Name: diagnosis of musculus gastrocnemius tightness - key factors for the clinical examination
Uploaded: 2016-07-07T10:00:00
Description: Watch this Scientific Journal Video about Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination at JoVE.com

We would like to thank Jakob Binder for his help in subject acquisition and organization of the examinations, Mrs. Hella Thun for designing Figure 1 and 2 and finally Mr. Dipl.-Ing. (FH), M.Sc. M. Saller for his statistical support.

Ethics statement: The study was approval by the local ethics committee of the University of Munich (# 007-14).
Note: An examination couch freely accessible on both sides and the foot end is required for non-weightbearing testing. A line (tape, approximately 2 meters) on the ground perpendicular the wall is needed for weightbearing testing. A standard goniometer with 2&#176; increments and 20 cm length is used. We recommend documenting the results using a standardized fourfold table containing ADF measurement for the knee extended and ...

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Examinations on 20 healthy individuals revealed, that 20 degrees of knee flexion already eliminates the influence of the M. gastrocnemius on ADF. Further knee flexion resulted in no significant ADF increase. The herein presented video description of a standardized non-weightbearing and weightbearing examination for MGT builds the prerequisite for future studies establishing physiological norm values.
The study has limitations. First, no custom-made measurement device was used to asses...

Limited ankle dorsiflexion (ADF) alters the gait kinematics and is held responsible for common foot pathologies including Achilles tendinopathy, stress fractures, metatarsalgia and plantar heel pain 1-5. The most common cause for limited ADF is isolated Musculus gastrocnemius tightness (MGT) 3,6.
The joint kinematics of the ankle are influenced by knee flexion as the M. gastrocnemius bridges both joints. The muscle is under tension when the knee is fully extended, as the origin of the muscle is then furthest proximal. The M. gastrocnemius then restrains ADF. Knee flexion approximates the muscle&#39;s origin, thereby reducing the tension of the M. gastrocnemius, and consequently increases ADF. Ankle dorsiflexion is then limited by other anatomical structures of the ankle joint. Figure 1 illustrates this principle. In the case of MGT, ADF is limited with the knee fully extended but substantially increases by flexion of the knee 4.
Clinical examination for MGT takes advantage of the above outlined principled and was first published in 1923 by Silfverski&#246;ld, a Swedish orthopedic surgeon 7. Since then, numerous examination techniques have been described, all of which compare the ADF with the knee extended and flexed. Published clinical tests can be categorized into non-weightbearing 5,8, weightbearing 9,10, and instrumented 11,12. Today, non-weightbearing examination is most commonly conducted 13. The patient is placed supine on the examination couch and ADF is assessed with the knee fully extended and typically 90&#176; flexed (Figure 2A). On the contrary, weightbearing ADF measurements are conducted with the subject upright in lunge stance. The rear knee is extended or bent and the subject is asked to lean forward just before heel liftoff (Figure 2B). For both tests MGT is diagnosed, if the ADF with the knee extended is impaired and knee flexion results in a significant increase of ADF.
Although non-weightbearing tests are frequently performed, weightbearing testing has several advantages. First, only one investigator is needed for weightbearing examination, whereas two examiners are required to achieve reliable non-weightbearing measurements. Second, the weightbearing examination more closely reflects the load during gait. Third, the force applied to the ankle is independent of the examiner. Fourth, weightbearing examination features a higher intra- and inter- rater reliability 9,10,13-15.
The major limitation of all tests for MGT is that&#160;the minimal degree of knee flexion needed to eliminate the restraining effect of the M. gastrocnemius on ADF is unknown 8,15. Whereas 90&#176; of knee flexion are typically applied in non-weightbearing testing 5,8,16, this is not feasible for the weightbearing examination. The broad population is unable to perform a Lunge with 90&#176; of knee flexion without lifting the heel off the ground. Consequently, most studies conducting weightbearing examinations did not control for knee flexion 8,15. In order to perform reliable weightbearing examinations it is essential to identify the minimal degree of knee flexion needed to eliminate the restraining effect of the M. gastrocnemius on ADF.
Overall, literature argues for weightbearing testing for the diagnosis of MGT. In order to provide a valid weightbearing examination procedure, the minimal degree of knee flexion required to eliminate the ADF restraining effect of the M. gastrocnemius must be known. The aim of this study was to investigate the influence of knee flexion on ADF in non-weightbearing and weightbearing testing and to provide a step-by-step guide to conduct non-weightbearing and weightbearing examination for MGT.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Examination couch</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Standard goniometer&#160;</td>
			<td colspan="2">MDF Instruments USA, Inc. Malibu, CA, USA</td>
			<td>2&#176; increments</td>
		</tr>
	</tbody>
</table>

diagnosis of musculus gastrocnemius tightness - key factors for the clinical examination

Common foot and ankle pathologies have been linked to isolated Musculus gastrocnemius tightness (MGT). Various examination techniques have been described to assess MGT. Still, a standardized examination procedure is missing. Literature argues for weightbearing examination but the degree of knee flexion needed to eliminate the restraining effect of the M. gastrocnemius on ankle dorsiflexion (ADF) is unknown. This manuscript investigates the effect of knee flexion on ankle dorsiflexion and provides a detailed description of a standardized examination protocol. Examination on 20 healthy individuals revealed, that 20&#176; of knee flexion is sufficient to fully eliminate the influence of the M. gastrocnemius on ADF. This builds the prerequisite for a standardized examination for MGT. Non-weightbearing and weightbearing examination of ADF has to be conducted with the knee fully extended and at least 20&#176; flexed. Two investigators should conduct non-weightbearing testing with the subject in supine position. In order to obtain reliable results, the axis of the fibula should be marked. One examiner can conduct weightbearing examination with the subject in lunge stance. Isolated MGT is present if ADF is impaired with the knee fully extended and knee flexion results in a significant ADF increase. The herein presented standardized examination is the prerequisite for future studies aiming at establishing norm values.

Both ankles of 20 healthy individuals (mean age of 27.1 &#177; 3.9 years), 50% female, were examined. Non-weightbearing and weightbearing tests at six different degrees of knee flexion (full extension, 20&#176;, 30&#176;, 45&#176;, 60&#176;, 75&#176;) and the Lunge Test (uncontrolled knee flexion) were conducted. A functional brace was used to control knee flexion. Measurements were performed by two investigators blinded to each other&#39;s results. Between each individual, the order of t...

Watch this Scientific Journal Video about Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination at JoVE.com

Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination

Isolated Musculus gastrocnemius tightness is a common cause for foot and ankle pathologies. Currently no standardized examination procedure exists. This manuscript demonstrates that 20 degree of knee flexion eliminates the restraining effect of the M. gastrocnemius on ankle dorsiflexion and presents a video description of a standardized examination protocol.

Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination

Common foot and ankle pathologies have been linked to isolated Musculus gastrocnemius tightness (MGT). Various examination techniques have been described ...

The overall goal of this diagnostic study is to define a standardized evidence-based examination procedure to assess ankle dorsiflexion with the knee extended and flexed. As there's no standardized test to assess ankle dorsiflexion, this examination method can help to define physiological and pathological values of ankle dorsiflexion. Clinicians will then be able to identify patients with impaired ankle dorsiflexion, which is the root of various pathologies to the foot and ankle.The main advantage of this technique is that it is a standardized evidence-based diagnostic approach. We first had the idea of this study when review literature on impaired ankle dorsiflexion. Many of the existing examination protocols vary significantly, therefore we aim to define a more standardized test to assess impaired ankle dorsiflexion.The visual demonstration of this examination technique is very beneficial, as it shows all of the key elements of this procedure in great detail.Mrs. Alena Sint is a member of our foot and ankle research group, and will help us to present the examination procedure. Begin by asking the subject to remove footwear and any clothing covering the legs.Have two investigators perform the measurement. One conducting the test, and the other measuring the degree of ankle dorsiflexion. Place the subject in a supine position on the examination couch.Mark the axis of the fibula by drawing a line connecting the center of the distal fibula at five centimeters and 15 centimeters above the tip of the distal fibula. Then have the first investigator place one hand at the level of the subtalar joint to ensure a neutral pronation supination position of the rear foot. And place the other hand around the mid-foot.Note that one hand stabilizes the subtalar joint and the other hand stabilizes the talonavicular joint, and applies force on the plantar aspect of the forefoot. To achieve maximum ankle dorsiflexion, ensure that the knee is fully extended. Next have the second investigator perform the measurement of the ankle dorsiflexion using a goniometer.Place one arm of the goniometer connecting the start and end point of the previously-marked axis of the fibula and align the other arm with the plantar aspect of the foot. Note this result on the documentation sheet. Then have the subject flex their knee 90 degrees and repeat this measurement procedure.Finally, repeat this entire procedure for the contralateral side. Note these results on the documentation sheet. Begin by asking the subject to stand opposite a wall.Ask the subject to get into a lunge position with the leg to be measured being the rear leg. Help the subject place their rear foot centered on the previously-marked line. Ensure that the heel and second toe of the rear leg are centered on the line.Have the subject hold on to the wall to stabilize the stance. Then ask the subject to fully extend their rear leg. Ensure that the knee is fully extended.Next have the subject move the hips toward the wall just before the heel of the rear leg lifts off the ground. The investigator has to ensure that the heel has not lifted off the ground. Place one hand on the dorsal aspect of the subtalar joint to ensure a neutral pronation supination position of the rear foot.With the other hand, perform the measurement of the ankle dorsiflexion by placing one arm of the goniometer on the floor, and align the other arm along the previously marked axis of the fibula. Note the result on the documentation sheet. Next have the subject move closer to the wall.Again, have the subject get into the lunge position, where their rear foot is centered on the previously-marked line. Have the subject hold on to the wall to stabilize their stance. Now ask the subject to flex their rear leg and move their hip toward the wall, just before the heel of the rear leg lifts off the ground.Use the goniometer to ensure that the rear knee is flexed at least 20 degrees. Then place one hand on the dorsal aspect of the subtalar joint to ensure a neutral pronation supination position of the rear foot. With the other hand, perform the measurement of the ankle dorsiflexion in the same manner as previously measured.Note the result. Repeat all steps for the contralateral side. Review the ankle dorsiflexion data on the documentation sheet, based on the subject's symptomatic side.Identify whether ankle dorsiflexion with the knee extended is less than 10 degrees on the symptomatic side. Then compare ADF with the knee extended and flexed. If ADF is greater than 10 degrees, compare ankle dorsiflexion with the knee extended between both legs.Consider musculus gastrocnemius tightness to be a possible cause, if ankle dorsiflexion is reduced on the symptomatic compared to the non-symptomatic side. This protocol demonstrates a standardized examination protocol for musculus gastrocnemius tightness, using a Non-weightbearing and a Weightbearing technique. The mean Ankle dorsiflexion of each step of Knee flexion indicated significantly higher values for Weightbearing measurements, compared to Non weightbearing measurements.Here, the difference of ADF between each step of knee flexion is shown. Significant differences were only observed between full knee extension and 20 degrees of knee flexion. After watching this video, you should have a good understanding of how to conduct this standardized examination to assess ankle dorsiflexion with the knee extended and flexed.Based on this, one can diagnose impaired ankle dorsiflexion as well as musculus gastrocnemius tightness. Once internalized, this technique can be done in a couple of minutes. While performing this procedure, it's important to keep the hind foot in neutral position, to stabilize the talonavicular joint and to ensure the appropriate knee position.This technique will pave the way for researchers in the field of foot and ankle disorders to finally agree on a consistent definition for physiological and pathological values of ankle dorsiflexion.

diagnosis-musculus-gastrocnemius-tightness-key-factors-for-clinical

Research

JoVE Journal

Medicine

Clinical Examination Protocol to Detect Atypical and Classical Scrapie in Sheep

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated prion protein by post mortem tests. Unless a random sample of the sheep or goat population is actively monitored for scrapie, identification of scrapie cases relies on the reporting of clinical suspects, which is dependent on the individual&#39;s familiarization with the disease and ability to recognize clinical signs associated with scrapie. Scrapie may not be considered in the differential diagnosis of neurological diseases in small ruminants, particularly in countries with low scrapie prevalence, or not recognized&#160;if it presents as nonpruritic form like atypical scrapie. To aid in the identification of clinical suspects, a short examination protocol is presented to assess the display of specific clinical signs associated with pruritic and nonpruritic forms of TSEs in sheep, which could also be applied to goats. This includes assessment of behavior, vision (by testing of the menace response), pruritus (by testing the response to scratching), and movement (with and without blindfolding). This may lead to a more detailed neurologic examination&#160;of reporting animals as scrapie suspects. It&#160;could also be used in experimental TSE studies of sheep or goats to evaluate disease progression or to identify clinical end-point.

A short clinical examination protocol is presented to aid in the detection of classical and atypical scrapie in sheep and goats.

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated ...

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed in mice is high. Phenotypic characterization of these mouse models also requires the examination of cardiac function and morphology. Echocardiography and magnetic resonance imaging (MRI) are commonly used approaches to characterize cardiac function and morphology in mice. Echocardiographic and MRI equipment specialized for use in small rodents is&#160;expensive and requires a dedicated space. This protocol describes cardiac measurements in mice using a clinical echocardiographic system with a 15 MHz human vascular probe. Measurements are performed on anesthetized adult mice. At least three image sequences are recorded and analyzed for each animal in M-mode in the parasternal short-axis view. Afterwards, cardiac histological examination is performed, and cardiomyocyte diameters are determined on hematoxylin-eosin- or wheat germ agglutinin (WGA)-stained paraffin sections. Vessel density is determined morphometrically after Pecam-1 immunostaining. The protocol has been applied successfully to pharmacological studies and different genetic animal models under baseline conditions, as well as after experimental myocardial infarction by the permanent ligation of the left anterior descending coronary artery (LAD). In our experience, echocardiographic investigation is limited to anesthetized animals and is feasible in adult mice weighing at least 25 g.

Echocardiographic examination is frequently used in mice. Expensive high-resolution ultrasound devices have been developed for this purpose. This protocol describes an affordable echocardiographic procedure combined with histological morphometric analyses to determine cardiac morphology.

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed ...

Observational Study Protocol for Repeated Clinical Examination and Critical Care Ultrasonography Within the Simple Intensive Care Studies

Longitudinal evaluations of critically ill patients by combinations of clinical examination, biochemical analysis and critical care ultrasonography (CCUS) may detect adverse events of interventions such as fluid overload at an early stage. The Simple Intensive Care Studies (SICS) is a research line that focuses on the prognostic and diagnostic value of combinations of clinical variables.
The SICS-I specifically focused on the use of clinical variables obtained within 24 h of acute admission for prediction of cardiac output (CO) and mortality. Its sequel, SICS-II, focuses on repeated evaluations during ICU admission. The first clinical examination by trained researchers is performed within 3 h after admission consisting of physical examination and educated guessing. The second clinical examination is performed within 24 h after admission and includes physical examination and educated guessing, biochemical analysis and CCUS assessments of heart, lungs, inferior vena cava (IVC) and kidney. This evaluation is repeated at days 3 and 5 after admission. CCUS images are validated by an independent expert, and all data is registered in an online secured database. Follow-up at 90 days includes registration of complications and survival status according to patient&#39;s medical charts and the municipal person registry. The primary focus of SICS-II is the association between venous congestion and organ dysfunction.
The purpose of publishing this protocol is to provide details on the structure and methods of this on-going prospective observational cohort study allowing answering multiple research questions. The design of the data collection of combined clinical examination and CCUS assessments in critically ill patients are explicated. The SICS-II is open for other centers to participate and is open for other research questions that can be answered with our data.

Structured protocols are necessary to provide answers on research questions in critically ill patients. The Simple Intensive Care Studies (SICS) provides an infrastructure for repeated measurements in critically ill patients including clinical examination, biochemical analysis and ultrasonography. SICS projects have specific focus but the structure is flexible to other investigations.

Longitudinal evaluations of critically ill patients by combinations of clinical examination, biochemical analysis and critical care ultrasonography (CCUS) ...

Transthoracic Echocardiographic Examination in the Rabbit Model

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to that of humans and that of other large animal models such as dogs and pigs. However, the rabbit model has the additional advantage of lower maintenance costs compared to other large animal models. The longitudinal evaluation of cardiac function using echocardiography, when appropriately implemented, is a useful methodology for preclinical assessment of novel therapies for heart failure with reduced ejection fraction (e.g. cardiac regeneration). The correct use of this non-invasive tool requires the implementation of a standardized examination protocol following international guidelines. Here we describe, step by step, a detailed protocol supervised by veterinary cardiologists for performing echocardiography in the rabbit model, and demonstrate how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Here we describe, step by step, a detailed protocol for performing echocardiography in the rabbit model. We show how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to ...

OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery

The use of in vitro maturation (IVM) before gynecological operation (OP-IVM) is an extension of conventional IVM that combines IVM following oocyte retrieval with routine gynecological surgery. OP-IVM is suitable for patients undergoing benign gynecological surgery who have the need for fertility preservation (FP) or infertility treatments such as in vitro fertilization and embryo transfer (IVF-ET). In the operating room, patients undergoing benign gynecological surgery are first anesthetized and receive ultrasound-guided immature follicle aspiration (IMFA) treatment. As the subsequent gynecological surgery is performed, the cumulus-oocyte complexes (COCs) are examined, and the immature COCs are transferred into the IVM medium and cultured for 28-32 hours in the IVF laboratory. After assessment, mature oocytes in the MII stage will be selected and cryopreserved in liquid nitrogen for FP or fertilized by intracytoplasmic sperm injection (ICSI) for IVF-ET. By combining IVM with gynecological surgery, immature oocytes that would have been discarded can be saved and used for assisted reproductive technology (ART). The procedure, significance and critical aspects of OP-IVM are described in this article.

In vitro maturation (IVM) before gynecological operation (OP-IVM) combines IVM following oocyte retrieval with routine gynecological surgery and serves as an extension of conventional IVM applications for fertility preservation.

The use of in vitro maturation (IVM) before gynecological operation (OP-IVM) is an extension of conventional IVM that combines IVM following oocyte ...

The Miniature Pig: A Large Animal Model for Cochlear Implant Research

Cochlear implants (CI) are the most effective method to treat people with severe-to-profound sensorineural hearing loss. Although CIs are used worldwide, no standard model exists for investigating the electrophysiology and histopathology in patients or animal models with a CI or for evaluating new models of electrode arrays. A large animal model with cochlea characteristics similar to those of humans may provide a research and evaluation platform for advanced and modified arrays before their use in humans. 
To this end, we established standard CI methods with Bama mini-pigs, whose inner ear anatomy is highly similar to that of humans. Arrays designed for human use were implanted into the mini pig cochlea through a round window membrane, and a surgical approach followed that was similar to that used for human CI recipients. Array insertion was followed by evoked compound action potential (ECAP) measurements to evaluate the function of the auditory nerve. This study describes the preparation of the animal, surgical steps, array insertion, and intraoperative electrophysiological measurements. 
The results indicated that the same CI used for humans could be easily implanted in mini-pigs via a standardized surgical approach and yielded similar electrophysiological outcomes as measured in human CI recipients. Mini-pigs could be a valuable animal model to provide initial evidence of the safety and potential performance of novel electrode arrays and surgical approaches before applying them to human beings.

Miniature pigs (mini-pigs) are an ideal large animal model for research into cochlear implants. Cochlear implantation surgery in mini-pigs can be utilized to provide initial evidence of the safety and potential performance of novel electrode arrays and surgical approaches in a living system similar to human beings.

Cochlear implants (CI) are the most effective method to treat people with severe-to-profound sensorineural hearing loss. Although CIs are used worldwide, ...

Subclavian Vein Puncture Method for Repeated Blood Collection in Conscious Rats

Subclavian Vein Blood Sampling in Conscious Rats

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein sampling without anesthesia and jugular vein sampling with anesthesia. However, most of these methods require assistance and anesthetic equipment and sometimes pose difficulties in terms of blood collection or the poor quality of blood samples. In addition, these methods of blood collection consume significant time and human resources when repeated blood sampling is required for a large number of rats. This study presents a technique for repetitive blood sampling in non-anesthetized rats by a single proficient individual. Highly satisfactory blood samples can be obtained by puncturing the subclavian vein. The method demonstrated an impressive overall success rate of 95%, with a median time of merely 2 min from rat restraint to the completion of blood collection. Furthermore, performing consecutive blood collections within the designated range does not inflict any harm on the rats. This method is worth promoting for blood collection, especially in large-scale pharmacokinetic studies.

Author Spotlight: Developing an Efficient Blood Collection Method in Non-Anesthetized Rats 

Here we present a combination of effective rat restriction and subclavian vein puncture methods that enable rapid, safe, and repeated blood collection in rats without anesthesia.

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein ...

Extraction of 3D Region of the Liver for Hepatic Fibrosis Diagnosis

To begin, open MATLAB software. Ensure that the IDEAL data have a horizontal resolution of 256 by 256 pixels with a pixel spacing of 1.5625 millimeters and a slice thickness of 10 millimeters. Copy all DICOM data to a customized working directory, and navigate to the directory containing the data in MATLAB's working directory.Then execute the description name function to add descriptive names to the folders for each sequence. Add a description name to each image sequence folder. To check images of IDEAL, change the directory of different phase folders.Execute the slice view function to view the impact sequences for each phase. Next, quickly browse the different sequences using the scroll bar at the bottom of the GUI. Select the MRI IDEAL outphase sequence to depict liver tissue boundaries.Initiate the Mimics software. Select new project, and in the ensuing dialogue box, navigate to the folder containing the IDEAL outphase images. Click next, followed by the convert button to enter into the sequence heading state.To create an empty mask, from the mask dialogue box, click on the new button and select the maximum threshold. Using the edit masks tool located beneath the segment label, delimit the area of the liver in all horizontal views. To generate the 3D spatial part of the liver, select the outlying liver mask and click on the calculate part for mask button.Next, click file, then export, and select the DICOM command. In the pop-up dialogue box, choose the liver mask, and set the file path and names, then click the OK button to complete the export of the 3D region of the liver to the specified DICOM files.

Advancing Cryostorage of Human Stem Cell-Derived Retinal Pigment Epithelial Cells

Cryopreservation of Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells at the Optimal Stage

Retinal pigment epithelial&#160;(RPE) cells derived from human embryonic stem cells (hESCs) are superior cell sources for cell replacement therapy in individuals with retinal degenerative diseases; however, studies on the stable and secure banking of these therapeutic cells are scarce. Highly variable cell viability and functional recovery of&#160;RPE cells after cryopreservation are the most commonly encountered issues. In the present protocol, we aimed to achieve the best cell recovery rate after thawing by selecting the optimal cell phase for freezing based on the original experimental conditions. Cells were frozen in the exponential phase determined by using the 5-ethynyl-2&#8242;-deoxyuridine labeling assay, which improved cell viability and recovery rate after thawing. Stable and functional cells were obtained shortly after thawing, independent of a long differentiation process. The methods described here allow the simple, efficient, and inexpensive preservation and thawing of hESC-derived RPE cells. Although this protocol focuses on RPE cells, this freezing strategy may be applied to many other types of differentiated cells.

Author Spotlight: Development of an Efficient and Feasible Method of Retinal Pigment Epithelial Cell Freezing

This study provides a detailed protocol for the efficient cryopreservation of human stem cell-derived retinal pigment epithelial cells.

Retinal pigment epithelial&#160;(RPE) cells derived from human embryonic stem cells (hESCs) are superior cell sources for cell replacement therapy in ...

Exploring Obliterative Airway Disease in Lung Transplants

Murine Intrapulmonary Tracheal Transplantation: A Model for Investigating Obliterative Airway Disease After Lung Transplantation

Murine intrapulmonary tracheal transplantation (IPTT) is used as a model of obliterative airway disease (OAD) following lung transplantation. Initially reported by our team, this model has gained use in the study of OAD due to its high technical reproducibility and suitability for investigating immunological behaviors and therapeutic interventions.
In the IPTT model, a rodent tracheal graft is directly inserted into the recipient's lung through the pleura. This model is distinct from the heterotopic tracheal transplantation (HTT) model, wherein grafts are transplanted into subcutaneous or omental sites, and from the orthotopic tracheal transplantation (OTT) model in which the donor trachea replaces the recipient's trachea. 
Successful implementation of the IPTT model requires advanced anesthetic and surgical skills. Anesthetic skills include endotracheal intubation of the recipient, setting appropriate ventilatory parameters, and appropriately timed extubation after recovery from anesthesia. Surgical skills are essential for precise graft placement within the lung and for ensuring effective sealing of the visceral pleura to prevent air leakage and bleeding. In general, the learning process takes approximately 2 months.
In contrast to the HTT and OTT models, in the IPTT model, the allograft airway develops airway obliteration in the relevant lung microenvironment. This allows investigators to study lung-specific immunological and angiogenic processes involved in airway obliteration after lung transplantation. Furthermore, this model is also unique in that it exhibits tertiary lymphoid organs (TLOs), which are also seen in human lung allografts. TLOs are comprised of T and B cell populations and characterized by the presence of high endothelial venules that direct immune cell recruitment; therefore, they are likely to play a crucial role in graft acceptance and rejection. We conclude that the IPTT model is a useful tool for studying intrapulmonary immune and profibrotic pathways involved in the development of airway obliteration in the lung transplant allograft.

Author Spotlight: Investigating the Key Factors of Obliterative Bronchiolitis After Lung Transplantation

The murine intrapulmonary tracheal transplantation (IPTT) model is valuable for studying obliterative airway disease (OAD) after lung transplantation. It offers insights into lung-specific immunological and angiogenic behavior in airway obliteration after allotransplantation with high reproducibility. Here, we describe the IPTT procedure and its expected results.

Murine intrapulmonary tracheal transplantation (IPTT) is used as a model of obliterative airway disease (OAD) following lung transplantation. Initially ...