Peng Yinbo, Song Chenlu, Yang Chuanfeng, Guo Qige, Yao Min
Department of Burns and Plastic Surgery, Shanghai Ninth People's Hospital, Institute of Traumatic Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Department of Burns and Plastic Surgery, Shanghai Ninth People's Hospital, Institute of Traumatic Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China; Department of Dermatology, Wellman Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, Boston, MA, USA.
Int J Nanomedicine. 2017 Jan 4;12:295-304. doi: 10.2147/IJN.S122357. eCollection 2017.
Silver nanoparticles (AgNPs) are being widely applied as topical wound materials; however, accumulated deposition of silver in the liver, spleen, and other main organs may lead to organ damage and dysfunction. We report here that low molecular weight chitosan-coated silver nanoparticles (LMWC-AgNPs) are effective against methicillin-resistant (MRSA), have better biocompatibility, and have lower body absorption characteristics when compared with polyvinylpyrrolidone-coated silver nanoparticles (PVP-AgNPs) and silver nanoparticles without surface stabilizer (uncoated-AgNPs) in a dorsal MRSA wound infection mouse model. LMWC-AgNPs were synthesized by reducing silver nitrate with low molecular weight chitosan as a stabilizer and reducing agent, while PVP-AgNPs were synthesized using polyvinylpyrrolidone as a stabilizer and ethanol as a reducing agent. AgNPs with different surface stabilizers were identified by UV-visible absorption spectrometry, and particle size was determined by transmission electron microscopy. UV-visible absorption spectra of LMWC-AgNPs, PVP-AgNPs and uncoated-AgNPs were similar and their sizes were in the range of 10-30 nm. In vitro experiments showed that the three types of AgNPs had similar MRSA-killing effects, with obvious effect at 4 μg/mL and 100% effect at 8 μg/mL. Bacteriostatic annulus experiments also showed that all the three types of AgNPs had similar antibacterial inhibitory effect at 10 μg/mL. Cell counting kit-8 assay and Hoechst/propidium iodide (PI) staining showed that LMWC-AgNPs were significantly less toxic to human fibroblasts than PVP-AgNPs and uncoated-AgNPs. Treatment of mice with MRSA wound infection demonstrated that the three types of AgNPs effectively controlled MRSA wound infection and promoted wound healing. After continuous application for 14 days, LMWC-AgNPs-treated mice showed significantly reduced liver dysfunction as demonstrated by the reduced alanine aminotransferase and aspartate aminotransferase levels and liver deposition of silver, in comparison to mice treated with uncoated-AgNPs or PVP-AgNPs. Our results demonstrated that LMWC-AgNPs had good anti-MRSA effects, while harboring a better biocompatibility and lowering the body's absorption characteristics.
银纳米颗粒（AgNPs）作为局部伤口敷料正被广泛应用；然而，银在肝脏、脾脏和其他主要器官中的累积沉积可能会导致器官损伤和功能障碍。我们在此报告，在背部耐甲氧西林金黄色葡萄球菌（MRSA）伤口感染小鼠模型中，与聚乙烯吡咯烷酮包被的银纳米颗粒（PVP-AgNPs）和无表面稳定剂的银纳米颗粒（未包被-AgNPs）相比，低分子量壳聚糖包被的银纳米颗粒（LMWC-AgNPs）对MRSA有效，具有更好的生物相容性，且体内吸收特性更低。LMWC-AgNPs通过用低分子量壳聚糖作为稳定剂和还原剂还原硝酸银来合成，而PVP-AgNPs则使用聚乙烯吡咯烷酮作为稳定剂和乙醇作为还原剂来合成。通过紫外-可见吸收光谱法鉴定具有不同表面稳定剂的AgNPs，并通过透射电子显微镜测定粒径。LMWC-AgNPs、PVP-AgNPs和未包被-AgNPs的紫外-可见吸收光谱相似，其粒径在10-30nm范围内。体外实验表明，这三种类型的AgNPs具有相似的杀灭MRSA的效果，在4μg/mL时效果明显，在8μg/mL时达到100%的效果。抑菌环实验还表明，这三种类型的AgNPs在10μg/mL时具有相似的抗菌抑制作用。细胞计数试剂盒-8检测和Hoechst/碘化丙啶（PI）染色表明，LMWC-AgNPs对人成纤维细胞的毒性明显低于PVP-AgNPs和未包被-AgNPs。对患有MRSA伤口感染的小鼠进行治疗表明，这三种类型的AgNPs均能有效控制MRSA伤口感染并促进伤口愈合。连续应用14天后，与用未包被-AgNPs或PVP-AgNPs治疗的小鼠相比，用LMWC-AgNPs治疗的小鼠肝功能障碍明显减轻，表现为丙氨酸转氨酶和天冬氨酸转氨酶水平降低以及肝脏银沉积减少。我们的结果表明，LMWC-AgNPs具有良好的抗MRSA效果，同时具有更好的生物相容性并降低了体内吸收特性。
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