany.
Chemistry Department, Saarland University, Saarbrücken, 66123, Germany.
Mater Today Bio. 2024 Dec 30;30:101437. doi: 10.1016/j.mtbio.2024.101437. eCollection 2025 Feb.
Engineered living materials (ELMs), which usually comprise bacteria, fungi, or animal cells entrapped in polymeric matrices, offer limitless possibilities in fields like drug delivery or biosensing. Determining the conditions that sustain ELM performance while ensuring compatibility with ELM hosts is essential before testing them in vivo. This is critical to reduce animal experimentation and can be achieved through investigations. Currently, there are no standards that ensure ELM compatibility with host tissues. Towards this goal, we designed a 96-well plate-based screening method to streamline ELM growth across culture conditions and determine their compatibility potential . We showed proliferation of three bacterial species encapsulated in hydrogels over time and screened six different cell culture media. We fabricated ELMs in bilayer and monolayer formats and tracked bacterial leakage as a measure of ELM biocontainment. After screening, an appropriate medium was selected that sustained growth of an ELM, and it was used to study cytocompatibility . ELM cytotoxicity on murine fibroblasts and human monocytes was studied by adding ELM supernatants and measuring cell membrane integrity and live/dead staining, respectively, proving ELM cytocompatibility. Our work illustrates a simple setup to streamline the screening of compatible environmental conditions of ELMs with the host.
工程化活材料（ELMs）通常由包裹在聚合物基质中的细菌、真菌或动物细胞组成，在药物递送或生物传感等领域提供了无限可能。在体内测试ELMs之前，确定维持其性能同时确保与ELM宿主兼容的条件至关重要。这对于减少动物实验至关重要，并且可以通过研究来实现。目前，尚无确保ELM与宿主组织兼容性的标准。为了实现这一目标，我们设计了一种基于96孔板的筛选方法，以简化ELM在不同培养条件下的生长，并确定它们的兼容潜力。我们展示了随着时间的推移，包裹在水凝胶中的三种细菌的增殖情况，并筛选了六种不同的细胞培养基。我们以双层和单层形式制备了ELMs，并跟踪细菌泄漏情况作为ELM生物安全性的衡量指标。筛选后，选择了一种能维持ELM生长的合适培养基，并用于研究细胞相容性。通过添加ELM上清液并分别测量细胞膜完整性和活/死染色，研究了ELM对小鼠成纤维细胞和人单核细胞的细胞毒性，证明了ELM的细胞相容性。我们的工作展示了一种简单的设置，可简化对ELMs与宿主兼容环境条件的筛选。
