Notch-enriched Mesenchymal Stromal Cells for Bone Allograft Repair

富含缺口的间充质基质细胞用于同种异体骨移植修复

• 批准号: 10543407
• 负责人: Richard Shane Barton
• 金额: $ 31.49万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10543407
• 关键词: Address; Affect; Allografting; Animals; Autologous Transplantation; Binding; Biological Assay; Blood Vessels; Bone Marrow; Bone Marrow Aspiration; Bone Tissue; Bone callus; Bromodeoxyuridine; Cell Aging; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cell Maintenance; Cells; Cellular Metabolic Process; Characteristics; Clinical Trials; Data; Defect; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Exhibits; Extracellular Matrix; Femur; Flow Cytometry; Future; Generations; Genes; Goals; Human; Immunohistochemistry; Impairment; Implant; Label; Ligands; Mediating; Mesenchymal Stem Cells; Methodology; Molecular; Monitor; Mus; Mutagenesis; Operative Surgical Procedures; Osteogenesis; Oxidative Stress; Pathway interactions; Patients; Periosteum; Population; Process; Recombinants; Regulation; Role; Signal Induction; Signal Transduction; Site; Skeletal Development; Stains; Stromal Cells; TWIST1 gene; Temperature; Testing; Therapeutic; Tissue Engineering; Transfection; Transplantation; United States; Up-Regulation; Vascularization; Work; allogenic bone transplantation; angiogenesis; biomechanical test; bone; bone healing; bone lead; bone marrow mesenchymal stem cell; bone repair; chromatin immunoprecipitation; experimental study; failure Implantation; gain of function; healing; histological stains; in vivo; laser capture microdissection; long bone; loss of function; mesenchymal stromal cell; microCT; mouse model; next generation; notch protein; novel; novel therapeutic intervention; osteogenic; pre-clinical; promoter; repaired; skeletal; stem cell proliferation; stem cells; stemness; success; tissue regeneration; tissue repair; transcription factor; transdifferentiation

Abstract: Segmental long bone defect surgeries require large devitalized allograft transplantations to replace missing host bone segments; however, significant problems often arise due to the impaired ability of the devitalized allograft to incorporate into the host bone. Recently, several discoveries in our lab have provided the basis for selecting, expanding and delivering of a substantial number of mesenchymal stem cells (MSCs), which retain their original stem cell characteristics, to be utilized in skeletal therapeutic applications: i) Notch2-selected and Notch activation by recombinant Jagged1 (JAG1) ligand expanded MSCs possess enhanced potentials for chondrogenic and osteogenic differentiation; ii) MSC sheets generated in temperature-responsive culture dishes exhibit a more robust effect on allograft incorporation into the host bone when transplanted to the site of the defect as a tissue-engineered periosteum surrounding the implanted graft; iii) Notch signaling induces rapid TWIST1 expression in MSCs. Based on these novel findings, we hypothesize that cell sheets generated with JAG1 expanded Notch2 positive MSCs on thermo-responsive culture dishes will significantly enhance critical segmented bone defects healing in a femoral allograft repair mouse model. To test this hypothesis, we will first to determine whether Notch2-selected and JAG1 expanded MSC sheets significantly enhance allograft healing and incorporation using our pre-clinical femoral allograft mouse model, as compared to cell sheets derived from traditionally isolated and expanded MSCs. Second, we will examine whether TWIST1 is require for JAG1-mediated MSC proliferation and maintenance in culture. Finally, we will determine how Notch activation by JAG1 involved in MSC sheet- induced angiogenesis. Data generated by this proposal will likely 1) identify optimized cell populations for generating tissue engineered periosteum (next generation MSC sheets), 2) validate a novel therapeutic approach for enhancing the repair of massive devitalized bone allograft and lead to advances in understanding the mechanism underlying MSC sheet-induced tissue repair, and 3) pave the path for large pre-clinical animal studies and subsequent clinical trials.

翻译后摘要：节段性长骨缺损手术需要大失活同种异体骨 移植以替换缺失的宿主骨段;然而， 由于失活的同种异体移植物结合到宿主骨中的能力受损而出现。 最近，我们实验室的几项发现为选择、扩展和 递送大量的间充质干细胞（MSC），其保持其原始的细胞分化。 用于骨骼治疗应用的干细胞特征：i）Notch 2选择的和 重组锯齿蛋白1（JAG 1）配体扩增的MSC具有增强的Notch激活 软骨形成和成骨分化的潜力; ii）在 温度响应性培养皿对同种异体移植物掺入 当作为组织工程骨膜移植到缺损部位时， iii）Notch信号传导诱导MSC中TWIST 1的快速表达。 基于这些新的发现，我们假设JAG 1产生的细胞片层扩增了， Notch 2阳性MSC在温敏培养皿上将显著提高临界细胞数。 股骨同种异体移植修复小鼠模型中的分段骨缺损愈合。为了验证这个假设， 我们将首先确定Notch 2选择的和JAG 1扩增的MSC片材是否显著 使用我们的临床前股骨同种异体移植小鼠模型增强同种异体移植物愈合和结合， 与来源于传统分离和扩增的MSC的细胞片相比。二是 将检测TWIST 1是否是JAG 1介导MSC增殖和维持所必需的 在文化中。最后，我们将确定Notch激活JAG 1如何参与MSC片层- 诱导血管生成。此建议生成的数据可能会1）识别优化的单元格 用于产生组织工程化骨膜的群体（下一代MSC片），2） 验证了一种新的治疗方法，用于增强大块失活骨的修复， 同种异体移植物，并导致理解MSC片诱导的机制的进展， 组织修复，以及3）为大型临床前动物研究和后续临床研究铺平道路 审判