Enhancement Of Bone Regeneration From Human Adipose-Derived Stromal Cells

增强人类脂肪源性基质细胞的骨再生

• 批准号: 7897643
• 负责人: Benjamin Levi
• 金额: $ 5.68万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7897643
• 关键词: Abdomen; Address; Adipocytes; Adipose tissue; Adult; Affect; Alkaline Phosphatase; Apatites; Bone Cements; Bone Marrow Cells; Bone Matrix; Bone Regeneration; Calvaria; Cell Therapy; Cell surface; Cells; Clinical; Data; Defect; Degenerative Disorder; Development; Disease; ENG gene; Ethics; Fluorescence-Activated Cell Sorting; Fluorescent in Situ Hybridization; Gene Expression; Glycolates; Goals; Harvest; Healed; Health Care Costs; Hereditary Disease; Histology; Human; Implant; In Vitro; Investigation; Knowledge; Mission; Modality; Modeling; Molecular Profiling; Mus; Natural regeneration; Neurosurgeon; Nude Mice; Operating Rooms; Operative Surgical Procedures; Orthopedics; Osteocalcin; Osteogenesis; Outcome; Patients; Plague; Plastics; Polymerase Chain Reaction; Population; Procedures; Skeleton; Staining method; Stains; Stem cells; Stromal Cells; Suction Lipectomy; Techniques; Thigh structure; Time; Tissue Engineering; Tissues; Translating; Trauma; United States National Institutes of Health; Visit; base; bone; cell type; cost; craniofacial; design; disability burden; embryonic stem cell; healing; implantation; in vivo; innovation; malformation; novel; novel strategies; osteogenic; osteopontin; osteoprogenitor cell; reconstruction; regenerative; repaired; response; scaffold; skeletal; skeletal regeneration; skeletal tissue; tomography

DESCRIPTION (provided by applicant): In 2006, data from the U.S. Health Cost & Utilization Project revealed over one million orthopedic-related procedures, at a cost in excess of 7 billion dollars making skeletal repair using tissue engineering an attractive target. The ability of adipose-derived stromal cells (ASCs) to differentiate into bone suggests that they may be used to fulfill the mounting needs of patients with genetic disorders, degenerative diseases, and traumatic or post-surgical tissue deficits of the skeleton. Though similar techniques can be performed with bone marrow cells and embryonic stem cells, adipose cells are easier to obtain in large quantities and lack the ethical concerns. The ultimate translational goal would be the ability within one visit to the operating room to harvest adipose tissue, obtain sufficient numbers of human ASCs, isolate those cells with a greater osteogenic potential and implant these cells into the skeletal defect. The proposal below will facilitate the development of a novel means to promote rapid and robust bone formation in patients with skeletal defects. The central hypothesis is that human adipose derived stromal cells represent a readily available population of cells with which to design cell-based therapies for skeletal regeneration. In the first specific aim, a human ASC subpopulation with enhanced osteogenic capacity in vitro will be identified. Preliminary studies have shown the cell surface marker CD105 to have an increased osteogenic potential in mice and this same marker will be used to identify humans cells with increased osteogenic capabilities. Human adipose tissue will be harvested using standard liposuction techniques. Human ASCs with enhanced osteogenic potential will be isolated based on cell surface markers (CD105high and CD105low) using fluorescence activated cell sorting (FACS). The osteogenic potential of these two subpopulations will be analyzed using QT-PCR and Alizarin red staining. In the second specific aim, skeletal healing will be accelerated in vivo by implanting the subpopulation of human ASCs with enhanced osteogenic potential from Specific Aim #1 onto a skeletal defect using a PLGA scaffold. Osseous healing will be assessed and quantified using microcomputed tomography (microCT), detailed histology, histomorphometry and fluorescent in situ hybridization(FISH). Data obtained from these aims will demonstrate an innovative strategy to repair and regenerate bone that has been affected by trauma, disease, surgery or malformations which is consistent with the NIH mission to reduce the burdens of disability.

描述（由申请人提供）：2006 年，美国健康成本与利用项目的数据显示，有超过 100 万例与骨科相关的手术，成本超过 70 亿美元，这使得利用组织工程进行骨骼修复成为一个有吸引力的目标。脂肪源性基质细胞（ASC）分化为骨骼的能力表明它们可用于满足患有遗传性疾病、退行性疾病以及骨骼外伤或术后组织缺陷的患者日益增长的需求。尽管可以用骨髓细胞和胚胎干细胞进行类似的技术，但脂肪细胞更容易大量获得，并且缺乏伦理问题。最终的转化目标是能够在一次进入手术室的时间内收获脂肪组织，获得足够数量的人类ASC，分离那些具有更大成骨潜力的细胞并将这些细胞植入骨骼缺陷中。以下提案将有助于开发一种新方法，以促进骨骼缺陷患者快速、稳健的骨形成。中心假设是，人类脂肪来源的基质细胞代表了一种容易获得的细胞群，可以用来设计基于细胞的骨骼再生疗法。第一个具体目标是鉴定体外成骨能力增强的人类 ASC 亚群。初步研究表明细胞表面标记物 CD105 在小鼠中具有增强的成骨潜力，并且该标记物将用于识别具有增强的成骨能力的人类细胞。将使用标准吸脂技术收获人体脂肪组织。将使用荧光激活细胞分选 (FACS) 根据细胞表面标记（CD105high 和 CD105low）分离具有增强成骨潜力的人类 ASC。将使用 QT-PCR 和茜素红染色分析这两个亚群的成骨潜力。在第二个具体目标中，通过使用 PLGA 支架将特定目标 #1 中具有增强成骨潜力的人类 ASC 亚群植入到骨骼缺陷上，将加速体内骨骼愈合。将使用显微计算机断层扫描 (microCT)、详细组织学、组织形态计量学和荧光原位杂交 (FISH) 对骨愈合进行评估和量化。从这些目标获得的数据将证明修复和再生受创伤、疾病、手术或畸形影响的骨骼的创新策略，这与 NIH 减轻残疾负担的使命是一致的。

项目成果

Deleterious effects of freezing on osteogenic differentiation of human adipose-derived stromal cells in vitro and in vivo.

冷冻对人脂肪基质细胞体外和体内成骨分化的有害影响。

• 发表时间: 2011-03
• 影响因子: 4
• 作者: James, Aaron W;Levi, Benjamin;Nelson, Emily R;Peng, Michelle;Commons, George W;Lee, Min;Wu, Benjamin;Longaker, Michael T
• 通讯作者: Longaker, Michael T

Concise Review: Intraoperative Strategies for Minimal Manipulation of Autologous Adipose Tissue for Cell‐ and Tissue‐Based Therapies

简评：基于细胞和组织的治疗中自体脂肪组织最小化操作的策略

• DOI: 10.3390/ijms241310982
• 发表时间: 2023-07-01
• 期刊: International Journal of Molecular Sciences
• 影响因子: 5.6
• 作者: A. Trivisonno;Robert;W.;Ale;er;er;S. Baldari;Steven;R.;Cohen;G. Rocco;P. Gentile;G. Magalon;J. Magalon;R;y;y;B.;Miller;Hayley Womack;Gabriele Toietta
• 通讯作者: Gabriele Toietta

Acute skeletal injury is necessary for human adipose-derived stromal cell-mediated calvarial regeneration.

急性骨骼损伤对于人类脂肪基质细胞介导的颅骨再生是必要的。

• 发表时间: 2011-03
• 影响因子: 3.6
• 作者: Levi, Benjamin;James, Aaron W;Nelson, Emily R;Peng, Michelle;Wan, Derrick C;Commons, George W;Lee, Min;Wu, Benjamin;Longaker, Michael T
• 通讯作者: Longaker, Michael T

CD105 protein depletion enhances human adipose-derived stromal cell osteogenesis through reduction of transforming growth factor β1 (TGF-β1) signaling.

CD105 蛋白耗竭通过减少转化生长因子β1 (TGF-β1) 信号传导增强人类脂肪源性基质细胞成骨。

• 发表时间: 2011-11-11
• 作者: Levi, Benjamin;Wan, Derrick C;Glotzbach, Jason P;Hyun, Jeong;Januszyk, Michael;Montoro, Daniel;Sorkin, Michael;James, Aaron W;Nelson, Emily R;Li, Shuli;Quarto, Natalina;Lee, Min;Gurtner, Geoffrey C;Longaker, Michael T
• 通讯作者: Longaker, Michael T

Molecular analysis and differentiation capacity of adipose-derived stem cells from lymphedema tissue.

淋巴水肿组织脂肪干细胞的分子分析和分化能力。

• 发表时间: 2013-09
• 影响因子: 3.6
• 作者: Levi, Benjamin;Glotzbach, Jason P;Sorkin, Michael;Hyun, Jeong;Januszyk, Michael;Wan, Derrick C;Li, Shuli;Nelson, Emily R;Longaker, Michael T;Gurtner, Geoffrey C
• 通讯作者: Gurtner, Geoffrey C