Extracorporeal Shock Wave-Stimulated Periosteum for Bone Reconstruction

体外冲击波刺激骨膜进行骨重建

• 批准号: 8838173
• 负责人: Myron Spector
• 金额: --
• 依托单位: VA BOSTON HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8838173
• 关键词: Aftercare; Anesthesia procedures; Animal Model; Animals; Autologous; Autologous Transplantation; Becaplermin; Bicuspid; Biological Assay; Bone Marrow; Bone Regeneration; Bone Surface; Cell Count; Cell Culture Techniques; Cells; Chemotactic Factors; Clinical; Defect; Devices; Dose; Edentulous Mouth; Evaluation; Future; Goals; Goat; Harvest; Human; Immunohistochemistry; Implant; In Situ; Injury; Investigation; Mandible; Marrow; Measurement; Mesenchymal Stem Cells; Minerals; Modality; Modeling; Operative Surgical Procedures; Osteogenesis; Outcome; Pathologist; Patients; Periosteal Cell; Periosteum; Procedures; Process; Proliferating; Rattus; Recombinants; Recruitment Activity; Sampling; Shock; Site; Source; Sterility; Stromal Cells; Surface; Surgical Management; System; Testing; Therapeutic; Thick; Time; Tissues; Titania; Titanium; Transplantation; United States Food and Drug Administration; Work; base; bone; bone growth factor; calcium phosphate; craniofacial; energy density; maxillofacial; novel; osteogenic; osteoprogenitor cell; platelet-derived growth factor BB; reconstruction; research study; response; scaffold; tibia

DESCRIPTION (provided by applicant): There are 4 specific aims for this study. Specific Aim #1 is to determine the effects of extracorporeal shock waves (ESWs) on the proliferation of osteogenic cells in the periosteum of the tibia and edentulous mandible of the same animals, by conducting a dose-response experiment. Three energy densities will be employed (0, 0.15, and 0.45 mJ/mm2; a one-time application of 3,000 shocks for each energy density), and the results compared after 2, 4, and 8 days. The outcome variables will be selected cell counts and periosteum thickness measurement from histomorphometry employing immunohistochemistry, and osteogenic cell culture assay. Our objective is to find the dose (0.15 vs 0.45 mJ/mm2) and time post-treatment that yields the greatest number of osteogenic cells prior to the formation of bone. The goal is to be able to elevate or harvest the proliferating cambium layer of cells; the presence of bony trabeculae would interfere with the harvest of the cells. Our initial goal, in this proposal, is to harvest the osteogenic cells; future studies may be directed toward harvesting the cancellous bone that forms in the ESW-stimulated periosteum. Specific Aim #2 is to determine the effects of ESWs on the osteogenic cells in the marrow of the same tibial and mandibular samples employed in Specific Aim #1. We have already found in prior work that ESWs induce changes in the marrow. We have been informed by a pathologist that the changes are consistent with those that would be expected to be reversible, and of no adverse clinical consequence. We would like to determine if the ESW-stimulated marrow contains a greater percentage of mesenchymal stem cells (MSCs; i.e., marrow stromal cells, or osteoprogenitor cells). We will make this determination by assaying the osteogenic potential of the marrow in the cell culture assay. If we find that ESWs do, in fact, stimulate osteogenic cellular activity in the marrow, this will be investigated in future studies. The findings could be of value in developing a simple, non-invasive procedure for enhancing an alternative autologous cell source (i.e., marrow) for bone reconstruction procedures. Specific Aim #3 is to determine the effects of ESW-stimulated periosteum on bone regeneration in porous calcium phosphate (CaP) blocks in defects in the mandible. The ESW dose that yields the greatest number of osteogenic cells in Specific Aim #1 will be applied to the periosteum of the edentulous premolar region of goats. After the prescribed period post-ESW, the periosteum will be elevated and a porous CaP block containing 2 threaded titanium implants placed into the surgically prepared site. The elevated periosteum will then be applied to the surface of the block. In another group of animals the ESW-stimulated periosteum of the tibia will be harvested as a free autologous graft to be applied to the CaP block. In Specific Aim #4, recombinant human (rh) PDGF-BB will be incorporated into the CaP block before it is implanted into the defect and covered with the ESW-stimulated periosteum as in Specific Aim #3.

描述（由申请人提供）： 这项研究有 4 个具体目标。 具体目标#1是通过进行剂量反应实验来确定体外冲击波（ESW）对同一动物的胫骨骨膜和无齿下颌骨中成骨细胞增殖的影响。将采用三种能量密度（0、0.15 和 0.45 mJ/mm2；每种能量密度一次性施加 3,000 次冲击），并在 2、4 和 8 天后比较结果。结果变量将是采用免疫组织化学和成骨细胞培养测定法从组织形态计量学中选择的细胞计数和骨膜厚度测量。我们的目标是找到在骨形成之前产生最大数量成骨细胞的治疗后剂量（0.15 与 0.45 mJ/mm2）和时间。目标是能够提升或收获细胞增殖形成层；骨小梁的存在会干扰细胞的收获。在这个提案中，我们的最初目标是收获成骨细胞；未来的研究可能会针对采集 ESW 刺激的骨膜中形成的松质骨。 具体目标 #2 是确定 ESW 对具体目标 #1 中使用的相同胫骨和下颌样本的骨髓中成骨细胞的影响。我们在之前的工作中已经发现 ESW 会引起骨髓的变化。病理学家告知我们，这些变化与预期可逆的变化一致，并且不会产生不良临床后果。我们想确定 ESW 刺激的骨髓是否含有更高比例的间充质干细胞（MSC；即骨髓基质细胞或骨祖细胞）。我们将通过在细胞培养测定中测定骨髓的成骨潜力来做出这一决定。如果我们发现 ESW 实际上能够刺激骨髓中的成骨细胞活性，我们将在未来的研究中对此进行研究。这些发现对于开发一种简单的非侵入性手术来增强骨重建手术的替代自体细胞来源（即骨髓）可能具有价值。 具体目标#3 是确定 ESW 刺激的骨膜对下颌骨缺损多孔磷酸钙 (CaP) 块中骨再生的影响。在特定目标 #1 中产生最大数量成骨细胞的 ESW 剂量将应用于山羊无牙颌前磨牙区域的骨膜。在 ESW 后规定的时间后，骨膜将被抬高，并将含有 2 个螺纹钛植入物的多孔 CaP 块放置到手术准备的部位。然后将升高的骨膜施加到块的表面。在另一组动物中，ESW 刺激的胫骨骨膜将作为游离自体移植物被收获，应用于 CaP 块。 在具体目标 #4 中，重组人 (rh) PDGF-BB 将被纳入 CaP 块中，然后将其植入缺损处并像具体目标 #3 中那样用 ESW 刺激的骨膜覆盖。