PDF(Pubmed)
Abstract：
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.
摘要：
急性复发性扁桃体炎是一种慢性，与生物膜相关的感染，对患者和医疗保健系统来说是一个巨大的负担。它通常用反复的抗生素治疗，这有助于抗菌素耐药性。研究生物膜是了解这种疾病的关键。使用3D生物打印的水凝胶进行体外建模是实现这一目标的有希望的方法。开发了一种新型的明胶-PEGDA荧光假单胞菌生物墨水，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。将生物打印的构建体在37°C下在溶生肉汤中培养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。成功制造了生物相容性3D打印的载有细菌的水凝胶构建体。使用光学荧光显微镜观察细菌生长。活/死细胞染色方案证明了细菌活力。在12天培养期后获得的结果显示,与0%对照相比,1%明胶浓度构建体中的细菌生长更高。这项研究表明，首次使用载有细菌的明胶-PEGDA水凝胶进行3D打印构建体的生物制造，该构建体设计用于模拟急性复发性扁桃体炎。启动一项与临床相关的离体扁桃体细菌的研究将是改善这种有影响力但尚未研究的疾病治疗的重要的下一步。
