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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要慢性、顽固性伤口和溃疡对治疗糖尿病、肥胖者和老年人构成重大挑战患者。需要新的治疗方案来解决发病率上升的问题；需要有针对性的方法来重新启动正常的愈合级联。组织粘合剂是钉书钉和缝线的广泛使用的替代品。这些迅速与标准凝胶相比，固化聚合物凝胶应用于伤口时可减少疤痕、住院时间和感染缝合，同时消除了针和拆线的需要。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，但粘合强度很小，并且依赖于动物提取物，这会降低功效生物相容性和生物活性。旨在扩大糖尿病和老年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新型基因功能化重组蛋白治疗生物活性作为可配置药物输送装置的基础；从建造一个生物活性、生物相容性组织粘合剂，用于伤口顽固高风险患者的早期伤口护理。目前，还没有工程化的 ECM 蛋白组织粘合剂。作为基础设计，我将采用建立了针对多个位点特异性的基因组记录生物聚合物合成技术掺入两种非标准氨基酸 (nsAA)、粘膜粘附剂 L-二羟基苯丙氨酸 (L-DOPA) 和光交联降冰片烯氨基酸（NorAA），各自形成单独的上皮化诱导，重组人发角蛋白异二聚体亚基，分别为 K85 和 K35。天然角蛋白和 nsAA 角蛋白将通过缓慢的硫醇介导的丝组装或快速的现场降冰片烯组装成支架交联，并进行结构表征和细胞活力测定。 NorAA-DOPA-角蛋白支架预计可在几秒钟内快速固化，并呈现出显着增强的粘合强度，可与可用的皮肤粘合剂。将进行设计的粘合支架变体的体内表征对 C57BL/6J 糖尿病小鼠；例如愈合率、粘合强度、形态分析和组织病理学化验；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顽固性皮肤伤口的粘合剂将显着提高愈合率、止血并减少糖尿病小鼠的疤痕。