Jarman Evan, Burgess Jordan, Sharma Ayushi, Hayashigatani Kate, Singh Amar, Fox Paige
Division of Plastic & Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, United States of America.
Division of Plastic & Reconstructive Surgery, Veterans Affairs Palo Alto Health Care System, Palo Alto, California, United States of America.
PLoS One. 2024 May 3;19(5):e0303039. doi: 10.1371/journal.pone.0303039. eCollection 2024.
The complexity of chronic wounds creates difficulty in effective treatments, leading to prolonged care and significant morbidity. Additionally, these wounds are incredibly prone to bacterial biofilm development, further complicating treatment. The current standard treatment of colonized superficial wounds, debridement with intermittent systemic antibiotics, can lead to systemic side-effects and often fails to directly target the bacterial biofilm. Furthermore, standard of care dressings do not directly provide adequate antimicrobial properties. This study aims to assess the capacity of human-derived collagen hydrogel to provide sustained antibiotic release to disrupt bacterial biofilms and decrease bacterial load while maintaining host cell viability and scaffold integrity. Human collagen harvested from flexor tendons underwent processing to yield a gellable liquid, and subsequently was combined with varying concentrations of gentamicin (50-500 mg/L) or clindamycin (10-100 mg/L). The elution kinetics of antibiotics from the hydrogel were analyzed using liquid chromatography-mass spectrometry. The gel was used to topically treat Methicillin-resistant Staphylococcus aureus (MRSA) and Clostridium perfringens in established Kirby-Bauer and Crystal Violet models to assess the efficacy of bacterial inhibition. 2D mammalian cell monolayers were topically treated, and cell death was quantified to assess cytotoxicity. Bacteria-enhanced in vitro scratch assays were treated with antibiotic-embedded hydrogel and imaged over time to assess cell death and mobility. Collagen hydrogel embedded with antibiotics (cHG+abx) demonstrated sustained antibiotic release for up to 48 hours with successful inhibition of both MRSA and C. perfringens biofilms, while remaining bioactive up to 72 hours. Administration of cHG+abx with antibiotic concentrations up to 100X minimum inhibitory concentration was found to be non-toxic and facilitated mammalian cell migration in an in vitro scratch model. Collagen hydrogel is a promising pharmaceutical delivery vehicle that allows for safe, precise bacterial targeting for effective bacterial inhibition in a pro-regenerative scaffold.
慢性伤口的复杂性给有效治疗带来困难，导致护理时间延长和严重的发病率。此外，这些伤口极易形成细菌生物膜，使治疗更加复杂。目前对浅表性感染伤口的标准治疗方法是用间歇性全身抗生素进行清创，这可能会导致全身副作用，而且往往无法直接针对细菌生物膜。此外，标准护理敷料不能直接提供足够的抗菌性能。本研究旨在评估人源胶原蛋白水凝胶提供持续抗生素释放以破坏细菌生物膜并降低细菌载量，同时维持宿主细胞活力和支架完整性的能力。从屈肌腱采集的人胶原蛋白经过处理制成可凝胶化液体，随后与不同浓度的庆大霉素（50 - 500 mg/L）或克林霉素（10 - 100 mg/L）混合。使用液相色谱 - 质谱法分析抗生素从水凝胶中的洗脱动力学。该凝胶用于在既定的 Kirby - Bauer 和结晶紫模型中局部治疗耐甲氧西林金黄色葡萄球菌（MRSA）和产气荚膜梭菌，以评估细菌抑制效果。对二维哺乳动物细胞单层进行局部处理，并对细胞死亡进行定量以评估细胞毒性。用嵌入抗生素的水凝胶处理细菌增强的体外划痕试验，并随时间成像以评估细胞死亡和迁移情况。嵌入抗生素的胶原蛋白水凝胶（cHG + abx）显示抗生素可持续释放长达48小时，成功抑制了MRSA和产气荚膜梭菌生物膜，同时在长达72小时内保持生物活性。发现在体外划痕模型中，给予浓度高达最低抑菌浓度100倍的抗生素的cHG + abx无毒，并促进了哺乳动物细胞迁移。胶原蛋白水凝胶是一种有前景的药物递送载体，它能够在促进再生的支架中实现安全、精确的细菌靶向，以有效抑制细菌。
