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.

摘要