Functional domains of bone sialoprotein in dentoalveolar development and healing

骨唾液蛋白在牙槽发育和愈合中的功能域

• 批准号: 10392346
• 负责人: Michael B. Chavez
• 金额: $ 4.67万
• 依托单位: OHIO STATE 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 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392346
• 关键词: Ablation; Adult; Affect; Age; Alveolar Bone Loss; Alzheimer' s Disease; Arginine; Aspartate; Atomic Force Microscopy; Binding; Biological; Biological Assay; Biology; Bone Diseases; Bone Regeneration; Bone remodeling; C-terminal; CRISPR/Cas technology; Caring; Cell Culture Techniques; Cell Line; Cells; Cementoblast; Collagen; Collagen Fiber; Collagen Fibril; Collagen Type I; Complex; Connective Tissue; Crystallization; Defect; Dental; Dental Cementum; Dental Implants; Deposition; Development; Diabetes Mellitus; Disease; Esthetics; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Family; Fibrillar Collagen; Fracture; Gel; Gene Expression; Genes; Genetic; Glycine; Glycoproteins; Goals; Health; Heart Diseases; Histology; Human body; Hydrophobicity; Hydroxyapatites; Implantation procedure; In Vitro; Individual; Injury; Integrin Binding; Integrins; Joints; Knock-in; Knock-out; Knowledge; Left; Ligand Binding; Link; Location; Mastication; Mechanics; Minerals; Modeling; Mus; Mutant Strains Mice; Mutation; N Domain; N-terminal; Natural regeneration; Odontogenesis; Oral health; Osteoblasts; Osteogenesis; Osteoporosis; Peptides; Periodontal Diseases; Periodontal Ligament; Phenotype; Physiologic calcification; Planet Earth; Play; Polyglutamic Acid; Population; Postmenopause; Premature Birth; Process; Proteins; Quality of life; Rattus; Role; Serum; Siblings; Signal Transduction; Skeletal Development; Skeleton; Speech; Structure; System; Testing; Therapeutic; Time; Tissues; Tooth Loss; Tooth structure; Transgenic Mice; Woman; alveolar bone; base; biomineralization; bone; bone fragility; bone healing; bone repair; bone sialoprotein; cost; craniofacial; efficacy outcomes; efficacy testing; fragility fracture; healing; human old age (65+); improved; in vivo; insight; long bone; member; microCT; mineralization; novel; novel therapeutics; outcome prediction; practical application; repaired; skeletal; substantia spongiosa; tissue regeneration; tissue repair

Abstract: Oral health is vital for overall health and quality of life, as exemplified by the importance of teeth in mastication, speech, and esthetics, and by recent connections made between oral health and diabetes, heart disease, preterm birth, and Alzheimer's disease. Periodontal disease, the breakdown of the connective tissues around the teeth, is one of the most prevalent diseases on earth, affecting 47% of adults and 70% of adults over the age of 65. The periodontal complex is a unique joint composed of two hard tissues, cementum and alveolar bone, and an intervening and unmineralized periodontal ligament (PDL). Periodontal disease leads to destruction of periodontal tissues and tooth loss if left untreated. Therapeutic approaches to regenerate or repair periodontal tissues are unpredictable at present, in part because of gaps in knowledge regarding molecules guiding dental and periodontal development. Our goal is to more successfully promote periodontal tissue repair, regeneration, and return to function. Factors directing cementum and alveolar bone mineralization are key for proper periodontal development and function, and likely play important roles in tissue repair. Bone sialoprotein (gene: Ibsp; protein: BSP) is an extracellular matrix protein highly expressed during cementum and alveolar bone formation. BSP has several putative biological roles based on its highly conserved functional domains involved in collagen binding (hydrophobic N-terminal domain), hydroxyapatite nucleation (polyglutamic acid sequences), and RGD-integrin cell signaling (C-terminal motif). BSP was demonstrated to be important in skeletal development, as genetic ablation in Ibsp knockout (Ibsp-/-) mice resulted in a skeletal phenotype marked by mildly delayed long bone mineralization and reduced trabecular bone remodeling. However, ablation of BSP causes even more dramatic effects in dentoalveolar tissues, where Ibsp-/- mice exhibited lack of cementum, severely hypomineralized alveolar bone, disrupted dental attachment, periodontal breakdown, and tooth loss. We hypothesize that BSP directs osteoblast function and mineralization activities and plays an important role in periodontal and alveolar bone repair. We will test this hypothesis in the following three aims: Aim 1: Define the binding location of BSP on type I collagen to define spatial mechanisms by which BSP may contribute to ECM mineralization. Aim 2: Elucidate the mechanistic roles of the RGD integrin-binding domain and the collagen- binding domain using newly generated cementoblast cell lines and mutant mice with a knock-in mutation inactivating the RGD motif. Aim 3: Investigate the efficacy of BSP to enhance alveolar bone healing using exogenous native rat BSP (nBSP) to investigate its use as a therapeutic in promoting alveolar bone repair. Importantly, insights gained will aid not only in regeneration of alveolar bone surrounding teeth or necessary for dental implant placement, but will also be potentially applicable towards healing critical size bone defects and fractures, and ameliorating or reversing systemic bone disorders such as osteoporosis.

抽象的： 口腔健康对于整体健康和生活质量至关重要，牙齿在咀嚼中的重要性就证明了这一点， 言语和美学，以及最近口腔健康与糖尿病、心脏病、 早产和阿尔茨海默病。牙周病，周围结缔组织的破坏 牙齿是地球上最常见的疾病之一，影响 47% 的成年人和 70% 的老年人 65.牙周复合体是由两种硬组织、牙骨质和牙槽骨组成的独特关节， 以及中间未矿化的牙周膜（PDL）。牙周病会导致牙周组织破坏 如果不及时治疗，牙周组织和牙齿会脱落。牙周再生或修复的治疗方法 目前，组织是不可预测的，部分原因是关于引导牙科的分子的知识存在差距。 和牙周发育。我们的目标是更成功地促进牙周组织修复、再生、 并返回功能。指导牙骨质和牙槽骨矿化的因素是适当的关键 牙周发育和功能，并可能在组织修复中发挥重要作用。骨唾液蛋白（基因： 磅；蛋白质：BSP）是一种在牙骨质和牙槽骨中高度表达的细胞外基质蛋白 形成。基于其高度保守的功能域，BSP 具有多种假定的生物学作用 胶原蛋白结合（疏水性 N 末端结构域）、羟基磷灰石成核（聚谷氨酸序列）、 和 RGD-整合素细胞信号传导（C 端基序）。 BSP 被证明对骨骼发育很重要 发育，因为 Ibsp 敲除 (Ibsp-/-) 小鼠的基因消除导致骨骼表型轻微 延迟长骨矿化并减少骨小梁重塑。然而，BSP 的消融会导致 对牙槽组织的影响甚至更为显着，其中 Ibsp-/- 小鼠表现出牙骨质缺乏，严重 牙槽骨矿化不足、牙齿附着破坏、牙周破坏和牙齿脱落。我们 假设 BSP 指导成骨细胞功能和矿化活动，并在 牙周和牙槽骨修复。我们将通过以下三个目标来检验这一假设： 目标 1：定义 BSP 在 I 型胶原蛋白上的结合位置，以确定 BSP 可能有助于 ECM 的空间机制 矿化。目标 2：阐明 RGD 整合素结合域和胶原蛋白的机制作用 使用新生成的成牙骨质细胞系和具有敲入突变的突变小鼠的结合域 使 RGD 基序失活。目标 3：利用 BSP 来研究 BSP 促进牙槽骨愈合的功效 外源性天然大鼠 BSP (nBSP)，以研究其作为促进牙槽骨修复的治疗剂的用途。 重要的是，获得的见解不仅有助于牙齿周围牙槽骨的再生，也有助于牙齿周围牙槽骨的再生。 牙种植体植入，但也可能适用于修复临界大小的骨缺损和 骨折，改善或逆转全身性骨骼疾病，例如骨质疏松症。