Spatiotemporal Modulation of Osteogenesis in a 3-D Stromal/Stem Cell Model
3-D 基质/干细胞模型中成骨的时空调节
基本信息
- 批准号:8979685
- 负责人:
- 金额:$ 35.75万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2015
- 资助国家:美国
- 起止时间:2015-01-01 至 2016-08-14
- 项目状态:已结题
- 来源:
- 关键词:3-DimensionalAdipose tissueAdultAffectAlamarBlueAlkaline PhosphataseAngiogenic FactorAnimalsBlood VesselsBone RegenerationCD-1 Nude MouseCalciumCalvariaCell SurvivalCell TherapyCell modelCellsComplexCuesCulture MediaDNADefectDevelopmentEnvironmentEvaluationFractureGene DeliveryGene ExpressionGoalsHealthHumanHydrogelsIn VitroLeadLifeLightLiteratureLocationMechanicsMesenchymal Stem CellsMetabolicMethodsMethylcelluloseMicroRNAsModelingMusMusculoskeletalNude MiceOperative Surgical ProceduresOsteocalcinOsteogenesisPECAM1 genePolymersPositioning AttributeProcessProductionQualifyingResearchRoleSignal TransductionSiteSpatial DistributionStaining methodStainsStem cellsStructureSystemTechniquesTestingThickTimeTissue EngineeringTissue ModelTissuesUntranslated RNAVariantVascular Endothelial Growth FactorsVascularizationX-Ray Computed Tomographyadult stem cellangiogenesisbaseboneclinically relevantcomparativegene delivery systemimprovedin vivoinnovationlaser capture microdissectionmineralizationnanoplasmonicosteogenicosteopontinphotoactivationplasmonicsrepairedscaffoldspatiotemporalstem cell differentiationvon Willebrand Factor
项目摘要
DESCRIPTION (provided by applicant): A substantial body of literature supports linkages between osteogenic and angiogenic signals both in vitro and in vivo, but little is known about how their spatial and temporal coordination impacts the differentiation of adult stromal/stem cells (ASC) and bone regeneration. This study explores the coordination of angiogenic and osteogenic signals utilizing two techniques, recently described by our labs; i) the production ASCs sheets that can be stacked to generate quasi three- dimensional (3-D) structures and ii) the photo-controlled differentiation of ASCs through tightly regulated miRNA mimic delivery. The combination of these techniques will allow the induction of spatiotemporal gradients of angiogenic and osteogenic factors in a 3-D model to study how the timing and magnitude of differentiation cues impact ASC cell fate in the complex fracture site environment. Thus, our overall goal for this project is the development of an in vitro, quasi 3-D model of ASC osteogenesis to establish the effect of spatiotemporally modulated osteogenic and angiogenic cues and correlation of these results in a nude mouse calvarial defect model. As noted above we have developed a method of ASC cell sheet production utilizing a thermally reversible methylcellulose hydrogel polymer and automated cell manipulation system enabling the development of quasi 3-D structures to serve as model tissues. We have also demonstrated a light activated Plasmonic Gene Delivery system (PGDs), which serves as a convenient inducible gene delivery vehicle. When combined with miRNA mimics, miR-148b and miR-132, PGDs have been demonstrated to induce de novo osteogenic and angiogenic differentiation in human adipose derived ASCs, respectively. Combining both these methods will allow the induction of spatiotemporal differentiation gradients within the quasi 3-D ASC sheets and improve our understanding of the interplay of osteogenic and angiogenic factors in ASC based bone repair. The hypothesis will be tested in the following three Aims: Aim 1 will assess the impact on cell sheet stacking on ASC viability and differentiation. ASC cell sheets will be stacked into 1, 2, 5, 10 and 20 layers and cultured for up to 28 days to determine the impact on ASC viability, proliferation and differentiation potential. Aim 2 will assess the impact of temporally and spatialy varying the light activation of PGDs with osteogenic and angiogenic miRNA mimics on the osteogenesis and angiogenesis of ASCs within the quasi 3-D stem cells sheets. In aim 3 we will correlate in vitro results with in vivo in a 12-week nude mouse calvarial defect model. Optimal 3-D stack sizes and spatiotemporal induction conditions determined in Aim 1&2 will be tested in the repair of a calvarial defect in a CD-1 nude mouse. Animals will be euthanized at 1,6 and 12 weeks post-surgery and undergo histological, x-ray and �CT evaluation for bone formation and vascularization of the defect site. The successful conduct of the project will provide a greater understanding of how spatiotemporally coordinated expression of osteo- and angiogenic factors impact ASC differentiation into bone and additionally, this research will lead to improved stromal/stem cell based therapies for critical sized bone defect repair.
描述(由申请人提供):大量文献支持体外和体内成骨和血管生成信号之间的联系,但关于它们的空间和时间协调如何影响成体基质/干细胞(ASC)的分化和骨再生,我们知之甚少。本研究利用两种技术探讨血管生成和成骨信号的协调,最近由我们的实验室描述;i)生产ASCs薄片,可以堆叠以产生准三维(3-D)结构;ii)通过严格调节miRNA模拟递送的ASCs光控分化。这些技术的结合将允许在三维模型中诱导血管生成和成骨因子的时空梯度,以研究分化线索的时间和大小如何影响复杂骨折部位环境中ASC细胞的命运。因此,我们这个项目的总体目标是建立一个体外ASC成骨的准三维模型,以建立时空调节的成骨和血管生成线索的影响,以及这些结果在裸鼠颅骨缺损模型中的相关性。如上所述,我们开发了一种利用热可逆甲基纤维素水凝胶聚合物和自动细胞操作系统生产ASC细胞片的方法,从而能够开发准3-D结构作为模型组织。我们还展示了一种光激活等离子体基因传递系统(PGDs),它是一种方便的诱导型基因传递载体。当与miRNA模拟物miR-148b和miR-132联合使用时,PGDs已被证明分别诱导人脂肪源性ASCs新生成骨和血管生成分化。结合这两种方法将允许在准三维ASC薄片内诱导时空分化梯度,并提高我们对基于ASC的骨修复中成骨因子和血管生成因子相互作用的理解。该假设将在以下三个目标中进行验证:目标1将评估细胞片堆叠对ASC活力和分化的影响。将ASC细胞片堆叠成1、2、5、10和20层,培养28天,以确定对ASC活力、增殖和分化潜力的影响。目的2将评估具有成骨和血管生成miRNA模拟物的PGDs的光激活在时间和空间上的变化对准3d干细胞片内ASCs的成骨和血管生成的影响。在目标3中,我们将把体外结果与体内12周的裸鼠颅骨缺损模型相关联。Aim 1和Aim 2中确定的最佳3d堆叠大小和时空诱导条件将在CD-1裸鼠颅骨缺损修复中进行测试。动物将在术后1周、6周和12周实施安乐死,并进行组织学、x线和CT评估骨形成和缺损部位的血管化情况。该项目的成功实施将使我们更好地了解骨和血管生成因子的时空协调表达如何影响ASC向骨分化,此外,这项研究将导致基于基质/干细胞的治疗方法的改进,用于临界大小的骨缺陷修复。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Daniel J. Hayes其他文献
Climate change and the future of natural disturbances in the central hardwood region
气候变化和中部硬木地区自然干扰的未来
- DOI:
10.1007/978-3-319-21527-3_13 - 发表时间:
2016 - 期刊:
- 影响因子:0
- 作者:
Virginia H. Dale;M. Joseph Hughes;Daniel J. Hayes - 通讯作者:
Daniel J. Hayes
An assessment of the carbon balance of Arctic tundra
北极苔原碳平衡评估
- DOI:
- 发表时间:
2016 - 期刊:
- 影响因子:0
- 作者:
A. D. McGuire;T. Christensen;Daniel J. Hayes;A. Héroult;E. Euskirchen;J. Kimball;C. Koven;P. Lafleur;Paul A. Miller;Walter Oechel;P. Peylin;Mathew Williams - 通讯作者:
Mathew Williams
Traitement des occlusions proximales de l’artère fémorale superficielle par endartériectomie à ciel ouvert et angioplastie fémoro-poplitée endovasculaire avec stent
股动脉近端闭塞术、股动脉内切除术和股腘血管内血管成形术(含支架)
- DOI:
- 发表时间:
2011 - 期刊:
- 影响因子:0
- 作者:
Daniel J. Hayes;M. J. Dougherty;K. D. Calligaro - 通讯作者:
K. D. Calligaro
Separation of rat pituitary secretory granules by continuous flow electrophoresis
连续流电泳分离大鼠垂体分泌颗粒
- DOI:
10.1002/elps.1150111118 - 发表时间:
1990 - 期刊:
- 影响因子:2.9
- 作者:
Daniel J. Hayes;Carrie Exton;T. Salada;Kathy Shellenberger;Jenny Waddle;W. Hymer - 通讯作者:
W. Hymer
Management of flush superficial femoral artery occlusions with combined open femoral endarterectomy and endovascular femoral-popliteal angioplasty and stent-grafting.
联合开放式股动脉内膜切除术和血管内股腘血管成形术和支架移植术治疗股浅动脉闭塞。
- DOI:
- 发表时间:
2011 - 期刊:
- 影响因子:1.5
- 作者:
Daniel J. Hayes;M. Dougherty;K. Calligaro - 通讯作者:
K. Calligaro
Daniel J. Hayes的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Daniel J. Hayes', 18)}}的其他基金
Ultrasound responsive hydrogels for stimulated combinatorial drug delivery
用于刺激组合药物递送的超声响应水凝胶
- 批准号:
10742110 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Biomedical Engineering Design: Integrating Simulation, Clinical Immersion, and Regulatory Training
生物医学工程设计:集成模拟、临床沉浸和监管培训
- 批准号:
10493117 - 财政年份:2021
- 资助金额:
$ 35.75万 - 项目类别:
Biomedical Engineering Design: Integrating Simulation, Clinical Immersion, and Regulatory Training
生物医学工程设计:集成模拟、临床沉浸和监管培训
- 批准号:
10204301 - 财政年份:2021
- 资助金额:
$ 35.75万 - 项目类别:
Spatio-temporal Modulation of Osteogenesis in a 3-D Stromal Stem Cell Model
3-D 基质干细胞模型中成骨的时空调节
- 批准号:
9393199 - 财政年份:2017
- 资助金额:
$ 35.75万 - 项目类别:
相似海外基金
Deciphering the role of adipose tissue in common metabolic disease via adipose tissue proteomics
通过脂肪组织蛋白质组学解读脂肪组织在常见代谢疾病中的作用
- 批准号:
MR/Y013891/1 - 财政年份:2024
- 资助金额:
$ 35.75万 - 项目类别:
Research Grant
ESTABLISHING THE ROLE OF ADIPOSE TISSUE INFLAMMATION IN THE REGULATION OF MUSCLE MASS IN OLDER PEOPLE
确定脂肪组织炎症在老年人肌肉质量调节中的作用
- 批准号:
BB/Y006542/1 - 财政年份:2024
- 资助金额:
$ 35.75万 - 项目类别:
Research Grant
Canadian Alliance of Healthy Hearts and Minds: Dissecting the Pathways Linking Ectopic Adipose Tissue to Cognitive Dysfunction
加拿大健康心灵联盟:剖析异位脂肪组织与认知功能障碍之间的联系途径
- 批准号:
479570 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Operating Grants
Determinants of Longitudinal Progression of Adipose Tissue Inflammation in Individuals at High-Risk for Type 2 Diabetes: Novel Insights from Metabolomic Profiling
2 型糖尿病高危个体脂肪组织炎症纵向进展的决定因素:代谢组学分析的新见解
- 批准号:
488898 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Operating Grants
Activation of human brown adipose tissue using food ingredients that enhance the bioavailability of nitric oxide
使用增强一氧化氮生物利用度的食品成分激活人体棕色脂肪组织
- 批准号:
23H03323 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Development of new lung regeneration therapies by elucidating the lung regeneration mechanism of adipose tissue-derived stem cells
通过阐明脂肪组织干细胞的肺再生机制开发新的肺再生疗法
- 批准号:
23K08293 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
A study on the role of brown adipose tissue in the development and maintenance of skeletal muscles
棕色脂肪组织在骨骼肌发育和维持中作用的研究
- 批准号:
23K19922 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Grant-in-Aid for Research Activity Start-up
Adipose Tissue T Cell Polarization and Metabolic Health in Persons Living with HIV
HIV 感染者的脂肪组织 T 细胞极化和代谢健康
- 批准号:
10619176 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Estrogen Signaling in the Ventromedial Hypothalamus Modulates Adipose Tissue Metabolic Adaptation
下丘脑腹内侧区的雌激素信号调节脂肪组织代谢适应
- 批准号:
10604611 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别:
Obesity and Childhood Asthma: The Role of Adipose Tissue
肥胖和儿童哮喘:脂肪组织的作用
- 批准号:
10813753 - 财政年份:2023
- 资助金额:
$ 35.75万 - 项目类别: