Recellularization mechanisms in mononuclear cell seeded tissue engineered valves

单核细胞接种的组织工程瓣膜的再细胞化机制

• 批准号: 8969308
• 负责人: Gabriel LeVerne Converse
• 金额: $ 7.5万
• 依托单位: CHILDREN'S MERCY HOSP (KANSAS CITY, MO)
• 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/8969308
• 关键词: Address; Adult; Biochemical; Biological; Biological Assay; Bioprosthesis device; Biopsy; Bioreactors; Bone Marrow; Cell Culture Techniques; Cell Fraction; Cell Line; Cell surface; Cells; Child; Clinical; Clinical Treatment; Collagen; Complex; Congenital Heart Defects; Consensus; Cues; Custom; Cytokine Signaling; Development; Disease; Elastin; Extracellular Matrix; Face; Gene Expression; Gene Expression Profiling; Growth; Heart Valves; Histological Techniques; Human; Immune response; Implant; In Vitro; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Intention; Investigation; Laboratories; Leukocytes; Life; Longevity; Measures; Mechanics; Mediating; Mesenchymal Stem Cells; Modeling; Mononuclear; Operative Surgical Procedures; Patients; Performance; Phase; Phenotype; Physiological; Population; Process; Production; Proliferating; Protocols documentation; Pulmonary valve structure; Regenerative Medicine; Repeat Surgery; Role; Safety; Services; Surface; System; Time; Tissue Engineering; Tissues; Translations; aortic valve; calcification; clinical practice; conditioning; congenital heart disorder; cytokine; heart valve replacement; implantation; improved; in vivo; interest; interstitial cell; macrophage; monocyte; paracrine; pressure; protein expression; public health relevance; restoration; scaffold; semilunar valve; success

 DESCRIPTION (provided by applicant): While tissue engineering offers the promise of improved clinical treatment for numerous disorders, obstacles that have limited the translation of laboratory successes into clinical practice must be overcome before this potential is realized. This holds true for the tissue engineered heart valve (TEHV), a living, growing valve substitute that would significantly improve the treatment of congenital heart disorders in children and degenerative valve disorders in adults. However, scientific and regulatory challenges limit the practicality and attractiveness of previous heart valve tissue engineering strategies. The use of complex bioreactors systems that are not disposable or patient-specific introduces a substantial regulatory obstacle that must be overcome. We have addressed this issue through the development of a fully disposable, single-use bioreactor for heart valve seeding and conditioning, permitting the investigation of clinically feasible TEHV processing strategies. Scaffold safety is also of major concern, as the implanted construct must be fully functional as a valve from the time of implantation forward. This concern can be mitigated through the use of decellularized heart valve scaffolds, which have a record of clinical safety; however, previous difficulties in introducing sub-surface cells into the leaflet of the decellularized heart valve hae dampened enthusiasm for this particular TEHV scaffold. Using our custom bioreactor, we have overcome this limitation, successfully creating a pilot population of bone marrow mononuclear cells (BM-MNCs) within the leaflet. The objectives of this project are to determine the potential for ex vivo maturation of this pilot BM-MNC population through extended bioreactor processing and to evaluate the subsequent in vitro inflammatory response to the seeded tissue. Two Specific Aims have been formulated to accomplish these objectives. In the first Aim, extended bioreactor processing will be utilized to determine the extent to which the pilot BM-MNC population proliferates and differentiates during the culture period. We will also evaluate the potential for remodeling of the decellularized scaffold. Aim 2 will utilize an in vitro macrophage cytokine signaling assay to assess the inflammatory response to the seeded tissue. Accomplishing these two Specific Aims will elucidate the recellularization mechanisms of the decellularized heart valve in the context of heart valve tissue engineering.

 描述（由申请人提供）：虽然组织工程为许多疾病提供了改善临床治疗的前景，但在实现这一潜力之前，必须克服限制将实验室成功转化为临床实践的障碍。这适用于组织工程心脏瓣膜（TEHV），这是一种活的、生长的瓣膜替代物，将显著改善儿童先天性心脏病和成人退行性瓣膜病的治疗。然而，科学和监管的挑战限制了以前的心脏瓣膜组织工程策略的实用性和吸引力。非一次性或患者特异性的复杂生物反应器系统的使用引入了必须克服的实质性监管障碍。我们已经通过开发用于心脏瓣膜接种和调节的完全一次性、一次性生物反应器解决了这一问题，从而可以研究临床可行的TEHV处理策略。支架的安全性也是主要问题，因为植入的结构必须从植入时起就像瓣膜一样完全发挥功能。这种担心可以通过使用脱细胞心脏瓣膜支架来缓解，这种支架有临床安全性的记录;然而，以前在将亚表面细胞引入脱细胞心脏瓣膜小叶中的困难已经抑制了对这种特殊TEHV支架的热情。使用我们的定制生物反应器，我们克服了这一限制，成功地在瓣叶内创建了骨髓单核细胞（BM-MNC）的中试群体。本项目的目的是通过延长的生物反应器处理确定该中试BM-MNC群体离体成熟的潜力，并评价随后对接种组织的体外炎症反应。为实现这些目标，制定了两个具体目标。在第一个目标中，将利用延长的生物反应器处理来确定中试BM-MNC群体在培养期间增殖和分化的程度。我们还将评估脱细胞支架重塑的潜力。目的2将利用体外巨噬细胞细胞因子信号传导测定来评估对接种组织的炎症反应。实现这两个具体目标将阐明心脏瓣膜组织工程背景下脱细胞心脏瓣膜的再细胞化机制。