Mekontso Joëlle Aurelie, Farrukh Usama, Trujillo Sara, Del Campo Aránzazu
INM-Leibniz Institute for New Materials, Saarbrucken, Germany; Chemistry Department, Saarland University, 66123 Saarbrucken, Germany.
INM-Leibniz Institute for New Materials, Saarbrucken, Germany.
Biomater Adv. 2025 Apr;169:214182. doi: 10.1016/j.bioadv.2025.214182. Epub 2025 Jan 10.
Living Therapeutic Materials (LTMs) are a promising alternative to polymeric drug carriers for long term release of biotherapeutics. LTMs contain living drug biofactories that produce the drug using energy sources from the body fluids. To clarify their application potential, it is fundamental to adapt biocompatibility and cytotoxicity assays applied from non-living biomaterials and therapeutics to evaluate how LTMs interact with host cells. Here, we have established a first step in this direction, by developing a practical workflow to parallelize in vitro assessment of minimal safety and cytocompatibility properties of bacterial LTMs. It allows systematic monitoring and quantification of the dynamic evolution of the bacterial population (growth, metabolic activity) in parallel to quantify the response of different mammalian cells to LTM supernatants with regards to cytotoxicity and release of pro-inflammatory cytokines over a period of 7 days using a maximum of 10 samples. The protocol was tested with a Pluronic-based thin film containing ClearColi. The results show no cytotoxic effects of ClearColi containing hydrogels in three mammalian cell lines, and no induction of pro-inflammatory cytokines under the tested conditions. This workflow represents a first step in establishing a roadmap for the safety assessment of LTMs, and investigation of biocompatibility potential of future living therapeutic devices.
活体治疗材料（LTMs）是聚合物药物载体的一种有前景的替代物，可用于生物治疗药物的长期释放。LTMs包含活体药物生物工厂，这些工厂利用体液中的能量来源生产药物。为了阐明它们的应用潜力，将适用于非活体生物材料和治疗药物的生物相容性和细胞毒性测定方法进行调整，以评估LTMs与宿主细胞如何相互作用，这是至关重要的。在此，我们朝着这个方向迈出了第一步，开发了一种实用的工作流程，以并行方式在体外评估细菌LTMs的最低安全性和细胞相容性特性。它允许对细菌群体的动态演变（生长、代谢活性）进行系统监测和量化，并同时量化不同哺乳动物细胞对LTMs上清液的反应，包括在7天内的细胞毒性和促炎细胞因子的释放情况，最多使用10个样本进行测试。该方案用含有ClearColi的基于普朗尼克（Pluronic）的薄膜进行了测试。结果表明含有ClearColi的水凝胶在三种哺乳动物细胞系中没有细胞毒性作用，并且在测试条件下没有诱导促炎细胞因子。这种工作流程是为LTMs安全性评估建立路线图以及研究未来活体治疗装置生物相容性潜力的第一步。
