Molecular Engineering of Cartilage PCM Mechanotransduction in Osteoarthritis Using Biomimetic Proteoglycans

使用仿生蛋白多糖进行骨关节炎软骨 PCM 机械转导的分子工程

• 批准号: 10344701
• 负责人: MICHELE S MARCOLONGO
• 金额: $ 31.17万
• 依托单位: VILLANOVA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-10 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10344701
• 关键词: 3-Dimensional; Adhesions; Affect; American; Anterior Cruciate Ligament; Arthritis; Attenuated; Binding; Biomimetics; Bovine Cartilage; Calcium Signaling; Cartilage; Cartilage Matrix; Catabolism; Cells; Cellular Metabolic Process; Chemicals; Chondrocytes; Collagen; Collagen Type VI; Degenerative polyarthritis; Deposition; Disease; Early treatment; Engineering; Enzymes; Event; Experimental Designs; Extracellular Matrix; Functional disorder; GAG Gene; Gene Expression; Health; Histology; Human; Hydrogels; Immunohistochemistry; In Situ; In Vitro; Individual; Inflammatory; Interleukin-1 beta; Intervention; Intra-Articular Injections; Knee; Knowledge; Life; Matrix Metalloproteinases; Mediating; Metabolic; Mission; Modulus; Molecular; Musculoskeletal Diseases; Nanostructures; Natural regeneration; Operative Surgical Procedures; Organism; Oryctolagus cuniculus; Pain; Physiological; Play; Property; Proteoglycan; Resistance; Role; Sepharose; Signal Transduction; Spectrum Analysis; Stimulus; Structure; Testing; Therapeutic Agents; Thick; Time; United States National Institutes of Health; Work; acrylic acid; aggrecan; aggrecanase; biglycan; biomechanical test; cartilage degradation; cartilage regeneration; chondroitin sulfate glycosaminoglycan; disability; in vivo; interest; mechanical properties; mechanical stimulus; mechanotransduction; minimally invasive; nanoarchitecture; perlecan; proteoglycan core protein; response; skeletal; three dimensional cell culture; tool

Abstract Regeneration of osteoarthritic cartilage has been a largely unmet biomedical challenge for the past fifty years. Numerous strategies are being employed to harness the synthetic power of cells to generate new extracellular matrix in the hope of reversing the pain and dysfunction associated with osteoarthritis (OA), in keeping with the mission of the NIH to seek fundamental knowledge about of living systems and the application of that knowledge to enhance health, lengthen life, and reduce illness and disability. Of particular interest is the emerging role of the pericellular matrix (PCM), the region immediately surrounding the chondrocyte, due to its demonstrated importance in mediating chondrocyte mechanotransduction in both healthy and OA cartilage. In OA, degeneration of the PCM is one leading event of disease initiation, contributing to disrupted chondrocyte mechanotransduction and irreversible cartilage degradation. Thus, if we can engineer the properties of the PCM, there is a potential for us to modulate chondrocyte mechanosensitive activities, and in turn, to promote cartilage regeneration and/or to attenuate osteoarthritic cartilage degeneration. Our biomimetic proteoglycans (BPGs) have the niche effect of engineering cartilage PCM. We chemically end-attached 7-8 chondroitin sulfate glycosaminoglycans (CS-GAGs) to a poly(acrylic acid) (PAA) core (Mw ~10 kDa), resulting in a biomimetic proteoglycan, BPG10, with a bottle-brush nanostructure mimicking the native aggrecan. When infiltrated into bovine cartilage explants in vitro or intra-articularly injected into rabbit knees in vivo, BPG10 was preferentially localized in the PCM. This localization led to a significant increase in the micromodulus of the PCM in vitro, and in turn, significantly enhanced chondrocyte intracellular calcium signaling activities. The role of BPG10 is also relevant to OA. When infiltrated into human OA cartilage, BPG10 was also localized in the PCM, and enhanced the local PCM modulus, indicating a potential for restoring degenerative PCM and rescuing disrupted chondrocyte mechanosensitive activities. Given that the synthetic PAA core is not susceptible to physiologic enzymes, as are natural proteoglycans, BPG10 could also be resistant to chondrocyte catabolism in vivo. Our central hypothesis is that biomimetic proteoglycans will molecularly engineer the PCM, increasing the micromodulus of the PCM through interactions with native PCM molecules, thus promoting chondrocyte mechanotransduction and attenuating OA-induced cartilage degeneration. To test this hypothesis, we will: (1) study the physical interactions between BPG10 and cartilage matrix biomolecules; (2) determine if BPG10 augments the neo-PCM of chondrocytes in 3D culture and the PCM of degrading cartilage explants, and thus, modulates chondrocyte mechanotransduction and metabolic activities and (3) test if intra-articular administration of BPG10 attenuates the progression of OA in rabbits in vivo. In these studies, individual CS-GAGs will be tested as a control to examine the role of BPG10's unique structure.

抽象的 在过去的五十年中，骨关节炎软骨的再生在很大程度上是未得到的生物医学挑战。 正在采用许多策略来利用细胞的合成能力来产生新的细胞外 基质希望扭转与骨关节炎（OA）相关的疼痛和功能障碍 NIH的使命是寻求有关生命系统的基本知识和知识的应用 为了增强健康，延长寿命并减少疾病和残疾。特别感兴趣的是新兴角色 周围的细胞基质（PCM），该区域立即围绕软骨细胞围绕它 在健康和OA软骨中介导软骨细胞机械转导的重要性。在OA， PCM的退化是疾病开始的一个主要事件，导致软骨细胞干扰 机械转导和不可逆的软骨降解。因此，如果我们可以设计PCM的属性， 我们有可能调节软骨细胞机械敏感活动，进而促进软骨 再生和/或减弱骨关节炎软骨变性。我们的仿生蛋白聚糖（BPGS） 具有工程软骨PCM的利基效应。我们化学连接的7-8个硫酸软骨素 糖胺聚糖（CS-GAG）到聚（丙烯酸）（PAA）核（MW〜10 kDa），导致仿生型 蛋白聚糖，BPG10，带有奶瓶纳米结构，模仿了本地脂肪。当渗入 BPG10在体外注入兔子膝盖的体外或关节内注射牛软骨外植体优先 位于PCM中。这种本地化导致PCM在体外的微单元显着增加，并且 反过来，软骨细胞内钙信号传导活性显着增强。 BPG10的作用也是 与OA有关。当浸入人OA软骨中时，BPG10也位于PCM中，并增强 当地的PCM模量，表明有可能恢复退行性PCM并营救中断 软骨细胞机械敏感活性。鉴于合成PAA核不容易受到生理的影响 酶和天然蛋白聚糖一样，BPG10在体内也可以抵抗软骨细胞分解代谢。我们的 中心假设是，仿生蛋白聚糖将分子设计PCM，从而增加 通过与天然PCM分子的相互作用，PCM的微单元，从而促进软骨细胞 机械转导和衰减OA诱导的软骨变性。为了检验这一假设，我们将：（1） 研究BPG10与软骨基质生物分子之间的物理相互作用； （2）确定BPG10是否 增强3D培养中软骨细胞的新PCM和降解软骨外植体的PCM，因此 调节软骨细胞机械转导和代谢活性，（3）测试关节内给药 BPG10的体内兔子中OA的进展减弱。在这些研究中，将测试单个CS-GAG 作为检查BPG10独特结构的作用的控制。