Name: a morphometric and cellular analysis method for the murine mandibular condyle
Uploaded: 2018-01-11T12:30:00
Description: Watch this Scientific Journal Video about A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle at JoVE.com

作者想感谢 Dr. 的好心提供了转基因小鼠和李 Chen 的组织学帮助。
该出版物的研究报告得到了国立卫生研究院的牙科 & 颅面研究协会的支持, 该研究所的奖项编号 K08DE025914 和美国正畸基金会素密亚达夫。

康涅狄格大学卫生中心的机构动物保育委员会批准了所有的动物程序。
1. 压缩静态颞下颌关节负荷: 口强行打开
注: 四周老的转基因小鼠窝藏荧光记者的胶原蛋白 (Col2a1XCol10a1), 亲切提供由 Dr. 大卫罗 (康涅狄格大学), 被用于本手稿描述的实验 (n = 8; 4 男性和4女性)。Col2a1 青色 (蓝色) 转基因是在 MCC 的 prehypertrophic 区的细胞中表达的, 而 Col10a1 樱桃 (红) 细胞存?...

<ol>
	<li>LeResche, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+Temporomandibular+Disorders:+Implications+for+the+Investigation+of+Etiologic+Factors.">Epidemiology of Temporomandibular Disorders: Implications for the Investigation of Etiologic Factors.</a> Crit Rev Oral Biol Med. 8 (3), 291-305 (1997).</li><li>Chen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+functional+loading+causes+differential+effects+in+the+subchondral+bone+and+condylar+cartilage+in+the+temporomandibular+joint+from+young+mice.">Altered functional loading causes differential effects in the subchondral bone and condylar cartilage in the temporomandibular joint from young mice.</a> Osteoarthr Cartil. 17 (3), 354-361 (2009).</li><li>Pirttiniemi, P., Kantomaa, T., Sorsa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+decreased+loading+on+the+metabolic+activity+of+the+mandibular+condylar+cartilage+in+the+rat.">Effect of decreased loading on the metabolic activity of the mandibular condylar cartilage in the rat.</a> Eur J Orthod. 26 (1), 1-5 (2004).</li><li>Chavan, S. J., Bhad, W. A., Doshi, U. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+temporomandibular+joint+changes+in+Twin+Block+and+Bionator+appliance+therapy:+a+magnetic+resonance+imaging+study.">Comparison of temporomandibular joint changes in Twin Block and Bionator appliance therapy: a magnetic resonance imaging study.</a> Prog Orthod. 15 (57), (2014).</li><li>Dutra, E. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+and+Matrix+Response+of+the+Mandibular+Condylar+Cartilage+to+Botulinum+Toxin.">Cellular and Matrix Response of the Mandibular Condylar Cartilage to Botulinum Toxin.</a> PLoS ONE. 11 (10), 0164599 (2016).</li><li>Benjamin, M., Ralphs, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biology+of+fibrocartilage+cells.">Biology of fibrocartilage cells.</a> Int Rev Cytol. 233, 1-45 (2004).</li><li>Shen, G., Darendeliler, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Adaptive+Remodeling+of+Condylar+Cartilage-+A+Transition+from+Chondrogenesis+to+Osteogenesis.">The Adaptive Remodeling of Condylar Cartilage- A Transition from Chondrogenesis to Osteogenesis.</a> J Dent Res. 84 (8), 691-699 (2005).</li><li>Utreja, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+and+matrix+response+of+temporomandibular+cartilage+to+mechanical+loading.">Cell and matrix response of temporomandibular cartilage to mechanical loading.</a> Osteoarthr Cartil. 24 (2), 335-344 (2016).</li><li>Kaul, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effect+of+Altered+Loading+on+Mandibular+Condylar+Cartilage.">The Effect of Altered Loading on Mandibular Condylar Cartilage.</a> PLoS ONE. 11 (7), 0160121 (2016).</li><li>Dyment, N. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-Throughput,+Multi-Image+Cryohistology+of+Mineralized+Tissues.">High-Throughput, Multi-Image Cryohistology of Mineralized Tissues.</a> J Vis Exp. , e54468 (2016).</li><li>Kawamoto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+new+adhesive+film+for+the+preparation+of+multi-purpose+fresh-frozen+sections+from+hard+tissues,+whole-animals,+insects+and+plants.">Use of a new adhesive film for the preparation of multi-purpose fresh-frozen sections from hard tissues, whole-animals, insects and plants.</a> Arch Histol Cytol. 66 (2), 123-143 (2003).</li><li>Hayman, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tartrate-resistant+acid+phosphatase+(TRAP)+and+the+osteoclast/immune+cell+dichotomy.">Tartrate-resistant acid phosphatase (TRAP) and the osteoclast/immune cell dichotomy.</a> Autoimmunity. 41 (3), 218-223 (2008).</li></ol>

本手稿描述了小鼠下颌髁和下颌骨的形态测量和细胞分析方法。射线照相测量也可以用来分析其他的骨骼从小实验动物。此外, 细胞分析 (细胞定量和软骨距离映射) 不限于啮齿动物的下颌骨髁突, 但可用于量化组织切片的众多组织上。
表达荧光的转基因小鼠模型是视觉理解基因表达变化的绝佳工具。本报告中使用的 double-collagen 荧光报道鼠模型 (Col2a1XCol10a1) 特别适合于 mcc 的?...

颞下颌关节是一个独特的 load-bearing 关节位于颅面地区, 是由软骨。颞下颌关节的 MCC 对于关节功能是必不可少的, 包括在说话和咀嚼时不受阻碍的颚运动, 但它通常受到退行性疾病的影响, 包括骨关节炎1。颞下颌关节有能力适应外部刺激和负荷改变, 导致结构和细胞的变化的组成部分的 MCC2,3,4,5。MCC 的 load-bearing 性能可以通过其成分之间的相互作用来解释, 包括水、胶原网络和密被包装的蛋白。MCC 有四独特的细胞区, 表达不同类型的胶原蛋白和 non-collagen 蛋白: 1) 表面或关节区;2) 增殖区, 由未分化的间充质细胞组成, 并对负荷需求作出反应;3) prehypertrophic 区, 由成熟的软骨细胞表达2型胶原;和 4) 肥厚区, 该地区的肥大软骨细胞表达型胶原10死亡, 并经历钙化。non-mineralized 地区是丰富的蛋白, 提供抗压缩力6。
在 MCC 的肥厚区有连续的矿化, 在那里从软骨到成骨的转变发生, 保证下颌髁突的下骨的健壮的矿物结构7。unmineralized 和矿化区的细胞变化最终导致下颌骨髁突和下颌骨的形态学和结构改变。维持 MCC 所有细胞区域的稳态和下部分的矿化, 对颞下颌关节的健康、load-bearing 能力和完整性至关重要。
多胶原蛋白转基因小鼠模型 (如 Utreja et al.所述)8是一个很好的工具, 可以用来理解胶原蛋白表达的变化, 因为所有的转基因都是在 MCC 中表达的。对于 in-depth 的组织学评价, 组织学染色用于研究基质沉积、矿化、细胞增殖和凋亡, 以及在 MCC 不同细胞层的蛋白质表达。
在这篇手稿中, 组织学和形态学分析被用来评价细胞和结构的变化, 在 MCC 和下骨的下颌髁突的小鼠。此外, 还描述了用于分析荧光组织学图像和绘制光学显微镜幻灯片的细胞定量方法。压缩静态颞下颌关节加载方法, 导致细胞和形态学的变化, 在 MCC 和下骨9, 也说明了验证我们的方法。
本文所述方法可用于确定鼠下颌骨髁突和下颌骨的形态和组织学变化, 或分析其他软骨骨化区和其他矿化组织的形态。

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a morphometric and cellular analysis method for the murine mandibular condyle

颞下颌关节具有适应外部刺激的能力, 负荷变化会影响髁的位置, 以及下颌髁突软骨的结构和细胞成分。本文介绍了分析这些变化的方法和改变小鼠颞下颌关节负荷的方法 (即,压缩静态颞下颌关节负荷)。这里的结构评估是一个简单的形态学方法, 使用 Digimizer 软件, 并在小骨骼的射线照相中进行。此外, 分析了细胞的变化导致胶原蛋白表达改变, 骨重塑, 细胞分裂, 和蛋白多糖分布的 MCC。在组织学切片的这些变化的量化-通过计数的阳性荧光像素使用图像软件和测量的距离测绘和染色面积与 Digimizer-也被证明。这里所示的方法不仅限于小鼠颞下颌关节, 还可用于小型实验动物和其他软骨骨化地区的骨骼。

进行描述性统计, 检查形态学测量的分布 (下颌长度, 髁突长度, 髁突宽度) 和组织学分析。结果被比较了被装载的小组 (即,小鼠受压缩装载与 beta 钛春天) 和控制小组 (即,匹配控制小鼠没有接受任何过程)。统计学意义上的差异之间的方法是由不配对 t 检验, 和 p 值的 < 0.05 被认为是有统计学意义的。
如步骤4和<strong ...

Watch this Scientific Journal Video about A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle at JoVE.com

A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle

本文介绍了分析啮齿动物下颌骨髁突内形态和细胞变化的方法。

小鼠下颌骨髁突的形态学及细胞分析方法

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