Effects of mechanical loading on energy production of intervertebral disc cells

机械负荷对椎间盘细胞能量产生的影响

• 批准号: 8088237
• 负责人: Chun-Yuh Huang
• 金额: $ 7.13万
• 依托单位: UNIVERSITY OF MIAMI CORAL GABLES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8088237
• 关键词: Adenosine Triphosphate; Affect; Anabolism; Applications Grants; Biochemical; Biological Assay; Cells; Chondrocytes; Consumption; Convection; Cytoskeleton; Data; Development; Diffusion; Energy Metabolism; Environment; Exhibits; Extracellular Matrix; Flowcharts; Foundations; Future; Glucose; Glycolysis; Goals; Grant; Human body; In Vitro; Individual; Intervertebral disc structure; Intrinsic factor; Low Back Pain; Malnutrition; Mechanics; Mediating; Metabolism; Motion; Nitric Oxide; Nutrient; Optics; Organ Culture Techniques; Outcome; Outcome Study; Oxygen; Pathway interactions; Play; Process; Production; Proteoglycan; Research; Research Technics; Role; Solid; Stimulus; System; Testing; Theoretical Studies; Tissue Engineering; Tissues; Transmembrane Transport; United States; Work; adult stem cell; articular cartilage; glucose production; improved; interdisciplinary approach; intervertebral disk degeneration; oxygen transport; prevent; public health relevance; solute; vertebra body

DESCRIPTION (provided by applicant): SUMMERY Degeneration of intervertebral disc (IVD) is closely associated with low back pain which afflicts 52 million individuals in the United States [21]. Since IVD is the largest avascular tissue in the human body, vital nutrients (e.g., oxygen and glucose) are delivered by diffusion and convection over a long distance through dense extracellular matrix to IVD cells. Therefore, poor nutrient supply has been suggested as a potential mechanism for disc degeneration [3,5,35]. The long term goals of our research are: (1) to understand mechanobiology of intervertebral disc and the mechanisms of disc degeneration and (2) to develop strategies to avoid or retard disc degeneration. Cells consume oxygen and glucose to produce energy that is an essential component in cellular matrix synthesis [61] for maintaining the integrity of tissue and preventing tissue degeneration. Adenosine triphosphate (ATP) is the major energy form which is mainly generated through glycolysis in the IVD [4]. During body motion, IVDs transmit large loads between bony vertebral bodies. Our recent theoretical study demonstrated that dynamic compression promoted glycolysis within the IVD by enhancing the transport of oxygen and lactate [42,43], suggesting that energy production of IVD cells can be promoted extrinsically by dynamic loading. Furthermore, previous studies showed that mechanical loading altered membrane transport of glucose and production of nitric oxide [8,16,17,62] which may affect cellular energy production intrinsically. The major objectives of this proposal are to examine (1) the overall effects of dynamic and static compression on ATP production of the IVD cells in the whole disc culture and (2) the intrinsic effect of mechanical loading on ATP production of the IVD cells. The hypotheses are proposed as the followings: (1) Cellular ATP production and transport of lactate and oxygen are promoted in the IVD under dynamic loading whereas static compression exhibits the reverse effect; and (2) Dynamic and static compression influence ATP production of IVD cells intrinsically. To test these hypotheses, we will (1) determine the concentrations of ATP, oxygen, and lactate in the IVD under static and dynamic compression (Specific Aim 1) and (2) determine the intrinsic effects of dynamic and static compression on the ATP production of IVD cells (Specific Aim 2). To achieve these specific aims, we will construct an organ culture system to provide an in-vitro culture environment and mechanical stimuli for IVD. Biochemical assays will be performed to assess concentrations of lactate and ATP in the IVD while oxygen concentration will be determined by an optic sensing system. The outcomes of the proposed studies will improve our understanding of cellular energy metabolism and nutrient transport in the IVD under mechanical loading and facilitate the development of new strategies to prevent disc degeneration. PUBLIC HEALTH RELEVANCE: Degeneration of intervertebral disc (IVD) is closely associated with low back pain which afflicts 52 million individuals in the United States. Poor nutrient supply has been suggested as a potential mechanism for disc degeneration. The outcomes of the proposed studies will improve our understanding of cellular energy metabolism and nutrient transport in the IVD under mechanical loading and facilitate the development of new strategies to prevent disc degeneration.

描述(由申请人提供)： 夏季椎间盘退行性变(IVD)与腰痛密切相关，在美国有5200万人受到这种疼痛的困扰[21]。由于IVD是人体内最大的无血管组织，重要的营养物质(如氧气和葡萄糖)通过致密的细胞外基质远距离扩散和对流输送到IVD细胞。因此，营养供应不足被认为是椎间盘退变的潜在机制[3，5，35]。我们研究的长期目标是：(1)了解椎间盘的机制生物学和退变的机制；(2)开发避免或延缓退变的策略。细胞消耗氧气和葡萄糖来产生能量，这是细胞基质合成[61]中维持组织完整性和防止组织退化的重要组成部分。三磷酸腺苷(ATP)是IVD的主要能量形式，主要通过糖酵解产生[4]。在身体运动过程中，IVD在骨性椎体之间传递大量载荷。我们最近的理论研究表明，动态压缩通过促进氧和乳酸的运输来促进IVD内的糖酵解[42，43]，这表明动态负荷可以促进IVD细胞的能量产生。此外，以前的研究表明，机械负荷改变了葡萄糖的膜运输和一氧化氮的产生[8，16，17，62]，这可能从本质上影响细胞能量的产生。本研究的主要目的是研究(1)动态和静态压缩对IVD细胞产生ATP的整体影响，以及(2)机械负荷对IVD细胞产生ATP的内在影响。提出的假设如下：(1)在动态负荷下，IVD细胞的ATP生成和乳酸和氧的运输被促进，而静态压缩则相反；(2)动态和静态压缩对IVD细胞的ATP产生有内在的影响。为了验证这些假设，我们将(1)确定静态和动态压缩下IVD中的ATP、氧气和乳酸的浓度(特定目标1)和(2)确定动态和静态压缩对IVD细胞产生ATP的内在影响(特定目标2)。为了实现这些特定的目标，我们将构建器官培养系统，为IVD提供体外培养环境和机械刺激。将进行生化分析，以评估IVD中的乳酸和ATP浓度，而氧浓度将通过光学传感系统确定。这些研究结果将加深我们对机械负荷下IVD中细胞能量代谢和营养物质运输的理解，并有助于开发新的预防椎间盘退变的策略。 公共卫生相关性： 椎间盘退行性变(IVD)与腰痛密切相关，在美国有5200万人受到腰痛的困扰。营养供应不足被认为是腰椎间盘退变的一个潜在机制。这些研究结果将加深我们对机械负荷下IVD中细胞能量代谢和营养物质运输的理解，并有助于开发新的预防椎间盘退变的策略。

项目成果

Optical ATP biosensor for extracellular ATP measurement.

• DOI: 10.1016/j.bios.2012.12.027
• 发表时间: 2013-05-15
• 期刊: BIOSENSORS & BIOELECTRONICS
• 影响因子: 12.6
• 作者: Wang, C.;Huang, C. -Y. C.;Lin, W-C
• 通讯作者: Lin, W-C

Measurement of ATP-Induced Membrane Potential Changes in IVD cells.

• DOI: 10.1007/s12195-014-0355-6
• 发表时间: 2014-12-01
• 期刊: CELLULAR AND MOLECULAR BIOENGINEERING
• 影响因子: 2.8
• 作者: Gonzales, Silvia;Rodriguez, Brittany;Barrera, Carlos;Huang, Chun-Yuh Charles
• 通讯作者: Huang, Chun-Yuh Charles