ADULT STEM CELL SUPPRESSION DUE TO ALCOHOL INDUCED MICROENVIRONMENT ALTERATIONS

酒精引起的微环境改变对成体干细胞的抑制

• 批准号: 7458223
• 负责人: MANDI J. LOPEZ
• 金额: $ 22.05万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-15 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7458223
• 关键词: Acceleration; Address; Adipose tissue; Affect; Alcohol abuse; Alcohols; American; Animals; Behavior; Biocompatible Materials; Biomedical Research; Cells; Chronic; Clinical Research; Complex; Compression Fracture; Congenital failure of fusion; Ethanol; Face; Failure; Fracture Healing; Functional disorder; Harvest; Healed; Histology; Immunohistochemistry; Implant; In Vitro; Incidence; Investigation; Knowledge; Louisiana; Medical; Modality; Modeling; Numbers; Operative Surgical Procedures; Osteoblasts; Osteogenesis; Osteoporosis; Outcome Measure; Patients; Procedures; Public Health; Rate; Rattus; Regenerative Medicine; Reverse Transcriptase Polymerase Chain Reaction; Scientist; Spinal Fractures; Spinal Fusion; Standards of Weights and Measures; Stem cells; Stromal Cells; System; Testing; Tissues; Universities; Veterinary Medicine; Veterinary Schools; Work; adult stem cell; alcohol exposure; alcoholism/alcohol abuse; base; chronic alcohol ingestion; cost; healing; in vivo; novel therapeutics; prevent

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.

描述（由申请人提供）：酗酒和酗酒会影响至少1400万美国人。慢性酒精消耗与骨质疏松症和腰椎骨折率增加有关。后外侧脊柱融合是乙醇诱导的骨质疏松症常见的腰压缩骨折的标准治疗方法。慢性酒精暴露会抑制骨折的愈合，并导致脊柱融合失败的发生率明显更高。 2003年，与修订脊柱融合手术相关的成本高达72,000美元。再生医学是一种新型的治疗方式，它采用干细胞来促进组织愈合。脂肪组织衍生的基质细胞（ASC）在体内和体外都会促进成骨。干细胞与微环境之间的复杂相互作用是增强成骨的必要条件，而慢性酒精暴露的变化可以限制其综合功效。基于这些知识，我们假设：（a）长期摄入饮酒可减少ASC的数量，膨胀率和全权能力； （b）受体中的慢性酒精暴露会抑制或通过在合适的生物材料载体中应用正常ASC来阻止脊柱融合的加速； （c）从慢性酒精暴露受试者中收获并在合适的生物材料载体中收获的ASC不会在正常受体中加速脊柱融合。这些假设将以以下特定目的进行测试：目标1。确定慢性酒精摄入对大鼠模型中ASC的影响。研究将对从正常大鼠和暴露于已建立的慢性酒精摄入模型的大鼠收获的细胞进行研究。在体外扩展和传递的干细胞的数量和行为将通过标准程序进行量化。目标2。确定正常ASC对大鼠有或没有慢性酒精暴露的大鼠脊柱融合的影响。研究将使用从没有酒精暴露的受试者中收获的ASC进行研究，并没有慢性酒精暴露。 X光片，Micro-CT，RT-PCR，组成分析，组织学和免疫组织化学的结果度量将提供围绕正常干细胞在正常微环境中诱导正常微环境与微观环境诱导成骨的能力的信息。目标3。确定从大鼠慢性酒精暴露的大鼠从大鼠中收获的ASC对大鼠有或没有慢性酒精暴露的影响。将对从慢性酒精暴露受试者中植入的慢性酒精暴露受试者收获的ASC进行研究。与AIM 2相同的结果度量将提供围绕慢性酒精暴露于诱导正常与微环境与经慢性酒精暴露改变的微环境诱导成骨的能力的信息。这项研究的结果将大大提高围绕酒精诱导的干细胞和抑制骨形成的基质改变的知识。这项研究的结果将大大促进围绕酒精引起的干细胞和抑制骨形成的基质改变的知识。公共卫生相关性：面对慢性酒精滥用而导致脊柱融合失败的病理生理学的阐明将提供解决这一重要医疗问题的机制。此外，ASC应用是一种有前途的治疗方法，可能会加速与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。