Department of Oral and Maxillofacial Surgery, Division of Regenerative Orofacial Medicine, University Hospital Hamburg-Eppendorf, 20251 Hamburg, Germany.
Department of Oral and Maxillofacial Surgery, Division of Regenerative Orofacial Medicine, University Hospital Hamburg-Eppendorf, 20251 Hamburg, Germany; Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, 20251 Hamburg, Germany.
Biomater Adv. 2022 Apr;135:212740. doi: 10.1016/j.bioadv.2022.212740. Epub 2022 Mar 17.
Oral wounds are among the most troublesome injuries which easily affect the patients' quality of life. To date, the development of functional antibacterial dressings for oral wound healing remains a challenge. In this regard, we investigated antibacterial silk protein-based membranes for the application as wound dressings in oral and maxillofacial surgery. The present study includes five variants of casted membranes, i.e., i) membranes-silver nanoparticles (CM-Ag), ii) membranes-gentamicin (CM-G), iii) membranes-control (without functionalization) (CM-C), iv) membranes-silk sericin control (CM-SSC), and v) membranes-silk fibroin/silk sericin (CM-SF/SS), and three variants of nonwovens, i.e., i) silver nanoparticles (NW-Ag), ii) gentamicin (NW-G), iii) control (without functionalization) (NW-C). The surface structure of the samples was visualized with scanning electron microscopy. In addition, antibacterial testing was accomplished using agar diffusion assay, colony forming unit (CFU) analysis, and qrt-PCR. Following antibacterial assays, biocompatibility was evaluated by cell proliferation assay (XTT), cytotoxicity assay (LDH), and live-dead assay on L929 mouse fibroblasts. Findings indicated significantly lower bacterial colony growth and DNA counts for CM-Ag with a reduction of bacterial counts by 3log levels (99.9% reduction) in CFU and qrt-PCR assay compared to untreated control membranes (CM-C and CM-SSC) and membranes functionalized with gentamicin (CM-G and NW-G) (p < 0.001). Similarly, NW-G yielded significantly lower DNA and colony growth counts compared to NW-Ag and NW-C (p < 0.001). In conclusion, CM-Ag represented 1log level better antibacterial activity compared to NW-G, whereas NW-G showed better cytocompatibility for L929 cells. As data suggest, these two membranes have the potential of application in the field of bacteria-free oral wound healing. However, provided that loading strategy and cytocompatibility are adjusted according to the antibacterial agents' characteristic and fabrication technique of the membranes.
口腔伤口是最麻烦的损伤之一,容易影响患者的生活质量。迄今为止,开发用于口腔伤口愈合的功能性抗菌敷料仍然是一个挑战。在这方面,我们研究了用于口腔颌面外科的抗菌丝蛋白基膜作为伤口敷料。本研究包括五种铸膜变体,即 i)载银纳米粒子的膜(CM-Ag)、ii)载庆大霉素的膜(CM-G)、iii)对照膜(无功能化)(CM-C)、iv)载丝胶对照的膜(CM-SSC)和 v)丝素/丝胶载丝素的膜(CM-SF/SS),以及三种无纺变体,即 i)载银纳米粒子的无纺(NW-Ag)、ii)载庆大霉素的无纺(NW-G)、iii)对照无纺(无功能化)(NW-C)。用扫描电子显微镜观察样品的表面结构。此外,采用琼脂扩散试验、集落形成单位(CFU)分析和 qrt-PCR 进行抗菌试验。在进行抗菌试验后,通过 XTT 细胞增殖试验、LDH 细胞毒性试验和 L929 小鼠成纤维细胞的死活试验评估生物相容性。结果表明,与未经处理的对照膜(CM-C 和 CM-SSC)和用庆大霉素功能化的膜(CM-G 和 NW-G)相比,CM-Ag 的细菌菌落生长和 DNA 计数明显降低,CFU 和 qrt-PCR 试验中细菌计数减少了 3 个对数级(减少 99.9%)(p<0.001)。同样,与 NW-Ag 和 NW-C 相比,NW-G 的 DNA 和菌落生长计数明显降低(p<0.001)。综上所述,CM-Ag 的抗菌活性比 NW-G 好 1 个对数级,而 NW-G 对 L929 细胞的细胞相容性更好。由于数据表明,这两种膜有可能应用于无细菌的口腔伤口愈合领域。然而,需要根据抗菌剂的特性和膜的制造技术调整载药策略和细胞相容性。