Human Embryonic Stem Cells in Calcium Phosphate Constructs for Bone Regeneration

磷酸钙结构中的人胚胎干细胞用于骨再生

基本信息

批准号：
8281745
负责人：
HUAKUN XU
金额：
$ 19.19万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8281745
关键词：
Adult Age Aging Alginates Animal Model Arthritis Blood Vessels Bone Density Bone Marrow Bone Matrix Bone Regeneration Bone Tissue Cell Communication Cell Proliferation Cell physiology Cells Cephalic Chitosan Complex Defect Dental Dentures Disease Encapsulated Esthetics Fiber Fibrin Fracture Gold Growth Factor Harvest Healed Health Human Implant In Vitro Injectable Knowledge Lesion Mandible Maryland Maxilla Mechanics Mesenchymal Stem Cells Microspheres Minerals Molds Musculoskeletal Operative Surgical Procedures Orthopedics Osteogenesis Osteoporosis Paste substance Patients Peptides Population Prevalence Procedures Property Psychological reinforcement Quality of life RGD (sequence)Regenerative Medicine Reporting Research Resistance Risk Shapes Side Source Stem cells Stress System Teenagers Testing Time Tissue Engineering Umbilical cord structure Universities Weight-Bearing state bone bone engineering calcium phosphate cell growth cost craniofacial density design healing human embryonic stem cell improved in vivo interdisciplinary approach knowledge base mandible/maxilla meetings minimally invasive novel oral surgery specialty reconstruction regenerative scaffold tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): Seven million people suffer bone fractures in the U.S. each year. Musculoskeletal conditions cost the U.S. more than $200 billion annually. These numbers are increasing rapidly as the population ages. Human embryonic stem cells (hESCs) offer unlimited supplies of stem cells with a high potential for bone regeneration. However, there has been no report on the use of hESCs for bone tissue engineering via injectable calcium phosphates. While human bone marrow mesenchymal stem cells (hBMSCs) are useful, their harvest requires an invasive procedure, and their proliferation and differentiation potential is lot due to aging and diseases. Therefore, the objectives of this project are to: (1) investigate hESCs in injectable calcium phosphate cement (CPC) for bone engineering, in comparison with human umbilical cord MSCs (hUCMSCs) and hBMSCs; (2) establish the first knowledge base on hESC interactions with CPC scaffolds to guide hESCs for proliferation and osteodifferentiation; (3) design a RGD-grafted CPC scaffold for hESC encapsulation to enhance bone regeneration. While the Arg-Gly-Asp (RGD) peptide has been used in other scaffolds, there has been no report on its use in CPC. Aim 1 will investigate in vitro the hESC encapsulation and differentiation in injectable and macroporous CPC-RGD constructs, and test these hypotheses: (1) hESC-derived MSCs encapsulated in degradable microbeads and incorporated into CPC will synthesize the most amount of bone matrix, followed by hUCMSCs. The gold-standard hBMSCs will make the least bone matrix; (2) RGD-grafted CPC will greatly enhance cell function and bone matrix synthesis, without compromising the CPC injectability and the mechanical properties, which will match the reported strength of cancellous bone. Aim 2 will investigate the hESC-CPC-RGD constructs for bone regeneration in animal model, and test these hypotheses: (1) New bone volume, mineral density, and blood vessel density generated by CPC-RGD with hESC-derived MSCs will be the highest, followed by hUCMSCs. Both of them will far exceed those generated by hBMSCs; (2) hESC-CPC- RGD will be completely replaced by new bone across the entire critical-sized cranial defect at six months; (3) hESC-CPC-RGD will induce much more new bone than that without RGD. This project will yield ground- breaking knowledge on hESC encapsulation in CPC, hESC interaction with RGD-CPC and the guidance of hESCs for osteodifferentiation, and bone regeneration in animal model comparing hESCs, hUCMSCs, and hBMSCs side-by-side. The new stem cell paste can be used in minimally-invasive surgeries, fill complex- shaped defects, and be easily shaped for esthetics in dental and craniofacial applications. The novel hESC- CPC-RGD construct is expected to have wide orthopedic and craniofacial applications, with greatly enhanced bone regeneration to improve the health and quality of life for millions of people. If indeed, hESCs delivered via CPC are superior in osteogenesis compared to the gold-standard hBMSCs, which is anticipated to be shown by this project for the first time, the results will broadly impact the regenerative medicine field. PUBLIC HEALTH RELEVANCE: Despite the high promise of human embryonic stem cells (hESCs), there has been no report on hESC seeding with injectable calcium phosphate constructs for bone regeneration. The proposed research will investigate hESCs for bone tissue engineering via novel injectable calcium phosphate scaffolds for the first time, and establish ground-breaking knowledge on hESC guidance via peptide-calcium phosphate cement scaffold to enhance hESC function and bone regeneration in animal model. The novel hESC-calcium phosphate construct is expected to have a wide range of craniofacial and orthopedic applications, with greatly enhanced bone regeneration capability to improve the health and quality of life for millions of people.

描述（由申请人提供）：美国每年有 700 万人遭受骨折。美国每年因肌肉骨骼疾病造成的损失超过 2000 亿美元。随着人口老龄化，这些数字正在迅速增加。人类胚胎干细胞 (hESC) 提供无限量的干细胞供应，具有很高的骨再生潜力。然而，目前还没有关于通过注射磷酸钙将hESC用于骨组织工程的报道。虽然人骨髓间充质干细胞（hBMSC）很有用，但它们的收获需要侵入性操作，并且由于衰老和疾病，它们的增殖和分化潜力很大。因此，本项目的目标是：（1）研究用于骨工程的可注射磷酸钙水泥（CPC）中的hESCs，并与人脐带间充质干细胞（hUCMSCs）和hBMSCs进行比较；（2）建立第一个hESC与CPC支架相互作用的知识库，指导hESC增殖和骨分化； (3)设计RGD移植的CPC支架用于hESC封装以增强骨再生。虽然Arg-Gly-Asp（RGD）肽已用于其他支架，但尚未有其在CPC中使用的报道。目标 1 将在体外研究可注射和大孔 CPC-RGD 构建体中 hESC 的封装和分化，并测试这些假设：(1) 封装在可降解微珠中并掺入 CPC 中的 hESC 衍生 MSC 将合成最多量的骨基质，随后通过 hUCMSC。金标准hBMSCs将产生最少的骨基质； (2)RGD移植的CPC将大大增强细胞功能和骨基质合成，而不影响CPC的可注射性和机械性能，这将与报道的松质骨强度相匹配。目标 2 将研究用于动物模型中骨再生的 hESC-CPC-RGD 构建体，并测试这些假设：（1）由 CPC-RGD 与 hESC 衍生的 MSC 产生的新骨体积、矿物质密度和血管密度将是最高，其次是 hUCMSC。两者都将远远超过 hBMSC 产生的量； (2) hESC-CPC-RGD将在六个月时在整个临界大小的颅骨缺损处完全被新骨替代； (3)hESC-CPC-RGD会比没有RGD诱导更多的新骨。该项目将在 CPC 中的 hESC 封装、hESC 与 RGD-CPC 的相互作用以及 hESC 对骨分化的指导以及动物模型中的骨再生方面产生突破性的知识，同时比较 hESC、hUCMSC 和 hBMSC。新型干细胞糊剂可用于微创手术、填充复杂形状的缺损，并易于在牙科和颅面应用中塑形以实现美观。新型 hESC-CPC-RGD 构建体预计将具有广泛的骨科和颅面应用，并大大增强骨再生，以改善数百万人的健康和生活质量。如果确实通过 CPC 输送的 hESC 与金标准 hBMSC 相比在成骨方面更优越（预计该项目将首次证明这一点），那么其结果将广泛影响再生医学场地。公共健康相关性：尽管人类胚胎干细胞 (hESC) 前景广阔，但尚未有关于用可注射磷酸钙结构接种 hESC 进行骨再生的报道。拟议的研究将首次通过新型可注射磷酸钙支架研究用于骨组织工程的hESC，并建立通过肽-磷酸钙水泥支架引导hESC以增强动物模型中hESC功能和骨再生的突破性知识。这种新型hESC-磷酸钙结构预计将在颅面和骨科领域具有广泛的应用，并大大增强骨再生能力，从而改善数百万人的健康和生活质量。