具有抑菌、纳米酶、抗炎、清除 ROS、生成氧和一氧化氮功能的水凝胶促进感染性糖尿病创面愈合。

Promoting the healing of infected diabetic wound by an anti-bacterial and nano-enzyme-containing hydrogel with inflammation-suppressing, ROS-scavenging, oxygen and nitric oxide-generating properties.

机构信息

MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou, 310027, China; Shanxi-Zheda Institute of Advanced Materials and Chemical Engineering, Taiyuan, 030000, China.

Department of Infectious Diseases, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, China.

出版信息

Biomaterials. 2022 Jul;286:121597. doi: 10.1016/j.biomaterials.2022.121597. Epub 2022 May 22.

DOI:10.1016/j.biomaterials.2022.121597

PMID:35688112

Abstract

The diabetic wound is easily to develop into a chronic wound because of the extremely serious and complex inflammatory microenvironment including biofilm formation, over-expressed reactive oxygen species (ROS), hypoxia and insufficiency of nitric oxide (NO) synthesis. In this work, a multifunctional hydrogel was designed and prepared by crosslinking hydrophilic poly(PEGMA-co-GMA-co-AAm) (PPGA) polymers with hyperbranched poly-L-lysine (HBPL)-modified manganese dioxide (MnO) nanozymes. Pravastatin sodium, which is supposed to participate in the synthesis of NO, was further loaded to obtain the HMP hydrogel. The capabilities of this hydrogel in scavenging different types of ROS, generating O, killing broad spectrum bacteria, and protecting cells against oxidative stress were confirmed in vitro. The transcriptome analysis revealed that HBPL inhibited bacterial quorum sensing (QS) system, downregulated virulent genes, and interfered bacterial metabolism. The HBPL-crosslinked hydrogels killed up to 94.1%-99.5% of methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli (E. coli) and Pseudomonas aeruginosa even at 10 CFU/mL. HBPL modification greatly increased the stability of MnO nanosheets in physiological environment. The MRSA-caused infection was effectively treated by the HBPL-crosslinked HMP hydrogel in vivo, and thereby the wound closure at inflammatory phase was promoted significantly. The treatment of HMP hydrogel reduced the ROS degree and relieved the inflammatory level significantly, accompanied by the decreased neutrophil infiltration and enhanced M2-type macrophage polarization in vivo. Significantly lower levels of inflammatory factors such as interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) and chemokines-1 (CXCL-1), and higher levels of anti-inflammatory cytokines such as IL-4 and IL-10 were also confirmed. Moreover, the HMP hydrogel could promote the secretion of transforming growth factor-β (TGF-β) and stimulate neovascularization, and deposition of collagen with a thicker skin and epithelium structure.

摘要

糖尿病创面极易发展成慢性创面，其复杂的炎症微环境包括生物膜形成、过量表达的活性氧（ROS）、缺氧和一氧化氮（NO）合成不足等。本工作通过将亲水性聚（PEGMA-co-GMA-co-AAm）（PPGA）聚合物与超支化聚-L-赖氨酸（HBPL）修饰的二氧化锰（MnO）纳米酶交联，设计并制备了一种多功能水凝胶。进一步负载具有参与 NO 合成作用的普伐他汀钠，得到 HMP 水凝胶。体外证实了该水凝胶清除不同类型 ROS、产生 O、广谱杀菌和保护细胞免受氧化应激的能力。转录组分析表明 HBPL 抑制了细菌群体感应（QS）系统，下调了毒力基因，并干扰了细菌代谢。HBPL 交联水凝胶在 10 CFU/mL 时对耐甲氧西林金黄色葡萄球菌（MRSA）、大肠杆菌（E. coli）和铜绿假单胞菌的杀灭率高达 94.1%-99.5%。HBPL 修饰极大地提高了 MnO 纳米片在生理环境中的稳定性。体内 HBPL 交联 HMP 水凝胶有效治疗 MRSA 感染，显著促进炎症期伤口闭合。HMP 水凝胶治疗显著降低了 ROS 程度，减轻了炎症水平，同时减少了中性粒细胞浸润，增强了体内 M2 型巨噬细胞极化。体内还证实了炎症因子如白细胞介素-1β（IL-1β）、IL-6、肿瘤坏死因子-α（TNF-α）和趋化因子-1（CXCL-1）的水平显著降低，抗炎细胞因子如 IL-4 和 IL-10 的水平显著升高。此外，HMP 水凝胶还可以促进转化生长因子-β（TGF-β）的分泌，刺激新血管形成，以及更厚的皮肤和上皮结构中胶原蛋白的沉积。