Programmable Keratinous Bio-adhesives for Recalcitrant Wound Recovery

用于顽固性伤口恢复的可编程角蛋白生物粘合剂

• 批准号: 10472487
• 负责人: Michael William Grome
• 金额: $ 6.98万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10472487
• 关键词: Address; Adhesives; Aging; American; Amino Acids; Amputation; Animals; Appearance; Binding; Biocompatible Materials; Biological Assay; Biopolymers; Cell Proliferation; Cell Survival; Cells; Chemical Engineering; Chemicals; Chronic; Cicatrix; Circular Dichroism; Codon Nucleotides; Collagen; Cosmetics; Cyanoacrylates; Cysteine; Data; Dermal; Devices; Diabetes Mellitus; Diabetic mouse; Dialysis procedure; Dopa; Drug Delivery Systems; Electron Microscopy; Engineering; Ensure; Eosine Yellowish; Epithelial; Excision; Extracellular Matrix; Extracellular Matrix Proteins; Fibrin Tissue Adhesive; Fibroblasts; Filament; Foot Ulcer; Foundations; Gel; Genes; Genetic; Glues; Hair; Hemorrhage; Hemostatic Agents; Hospitals; Human; Hydrogels; In Vitro; Individual; Infection; Injectable; Keratin; Levodopa; Lower Extremity; Mediating; Medical; Nature; Needles; Obesity; Organism; Organism Strains; Patients; Peptides; Pharmaceutical Preparations; Polymers; Production; Protocols documentation; Recombinant Proteins; Recombinants; Recovery; Recurrence; Research; Sense Codon; Severities; Site; Sterile coverings; Structural Models; Structure; Sulfhydryl Compounds; Surgical sutures; Technology; Testing; Therapeutic; Time; Tissue Adhesives; Transgenic Organisms; Translations; Treatment Effectiveness; Tyrosine; Ulcer; Variant; Visible Radiation; analog; antimicrobial; biomaterial compatibility; chronic wound; cost; cost estimate; crosslink; design; design-build-test; diabetic; diabetic patient; diabetic wound healing; disability; effective therapy; efficacy testing; emotional distress; expression vector; healing; high risk; in vivo; injured; innovation; keratinocyte; migration; mouse model; non-healing wounds; novel; obese patients; older patient; porcine model; programs; pyrrolysine; recombinant peptide; scaffold; skin wound; wound; wound care; wound closure; wound healing; wound treatment

PROJECT SUMMARY Chronic, recalcitrant wounds and ulcers pose significant challenges to treating diabetic, obese, and elderly patients. New treatment options are needed to address rising rates; requiring a targeted approach to re-initiate the normal healing cascade. Tissue adhesives are widely used alternatives to staples and sutures. These rapidly curing polymer gels, when applied to wounds, reduce scarring, hospital time, and infection compared to standard sutures, while eliminating the need for needles and suture removal. Unfortunately, these wound treatment options offer little bioactivity; unsuitable for treating chronic wounds. Extracellular matrix (ECM) dressings (e.g. keratin) are bioactive, but offer little adhesive strength and rely on animal extractions that reduce efficacy in biocompatibility and bioactivity. Aimed at broadening available treatment options for diabetic and aging patients, this research seeks to design, build, and test novel genetically functionalized recombinant proteins with innate therapeutic bioactivity as a foundation for configurable drug delivery devices; starting with the construction of a bioactive, biocompatible tissue adhesive for early wound care in patients at high risk of wound recalcitrance. Currently, there are no engineered ECM protein tissue adhesives. As a foundational design, I will employ established genomically recoded organism polymer synthesis technologies for multiple site-specific incorporations of two non-standard amino acids (nsAAs), muco-adhesive L-dihydroxyphenylalanine (L-DOPA) and photo-cross-linkable norbornene amino acid (NorAA), each into separate epithelialization-inducing, recombinant human hair keratin heterodimer subunits, K85 and K35, respectively. Native and nsAA-keratins will be assembled into scaffolds, either via slow thiol-mediated filament assembly or rapid, on-site norbornene crosslinking, and subjected to structural characterization and cell viability assays. NorAA-DOPA-keratin scaffolds are expected to rapidly cure in seconds and present significantly enhanced adhesive strength, comparable to available dermal adhesives. In vivo characterizations of designed adhesive scaffold variants will be performed on C57BL/6J diabetic mice; e.g. healing rates, adhesive strength, morphometric analyses, and histopathological assays; comparing results to currently available tissue adhesives. I hypothesize that applying these novel keratin adhesives to recalcitrant dermal wounds will significantly enhance healing rates, block bleeding, and reduce scarring in diabetic mice.

项目摘要 慢性，顽固的伤口和溃疡对治疗糖尿病，肥胖和老年人构成重大挑战 患者。需要新的治疗选择来解决上升的利率；需要针对性的方法重新启动 正常的治愈级联。组织粘合剂是钉书钉和缝合线的广泛使用的替代方法。这些迅速 与标准相比 缝合线，同时消除了对针和缝合线去除的需求。不幸的是，这些伤口治疗 选项几乎没有生物活性；不适合治疗慢性伤口。细胞外基质（ECM）敷料（例如 角蛋白）具有生物活性，但几乎没有粘合力，并依赖于降低功效的动物提取物 生物相容性和生物活性。旨在扩大糖尿病和衰老患者的可用治疗选择， 这项研究旨在设计，构建和测试具有先天性的遗传功能化重组蛋白 治疗性生物活性作为可构型药物输送设备的基础；从建造 生物活性，生物相容性的组织粘合剂，可用于早期伤口重新耐受风险的患者早期伤口护理。 目前，没有工程的ECM蛋白质组织粘合剂。作为基础设计，我将使用 已建立的基因组重建生物聚合物合成技术，用于多种位点特异性 两种非标准氨基酸（NSAA），粘粘性L-二羟基苯基丙氨酸（L-DOPA）的结合 以及可与光的链链氨基烯氨基酸（Noraa），每个氨基酸（NORAA），分别为单独的上皮化诱导 重组人发角质异二聚体亚基，K85和K35。本地和nsaa-keratins将 通过缓慢的硫醇介导的细丝组件或快速的现场悬烯烯烯烃组装成脚手架 交联，并经过结构表征和细胞活力测定。 Noraa-dopa-keratin脚手架 预计将在几秒钟内快速固化，并具有显着增强的粘合强度，可与 可用的皮肤粘合剂。将执行设计的粘合脚手架变体的体内特征 在C57BL/6J糖尿病小鼠上；例如愈合率，粘合强度，形态分析和组织病理学 测定；将结果与当前可用的组织粘合剂进行比较。我假设应用这些新型角蛋白 对顽固性皮肤伤口的粘合剂将显着提高愈合率，阻塞出血并减少 在糖尿病小鼠中疤痕。