作者要感谢杰米翠的技术工作。这项工作是支持的，部分由研究资助SC CTSI NIH / NCRR / NCATS - UL1TR000130（DLD），AA022448（DLD）和药房的南加州大学。此外，含IVM快速溶解口服片经固化药业（奥克斯纳德，CA）的友情提供。

描述的所有程序已经批准了南加州大学的机构动物护理和使用委员会（IACUC）。 1.动物处理和习惯化<ol><li>介绍<ol><li>允许小鼠适应的饲养条件和处理实验技术，在随后的给药方法减少压力。在此期间，记录体重和各小鼠的食物摄入;如果需要的话，它们可以在以后用作非侵入点比较，以评估药物毒性（可选）的。前者也可用于确定基于重量的药物剂量。 注意:有趣的是，应根据实验者与动物相互作用的数目来确定的习惯期的长短。根据我们的经验，在C57BL / 6J品系小鼠已观察到有麻烦我们典型的为期一周的习惯时间内超过〜5人habituating搬运。如果可行的话，我们也鼓励实验者（S）使用香味乳液，香皂，香水和处理动物之前与以前的报告表明新的刺激，如气味，可能会阻碍习惯11期不要。 </li></ol></li><li>脚步<ol><li>研究开始前至少5天开始，将动物转移到在其中随后的实验将发生室。 </li><li>牢牢抓住尾巴的中间，轻轻提起动物出笼子，进入重船记录鼠标的重量。 <ol><li>一旦放置在标尺上的鼠标，因为它是在记提升出cagekeeping的适当掌握尾部的内侧/近侧方面中，以避免脱套伤，抓尾部12的远侧方面时可能发生。 </li><li>记录的权重值，一旦老鼠已被安全地放置在权衡船。 </li></ol></li><li>记录食物的重量为RECOM谁料连续2天，以建立基线的食品消费水平将会在实验过程中13帮助捕获潜在的毒性。 </li></ol></li></ol> 2.准备喂食针的，蔗糖溶液，与个人ODS <ol><li>使用高压釜灭菌的不锈钢管饲针头在用于实验的准备。准备2根/基和1个另外的针被用作后备如果需要的话。 注意:此研究利用1.5英寸，弯曲的，18 G，不锈钢在针的末端馈送带有2.25毫米球针。 </li><li> 20ml水加至0.85克蔗糖以创建一个4.25％（重量/体积）的蔗糖溶液。 注:根据正在评估其他低剂量蔗糖的参数可能也可使用14。 </li><li>通过切割标准的矩形试片成片是在直径为0.5厘米英寸制备每个动物的口腔溶解条（ODS）我们ING市售的单季英寸的孔，打孔机。在此演示中使用矩形试验片制成含有6毫克伊维菌素（IVM），并切成此大小时，分配一个0.21毫克剂量将每个独立的圆形片近似10毫克/公斤的剂量为每只动物（我们的期望的剂量/动物）。此剂量可通过浓缩/在矩形条稀释整体制剂和/或切断所述带状成不同尺寸来调整。 </li></ol> 3.药物 - 送货<ol><li>如先前在步骤1.1-1.2中所述记录重量值。 </li><li>记录重量后，将鼠标移动到金属网格笼子顶部。一旦出现，使用占主导地位的拇指，食指，和第三的手指，轻轻地拉回来的尾巴。这将导致鼠标无意中夹住金属丝笼顶部和伸出它的后面，伸展它的身体从实验者路程。 </li><li>把握颈背与非待办事项minant拇指，食指，和第三手指舒适地抑制所述动物用把手是安全。 注意:用较小的手的实验者也可以使用4 个手指，同一只手的，拉回背面的颈背，将获得的小鼠的更安全的保持状态。另外，根据个人喜好，无名指和/或小指也可以用于固定尾巴然而这是没有必要的12。 </li><li>按住鼠标在直立位置。 </li><li>注意:观察小鼠的粘膜的颜色。紫色/蓝色的外观是一个迹象表明实验者得太紧收紧颈背和鼠标无法呼吸。 <ol><li>如果发生这种情况，立即把鼠标放在铁笼顶部回落，并允许它重新尝试抑制动物之前休息2-3〜分钟。 </li></ol></li><li>浸没灌胃针的灯泡尖端4.25％蔗糖溶液以用作促味剂粘合剂的消耗臭氧层物质。 </li><li>向下按压与灌胃针的尖泡一个预切的消耗臭氧层物质的口腔溶解条连接。 </li><li>通过将其鼻孔和/或口附近目前的消耗臭氧层物质的老鼠，并让动物消耗薄膜。可替代地，ODS可以通过直接放置在薄带到舌头的小鼠吞局部递送到口腔。 </li><li>药物剂量会期间更换针头中期组，以防止感染扩散。 </li></ol>

<ol>
	<li>Brown, A. P., Dinger, N., Levine, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress+produced+by+gavage+administration+in+the+rat.">Stress produced by gavage administration in the rat.</a> Contemp Top Lab Anim Sci. 39 (1), 17-21 (2000).</li><li>Gonzales, C., 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=Alternative+method+of+oral+administration+by+peanut+butter+pellet+formulation+results+in+target+engagement+of+BACE1+and+attenuation+of+gavage-induced+stress+responses+in+mice.">Alternative method of oral administration by peanut butter pellet formulation results in target engagement of BACE1 and attenuation of gavage-induced stress responses in mice.</a> Pharmacol Biochem Behav. 126, 28-35 (2014).</li><li>Walker, M. 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DD已经申请了专利作为这个方法的共同发明人，并有兴趣与科学委员会分享这一方法，使这项技术众所周知的，随时可用于慢性动物试验（ 在实验动物药物活性成分的口服给药使用Fast-溶解口服电影 （戴维斯，共同发明人，专利申请提交2014年6月23日）。 没有其他竞争经济利益的说明。

口管饲法是与导致潜在的损害的数据收集和动物减员1的高速率许多并发症。此处的口服药物递送的一个简单的方法被引入，通过该小鼠（和潜在的其他动物）可容易且可靠地通过口服溶解条（ODS）消耗的首选药物。值得注意的是，使用这种方法的代表安全，方便，和人性化的替代口服管饲法。 其他流动资产替代口服灌胃，虽然有点希望的，与该妨碍他们覆盖了全方?...

为了使药物从临床前（动物试验）成功翻译到临床发展，一系列的明确定义，急性和慢性的药理学和毒理学研究需要在动物模型中也可以执行使用管理的预期临床路径（这是通常口服途径）。为了实现这个目标，最临床前研究目前利用给定的口服管饲法程序，该方法在测试化合物对动物的一致和精确的递送的效果。然而，在许多情况下，经口管饲法不能很好被动物耐受，越来越多的证据表明，该方法是用应力感应1的显著量相关联。 这是在用于需要长期的临床前研究慢性终身的条件下，如治疗的发展尤其如此与的情况下，但不限于，几乎每一个神经精神障碍。对于插件tance，在阿尔茨海默病的转基因小鼠模型，最近建议，强饲法本身可以由于加药引起的应力2混淆的实验结果。同样地，在酒精消费的小鼠模型中，我们的实验室未发表的研究结果已经发现，强饲管的重复插入 - 在其本身和 - 可以显著酒精摄入的水平减少到危及实验的完整性的点范例（ 见图1）。 定与口腔管饲过程相关联的上述限制，相当大的努力已投入朝着显影临床前口服药物递送的新方法。当前备选的方法包括掺入试验化合物成花生酱的混合物3，凝胶状模具4-5，巧克力小球6，饮用水7和晶片饼干8，所有这些都是associ有不同程度的问题，包括无法ated在动物模型可靠地使用，缺乏通过试验化合物的摄取的被测试受试者或风味个人偏好，使药物不易消耗9。 本文所介绍的过程描述了在其中需要的药物掺入调味，口腔溶解条（ODS），可以很容易地和很容易地被用于口服施用测试化合物对测试动物的方法。本文侧重于展示利用老鼠进行的ODS方法的有效性。然而，没有理由认为同样的方法将不会也有在其他啮齿动物，以及较大的哺乳动物受试者是有用的。临床上，使用消耗臭氧层物质的开始在儿童和老年患者被采纳，以此来克服吞咽困难10。我们建议，消耗臭氧层物质，也可以成功地在临床前药物研究计划作为一种安全，方便使用和人道的替代口服灌胃。

<table border="0" cellpadding="0" cellspacing="0" width="948">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Orally Dissolving Strip&#160;</td>
			<td style="width:185px;">Cure Pharmaceutical</td>
			<td style="width:89px;"></td>
			<td style="width:471px;">Available upon special request. Alternatively, ODS may also be prepared in-house.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Gavage Feeding Needle</td>
			<td style="width:185px;">VWR</td>
			<td align="right" style="width:89px;">7912</td>
			<td>Our rodent studies utilized a 18-gauge feeding needle. This size may need to be adjusted for use in other mammilian species.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Sucrose</td>
			<td style="width:185px;">Sigma Aldrich</td>
			<td style="width:89px;">S0389-500G</td>
			<td></td>
		</tr>
	</tbody>
</table>

utilizing an orally dissolving strip for pharmacological and toxicological studies: a simple and humane alternative to oral gavage for animals

之前在人类中测试新的治疗，短期和长期的临床前（ 即动物），重复的药理和毒理试验是必需的。在大多数情况下，给药的优选途径是通过口服给药。在当前时刻，口服递送是使用口服管饲法过程，部分是因为它可以实现在动物模型一致和精确的定量给料大多完成。虽然这种方法是公认它确实有并发症，可能导致动物的高损耗率。为此，这里所介绍的过程描述，可以简单地呈现给测试动物在那里，然后迅速采取与替代，其中所希望的药物被掺入一个促味剂，口腔溶解条（ODS）的经口管饲法被检者的最小操纵。在此，我们证明，使用消耗臭氧层物质的方法临床前，口服给药代表了安全，便捷，人性化和阿尔特rnative口腔灌胃。

在下面的代表调查，社交饮酒是使用24小时双瓶选择（TBC）模式如前所述15建模。简言之，将小鼠获得的溶液两瓶，其中一个含有水，另10％（体积/体积）乙醇溶液中。受试者被随后分配到药物处理组，使得平均10E（10％乙醇/ 90％水体积/体积）的摄入量是跨组相似。 图1示出了当使用一个管饲车辆口服偏好...

Watch this Scientific Journal Video about Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals at JoVE.com

Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals

以下方案描述了使用代替更常用的经口管饲法口服溶解条（ODS）慢性口服药物给药的新方法。我们表明，使用消耗臭氧层物质的方法临床前，口服给药代表了安全，方便，和人性化的替代口服灌胃。

利用一种口腔溶解地带的药理毒理研究:一个简单的和人道的替代口服灌胃动物

Prior to testing novel therapeutics in humans, short and long term preclinical (i.e., animal), repetitive pharmacological and toxicological testing is ...

utilizing-an-orally-dissolving-strip-for-pharmacological

Research

JoVE Journal

Medicine

12.6K 次观看.  University of Southern California School of Pharmacy. 以下方案描述了使用代替更常用的经口管饲法口服溶解条（ODS）慢性口服药物给药的新方法。我们表明，使用消耗臭氧层物质的方法临床前，口服给药代表了安全，方便，和人性化的替代口服灌胃。 

视频: Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

多光子成像在口腔鳞状细胞癌原位小鼠模型的肿瘤细胞的侵袭

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

科研

医学

A Simple Guide Screw Method for Intracranial Xenograft Studies in Mice

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma (glioma) and medulloblastoma. The widely used stereotactic approach only allows for the injection of a single animal at a time, is labor intensive and requires highly specialized equipment. The guide screw method, initially developed by Lal et al.,1 was developed to eliminate cumbersome stereotactic procedures. We now describe a modified guide screw approach that is rapid and exceptionally safe; both of which are critical ethical considerations. Notably, our procedure now incorporates an infusion pump that allows up to 10 animals to be simultaneously injected with tumor cells.
To demonstrate the utility of this procedure, we established human U87MG glioma cells as intracranial xenografts in mice, which were then treated with AMG102; a fully human antibody directed to HGF/scatter factor currently undergoing clinical evaluation2-5. Systemic injection of AMG102 significantly prolonged the survival of all mice with intracranial U87MG xenografts and resulted in a number of complete cures. 
This study demonstrates that the guide screw method is an inexpensive, highly reproducible approach for establishing intracranial xenografts. Furthermore, it provides a relevant physiological model for validating novel therapeutic strategies for the treatment of brain cancers.

In order to evaluate novel therapeutic paradigms for the treatment of glioma, physiological relevant models are essential. We utilize an implantable guide screw procedure for establishment of intracranial xenograft models that is more rapid and safer than stereotactic approaches.

一个简单的指南螺杆颅内移植瘤小鼠的研究方法

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma ...

Sublingual Immunotherapy as an Alternative to Induce Protection Against Acute Respiratory Infections

Sublingual route has been widely used to deliver small molecules into the bloodstream and to modulate the immune response at different sites. It has been shown to effectively induce humoral and cellular responses at systemic and mucosal sites, namely the lungs and urogenital tract. Sublingual vaccination can promote protection against infections at the lower and upper respiratory tract; it can also promote tolerance to allergens and ameliorate asthma symptoms. Modulation of lung&#8217;s immune response by sublingual immunotherapy (SLIT) is safer than direct administration of formulations by intranasal route because it does not require delivery of potentially harmful molecules directly into the airways. In contrast to intranasal delivery, side effects involving brain toxicity or facial paralysis are not promoted by SLIT. The immune mechanisms underlying SLIT remain elusive and its use for the treatment of acute lung infections has not yet been explored. Thus, development of appropriate animal models of SLIT is needed to further explore its potential advantages. 
This work shows how to perform sublingual administration of therapeutic agents in mice to evaluate their ability to protect against acute pneumococcal pneumonia. Technical aspects of mouse handling during sublingual inoculation, precise identification of sublingual mucosa, draining lymph nodes and isolation of tissues, bronchoalveolar lavage and lungs are illustrated. Protocols for single cell suspension preparation for FACS analysis are described in detail. Other downstream applications for the analysis of the immune response are discussed. Technical aspects of the preparation of Streptococcus pneumoniae inoculum and intranasal challenge of mice are also explained.
SLIT is a simple technique that allows screening of candidate molecules to modulate lungs&#8217; immune response. Parameters affecting the success of SLIT are related to molecular size, susceptibility to degradation and stability of highly concentrated formulations.

The present work illustrates the convenience of using sublingual immunotherapy to boost the innate immune response in the lungs and confer protection against acute pneumococcal pneumonia in mouse.

舌下免疫治疗作为一种替代起到保护作用对急性呼吸道感染

Sublingual route has been widely used to deliver small molecules into the bloodstream and to modulate the immune response at different sites. It has been ...

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 PtenL/L p53L/L LSL-mTERTL/L and thymic T-cell lymphoma model G4 Atm-/- mTERTER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.

Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

在组织变性/再生和癌症的研究利用小鼠诱导端粒酶等位基因

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

一个简单的设备，迅速准备小鼠肠的全坐骑

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Development of an Alpha-synuclein Based Rat Model for Parkinson's Disease via Stereotactic Injection of a Recombinant Adeno-associated Viral Vector

In order to study the molecular pathways of Parkinson&#39;s disease (PD) and to develop novel therapeutic strategies, scientific investigators rely on animal models. The identification of PD-associated genes has led to the development of genetic PD models. Most transgenic &#945;-SYN mouse models develop gradual &#945;-SYN pathology but fail to display clear dopaminergic cell loss and dopamine-dependent behavioral deficits. This hurdle was overcome by direct targeting of the substantia nigra with viral vectors overexpressing PD-associated genes. Local gene delivery using viral vectors provides an attractive way to express transgenes in the central nervous system. Specific brain regions can be targeted (e.g. the substantia nigra), expression can be induced in the adult setting and high expression levels can be achieved. Further, different vector systems based on various viruses can be used. The protocol outlines all crucial steps to perform a viral vector injection in the substantia nigra of the rat to develop a viral vector-based alpha-synuclein animal model for Parkinson&#39;s disease.

This manuscript describes how viral vector-mediated local gene delivery provides an attractive way to express transgenes in the central nervous system. The protocol outlines all crucial steps to perform a viral vector injection in the substantia nigra of the rat to develop a viral vector-based animal model for Parkinson's disease.

通过重组腺相关病毒载体注射立体的α-突触核蛋白基于大鼠模型为帕金森氏病的发展

In order to study the molecular pathways of Parkinson&#39;s disease (PD) and to develop novel therapeutic strategies, scientific investigators rely on ...

Isolation and Culture of Primary Endothelial Cells from Canine Arteries and Veins

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study vasculogenesis, (tumor) angiogenesis, and atherosclerosis. The current standard for in vitro research on human endothelial cells (ECs) is the use of Human Umbilical Vein Endothelial Cells (HUVECs) and Human Umbilical Artery Endothelial Cells (HUAECs). For canine endothelial research, only one cell line (CnAOEC) is available, which is derived from canine aortic endothelium. Although currently not completely understood, there is a difference between ECs originating from either arteries or veins. For a more direct approach to in vitro functionality studies on ECs, we describe a new method for isolating Canine Primary Endothelial Cells (CaPECs) from a variety of vessels. This technique reduces the chance of contamination with fast-growing cells such as fibroblasts and smooth muscle cells, a problem that is common in standard isolation methods such as flushing the vessel with enzymatic solutions or mincing the vessel prior to digestion of the tissue containing all cells. The technique we describe was optimized for the canine model, but can easily be utilized in other species such as human.

Novel isolation methods of primary endothelial cells from blood vessels are needed. This protocol describes a new technique that completely inverts blood vessels of interest, exposing only the endothelial side to enzymatic digestion. The resulting pure endothelial cell culture can be used to study cardiovascular diseases, disease modelling, and angiogenesis.

分离培养犬从动脉和静脉主血管内皮细胞

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study ...

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition leads to a better understanding of cariogenic and periopathogenic mechanisms. Microbial changes taking place in the oral cavity during childhood are of interest for several reasons. The evolution of the child oral microbiota and shifts in its composition need to be analyzed further to understand and possibly prevent the onset of disease. At the same time, advanced knowledge of the natural composition of oral biofilm is needed. Early stages of caries-free permanent dentition with healthy gums provide a widely unaffected subgingival habitat that can serve as an in situ baseline for studying features of oral health and disease. Analysis of children's oral biofilm during different stages in life is thus an important theme in the field. Modern molecular analysis methods can provide comprehensive information about the bacterial diversity of such biofilms. To enable microbiota data comparison, it is important to standardize each step in the procedure for molecular data generation. This procedure spans from clinical sampling, Next Generation Sequencing (NGS), bioinformatic data processing, to taxonomic interpretation. One of the most critical factors here is biofilm sampling. Sampling in children is even more challenging in particular due to limited space in subgingival areas. We thus focus on the use of paper points for subgingival sampling. This article provides a detailed protocol for oral biofilm sampling of the subgingival sulcus, the mucosa, and saliva in children.

Changes in the oral microbiome throughout childhood are of growing interest. Comparison of different microbiome studies reveals a lack of standardized sampling protocols. Limited space makes sampling the sound subgingival sulcus of children challenging. Paper point sampling is presented here in detail as the method of choice for this area.

口腔生物膜取样在健康儿童中的微生物分析

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition ...

Oral Health Assessment by Lay Personnel for Older Adults

Oral health is an often-undervalued contributor to overall health. The literature, however, underscores the myriad of systemic diseases influenced by oral health, including type II diabetes, heart disease, and atherosclerosis. Thus, assessments of oral health, called oral screenings, have a significant role in assessing risk of disease, managing disease, and even improving disease by oral care. Here we present a method to assess oral health quickly and consistently across time. The protocol is simple enough for non-oral health professionals such as students, family, and caregivers. Useful for any age of patient, the method is particularly key for older individuals who are often at risk of inflammation and chronic disease. Components of the method include existing oral health assessment scales and inventories, which are combined to produce a comprehensive assessment of oral health. Thus, oral characteristics assessed include intraoral and extraoral structures, soft and hard tissues, natural and artificial teeth, plaque, oral functions such as swallowing, and the impact this oral health status has on the patient&#39;s quality of life. Advantages of this method include its inclusion of measures and perceptions of both the observer and patient, and its ability to track changes in oral health over time. Results acquired are quantitative totals of questionnaire and oral screening items, which can be summed for an oral health status score. The scores of successive oral screenings can be used to track the progression of oral health across time and guide recommendations for both oral and overall health care.

The goal of this protocol is to enable non-dental professionals to assess oral health status for research or health-screening purposes. Aspects assessed include lips, tongue, soft and hard tissues, natural and artificial teeth, oral cleanliness, plaque, swallowing, and impact of oral health on quality of life.

老年人外行人员口腔健康评估

Oral health is an often-undervalued contributor to overall health. The literature, however, underscores the myriad of systemic diseases influenced by oral ...

A Standardized Approach to Extra-Oral and Intra-Oral Digital Photography

Contemporary dentistry mandates a more comprehensive and personalized analysis of each patient. Technological advances in digital photography have played vital roles in diagnostic accuracy, treatment planning, execution of therapies, and outcome evaluations, including esthetic enhancement. Digital photography also provides an excellent platform for patient education, communication, and co-management of cases with other healthcare providers. However, intra-oral photography often faces challenges such as inaccessibility of areas to be captured, different moveable vs. fixed tissues involved, contamination with saliva or blood, and differing illumination needs on various locations. Thus, a more standardized and systematic approach is proposed for intra- and extra-oral documentation via digital photography to overcome the existing technical challenges. The current work will outline the appropriate equipment specifications (camera bodies, macro lens, and flashes), positions and postures of the operator and patients, proper techniques of tissue retraction, the use of appropriate intra-oral mirrors, and the essential elements such as aperture settings (F-stop), ISO, shutter speed, and white balance. This article aims to provide all dental professionals with an approachable linear array of guidelines to produce simplified and standardized visual tools for more efficient and effective documentation.

The present protocol describes a method to capture intra- and extra-oral photographs for digital documentation in the clinical practice of dentistry. The protocol highlights all the critical components involved and the commonly utilized settings to capture dental photographs for clinical documentation.

口腔外和口腔内数码摄影的标准化方法

Contemporary dentistry mandates a more comprehensive and personalized analysis of each patient. Technological advances in digital photography have played ...