Denton Oliver, Wan Yifei, Beattie Laura, Jack Téa, McGoldrick Preston, McAllister Holly, Mullan Cara, Douglas Catriona M, Shu Wenmiao
Department of Biomedical Engineering, University of Strathclyde, Glasgow G1 1XQ, UK.
Department of Otolaryngology/ENT Surgery, NHS Greater Glasgow and Clyde, Glasgow G51 4TF, UK.
Bioengineering (Basel). 2024 Feb 21;11(3):202. doi: 10.3390/bioengineering11030202.
Acute recurrent tonsillitis is a chronic, biofilm-related infection that is a significant burden to patients and healthcare systems. It is often treated with repeated courses of antibiotics, which contributes to antimicrobial resistance. Studying biofilms is key to understanding this disease. In vitro modelling using 3D bioprinted hydrogels is a promising approach to achieve this. A novel gelatin-PEGDA pseudomonas fluorescens-laden bioink was developed and bioprinted in a 3D hydrogel construct fabricated using computer-aided design to mimic the tonsillar biofilm environment. The bioprinted constructs were cultured at 37 °C in lysogeny broth for 12 days. Bacterial growth was assessed by spectrophotometry. Cellular viability analysis was conducted using optical fluorescence microscopy (FDA/PI staining). A biocompatible 3D-printed bacteria-laden hydrogel construct was successfully fabricated. Bacterial growth was observed using optical fluorescence microscopy. A live/dead cellular-staining protocol demonstrated bacterial viability. Results obtained after the 12-day culture period showed higher bacterial growth in the 1% gelatin concentration construct compared to the 0% control. This study demonstrates the first use of a bacteria-laden gelatin-PEGDA hydrogel for biofabrication of a 3D-printed construct designed to model acute recurrent tonsillitis. Initiating a study with clinically relevant ex vivo tonsil bacteria will be an important next step in improving treatment of this impactful but understudied disease.
急性复发性扁桃体炎是一种与生物膜相关的慢性感染，给患者和医疗系统带来了沉重负担。它通常采用反复使用抗生素疗程进行治疗，这加剧了抗菌药物耐药性。研究生物膜是理解这种疾病的关键。使用3D生物打印水凝胶进行体外建模是实现这一目标的一种有前景的方法。一种新型的载有荧光假单胞菌的明胶 - 聚乙二醇二丙烯酸酯生物墨水被开发出来，并在使用计算机辅助设计制造的3D水凝胶构建体中进行生物打印，以模拟扁桃体生物膜环境。将生物打印构建体在37°C的溶菌肉汤中培养12天。通过分光光度法评估细菌生长。使用光学荧光显微镜（FDA/PI染色）进行细胞活力分析。成功制造出了一种生物相容性的3D打印载细菌水凝胶构建体。使用光学荧光显微镜观察到细菌生长。活/死细胞染色方案证明了细菌的活力。12天培养期后获得的结果显示，与0%的对照组相比，1%明胶浓度构建体中的细菌生长更高。本研究展示了首次使用载细菌的明胶 - 聚乙二醇二丙烯酸酯水凝胶对旨在模拟急性复发性扁桃体炎的3D打印构建体进行生物制造。启动一项使用临床相关的离体扁桃体细菌的研究将是改善这种有重大影响但研究不足的疾病治疗的重要下一步。
