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: 10631878
• 负责人: Michael B. Chavez
• 金额: $ 4.76万
• 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 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10631878
• 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; Cephalic; Collagen; Collagen Fiber; Collagen Fibril; Collagen Type I; Complex; Connective Tissue; Defect; Dental; Dental Cementum; Dental Implants; Dentin; Deposition; Development; Diabetes Mellitus; Disease; Esthetics; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Family; Fibrillar Collagen; Fracture; Gel; Gene Expression; Genes; Genetic; Glycine; Glycoproteins; Goals; Growth; Health; Heart Diseases; Histology; Human body; Hydrophobicity; Hydroxyapatites; Implantation procedure; In Vitro; Individual; Injury; Integrin Binding; Integrins; Joints; Knock-in; Knock-out; Knockout Mice; Knowledge; Left; Ligand Binding; Link; Location; Mastication; Mechanics; Modeling; Mus; Mutant Strains Mice; Mutation; N-terminal; Natural regeneration; Odontogenesis; Oral health; Osteoblasts; Osteogenesis; Osteoporosis; Outcome; Peptides; Periodontal Diseases; Periodontal Ligament; Phenotype; Physiologic calcification; Planet Earth; Play; Polyglutamic Acid; Population; Postmenopause; Premature Birth; Process; Proliferating; Proteins; Quality of life; RGD (sequence); Rattus; Role; Serum; Signal Transduction; Skeletal Development; Skeleton; Speech; Structure; System; Testing; Therapeutic; Time; Tissues; Tooth Loss; Tooth structure; Transgenic Mice; Woman; alveolar bone; biomineralization; bone; bone fragility; bone healing; bone repair; bone sialoprotein; cost; craniofacial; efficacy testing; fragility fracture; healing; human old age (65+); improved; in vivo; insight; long bone; member; microCT; mineralization; novel; novel therapeutics; 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.

摘要： 口腔健康对整体健康和生活质量至关重要，牙齿在咀嚼中的重要性就是例证。 言语和美学，以及最近口腔健康与糖尿病、心脏病、 早产和阿尔茨海默氏症。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一，47%的成年人和70%的老年人受到影响。 65岁。牙周复合体是一个独特的关节，由牙骨质和牙槽骨两个硬组织组成， 以及中间的未矿化的牙周膜(PDL)。牙周病导致牙周炎的破坏 如果不治疗，牙周组织和牙齿会脱落。再生或修复牙周的治疗方法 组织目前是不可预测的，部分原因是关于引导牙齿的分子的知识空白。 和牙周发育。我们的目标是更成功地促进牙周组织的修复，再生， 并回到正常工作状态。引导牙骨质和牙槽骨矿化的因素是 牙周的发育和功能，在组织修复中可能起着重要的作用。骨涎蛋白(基因： Ibsp；Protein：BSP)是一种细胞外基质蛋白，在牙骨质和牙槽骨中高表达 队形。BSP基于其高度保守的功能结构域，具有几种假定的生物学作用 在胶原结合(疏水N-末端结构域)、羟基磷灰石成核(聚谷氨酸序列)中， 和RGD-整合素细胞信号(C-末端基序)。BSP在骨骼中被证明是重要的 发育，因为IBSP基因敲除(IBSP-/-)小鼠的遗传消融导致了轻微的 长骨矿化延迟，骨小梁重塑减少。然而，BSP的消融导致 在牙槽组织中，IBSP-/-小鼠表现出严重的牙骨质缺乏，这种影响甚至更显著 牙槽骨矿化不足，牙齿附着中断，牙周破裂，牙齿脱落。我们 假设BSP指导成骨细胞的功能和矿化活动，并在 牙周和牙槽骨修复。我们将在以下三个目标中检验这一假设：目标1：定义 BSP在I型胶原上的结合定位以确定BSP参与细胞外基质的空间机制 矿化作用。目的2：阐明RGD整合素结合域和胶原蛋白的作用机制。 利用新产生的成牙骨质细胞系和带有敲入突变的突变小鼠的结合结构域 使RGD基序失活。目的3：探讨补骨方促进牙槽骨愈合的疗效。 目的：探讨外源性天然大鼠BSP(NBSP)促进牙槽骨修复的治疗作用。 重要的是，所获得的洞察力不仅有助于牙齿周围牙槽骨的再生，也有助于 牙种植体植入，但也将潜在地适用于修复临界大小的骨缺损和 骨折，以及改善或逆转系统性骨骼疾病，如骨质疏松症。