Name: ex vivo mechanical loading of tendon
Uploaded: 2007-05-28T00:00:00
Description: Watch this Scientific Journal Video about Ex vivo Mechanical Loading of Tendon at JoVE.com

<ol>
<li>Supplement &nbsp;media CO2-independent media with Fetal Bovine serum, l-glutamine, antibiotics, and ascorbic acid</li>
<li>Aliquot 40 ml into 50 ml centrifuge tubes (1 for each tendon)</li>
<li>Aliquot 10 ml into 15 ml centrifuge tubes (1 for each tendon)</li>
<li>Sterilize surgical tools, autoclave sheets, tweezers and torque wrench in autoclave</li>
<li>Sterilize clamps and force micrometer tips by soaking in 70% ethanol overnight</li>
<li>Harvest tendons from the rear paws of the rabbit. Rabbit should have been euthanized...

<ol>
	<li>Asundi, K., Kursa, K., Lotz, J. C., Rempel, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+system+for+applying+cyclic+loads+to+connective+tissues+under+displacement+or+force+control.">In vitro system for applying cyclic loads to connective tissues under displacement or force control.</a> Ann Biomed Eng. 35, 1188-1195 (2007).</li><li>Asundi, K. R., Rempel, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+loading+inhibits+expression+of+MMP-3+but+not+MMP-1+in+an+in+vitro+rabbit+flexor+tendon+model.">Cyclic loading inhibits expression of MMP-3 but not MMP-1 in an in vitro rabbit flexor tendon model.</a> Clin Biomech. , .</li><li>Asundi, K. R., Rempel, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MMP-1,+IL-1&#946;+and+COX-2+mRNA+Expression+is+Modulated+by+Static+Load+in+Rabbit+Flexor+Tendon.">MMP-1, IL-1&#946; and COX-2 mRNA Expression is Modulated by Static Load in Rabbit Flexor Tendon.</a> Ann Biomed Eng. , .</li></ol>

The video presents a new system for the simultaneous loading of up to four live tendons in tissue culture. Typical loading durations are 2 to 48 hours. After loading, the tendons are removed and processed for gene expression. The system allows for the investigation of the effect of loading pattern on changes in genes associated with degenerative changes in the tendon. These experiments can provide some insight into mechanisms of injury associated with repetitive loading. They also provide insight into loads that can...

Custom 4 Linear Actuator loading system
Warm water bath
50 ml tubes
tendons (rabbit)
Sterile media 

ex vivo mechanical loading of tendon

Injuries to the tendon (e.g., wrist tendonitis, epicondyltis) due to overuse are common in sports activities and the workplace. Most are associated with repetitive, high force hand activities. The mechanisms of cellular and structural damage due to cyclical loading are not well known. The purpose of this video is to present a new system that can simultaneously load four tendons in tissue culture. The video describes the methods of sterile tissue harvest and how the tendons are loaded onto a clamping system that is subsequently immersed into media and maintained at 37&deg;C. One clamp is fixed while the other one is moved with a linear actuator. Tendon tensile force is monitored with a load cell in series with the mobile clamp. The actuators are controlled with a LabView program. The four tendons can be repetitively loaded with different patterns of loading, repetition rate, rate of loading, and duration. Loading can continue for a few minutes to 48 hours. At the end of loading, the tendons are removed and the mid-substance extracted for biochemical analyses. This system allows for the investigation of the effects of loading patterns on gene expression and structural changes in tendon. Ultimately, mechanisms of injury due to overuse can be studies with the findings applied to treatment and prevention.

Watch this Scientific Journal Video about Ex vivo Mechanical Loading of Tendon at JoVE.com

Ex vivo Mechanical Loading of Tendon

A new in vitro system for simultaneously loading four tendons in culture is described.

Injuries to the tendon (e.g., wrist tendonitis, epicondyltis) due to overuse are common in sports activities and the workplace.  Most are associated with ...

Research

JoVE Journal

Biology

Targeted Expression of GFP in the Hair Follicle Using Ex Vivo Viral Transduction

There are many cell types in the hair follicle, including hair matrix cells which form the hair shaft and stem cells which can initiate the hair shaft ...

Method for Culture of Early Chick Embryos ex vivo (New Culture)

Method for Culture of Early Chick Embryos ex vivo New Culture

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

An ex-ovo Chicken Embryo Culture System Suitable for Imaging and Microsurgery Applications

An ex-ovo Chicken Embryo Culture System Suitable for Imaging and Microsurgery Applications

Understanding the relationships between genetic and microenvironmental factors that drive normal and malformed embryonic development is fundamental for ...

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells

Epithelial ovarian cancer is a leading cause of female cancer mortality in the United States. In contrast to other women-specific cancers, like breast and ...

A System for ex vivo Culturing of Embryonic Pancreas

A System for ex vivo Culturing of Embryonic Pancreas

The pancreas controls vital functions of our body,  including the production of digestive enzymes and regulation of blood sugar  levels1. Although in the ...

Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles

Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles

 
Described here is a method to measure contractility of isolated skeletal muscles. Parameters such as muscle force, muscle power, contractile kinetics, ...

Ex vivo Culture of Mouse Embryonic Skin and Live-imaging of Melanoblast Migration

Ex vivo Culture of Mouse Embryonic Skin and Live-imaging of Melanoblast Migration

Melanoblasts are the neural crest derived precursors of melanocytes; the cells responsible for producing the pigment in skin and hair. Melanoblasts ...

An Ex vivo Culture System to Study Thyroid Development

An Ex vivo Culture System to Study Thyroid Development

The thyroid is a bilobated endocrine gland localized at the base of the neck, producing the thyroid hormones T3, T4, and calcitonin. T3 and T4 are ...

Spatiotemporal Mapping of Motility in Ex Vivo Preparations of the Intestines

Spatiotemporal Mapping of Motility in Ex Vivo Preparations of the Intestines

Multiple approaches have been used to record and evaluate gastrointestinal motility including: recording changes in muscle tension, intraluminal pressure, ...

The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

Many studies seek to identify and map the brain regions involved in specific physiological regulations. The proto-oncogene c-fos, an immediate early gene, ...