ADULT STEM CELL SUPPRESSION DUE TO ALCOHOL INDUCED MICROENVIRONMENT ALTERATIONS

描述（由申请人提供）：酗酒和酒精滥用影响着至少1400万美国人。慢性饮酒与骨质疏松症和腰椎骨折发生率增加有关。后外侧脊柱融合术是乙醇性骨质疏松症腰椎压缩性骨折的标准治疗方法。慢性酒精暴露抑制骨折愈合，导致脊柱融合失败的发生率显著升高。2003年脊柱融合翻修手术的费用高达72000美元。再生医学是一种利用干细胞促进组织愈合的新型治疗方式。脂肪组织源性基质细胞（ASCs）在体内和体外均能促进成骨。干细胞和微环境之间复杂的相互作用是促进成骨所必需的，而慢性酒精暴露导致的任何一种改变都可能限制它们的联合功效。基于这些知识，我们假设：(A)长期饮酒会减少ASCs的数量、扩张速度和胸膜电位容量；(B)受体的慢性酒精暴露抑制或阻止通过在合适的生物材料载体中应用正常ASCs来加速脊柱融合；(C)从慢性酒精暴露受试者身上采集的ASCs并将其应用于合适的生物材料载体中，不会加速正常受体的脊柱融合。这些假设将通过以下具体目标进行检验：目的1。目的：探讨慢性酒精摄入对大鼠ASCs的影响。研究将在从正常大鼠和暴露于已建立的慢性酒精摄入模型的大鼠身上采集的细胞上进行。干细胞在体外扩增和传代的数量和行为将用标准程序进行量化。目标2。目的：探讨正常ASCs对慢性酒精暴露和非慢性酒精暴露大鼠脊柱融合的影响。研究将从没有酒精暴露的受试者身上采集ASCs植入有和没有慢性酒精暴露的受试者体内。x线片、微型ct、RT-PCR、成分分析、组织学和免疫组织化学的结果测量将提供有关正常干细胞在正常微环境与慢性酒精暴露改变的微环境中诱导成骨能力的信息。目标3。确定慢性酒精暴露大鼠的ASCs对慢性酒精暴露和非慢性酒精暴露大鼠脊柱融合的影响。研究将从慢性酒精暴露者身上采集ASCs植入慢性酒精暴露者和非慢性酒精暴露者体内。与Aim 2相同的结果测量将提供有关慢性酒精暴露改变的干细胞在正常环境与慢性酒精暴露改变的微环境中诱导成骨的能力的信息。这项研究的结果将大大提高对酒精诱导的抑制骨形成的干细胞和基质改变的认识。这项研究的结果将大大有助于当前有关酒精诱导的干细胞和基质改变抑制骨形成的知识。公共卫生相关性：阐明慢性酒精滥用导致脊柱融合失败的病理生理学将为解决这一重要医学问题提供机制。此外，ASC应用是一种很有希望的治疗方法，可以加速与酒精暴露无关的天然成骨细胞前体数量减少的患者的骨折愈合。

DESCRIPTION (provided by applicant): Alcoholism and alcohol abuse affect at least 14 million Americans. Chronic alcohol consumption is associated with osteoporosis and an increased rate of lumbar spinal fractures. Posterolateral spinal fusion is the standard treatment for lumbar compression fractures common in ethanol-induced osteoporosis. Chronic alcohol exposure inhibits fracture healing and results in a significantly higher incidence of spinal fusion failures. Costs associated with revision spinal fusion surgery were as high as $72,000 each in 2003. Regenerative medicine is a novel therapeutic modality that employs stem cells to promote tissue healing. Adipose tissue-derived stromal cells (ASCs) promote osteogenesis both in vivo and in vitro. Complex interactions between stem cells and the microenvironment are necessary to enhance osteogenesis, and alterations in either from chronic alcohol exposure can limit their combined efficacy. Based on this knowledge we postulate: (A) That chronic alcohol ingestion reduces the number, rate of expansion, and pleuripotential capacity of ASCs; (B) That chronic alcohol exposure in the recipient inhibits or prevents the acceleration of spinal fusion by application of normal ASCs in a suitable biomaterial carrier; and (C) That ASCs harvested from subjects with chronic alcohol exposure and applied in a suitable biomaterial carrier do not accelerate spinal fusion in normal recipients. These hypotheses will be tested with the following Specific Aims: Aim 1. To determine the effect of chronic alcohol ingestion on ASCs in a rat model. Studies will be performed on cells harvested from normal rats and rats exposed to an established model of chronic alcohol ingestion. The number and behavior of stem cells expanded and passaged in vitro will be quantified with standard procedures. Aim 2. To determine the effect of normal ASCs on spinal fusion in rats with and without chronic alcohol exposure. Studies will be performed with ASCs harvested from subjects with no alcohol exposure implanted into subjects with and without chronic alcohol exposure. Outcome measures of radiographs, micro-CT, RT-PCR, compositional analysis, histology, and immunohistochemistry will provide information surrounding the ability of normal stem cells to induce osteogenesis in normal microenvironments versus microenvironments altered by chronic alcohol exposure. Aim 3. To determine the effect of ASCs harvested from rats with chronic alcohol exposure on spinal fusion in rats with and without chronic alcohol exposure. Studies will be performed with ASCs harvested from subjects with chronic alcohol exposure implanted into subjects with and without chronic alcohol exposure. Outcome measures identical to those of Aim 2 will provide information surrounding the ability of stem cells altered by chronic alcohol exposure to induce osteogenesis in normal versus microenvironments altered by chronic alcohol exposure. Results from this investigation will substantially advance knowledge surrounding alcohol-induced stem cell and matrix alterations that inhibit bone formation. Results from this investigation will contribute substantially to current knowledge surrounding alcohol-induced stem cell and matrix alterations that inhibit bone formation. PUBLIC HEALTH RELEVANCE: Elucidation of the pathophysiology contributing to failure of spinal fusion in the face of chronic alcohol abuse will provide mechanisms to address this important medical problem. Additionally, ASC application is a promising treatment that may accelerate fracture healing in patients with reduced numbers of native osteoblast precursors unrelated to alcohol exposure.