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卡拉胶固定化裂解大肠埃希噬菌体 vB_Eco2571-YU1 水凝胶的研制及其在伤口敷料中应用的研究。

Development of carrageenan-immobilized lytic coliphage vB_Eco2571-YU1 hydrogel for topical delivery of bacteriophages in wound dressing applications.

机构信息

School of Chemical Engineering, Yeungnam University, 280 Daehak-Ro, Gyeongsan, Gyeongbuk 38541, South Korea; Research Institute of Cell Culture, Yeungnam University, 280 Daehak-Ro, Gyeongsan, Gyeongbuk 38541, South Korea.

School of Chemical Engineering, Yeungnam University, 280 Daehak-Ro, Gyeongsan, Gyeongbuk 38541, South Korea; Research Institute of Cell Culture, Yeungnam University, 280 Daehak-Ro, Gyeongsan, Gyeongbuk 38541, South Korea.

出版信息

Int J Biol Macromol. 2024 Feb;259(Pt 2):129349. doi: 10.1016/j.ijbiomac.2024.129349. Epub 2024 Jan 12.

DOI:10.1016/j.ijbiomac.2024.129349
PMID:38219934
Abstract

Bacteriophages are employed as cost-effective and efficient antibacterial agents to counter the emergence of antibiotic-resistant bacteria and other host bacteria in phage therapy. The increasing incidence of skin wounds is a significant concern in clinical practice, especially considering the limitations of antibiotic therapy. Furthermore, the lack of an effective delivery system that preserves the stability of bacteriophages hampers their clinical implementation. In recent years, there has been a growing amount of research on bacteriophage applications in veterinary and biomedical sciences. In our study, lytic coliphage vB_Eco2571-YU1 was isolated against pathogenic Escherichia coli host bacteria, and hydrogel wound dressing materials were fabricated with marine polysaccharide carrageenan (carr-vB_Eco2571-YU1) for their antibacterial activity. Transmission electron microscopy (TEM) morphology identified it as a Myoviridae coliphage with an icosahedral head length and width of approximately 60 and 56.8 nm, respectively, and a tail length of 119.7 nm. The one-step growth curve of coliphage revealed a latent period of 10 min, a rise period of 15 min, and a burst size of 120 virions per cell. The bacteriolytic activity of unimmobilized coliphages was observed within 2 h; however, strain-specific phage resistance was acquired after 9 h. In contrast, carr-vB_Eco2571-YU1 showed a sharp decline in the growth of bacteria in the log phase after 2 h and did not allow for the acquisition of phage resistance by the E. coli strain. The stability of coliphage under different pH, temperature, osmolarity, detergents, and organic solvents was evaluated. We also studied the long-term storage of carr-vB_Eco2571-YU1 hydrogels at 4 °C and found that the titer value decreased during a time-dependent period of 28 days. These hydrogels were also found to be hemocompatible using a hemolysis assay. The addition of plasticizer (0.6 % (w/v)) to the carrageenan (2 % (w/v)) to prepare carr-vB_Eco2571-YU1 hydrogels showed a decrease in compressive strength with enhanced elasticity. This phage therapy using polymeric immobilization of bacteriophages is a promising next-generation wound dressing biomaterial alternative to conventional wound and skin care management.

摘要

噬菌体被用作具有成本效益和高效率的抗菌剂,以对抗噬菌体治疗中抗生素耐药菌和其他宿主菌的出现。皮肤伤口的发病率不断增加是临床实践中的一个重大问题,特别是考虑到抗生素治疗的局限性。此外,缺乏能够保持噬菌体稳定性的有效输送系统也阻碍了它们的临床应用。近年来,越来越多的研究关注噬菌体在兽医和生物医学科学中的应用。在我们的研究中,分离出了针对致病性大肠杆菌宿主菌的裂解性大肠杆菌噬菌体 vB_Eco2571-YU1,并制备了水凝胶伤口敷料材料,其中含有海洋多糖卡拉胶(carr-vB_Eco2571-YU1)以发挥其抗菌活性。透射电子显微镜(TEM)形态学鉴定其为肌尾噬菌体科噬菌体,具有二十面体头部长度和宽度分别约为 60 和 56.8nm,以及 119.7nm 的尾部长度。噬菌体的一步生长曲线显示潜伏期为 10min,上升期为 15min,每个细胞爆发大小为 120 个病毒粒子。未固定噬菌体的杀菌活性在 2h 内即可观察到;然而,在 9h 后获得了针对特定菌株的噬菌体抗性。相比之下,carr-vB_Eco2571-YU1 在 2h 后对数期细菌的生长急剧下降,并且大肠杆菌菌株无法获得噬菌体抗性。评估了噬菌体在不同 pH 值、温度、渗透压、清洁剂和有机溶剂下的稳定性。我们还研究了 carr-vB_Eco2571-YU1 水凝胶在 4°C 下的长期储存,发现在 28 天的时间依赖性期间,效价值下降。通过溶血试验发现这些水凝胶也具有血液相容性。向卡拉胶(2%(w/v))中添加增塑剂(0.6%(w/v))来制备 carr-vB_Eco2571-YU1 水凝胶,会降低抗压强度,同时提高弹性。这种使用噬菌体聚合物固定化的噬菌体治疗是一种很有前途的下一代伤口敷料生物材料,可替代传统的伤口和皮肤护理管理。

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