Cranial Bone Repair with Adipose-Derived Stem Cells

用脂肪干细胞修复颅骨

• 批准号: 6588557
• 负责人: Roy Clinton Ogle
• 金额: $ 14.8万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-15 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6588557
• 关键词: adipose tissue; bone development; bone regeneration; cell differentiation; clinical research; collagen; computed axial tomography; craniofacial; histology; human embryonic stem cell line; human tissue; immunocytochemistry; laboratory rat; oral facial restoration; osteoblasts; osteogenesis; polymerase chain reaction; stem cells; technology /technique development; telomerase; terminal nick end labeling; tissue /cell culture; tissue engineering; tissue support frame

DESCRIPTION (provided by applicant): The long-term goal of this proposal is to develop an effective therapy to repair bony defects in the craniofacial skeleton employing adult stem cells derived from adipose tissue. Reconstructive surgery for developmental anomalies, trauma and resection following tumor removal requires materials to replace or induce the regeneration of bone. Currently applied methods, such as use of autologous grafts and banked bone, are limited in efficacy and carry the risk of donor site morbidity and infection. Tissue engineering of replacement bone lacks the limitations of current approaches while offering distinct advantages. Adipose-derived adult stem cells (ADASC) are easily isolated and can be induced to differentiate ex vivo into a variety of neural and mesodermal cell lineages including osteoprogenitors. The central hypothesis of this study is that ADASCs can be maintained as replicating multipotent stem cells, which can be induced to differentiate into osteoblasts in vitro and, when delivered appropriately, will repair bony defects in vivo. Two Specific Aims are proposed to test the hypothesis: 1) to characterize and optimize the isolation, growth and differentiation of ADASCs in vitro; and 2) to test the capacity of human ADASCs to mediate repair of critical-sized bone defects in the skulls of athymic rats. In Aim 1 the clonal nature of the osteoprogenitor population within the total ADASC isolate and the timing of expression of bone specific genes will be determined. Using early and late markers of osteogenic differentiation, conditions for differentiation will be optimized and compared with primary human osteoblasts and osteoprogenitors derived from a human embryonic stem cell line. In Aim 2, ADASCs will be delivered in collagen sponges. Then de novo bone formation will be measured by three-dimensional reconstruction of computed tomography (3DCT) at intervals over the healing process to determine the optimal cell and passage number, stage of differentiation, and seeding density for bone repair. These studies should indicate the potential of the ADASC to contribute to bony tissue engineering and are a prerequisite to testing of the many tissue scaffold systems being developed and methods for using stem cells as gene delivery vehicles. The characterization studies may also provide the basis to use this stem cell system as a model for study of osteogenic differentiation.

描述（由申请人提供）：该提案的长期目标是开发一种有效的疗法来修复采用源自脂肪组织的成年干细胞的颅面骨骼中的骨缺陷。切除肿瘤后的发育异常，创伤和切除的重建手术需要替代或诱导骨骼再生的材料。目前应用的方法（例如使用自体移植物和库骨）在功效上受到限制，并承受供体现场发病率和感染的风险。替代骨的组织工程缺乏当前方法的局限性，同时提供了不同的优势。脂肪来源的成年干细胞（ADASC）很容易分离，可以诱导将离体分化为包括骨化剂在内的各种神经和中胚层细胞谱系。这项研究的中心假设是，ADASC可以保持为复制的多能干细胞，可以在体外诱导分化为成骨细胞，并在适当的情况下递送，将在体内修复骨缺陷。提出了两个具体的目的来检验假设：1）表征和优化体外ADASC的隔离，生长和分化； 2）测试人类ADASC的能力介导无胸骨大鼠头骨中关键尺寸骨缺损修复的能力。在AIM 1中，将确定全肌分离株内的骨基因生成剂种群的克隆性质，并确定骨特异性基因表达的时间。使用成骨分化的早期和晚期标记，将优化分化的条件，并将其与源自人类胚胎干细胞系的原代人成骨细胞和骨基因生成剂进行比较。在AIM 2中，ADASC将在胶原蛋白海绵中交付。然后，将通过在愈合过程中间隔对计算机断层扫描（3DCT）的三维重建（3DCT）进行测量，以确定最佳细胞和通过数量，分化阶段以及骨修复的播种密度。这些研究应表明ADASC有可能有助于骨组织工程，并且是测试开发许多组织支架系统的先决条件以及将干细胞用作基因递送车的方法。表征研究还可以提供使用此干细胞系统作为研究成骨分化的模型的基础。