Tissue-Engineered Constructs for Treatment of Intervertebral Disc Degeneration

描述 本项目的目的是开发一种组织工程化结构，包括工程化髓核和纤维环，以治疗纤维环（AF）缺陷和椎间盘（IVD）退行性变化。追求三个具体目标：“具体目标1将包括在透明质酸（HA）为基础的水凝胶构建体中建立来自兔和人NP和MSC细胞的体外组织工程NP制造，并评估细胞活力、基因调控、ECM产生和生物力学特性，作为凝胶生物物理特性、细胞接种密度和在盘状环境中扩展培养的函数。“具体目标2将需要建立和评估细胞浸润和功能成熟工程化的AF样结构，从兔和人AF和MSC接种的纳米纤维支架开始体外。“具体目标3将联合收割机结合目标1和2中开发的组织工程NP和AF，形成复合椎间盘样结构，然后将该结构应用于全椎间盘切除术的体内兔椎间盘模型。将通过MRI T2标测和随后的运动节段机械、生物化学和组织学特性的离体分析来阿塞塞评估疗效。研究设计：本研究将使用体外组织培养物进行特定目的1-2;特定目的3将使用兔动物模型和磁共振成像（MRI）。方法学：特定目的1和2是基于实验室的研究，将利用体外组织培养。Specific Aim 3将使用兔模型在体内进行椎间盘切除术和组织工程椎间盘植入，随后进行脊柱离体MRI和组织分析。特定目标1-3的主要研究结局指标是组织培养的生化分析和组织工程构建体的生物力学分析。此外，Special Aim 3将利用这些相同的结局，并对植入的组织工程结构进行定量MRI T2标测，并与对照（未经处理的椎间盘切除术）和相邻正常椎间盘进行比较。结果：此时不适用。临床关系：该项目的目标是开发组织工程结构来治疗退行性椎间盘疾病，以及验证放射学方法来无创评估退行性椎间盘疾病的潜在治疗方法。影响/意义：预计拟定的研究将增加对退行性椎间盘疾病的了解，并提供潜在的新型治疗方法，以逆转退行性病变并降低复发性椎间盘突出症的风险。此外，它将为申请人提供关键支持，因为他在费城VA医学中心和宾夕法尼亚大学开展脊柱外科研究密集型临床实践。

DESCRIPTION The objective of this project is to develop a tissue-engineered construct including an engineered nucleus pulposus and annulus fibrosus to treat defects of the annulus fibrosus (AF) and degenerative changes of the intervertebral disc (IVD). Three Specific Aims are pursued: " Specific Aim 1 will consist of establishing in vitro tissue engineered NP fabrication from rabbit and human NP and MSC cells in hyaluronic acid (HA)-based hydrogel constructs, and evaluation of cell viability, gene regulation, ECM production, and biomechanical properties as a function of gel biophysical properties, cell seeding density, and extended culture in a disc-like environment. " Specific Aim 2 will entail establishing and evaluating cellular infiltration and functional maturation engineered AF-like structures starting from rabbit and human AF and MSC seeded nanofibrous scaffolds in vitro. " Specific Aim 3 will combine the tissue-engineered NP and AF developed in Aims 1 and 2 to form a composite disc-like construct, and to then apply this construct to an in vivo rabbit disc model of total discectomy. Efficacy will be assesse via MRI T2 mapping and subsequent ex vivo analysis of motion segment mechanical, biochemical, and histologic properties. Research Design: This study will utilize in vitro tissue culture for Specific Aims 1-2; Specific Aim 3 will utilize a rabbit animal model and magnetic resonance imaging (MRI). Methodology: Specific Aims 1 and 2 are laboratory-based studies that will utilize in vitro tissue culture. Specific Aim 3 will be performed in vivo with a rabbit mdel for discectomy and tissue-engineered disc implantation with subsequent ex vivo MRI of the spine and tissue analysis. The primary study outcome measures for Specific Aims 1-3 are biochemical analysis of tissue culture and biomechanical analysis of tissue-engineered constructs. Additionally, Specific Aim 3 will utilize these same outcomes, as well as implement quantitative MRI T2 maps of the implanted tissue-engineered constructs with comparisons made to control (non-treated discectomy) and adjacent normal discs. Findings: n/a at this time. Clinical Relationships: The goal of this project is to develop tissue-engineered constructs to trea degenerative disc disease as well as to validate radiologic methods to noninvasively assess potential therapeutic treatments of degenerative disc disease. Impact/Significance: It is anticipated that the proposed study will offer an increased understanding of degenerative disc disease and potential novel therapeutic treatment to both reverse degenerative changes as well as decrease risk of recurrent disc herniation. Further, it will provide critical support for the applicant as he develops a research-intensive clinical practice in spine surgery at the Philadelphia VA Medical Center and the University of Pennsylvania.