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National Clinical Research Center of Stomatology

12 ARTICLES PUBLISHED IN JoVE

Medicine

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis (ALS)

Yana Yunusova^1,2, Jordan R. Green³, Jun Wang³, Gary Pattee⁴, Lorne Zinman^2,5

¹Department of Speech-Language Pathology, University of Toronto, ²ALS/ MN Clinic, Sunnybrook Health Science Centre, ³Department of Special Education and Communication Disorders, University of Nebraska-Lincoln, ⁴Department of Neurology, Munroe-Meyer Institute, University of Nebraska Medical Center, ⁵Department of Neurology, University of Toronto

Objective assessments of the physiological mechanisms that support speech are needed to monitor disease onset and progression in persons with ALS and to quantify treatment effects in clinical trials. In this video, we present a comprehensive, instrumentation-based protocol for quantifying speech motor performance in clinical populations.

Chemistry

Flow-pattern Guided Fabrication of High-density Barcode Antibody Microarray

Lisa S. Ramirez^1,2, Jun Wang^1,2

¹Department of Chemistry, University at Albany, State University of New York, ²Multiplex Biotechnology Laboratory, Cancer Research Center

This protocol outlines the fabrication of a large-scale, multiplexed two-dimensional DNA or antibody array, with potential applications in cell signaling studies and biomarker detection.

Medicine

Construction and Evaluation of a Murine Calvarial Osteolysis Model by Exposure to CoCrMo Particles in Aseptic Loosening

Hui Jiang^*1, Yicun Wang^*1, Zhantao Deng^*1,2,3, Jiewen Jin^2,3, Jia Meng¹, Shuo Chen¹, Jun Wang¹, Yang Qiu¹, Ting Guo¹, Jianning Zhao¹

¹Department of Orthopedics, Jinling Hospital, School of Medicine, Nanjing University, ²Center for Translational Medicine, Nanjing University Medical School, ³Jiangsu Key Laboratory for Molecular Medicine, Nanjing University Medical School

This manuscript describes a murine calvarial osteolysis model by exposure to CoCrMo particles, which constitutes an ideal animal model for assessing the interactions between wear particles and various cells in aseptic loosening.

Developmental Biology

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation

Qinggang Dai^*1, Yujiao Han^*2, Furong Xie^*1, Xuhui Ma³, Zhan Xu², Xiao Liu¹, Weiguo Zou², Jun Wang¹

¹Department of Pediatric Dentistry, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research Center of Stomatology, ²State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, ³Department of Oral and Maxillofacial-Head and Neck Oncology, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine

This manuscript describes a protocol to isolate and culture osteoclasts in vitro from mouse bone marrow, and to study the role of the mammalian/mechanistic target of rapamycin complex 1 in osteoclast formation.

Chemistry

Fabrication of Spherical and Worm-shaped Micellar Nanocrystals by Combining Electrospray, Self-assembly, and Solvent-based Structure Control

Xinyi Ding^1,2,3, Yuxiang Sun^1,2,3, Yanming Chen^1,2,3, Wanchuan Ding^1,2,3, Steven Emory^1,2,3,4, Tianhao Li^1,2,3, Zixing Xu^1,2,3, Ning Han^1,2,3, Jun Wang^1,2,3, Gang Ruan^1,2,3

¹Department of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing University, ²Institute of Materials Engineering, College of Engineering and Applied Sciences, Nanjing University, ³Collaborative Innovation Center of Chemistry for Life Sciences, Nanjing University, ⁴Department of Chemistry, Western Washington University

The present work describes a method to fabricate micellar nanocrystals, an emerging major class of nanobiomaterials. This method combines top-down electrospray, bottom-up self-assembly, and solvent-based structure control. The fabrication method is largely continuous, can produce high quality products, and possesses an inexpensive means of structure control.

Genetics

Culturing and Manipulation of O9-1 Neural Crest Cells

Bao H. Nguyen¹, Mamoru Ishii², Robert E. Maxson², Jun Wang¹

¹Molecular Physiology and Biophysics, Baylor College of Medicine, ²Department of Biochemistry and Molecular Biology, University of Southern California

O9-1 is a multipotent mouse neural crest cell line. Here we describe detailed step-by-step protocols for culturing O9-1 cells, differentiating O9-1 cells into specific cell types, and genetically manipulating O9-1 cells by using siRNA-mediated knockdown or CRISPR-Cas9 genome editing.

Behavior

Application of 3D Printing in the Construction of Burr Hole Ring for Deep Brain Stimulation Implants

Jiazhi Chen^*1,2, Xinyu Chen^*3, Siyuan Lv², Yuzhen Zhang¹, Hao Long¹, Kaijun Yang¹, Songtao Qi¹, Wangming Zhang², Jun Wang¹

¹Department of Neurosurgery, Nanfang Hospital, Southern Medical University, ²Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, ³Faculty of Brain Sciences, University of College London

Here, we present a protocol to demonstrate 3D printing in the construction of deep brain stimulation implants.

Biology

Skeletal Phenotype Analysis of a Conditional Stat3 Deletion Mouse Model

Yiling Yang^*1, Qianye Chen^*2, Siru Zhou¹, Xinyi Gong¹, Hongyuan Xu¹, Yueyang Hong¹, Qinggang Dai³, Lingyong Jiang¹

¹Center of Craniofacial Orthodontics, Department of Oral and Cranio-maxillofacial Science, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research center of Stomatology, ²Department of Stomatology, Dalian Medical University, ³The 2nd Dental Center, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research center of Stomatology

This protocol describes a canonical method to understand the critical genes controlling osteoclast activity in vivo. This method uses a transgenic mouse model and some canonical techniques to analyze skeletal phenotype.

Medicine

Isolation and Cultivation of Mandibular Bone Marrow Mesenchymal Stem Cells in Rats

Yueyang Hong^*1, Hongyuan Xu^*1, Yiling Yang¹, Siru Zhou¹, Anting Jin¹, Xiangru Huang¹, Qinggang Dai², Lingyong Jiang¹

¹Center of Craniofacial Orthodontics, Department of Oral and Cranio-maxillofacial Science, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research center of Stomatology, ²The 2nd Dental Center, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research center of Stomatology

This article presents a method that combines whole bone marrow adherence and flow cytometry sorting for isolating, cultivating, sorting, and identifying bone marrow mesenchymal stem cells from rat mandibles.

Bioengineering

Microfluidic Model to Mimic Initial Event of Neovascularization

Ping Zhao¹, Xing Zhang¹, Xiao Liu¹, Li Wang⁴, Haoran Su¹, Liyi Wang¹, Dongrui Zhang¹, Xiaoyan Deng³, Yubo Fan^1,2

¹Beijing Advanced Innovation Centre for Biomedical Engineering, Key Laboratory for Biomechanics and Mechanobiology of Chinese Education Ministry, School of Biological Science and Medical Engineering, Beihang University, ²School of Engineering Medicine, Beihang University, ³Artificial Intelligence Key Laboratory of Sichuan Province, School of Automation and Information Engineering, Sichuan University of Science and Engineering, ⁴Beijing Research Center of Urban System Engineering

Here, we provide a microfluidic chip and an automatically controlled, highly efficient circulation microfluidic system that recapitulates the initial microenvironment of neovascularization, allowing endothelial cells (ECs) to be stimulated by high luminal shear stress, physiological level of transendothelial flow, and various vascular endothelial growth factor (VEGF) distribution simultaneously.

Medicine

Real-Time Dynamic Navigation System for the Precise Quad-Zygomatic Implant Placement in a Patient with a Severely Atrophic Maxilla

Yihan Shen^*1, Qinggang Dai^*1, Baoxin Tao^*1, Wei Huang², Feng Wang², Kengliang Lan¹, Yuanyuan Sun¹, Xiaowan Ling¹, Lijun Yan¹, Yueping Wang¹, Yiqun Wu¹

¹The 2nd Dental Center, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology, ²Department of Dental Implantology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology

Here, we present a protocol to achieve precise quad-zygomatic implant placement in patients with severely atrophic maxilla using a real-time dynamic navigation system.

Developmental Biology

An Optimized O9-1/Hydrogel System for Studying Mechanical Signals in Neural Crest Cells

Tram P. Le^*1, Xiaolei Zhao^*1, Shannon Erhardt^1,2, Jianhua Gu³, Huie Wang³, Tina O. Findley¹, Jun Wang^1,2

¹Department of Pediatrics, McGovern Medical School, The University of Texas Health Science Center at Houston, ²The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, The University of Texas Health Science Center at Houston, ³SEM AFM Core in the Houston Methodist Hospital Research Institute

Detailed step-by-step protocols are described here for studying mechanical signals in vitro using multipotent O9-1 neural crest cells and polyacrylamide hydrogels of varying stiffness.