Self Assembly of Cartilage by Mesenchymal Stem Cells on Porous Chitosan/CaP

间充质干细胞在多孔壳聚糖/CaP上自组装软骨

• 批准号: 7980852
• 负责人: Steven Howard Elder
• 金额: $ 34.68万
• 依托单位: MISSISSIPPI STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7980852
• 关键词: Address; Affect; Agaricales; Allogenic; American; Animals; Biochemical; Biological Assay; Biomechanics; Bioreactors; Bone Marrow; Caliber; Cartilage; Cell-Matrix Junction; Cells; Centrifugation; Ceramics; Chitosan; Chondrocytes; Chondrogenesis; Custom; Defect; Degenerative Disorder; Degenerative polyarthritis; Dimensions; Engineering; Euthanasia; Extracellular Matrix; Face; Femur; Fibronectins; Flushing; Friction; Glass; Growth; Growth Factor; Head; Human; Hyaline Cartilage; Image; In Vitro; Joints; Life; Marrow; Measures; Mechanics; Mesenchymal Stem Cells; Metals; Methods; Microscopic; Microspheres; Modeling; Molds; New Zealand; Oryctolagus cuniculus; Osteoarthrosis Deformans; Perfusion; Permeability; Plastics; Porosity; Precipitation; Process; Productivity; Property; Public Health; Replacement Arthroplasty; Research; Scanning Electron Microscopy; Seeds; Sepharose; Shapes; Shear Strength; Slide; Staging; Staining method; Stains; Surface; System; Techniques; Testing; Thick; Tissue Engineering; Tissues; Transplantation; Wages; alternative treatment; articular cartilage; base; bone; bone cell; calcium phosphate; cartilage repair; cell growth; density; interfacial; osteochondral tissue; public health relevance; repaired; research study; scaffold; self assembly; therapy development

DESCRIPTION (provided by applicant): The objective of this research is to begin developing a method for total joint resurfacing with engineered cartilage. The approach is based on generating a biphasic construct of porous chitosan/calcium phosphate and hyaline cartilage, the cartilage having been produced by mesenchymal stem cells (MSCs) attached to and piled atop the scaffold. This proposal encompasses four aims: 1) Investigate the effects of cell seeding density and growth factor on the biochemical and mechanical properties of cartilage produced from human marrow-derived MSCs by the self-assembling process; 2) Optimize integration and attachment of engineered cartilage to a porous chitosan-nanocrystalline calcium phosphate (chitosan/CaP) scaffold; 3) Engineer a femoral head-sized hemispherical shell of cartilage attached to a mushroom-shaped chitosan/CaP scaffold; 4) Use biphasic cartilage/CaP-chitosan constructs derived from allogenic bone marrow MSCs to repair critical size defects surgically created in the trochlear groove of New Zealand White rabbit femora. Specific Aim 1 deals just with enhancing chondrogenesis by MSCs in a scaffold-free model. Since MSCs are a scarce commodity it is important to understand how many will be needed for optimal chondrogenesis. We will also assess the effects of TGF-21, TGF-23, and BMP-13, alone and in combination, for accelerated chondroinduction and rapid accumulation of extracellular matrix. Specific Aim 2 is focused on bonding of the engineered cartilage to a rigid, porous, osteoconductive scaffold to which native bone could integrate to anchor the engineered cartilage. We will determine how scaffold microstructure and coating with fibronectin affect chondrogenesis and the cartilage-scaffold interfacial strength. Engineered cartilage produced as part of Specific Aims 1 and 2 will be evaluated based on histological, biochemical, and biomechanical properties. Specific Aim 3 adds the complexity of molding the engineered cartilage and supporting scaffold into a hemispherical shape as would be required for femoral head resurfacing. We will attempt to produce a uniform thickness layer of cartilage in a hemispherical shell attached to the porous chitosan/CaP scaffold. The thickness, sphericity, and coefficient of friction of this cartilage will be measured to assess its suitability for joint resurfacing. If successful, Specific Aim 4 will provide a proof-of-concept by restoring hyaline cartilage to a full-thickness defect in a living animal. This project will impact public health by demonstrating a feasible approach to joint resurfacing with engineered cartilage. This proposal targets a specific challenge, femoral head resurfacing, and represents the first steps necessary to overcome this challenge. PUBLIC HEALTH RELEVANCE: This project addresses important issues in the development of treatment alternatives for osteoarthritis, a degenerative disease that affects millions of Americans and is responsible for billions of dollars annually in lost wages and productivity. Cartilage tissue engineering has the potential for resurfacing an osteoarthritic joint with healthy, functional tissue so that joint replacement with metal, plastic, or ceramic components is unnecessary. This project develops a joint resurfacing strategy based on a biphasic construct of self-assembled cartilage attached to a porous calcium phosphate-chitosan composite scaffold.

描述（由申请人提供）：这项研究的目的是开始开发一种与工程软骨进行全面关节重铺的方法。该方法基于生成多孔壳聚糖/钙和透明软骨的双相构建体，该软骨是由与附着在支架上并堆积在脚手架上的间充质干细胞（MSC）产生的。该建议包括四个目的：1）研究细胞播种密度和生长因子对通过自组装过程从人类衍生的MSC产生的软化和机械特性的影响； 2）优化工程软骨在多孔壳聚糖纳米晶钙磷酸钙（壳聚糖/CAP）支架上的整合和附着； 3）工程师在蘑菇形壳聚糖/帽子支架上连接的股头大小软骨半球形壳； 4）使用源自同源性骨髓MSC的双相软骨/帽壳构建体来修复新西兰白兔股骨的Trochlear凹槽中产生的临界尺寸缺陷。特定的目标1仅在无脚手架模型中增强MSC的软骨生成。由于MSC是一种稀缺的商品，因此重要的是要了解最佳软骨形成需要多少。我们还将单独和组合TGF-21，TGF-23和BMP-13的效果，以加速软骨诱导和细胞外基质的快速积累。特定目标2的重点是将工程软骨粘合到刚性，多孔，破骨的支架上，而天然骨可以整合到该脚手架上以锚定工程软骨。我们将确定支架微观结构和纤维蛋白涂层如何影响软骨生成和软骨型界面强度。将根据组织学，生化和生物力学特性评估作为特定目标1和2的一部分生产的工程软骨。特定目标3增加了将工程软骨和支撑支架塑造成半球形形状的复杂性，这是股骨头重新铺面所必需的。我们将尝试在附着在多孔壳聚糖/帽支架上的半球形壳中产生均匀的软骨层。将测量该软骨摩擦的厚度，球形和系数，以评估其对关节重外面的适用性。如果成功的话，特定的目标4将通过将透明软骨恢复到活动物中的全厚度缺陷来提供概念验证。该项目将通过证明与工程软骨共同重新铺面的可行方法来影响公共卫生。该提案针对特定的挑战，股骨头重新铺面，代表了克服这一挑战所需的第一步。 公共卫生相关性：该项目解决了开发骨关节炎治疗替代方案的重要问题，骨关节炎是一种退化性疾病，影响了数百万美国人，并负责每年数十亿美元的工资和生产力。软骨组织工程有可能用健康的功能性组织重塑骨关节关节，因此不必要用金属，塑料或陶瓷成分的关节替换。该项目基于与多孔磷酸钙 - 吉他式复合材料支架相连的自组装软骨的双相构造制定了联合重铺策略。