Graduate Institute of Chemical Engineering, National Taipei University of Technology, 3, Zhongxiao E Rd, Taipei, 106, Taiwan.
Graduate Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, 3, Zhongxiao E Rd, Taipei, 106, Taiwan.
Tissue Eng Regen Med. 2022 Oct;19(5):987-1000. doi: 10.1007/s13770-022-00462-4. Epub 2022 Jun 1.
Although non-lytic filamentous bacteriophages have been made into biomaterial to guide tissue growth, they had limited ability to prevent bacterial infection. In this work a lytic bacteriophage was used to make an antibacterial biomaterial for neural tissue repair.
Lytic phages were chemically bound to the surface of a chitosan film through glutaraldehyde crosslinking. After the chemical reaction, the contact angle of the sample surface and the remaining lytic potential of the phages were measured. The numbers of bacteria on the samples were measured and examined under scanning electron microscopy. Transmission electron microscopy (TEM) was used to observe the phages and phage-infected bacteria. A neuroblast cell line was cultured on the samples to evaluate the sample's biocompatibility.
The phages conjugated to the chitosan film preserved their lytic potential and reduced 68% of bacterial growth on the sample surface at 120 min (p < 0.001). The phage-linked surface had a significantly higher contact angle than that of the control chitosan (p < 0.05). After 120 min a bacterial biofilm appeared on the control chitosan, while the phage-linked sample effectively prevented biofilm formation. The TEM images demonstrated that the phage attached and lysed the bacteria on the phage-linked sample at 120 min. The phage-linked sample significantly promoted the neuroblast cell attachment (p < 0.05) and proliferation (p < 0.01). The neuroblast on the phage-linked sample demonstrated more cell extensions after day 1.
The purified lytic phages were proven to be a highly bioactive nanomaterial. The phage-chitosan composite material not only promoted neural cell proliferation but also effectively prevent bacterial growth, a major cause of implant failure and removal.
虽然非溶菌丝状噬菌体已被制成生物材料来引导组织生长，但它们预防细菌感染的能力有限。在这项工作中，溶菌噬菌体被用于制造一种用于神经组织修复的抗菌生物材料。
通过戊二醛交联将溶菌噬菌体化学结合到壳聚糖膜的表面。化学反应后，测量样品表面的接触角和噬菌体的剩余溶菌潜力。测量样品上细菌的数量，并在扫描电子显微镜下检查。透射电子显微镜（TEM）用于观察噬菌体和噬菌体感染的细菌。在样品上培养神经母细胞瘤系以评估样品的生物相容性。
连接到壳聚糖膜上的噬菌体保留了它们的溶菌潜力，并在 120 分钟时减少了样品表面 68%的细菌生长（p<0.001）。噬菌体连接的表面比对照壳聚糖的接触角显著更高（p<0.05）。在 120 分钟时，对照壳聚糖上出现了细菌生物膜，而噬菌体连接的样品有效地阻止了生物膜的形成。TEM 图像表明，噬菌体在 120 分钟时附着并裂解了噬菌体连接的样品上的细菌。噬菌体连接的样品显著促进了神经母细胞瘤的附着（p<0.05）和增殖（p<0.01）。噬菌体连接的样品上的神经母细胞瘤在第 1 天后表现出更多的细胞延伸。
纯化的溶菌噬菌体被证明是一种高度生物活性的纳米材料。噬菌体-壳聚糖复合材料不仅促进了神经细胞的增殖，而且有效地防止了细菌的生长，这是植入物失效和去除的主要原因。
