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Inactivation of Problematic Micro-organisms in Collagen Based Media by Pulsed Electric Field Treatment (PEF)

机译:脉冲电场处理(PEF)对基于胶原基培养基的问题微生物的灭活

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The potential of pulsed electric field (PEF) treatment as a novel/safe method of decontaminating collagen based biomat rices was investigated. Collagen gels (0.3% w/v) were seeded at a concentration of 10~6 CFU/ml with either Escherichia coli or Staphylococcus epidertnidis and then treated with 100 pulses at ~45 kV/cm, using a static PEF chamber. To investigate if the collagen gel had a "protective" effect on the bacteria, PEF inactivation was also investigated in phosphate buffered saline (PBS) and melted collagen gel (heat-treated, 10 min at 80°C, and as a result not set as a gel). When subjected to PEF, different levels of inactivation were observed with E. coli and S. epidermidis seeded in the different media. In all three media E. coli was more susceptible to PEF treatment than S. epidermidis. E. coli cell counts were reduced by ~3.3 logs in PBS, 2.6 logs in collagen gel and least inactivation (~1.5 log reduction) was observed in melted gel. S. epidermidis also showed higher inactivation levels in PBS compared to native collagen gel and melted collagen gel. Current results have established that PEF treatment can inactivate different types of bacteria seeded in collagen gels. However, complete inactivation was not achieved in any of the different types of suspending media, possibly due to the high seeding densities utilized. Previous results have established that PEF does not adversely affect the collagen structure. Future work is required to optimize microbial inactivation levels and to fully establish whether PEF treatment may offer a novel biomatrix decontamination method, which achieves sterility without damaging labile biological components.
机译:研究了脉冲电场(PEF)治疗作为一种新的/安全方法的脱落基于胶原基的生物培养率的潜力。用大肠杆菌或葡萄球菌的浓度为10〜6 CFU / ml的浓度为胶原凝胶(0.3%w / v),然后使用静止PEF室用100℃的〜45kV / cm的脉冲处理。为了研究胶原凝胶对细菌的“保护性”作用,在磷酸盐缓冲盐水(PBS)中还研究了PEF失活,并熔化胶原凝胶(在80℃下进行热处理,10分钟,并且不设定作为凝胶)。当受到PEF时,用在不同培养基中播种的大肠杆菌和S.表皮的大肠杆菌和S.表皮的不同水平。在所有三种培养基中,大肠杆菌更容易受到PEF治疗的影响,而不是S.表皮。大肠杆菌细胞计数通过〜3.3在PBS中减少〜3.3,2.6胶原凝胶的原木,在熔化的凝胶中观察到最少的灭活(〜1.5降低)。与天然胶原凝胶和熔化的胶原凝胶相比,S. Epidermidis还显示出PBS的更高的灭活水平。目前的结果已经确定PEF治疗可以在胶原凝胶中灭活不同类型的细菌。然而,在任何不同类型的悬浮媒体中没有实现完全失活,可能是由于所使用的高种子密度。以前的结果已经确定PEF不会对胶原蛋白结构产生不利影响。未来的工作是优化微生物灭活水平,并完全建立PEF治疗是否可以提供一种新的生物裂解净化方法,其在不损害不稳定生物成分的情况下实现无菌性。

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