Bioactive Resources for Innovative Clinical Applications Research Unit and Department of Pharmacy Practice, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Phatumwan, Bangkok 10330, Thailand.
Sensor Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn University, Phatumwan, Bangkok 10330, Thailand.
J Wound Care. 2020 Apr 1;29(Sup4):S25-S35. doi: 10.12968/jowc.2020.29.Sup4.S25.
To investigate the potential of sericin extracted by different methods to inhibit biofilm formation (prevention) and disrupt already formed biofilm (treatment).
In this study, sericin was extracted by heat, acid, alkali and urea. bacteria were cultivated in the presence of various concentrations of sericin to evaluate antibiofilm formation using cell density assay (inhibition effect before biofilm formed). Conversely, various concentrations of sericin were added to a biofilm already formed by bacteria, and the viability of bacteria assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (disruption effects after biofilm formed). Structures of extracted sericin were evaluated using circular dichroism and Fourier-transform infrared spectrometer.
The urea-extracted sericin at all concentrations (12.5mg/ml, 25mg/ml, 50mg/ml and 100mg/ml) showed the highest potential antibiofilm activity in terms of both inhibition and disruption effects, compared with sericin extracted by heat, acid or alkali. The heat-extracted and acid-extracted sericin were found to reduce the biofilm formation dose-dependently, while the alkali-extracted sericin did not show either inhibition or disruption effect on the bacterial biofilm. The urea-extracted sericin also killed the bacteria residing within the biofilm, possibly due to its modified structure which may destabilise the bacterial cell wall, leading to membrane disintegration and, finally, cell death.
Our results demostrated the antibiofilm activity of sericin. This could form the basis of further research on the mechanism and application of sericin as a novel antibiofilm agent.
研究不同方法提取的丝胶抑制生物膜形成(预防)和破坏已形成生物膜(治疗)的潜力。
在这项研究中,通过热法、酸法、碱法和尿素法提取丝胶。在不同浓度的丝胶存在下培养细菌,通过细胞密度测定法评估抗生物膜形成(在生物膜形成之前的抑制效果)。相反,向已经形成的细菌生物膜中添加不同浓度的丝胶,并通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物(MTT)测定法评估细菌的活力(在生物膜形成之后的破坏效果)。使用圆二色性和傅里叶变换红外光谱仪评估提取丝胶的结构。
与通过热法、酸法或碱法提取的丝胶相比,在抑制和破坏效果方面,所有浓度(12.5mg/ml、25mg/ml、50mg/ml 和 100mg/ml)的尿素提取丝胶均显示出最高的潜在抗生物膜活性。热提取和酸提取的丝胶被发现能够剂量依赖性地减少生物膜形成,而碱提取的丝胶对细菌生物膜既没有抑制作用也没有破坏作用。尿素提取的丝胶也能杀死生物膜内的细菌,可能是由于其结构的改变,这可能会使细菌细胞壁不稳定,导致膜破裂,最终导致细胞死亡。
我们的结果表明了丝胶的抗生物膜活性。这可以为进一步研究丝胶作为新型抗生物膜剂的机制和应用奠定基础。