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界面活性脂肪酶固定化多孔聚己内酯的抗菌膜性能制备通过 LB 技术。

Antibiofilm properties of interfacially active lipase immobilized porous polycaprolactam prepared by LB technique.

机构信息

Department of Biotechnology, Indian Institute of Technology Madras, Chennai, India.

出版信息

PLoS One. 2014 May 5;9(5):e96152. doi: 10.1371/journal.pone.0096152. eCollection 2014.

Abstract

Porous biomaterial is the preferred implant due to the interconnectivity of the pores. Chances of infection due to biofilm are also high in these biomaterials because of the presence of pores. Although biofilm in implants contributes to 80% of human infections, there are no commercially available natural therapeutics against it. In the current study, glutaraldehyde cross linked lipase was transferred onto a activated porous polycaprolactam surface using Langmuir-Blodgett deposition technique, and its thermostability, slimicidal, antibacterial, biocompatibility and surface properties were studied. There was a 20% increase in the activity of the covalently crosslinked lipase when compared to its free form. This immobilized surface was thermostable and retained activity and stability until 100°C. There was a 2 and 7 times reduction in carbohydrate and 9 and 5 times reduction in biofilm protein of Staphylococcus aureus and Escherichia coli respectively on lipase immobilized polycaprolactam (LIP) when compared to uncoated polycaprolactam (UP). The number of live bacterial colonies on LIP was four times less than on UP. Lipase acted on the cell wall of the bacteria leading to its death, which was confirmed from AFM, fluorescence microscopic images and amount of lactate dehydrogenase released. LIP allowed proliferation of more than 90% of 3T3 cells indicating that it was biocompatible. The fact that LIP exhibits antimicrobial property at the air-water interface to hydrophobic as well as hydrophilic bacteria along with lack of cytotoxicity makes it an ideal biomaterial for biofilm prevention in implants.

摘要

多孔生物材料因其孔隙的连通性而成为首选植入物。由于孔隙的存在,这些生物材料也容易发生生物膜感染。尽管植入物中的生物膜导致了 80%的人类感染,但目前还没有针对它的商业上可用的天然疗法。在当前的研究中,戊二醛交联脂肪酶通过 Langmuir-Blodgett 沉积技术被转移到活化的多孔聚己内酯表面上,并研究了其热稳定性、抑菌、抗菌、生物相容性和表面性质。与自由形式相比,共价交联脂肪酶的活性增加了 20%。这种固定化表面具有热稳定性,在 100°C 下仍保持活性和稳定性。与未涂层的聚己内酯 (UP) 相比,固定化脂肪酶的聚己内酯 (LIP) 可分别减少金黄色葡萄球菌和大肠杆菌的碳水化合物和生物膜蛋白 2 倍和 7 倍,9 倍和 5 倍。LIP 上的活细菌菌落数量比 UP 少 4 倍。脂肪酶作用于细菌的细胞壁,导致其死亡,这从原子力显微镜、荧光显微镜图像和释放的乳酸脱氢酶数量得到证实。LIP 允许超过 90%的 3T3 细胞增殖,表明其具有生物相容性。LIP 在空气-水界面表现出抗菌特性,对疏水和亲水细菌均有效,同时没有细胞毒性,这使其成为植入物中生物膜预防的理想生物材料。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/355b/4010425/8a38a604764a/pone.0096152.g001.jpg

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