Engineering the open porous nanofibrous microsphere integrated fibrillar hydrogel for the co-delivery of antibacterial and angiogenic agents aimed at the rapid diabetic wound repair

设计开放多孔纳米纤维微球集成纤维水凝胶，用于共同递送抗菌剂和血管生成剂，旨在快速修复糖尿病伤口

• 批准号: 10737115
• 负责人: Johnson Vitharikunnil John
• 金额: $ 25.62万
• 依托单位: TERASAKI INSTITUTE FOR BIOMEDICAL INNOVATION
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10737115
• 关键词: Acceleration; Affect; Amputation; Angiogenic Peptides; Anti-Bacterial Agents; Anti-Inflammatory Agents; Architecture; Bacterial Infections; Biocompatible Materials; Cell Migration Induction; Cells; Cessation of life; Chemistry; Cicatrix; Complication; Diabetic Foot Ulcer; Diabetic mouse; Disulfides; Engineering; Extracellular Matrix; Family suidae; Flow Cytometry; Gel; Gelatin; Glucose; Glycolates; Goals; Granulation Tissue; Growth Factor; Histology; Human; Hybrids; Hydrogels; Immune response; In Vitro; Individual; Infection Control; Infection prevention; Injectable; Matrix Metalloproteinases; Methacrylates; Microbial Biofilms; Microspheres; Modality; Modeling; Monitor; Mus; Nanofiber Scaffold; Natural regeneration; Nutrient; Oxidation-Reduction; Oxygen; Patients; Peptides; Photochemistry; Play; Porosity; Process; Production; Proliferating; Property; Protease Inhibitor; Quality of life; Role; Site; Skin; Splint Device; Sterile coverings; Sterility; Stimulus; Structure; Techniques; Technology; Testing; Therapeutic; Therapeutic Agents; Tissues; Type 2 diabetic; Vascular Endothelial Growth Factors; Vascularization; Wound Infection; Wound models; angiogenesis; antimicrobial; cathelicidin; cathelicidin antimicrobial peptide; cell motility; combinatorial; controlled release; cytotoxicity; design; diabetic patient; diabetic ulcer; diabetic wound healing; effective therapy; efficacy evaluation; healing; improved; in vivo; infection rate; innovation; invention; methicillin resistant Staphylococcus aureus; migration; mimetics; minimally invasive; nanofiber; neovascularization; novel; prevent; response; treatment strategy; wound; wound closure; wound dressing; wound environment; wound healing

PROJECT SUMMARY This proposal aims to engineer a multifactorial wound dressing composed of open porous nanofibrous microspheres (NMs) integrated with a stimuli-responsive fibrillar hydrogel (FHG) to release therapeutics to accelerate diabetic wound healing. The open porous structure in the proposed wound dressing can enhance cell migration and granulation tissue formation for rapid wound closure. Initially, the porous NMs will be fabricated from electrospun short nanofiber segments using our newly invented core-shell electrospray technique with bubble technology. Afterward, the peptide-tethered matrix metalloproteinases (MMP) responsive FHG will be integrated with porous NMs to develop an injectable hybrid gel followed by photocrosslinking. Previously, we demonstrated that porous nanofiber scaffolds show tremendous improvements in cell migration and cell/tissue integration. Inspired by the results, we will develop a new wound dressing platform that improves the wound healing process in several aspects; (i) The extracellular matrix (ECM) mimetic porous structure of the NMs can accelerate the cell migration during the healing. (ii) Engineering an ECM mimetic FHG with methacrylate gelatin (GelMA) and MMP responsive peptide linker can offer to maintain the moist conditions and sustained delivery of therapeutic agents locally. (iii) The engineered peptide conjugated NMs integrated FHG can prevent diabetic wound infection and tune the angiogenesis by sustained release of antibacterial and angiogenic peptides. To achieve these goals, we will incorporate antibacterial LL-37 mimic W379 peptide, and vascular endothelial growth factor (VEGF) mimetic QK peptide to the NMs integrated FHG. Specifically, we have identified that porous NMs properties can modulate cell migration through the porous microarchitecture and improve wound healing compared with nonporous NMs composed gel. The fibrillar hydrogel network will provide moist conditions in the wound similar to ECM and the controlled release of peptides in the wound milieu. We will evaluate and optimize material properties using the following characterization workflow: the porosity of NMs, the injectability of hybrid gel, in vitro cytotoxicity quantification, in vitro gel degradation with different MMP cell responses (survival, proliferation, and migration), in vitro antibacterial efficacy of W379 peptide, in vitro angiogenic properties of QK peptide, in vivo immune response (analysis by Flow cytometry), in vivo material degradation (analysis by histology), and in vivo tissue healing/regeneration (analysis by immunohistology). Afterward, we will focus on an infected diabetic mouse (db/db) splinted wound healing model to evaluate the efficacy of the dressing with and without peptides . We expect to see accelerated wound healing through the open porous NMs composed of the injectable hydrogel compared with commercially available Geistlich Derma gideTM composed of the injectable hydrogel. This project has the potential to develop a new class of biomaterial and create an inexpensive and effective treatment for diabetic wounds by increasing our understanding of how to inhibit biofilm formation and porosity-induced cell migration in diabetic wound closure.

项目概要 该提案旨在设计一种由开孔纳米纤维组成的多因素伤口敷料 微球（NM）与刺激响应纤维状水凝胶（FHG）集成，以释放治疗剂 加速糖尿病伤口愈合。所提出的伤口敷料中的开孔结构可以增强细胞 迁移和肉芽组织形成以快速伤口闭合。最初，将制造多孔纳米材料 使用我们新发明的核壳电喷雾技术从静电纺短纳米纤维段中提取 气泡技术。之后，肽系基质金属蛋白酶 (MMP) 响应的 FHG 将被 与多孔纳米材料集成，开发可注射的混合凝胶，然后进行光交联。此前，我们 证明多孔纳米纤维支架在细胞迁移和细胞/组织方面显示出巨大的改善 一体化。受研究结果的启发，我们将开发一种新的伤口敷料平台，以改善伤口 几个方面的愈合过程； (i) NM 的细胞外基质 (ECM) 模拟多孔结构可以 加速愈合过程中的细胞迁移。 (ii) 用甲基丙烯酸明胶设计 ECM 模拟 FHG (GelMA) 和 MMP 响应肽接头可以提供维持湿润条件和持续递送 局部治疗剂。 (iii) 集成 FHG 的工程肽缀合 NM 可以预防糖尿病 通过持续释放抗菌肽和血管生成肽来抑制伤口感染并调节血管生成。到 为了实现这些目标，我们将结合抗菌 LL-37 模拟 W379 肽和血管内皮细胞 生长因子 (VEGF) 模拟 QK 肽与 NM 集成 FHG。具体来说，我们已经确定了多孔 NMs 特性可以调节细胞通过多孔微结构的迁移并改善伤口愈合 与无孔纳米材料组成的凝胶相比。纤维状水凝胶网络将提供潮湿的条件 类似于 ECM 的伤口以及伤口环境中肽的受控释放。我们将评估并优化 使用以下表征工作流程测量材料属性：NM 的孔隙率、混合材料的可注射性 凝胶、体外细胞毒性定量、不同 MMP 细胞反应的体外凝胶降解（存活、 增殖和迁移）、W379 肽的体外抗菌功效、QK 的体外血管生成特性 肽、体内免疫反应（通过流式细胞术分析）、体内材料降解（通过流式细胞术分析） 组织学）和体内组织愈合/再生（免疫组织学分析）。之后， 我们将重点关注 感染糖尿病小鼠（db/db）夹板伤口愈合模型，以评估敷料的功效 不含肽 。我们期望通过由以下物质组成的开孔纳米材料加速伤口愈合 可注射水凝胶与市售的由可注射水凝胶组成的 Geistlich Derma gideTM 进行比较 水凝胶。该项目有潜力开发一类新型生物材料，并创造一种廉价且 通过增加我们对如何抑制生物膜形成的了解，有效治疗糖尿病伤口 糖尿病伤口闭合中孔隙度诱导的细胞迁移。