Functional Tissue Engineering of Cartilage Using Induced Pluripotent Stem Cells

使用诱导多能干细胞进行软骨功能组织工程

• 批准号: 9114384
• 负责人: Shannon Kathleen O'Connor
• 金额: $ 3.74万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9114384
• 关键词: 3-Dimensional; Activities of Daily Living; Adipose tissue; Adult; Affect; Aging; Area; Behavior Therapy; Biochemical; Biocompatible; Biocompatible Materials; Biomechanics; Body Temperature; Cartilage; Cells; Centers for Disease Control and Prevention (U.S.); Chondrocytes; Chondrogenesis; Collagen; Conditioned Culture Media; Data; Defect; Degenerative polyarthritis; Disease; Disease model; Drug Delivery Systems; Elastin; Encapsulated; Endotoxins; Engineering; Extracellular Matrix; FDA approved; Fibroblasts; Future; GAG Gene; Gene Delivery; Gene Expression; Glycerophosphates; Glycosaminoglycans; Goals; Growth; Histologic; Human Activities; Hydrogels; Immune; Implant; In Vitro; Injection of therapeutic agent; Joints; Life; Liquid substance; Longevity; Mechanics; Mediating; Mentors; Mesenchymal Stem Cells; Metals; Methods; Mus; Natural regeneration; Operative Surgical Procedures; Osteoblasts; Osteocalcin; Osteogenesis; Pain; Patients; Ploidies; Polymers; Population; Preclinical Drug Evaluation; Property; Prosthesis; Proteins; Quality of life; Replacement Arthroplasty; Risk; Role; Seeds; Signal Transduction; Signaling Molecule; Somatic Cell; Source; Staging; Stem cells; Subfamily lentivirinae; Supplementation; Tail; Techniques; Temperature; Time; Tissue Engineering; Tissues; Training; Transduction Gene; Transforming Growth Factors; Transition Temperature; United States; Viral; Work; X-Ray Computed Tomography; adult stem cell; articular cartilage; base; biocompatible polymer; bone; bone morphogenetic protein 2; bone morphogenic protein; caprolactone; cell type; design; disability; induced pluripotent stem cell; lentiviral-mediated; microCT; mineralization; novel; osteochondral tissue; osteogenic; osteopontin; polypeptide; programs; public health relevance; repaired; scaffold; small molecule; transforming growth factor beta3; transgene expression

 DESCRIPTION (provided by applicant): Osteoarthritis is a painful, degenerative joint disease that leads to significant disability and reduced quality of life, especially in the aging populatio. This affected population numbers 53 million in the United States according to newly-released data from the CDC, 23 million of whom suffer during activities of daily living. Treatment options remain limited; after implementation of lifestyle modifications, patients are given the option of joint replacement surgeries, but wear of implanted metal and polymer-based prosthetics yields an efficacy period of only 10-15 years. We intend to engineer a living tissue-based replacement that will avoid immune rejection, maintain the mechanical properties of normal cartilage tissue, be as durable as the host's original joints, and integrate into an osteochondral defect. To do so, we have created a three-dimensional scaffold woven with the biocompatible, FDA-approved, and slowly-degrading polymer poly(e-caprolactone) (PCL), with mechanical strength equivalent to native cartilage. With this scaffold, we have also demonstrated scaffold- mediated lentiviral transduction of chondrogenesis-inducing transforming growth factor ß3 (TGF-ß3), and separately with osteogenesis-inducing bone morphogenic protein 2 (BMP-2), both yielding sustained transgene expression in mesenchymal stem cells (MSCs). Induced pluripotent stem cells (iPSCs) are genetically matched to the donor somatic cells from which they are derived and represent a plentiful source of cells with potential to form all tissues. Our lab has shown tha iPSCs from mouse tail fibroblasts can be re-engineered to be chondrogenic, suggesting the possibility of turning patient-specific adult cells into chondrocytes and osteoblasts for osteochondral tissue engineering. However, to stimulate osteogenesis, additional small molecules are necessary usually as media supplements. In order to deliver these small molecules only to the osteogenic part of the construct, we will use a biosynthetic polymer, Elastin-Like Polypeptide (ELP), which has been engineered to have a reverse transition temperature at 35ºC (just below the temperature of the body), lending itself to being injected as a liquid at lower room temperatures, with solidification occurring above this temperature. This biocompatible hydrogel has been shown to induce chondrogenesis of adult stem cells and shown to be an excellent drug delivery vehicle; it may prove the key to creating an osteochondral tissue construct. The goals of this study are to develop functionalized scaffolds for chondrogenic and osteogenic differentiation of iPSCs, and to deliver local osteogenic factors using ELP as a carrier.

 描述（由申请人提供）：骨关节炎是一种疼痛的退行性关节疾病，可导致严重残疾和生活质量下降，尤其是在老年人群中。根据疾病预防控制中心最新公布的数据，美国有5300万受影响的人口，其中2300万人在日常生活活动中受到影响。治疗选择仍然有限;在改变生活方式后，病人可以选择关节置换手术，但植入金属和聚合物假体的有效期只有10-15年。我们打算设计一种基于活组织的替代物，这种替代物将避免免疫排斥，保持正常软骨组织的机械特性，与宿主的原始关节一样耐用，并整合到骨软骨缺损中。为了做到这一点，我们已经创建了一个三维支架编织与生物相容性，FDA批准，和缓慢降解的聚合物聚（ε-己内酯）（PCL），具有相当于天然软骨的机械强度。使用该支架，我们还证明了软骨形成诱导转化生长因子β 3（TGF-β 3）的支架介导的慢病毒转导，以及分别使用骨形成诱导骨形态发生蛋白2（BMP-2）的支架介导的慢病毒转导，两者都在间充质干细胞（MSC）中产生持续的转基因表达。诱导多能干细胞（iPSC）与它们所来源的供体体细胞在遗传上相匹配，并且代表了具有形成所有组织潜力的丰富细胞来源。我们的实验室已经证明，来自小鼠尾部成纤维细胞的iPSC可以被重新工程化为软骨细胞，这表明将患者特异性成体细胞转化为软骨细胞和成骨细胞用于骨软骨组织工程的可能性。然而，为了刺激骨生成，额外的小分子通常作为培养基补充剂是必要的。为了将这些小分子仅输送到构建体的成骨部分，我们将使用生物合成聚合物，弹性蛋白样多肽（ELP），其已被设计为具有35ºC的反向转变温度（略低于体温），使其在较低的室温下以液体形式注射，在此温度以上发生固化。这种生物相容性水凝胶已被证明可以诱导成体干细胞的软骨形成，并被证明是一种出色的药物递送载体;它可能是创建骨软骨组织构建体的关键。本研究的目的是开发用于诱导多能干细胞向软骨细胞和成骨细胞分化的功能化支架，并以ELP为载体在局部递送成骨因子。