Stable and Active bFGF-Polymer Conjugates for Wound Healing

用于伤口愈合的稳定且活性强的 bFGF-聚合物缀合物

基本信息

批准号：
8776294
负责人：
Heather D Maynard
金额：
$ 33.37万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-12-15 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8776294
关键词：
Acute Alkanesulfonates Biochemical Biochemistry Cell Line Cell Proliferation Cells Cellular Assay Chemistry Chronic Coupled Cutaneous Dermal Dermatology Detection Diabetic mouse Diabetic wound Dimerization Enzymes Fibroblast Growth Factor 2 Fibroblasts Goals Growth Factor Healed Health Heating Heparin Human In Vitro Infection Inflammation Methacrylates Migration Assay Modeling Molecular Molecular Weight Morbidity - disease rate Mus Outcomes Research Pain Pathogenesis Polyethylene Glycols Polymers Polysaccharides Process Proteins Receptor Activation Relative (related person)Research Risk Role Site Skin Stimulation of Cell Proliferation Testing Therapeutic Therapeutic Uses Trypsin Work Wound Healing cell growth cell motility cell type cytotoxicity dimer healing heparin receptor in vivo keratinocyte medical schools member migration multidisciplinary novel novel therapeutics polymerization pre-clinical preclinical efficacy prevent protector protein protein degradation receptor repaired skin disorder therapeutic protein

项目摘要

DESCRIPTION (provided by applicant): Acute and chronic wounds cause pain and suffering, in addition to significant morbidity and risk of infection, to a large number of people each year. As a result, there is considerable need for new therapeutics that induces and accelerate wound repair. Basic fibroblast growth factor (bFGF) stimulates the proliferation and migration of several cell types that have crucial roles in wound healing. However, the protein is rapidly degraded when delivered or upon storage, thus far preventing its therapeutic use. One objective of this research is to stabilize bFGF by covalently conjugating a synthetic polymer that mimics a natural protector of the protein, heparin. bFGF induces dimerization of its receptors and heparin aids in receptor activation. Thus, a second objective on this research is to produce dimeric polymer conjugates of bFGF in order to increase the mitogenic and migratory cellular activity of the protein relative to unmodified bFGF. The third objective is to test preclinical efficacy of the bFGF conjugates to heal wounds. Three specific aims are proposed to reach these objectives. The first aim is to evaluate the stability of bFGF-p (SS)-co-PEGMA polymer conjugates compared to controls and cytotoxicity of the polymers. It is hypothesized that bFGF- p (SS)-co-PEGMA will be more stable than bFGF-pPEGMA, bFGF-PEG and the unmodified protein, and the polymers will be nontoxic at therapeutically useful concentrations. To accomplish this, stability of the bFGF conjugate against trypsin, acidic pH, heat, stirring, and storage will be quantified by standard biochemical and cellular assays. The second aim is to determine in vitro bioactivity of bFGF-p (SS)-co-PEGMA-bFGF conjugates. It is hypothesized that dimeric bFGF p (SS)-co-PEGMA conjugates will be more active in human dermal fibroblast (HDF) and human dermal keratinocyte (HDK) cell proliferation and migration assays than the monomeric conjugates and bFGF alone. To investigate this, conjugate-induced stimulation of cell proliferation and migration of cells in vitro will be ascertained. Receptor activation will be verified. The third aim is to determine the ability of dimeric and monomeric bFGF sulfonated polymer conjugates to enhance wound healing. It is hypothesized that dimeric bFGF p(SS)-co-PEGMA conjugates will be more effective at healing normal and diabetic wounds in vitro and in vivo than the monomeric conjugate or bFGF alone. To test this, organotypic cultures and superficial wounding models in mice will be employed. One potential outcome of this research is to develop an active agent that promotes skin repair. The long-term goal of this research is to produce efficacious and stable therapeutics to treat acute and chronic wounds.

描述（由申请人提供）：除了明显的发病率和感染风险外，急性和慢性伤口每年都会导致疼痛和痛苦。结果，需要诱导和加速伤口修复的新疗法。碱性成纤维细胞生长因子（BFGF）刺激了在伤口愈合中具有至关重要作用的几种细胞类型的增殖和迁移。但是，该蛋白在递送或储存后迅速降解，迄今为止阻止其治疗用途。这项研究的一个目的是通过共价结合一种合成聚合物来稳定BFGF，该聚合物模仿了蛋白质肝素的天然保护因子。 BFGF诱导其受体的二聚化和肝素辅助受体激活。因此，这项研究的第二个目标是产生BFGF的二聚体聚合物偶联物，以增加蛋白质相对于未改性BFGF的有丝分裂和迁移性细胞活性。第三个目标是测试BFGF偶联物治愈伤口的临床前功效。提出了三个具体目标来达到这些目标。第一个目的是评估与聚合物的对照和细胞毒性相比，BFGF-P（SS）-Co-Pegma聚合物共轭物的稳定性。假设BFGF-P（SS）-Co-Pegma将比BFGF-PPEGMA，BFGF-PEG和未修饰的蛋白质更稳定，并且在治疗浓度下，聚合物将是无毒的。为此，通过标准的生化和细胞测定法对BFGF共轭物，酸性pH，热，搅拌和储存的稳定性将量化。第二个目的是确定BFGF-P（SS）-Co-Pegma-BFGF结合物的体外生物活性。假设二聚体BFGF P（SS）-Co-Pegma结合物将在人类皮肤成纤维细胞（HDF）和人类真皮角质形成细胞（HDK）细胞增殖和迁移分析中比单独单独的偶联物和BFGF更活跃。为了研究这一点，将确定结合诱导的细胞增殖刺激和体外细胞的迁移。受体激活将得到验证。第三个目的是确定二聚体和单体BFGF磺化聚合物结合物增强伤口愈合的能力。假设二聚体BFGF P（SS）-Co-Pegma结合物将比单独的单体结合物或BFGF更有效地在体外和体内愈合正常和糖尿病伤口。为了测试这一点，将采用小鼠的器官培养物和浅表损伤模型。这项研究的一个潜在结果是开发一种促进皮肤修复的活性剂。这项研究的长期目标是产生有效且稳定的治疗剂来治疗急性伤口。