<meta charset="utf8"></head><body>治療用包帯を評価するためのマウスにおける一貫した熱傷モデル

<ol>
	<li>Mofazzal Jahromi, M. 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=Nanomedicine+and+advanced+technologies+for+burns:+Preventing+infection+and+facilitating+wound+healing.">Nanomedicine and advanced technologies for burns: Preventing infection and facilitating wound healing.</a> Adv Drug Deliv Rev. 123, 33-64 (2018).</li><li>Burgess, M., Valdera, F., Varon, D., Kankuri, E., Nuutila, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+immune+and+regenerative+response+to+burn+injury.">The immune and regenerative response to burn injury.</a> Cells. 11 (19), 3073 (2022).</li><li>Arbuthnot, M. K., Garcia, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+resuscitation+and+management+of+severe+pediatric+burns.">Early resuscitation and management of severe pediatric burns.</a> Sem Pediatr Surg. 28 (1), 73-78 (2019).</li><li>Smolle, 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=Recent+trends+in+burn+epidemiology+worldwide:+A+systematic+review.">Recent trends in burn epidemiology worldwide: A systematic review.</a> Burns. 43 (2), 249-257 (2017).</li><li>Jeschke, M. G., 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=Burn+injury.">Burn injury.</a> Nat Rev Dis Primers. 6, 11 (2020).</li><li>Nomellini, V., Gomez, C. R., Gamelli, R. L., Kovacs, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+and+animal+models+of+systemic+insult:+Trauma,+burn,+and+sepsis.">Aging and animal models of systemic insult: Trauma, burn, and sepsis.</a> Shock. 31 (1), 11-20 (2009).</li><li>Wong, V. W., Sorkin, M., Glotzbach, J. P., Longaker, M. T., Gurtner, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+approaches+to+create+murine+models+of+human+wound+healing.">Surgical approaches to create murine models of human wound healing.</a> J Biomed Biotechnol. 2011, 969618 (2011).</li><li>Abdullahi, A., Amini-Nik, S., Jeschke, M. 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N., Suzuki, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunological+control+of+methicillin-resistant+Staphylococcus+aureus+(MRSA)+infection+in+an+immunodeficient+murine+model+of+thermal+injuries.">Immunological control of methicillin-resistant Staphylococcus aureus (MRSA) infection in an immunodeficient murine model of thermal injuries.</a> Clin Exp Immunol. 142 (3), 419-425 (2005).</li><li>Dai, T., 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=Animal+models+of+external+traumatic+wound+infections.">Animal models of external traumatic wound infections.</a> Virulence. 2 (4), 296-315 (2011).</li><li>Tavares Pereira, D. D. S., Lima-Ribeiro, M. H. M., De Pontes-Filho, N. T., Carneiro-Le&#227;o, A. M. D. A., Correia, M. T. D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+animal+model+for+studying+deep+second-degree+thermal+burns.">Development of animal model for studying deep second-degree thermal burns.</a> J Biomed Biotechnol. 2012, 460841 (2012).</li><li>Gong, Y., 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=Exudate+absorbing+and+antimicrobial+hydrogel+integrated+with+multifunctional+curcumin-loaded+magnesium+polyphenol+network+for+facilitating+burn+wound+healing.">Exudate absorbing and antimicrobial hydrogel integrated with multifunctional curcumin-loaded magnesium polyphenol network for facilitating burn wound healing.</a> ACS Nano. 17 (22), 22355-22370 (2023).</li><li>Dahiya, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burns+as+a+model+of+sirs.">Burns as a model of sirs.</a> Front Biosci. 14 (13), 4962-4967 (2009).</li><li>Yergoz, F., 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=Heparin+mimetic+peptide+nanofiber+gel+promotes+regeneration+of+full-thickness+burn+injury.">Heparin mimetic peptide nanofiber gel promotes regeneration of full-thickness burn injury.</a> Biomaterials. 134, 117-127 (2017).</li><li>Rao, G., 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=Rat+burn+model+to+study+full-thickness+cutaneous+thermal+burn+and+infection.">Rat burn model to study full-thickness cutaneous thermal burn and infection.</a> J Vis Exp. 186, e64345 (2022).</li><li>Yampolsky, M., Bachelet, I., Fuchs, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproducible+strategy+for+excisional+skin-wound-healing+studies+in+mice.">Reproducible strategy for excisional skin-wound-healing studies in mice.</a> Nat Protoc. 19 (1), 184-206 (2024).</li><li>Wang, 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=Engineering+bioactive+self-healing+antibacterial+exosomes+hydrogel+for+promoting+chronic+diabetic+wound+healing+and+complete+skin+regeneration.">Engineering bioactive self-healing antibacterial exosomes hydrogel for promoting chronic diabetic wound healing and complete skin regeneration.</a> Theranostics. 9 (1), 65-76 (2019).</li><li>Papp, 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=The+progression+of+burn+depth+in+experimental+burns:+A+histological+and+methodological+study.">The progression of burn depth in experimental burns: A histological and methodological study.</a> Burns. 30 (7), 684-690 (2004).</li><li>Barrett, J. P., 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=An+in+vitro+study+into+the+antimicrobial+and+cytotoxic+effect+of+Acticoat+dressings+supplemented+with+chlorhexidine.">An in vitro study into the antimicrobial and cytotoxic effect of Acticoat dressings supplemented with chlorhexidine.</a> Burns. 48 (4), 941-951 (2022).</li><li>Lou, P., 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=Injectable+self-assembling+peptide+nanofiber+hydrogel+as+a+bioactive+3D+platform+to+promote+chronic+wound+tissue+regeneration.">Injectable self-assembling peptide nanofiber hydrogel as a bioactive 3D platform to promote chronic wound tissue regeneration.</a> Acta Biomater. 135, 100-112 (2021).</li><li>Gurtner, G. C., Werner, S., Barrandon, Y., Longaker, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+repair+and+regeneration.">Wound repair and regeneration.</a> Nature. 453 (7193), 314-321 (2008).</li><li>Bielefeld, K. A., Amini-Nik, S., Alman, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cutaneous+wound+healing:+Recruiting+developmental+pathways+for+regeneration.">Cutaneous wound healing: Recruiting developmental pathways for regeneration.</a> Cell Mol Life Sci. 70 (12), 2059-2081 (2013).</li><li>Yildiz, S. C., Demir, C., Ayhanci, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Examination+of+the+effects+of+kefir+on+healing+factors+in+a+mice+burn+model+infected+with+E.+coli,+S.+aureus+and+P.+aeruginosa+using+qRT-PCR.">Examination of the effects of kefir on healing factors in a mice burn model infected with E. coli, S. aureus and P. aeruginosa using qRT-PCR.</a> Burns. 49 (2), 425-431 (2023).</li><li>Wang, Y., 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=Burn+injury:+Challenges+and+advances+in+burn+wound+healing,+infection,+pain+and+scarring.">Burn injury: Challenges and advances in burn wound healing, infection, pain and scarring.</a> Adv Drug Deliv Rev. 123, 3-17 (2018).</li><li>Chen, Y., 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=Research+advances+in+smart+responsive-hydrogel+dressings+with+potential+clinical+diabetic+wound+healing+properties.">Research advances in smart responsive-hydrogel dressings with potential clinical diabetic wound healing properties.</a> Mil Med Res. 10 (1), 37 (2023).</li><li>Ito, M., 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=Stem+cells+in+the+hair+follicle+bulge+contribute+to+wound+repair+but+not+to+homeostasis+of+the+epidermis.">Stem cells in the hair follicle bulge contribute to wound repair but not to homeostasis of the epidermis.</a> Nat Med. 11 (12), 1351-1354 (2005).</li><li>Ito, M., 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=Wnt-dependent+de+novo+hair+follicle+regeneration+in+adult+mouse+skin+after+wounding.">Wnt-dependent de novo hair follicle regeneration in adult mouse skin after wounding.</a> Nature. 447 (7142), 316-320 (2007).</li><li>Chen, K., 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=Pullulan-collagen+hydrogel+wound+dressing+promotes+dermal+remodeling+and+wound+healing+compared+to+commercially+available+collagen+dressings.">Pullulan-collagen hydrogel wound dressing promotes dermal remodeling and wound healing compared to commercially available collagen dressings.</a> Wound Repair Regen. 30 (3), 397-408 (2022).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 mL pipette tip</td>
			<td>KIRGEN,USA</td>
			<td>KG1333</td>
			<td>Used to locate burn wound sites</td>
		</tr>
		<tr>
			<td>3 M Tegaderm film</td>
			<td>3M,USA</td>
			<td>1624WCN 6 cm x 7 cm</td>
			<td>For the wound cover after burn induction</td>
		</tr>
		<tr>
			<td>4% paraformaldehyde(PFA)</td>
			<td>Biosharp,China</td>
			<td>BL539A</td>
			<td>Used to fix the skin samples</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Sigma-Aldrich,USA</td>
			<td>PHR8955-50MG</td>
			<td>For the pain management of the mice</td>
		</tr>
		<tr>
			<td>C57BL/6 mice</td>
			<td>Chengdu Dashuo experimental animal company,China</td>
			<td>none</td>
			<td>For the establishment of burn model</td>
		</tr>
		<tr>
			<td>Depilatory cream</td>
			<td>Veet,China</td>
			<td>&#8212;</td>
			<td>For the dorsum hair removal of the mice</td>
		</tr>
		<tr>
			<td>Digital Heating Device</td>
			<td>Shenzhen Kapper Technology Company,China</td>
			<td>No.936D</td>
			<td>For the burn induction of the mice</td>
		</tr>
		<tr>
			<td>Electric shaver</td>
			<td>AUX,China</td>
			<td>AUX-A5</td>
			<td>For the dorsum hair removal of the mice</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>&#8212;</td>
			<td>&#8212;</td>
			<td>Used to unfold of the skin samples</td>
		</tr>
		<tr>
			<td>GraphPad software</td>
			<td>&#8212;</td>
			<td>GraphPad prism 9.5.0</td>
			<td>For the analysis of burn wound area</td>
		</tr>
		<tr>
			<td>Heat-resistant gloves</td>
			<td>&#8212;</td>
			<td>&#8212;</td>
			<td>Used to hold the digital heating device tip</td>
		</tr>
		<tr>
			<td>Hematoxylin and Eosin Stain kit</td>
			<td>Solarbio,China</td>
			<td>G1120</td>
			<td>For the histological analysis of the slides</td>
		</tr>
		<tr>
			<td>ImageJ software</td>
			<td>&#8212;</td>
			<td>ImageJ 1.54f</td>
			<td>For the analysis of burn wound area</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD,China</td>
			<td>R510-22-10</td>
			<td>For the anesthesia of the mice</td>
		</tr>
		<tr>
			<td>Masson&#39;s Trichrome Stain Kit</td>
			<td>Solarbio,China</td>
			<td>G1340</td>
			<td>For the histological analysis of the slides</td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Olympus,Japan&#160;</td>
			<td>VS200 ASW</td>
			<td>Used to scan the H&#38;E and Masson stained slides</td>
		</tr>
		<tr>
			<td>Tissue cassette</td>
			<td>CITOTEST LABWARE MANUFACTURING Co., LTD,China</td>
			<td>31050102W</td>
			<td>For tissue paraffin dehydration and paraffin embedding</td>
		</tr>
	</tbody>
</table>

chessboard-like burn wound healing model of mice based on digital heating device

<meta charset="utf8"></head><body>デジタル加熱装置に基づくマウスのチェス盤状火傷創傷治癒モデル

デジタル加熱装置に基づくマウスのチェス盤状火傷創傷治癒モデル

Severe burn injuries are among the most traumatic and physically debilitating conditions, impacting nearly every organ system and resulting in ...

chessboard-like-burn-wound-healing-model-mice-based-on-digital

Research

JoVE Journal

Biology

Chessboard-like Burn Wound Healing Model of Mice Based on Digital Heating Device

217 ビュー.  Sichuan University. デジタル加熱装置を使用してマウスの熱傷創傷治癒モデルを確立するための合理化されたプロトコルが提示されます。皮膚上に作成されたチェス盤のような実験部位は、創傷治癒アッセイのためのさらなる機能分析を容易にします。

ビデオ: デジタル加熱装置に基づくマウスのチェス盤状火傷創傷治癒モデル

Visualizing Cytoplasmic Flow During Single-cell Wound Healing in Stentor coeruleus

Although wound-healing is often addressed at the level of whole tissues, in many cases individual cells are able to heal wounds within themselves, repairing broken cell membrane before the cellular contents leak out. The giant unicellular organism Stentor coeruleus, in which cells can be more than one millimeter in size, have been a classical model organism for studying wound healing in single cells. Stentor cells can be cut in half without loss of viability, and can even be cut and grafted together. But this high tolerance to cutting raises the question of why the cytoplasm does not simply flow out from the size of the cut. Here we present a method for cutting Stentor cells while simultaneously imaging the movement of cytoplasm in the vicinity of the cut at high spatial and temporal resolution. The key to our method is to use a &#34;double decker&#34; microscope configuration in which the surgery is performed under a dissecting microscope focused on a chamber that is simultaneously viewed from below at high resolution using an inverted microscope with a high NA lens. This setup allows a high level of control over the surgical procedure while still permitting high resolution tracking of cytoplasm.

Visualizing Cytoplasmic Flow During Single-cell Wound Healing in Stentor coeruleus

The giant ciliate Stentor coeruleus is a classical system for studying regeneration and wound healing in single cells.&#160;By imaging Stentor cells simultaneously at low and high magnification it is possible to measure cytoplasmic flows before, during, and after wounding.

Although wound-healing is often addressed at the level of whole tissues, in many cases individual cells are able to heal wounds within themselves, ...

生物学

Micron-scale Phenotyping Techniques of Maize Vascular Bundles Based on X-ray Microcomputed Tomography

It is necessary to accurately quantify the anatomical structures of maize materials based on high-throughput image analysis techniques. Here, we provide a 'sample preparation protocol' for maize materials (i.e., stem, leaf, and root) suitable for ordinary microcomputed tomography (micro-CT) scanning. Based on high-resolution CT images of maize stem, leaf, and root, we describe two protocols for the phenotypic analysis of vascular bundles: (1) based on the CT image of maize stem and leaf, we developed a specific image analysis pipeline to automatically extract 31 and 33 phenotypic traits of vascular bundles; (2) based on the CT image series of maize root, we set up an image processing scheme for the three-dimensional (3-D) segmentation of metaxylem vessels, and extracted two-dimensional (2-D) and 3-D phenotypic traits, such as volume, surface area of metaxylem vessels, etc. Compared with traditional manual measurement of vascular bundles of maize materials, the proposed protocols significantly improve the efficiency and accuracy of micron-scale phenotypic quantification.

We provide a novel method to improve the X-ray absorption contrast of maize tissue suitable for ordinary microcomputed tomography scanning. Based on CT images, we introduce a set of image-processing workflows for different maize materials to effectively extract microscopic phenotypes of vascular bundles of maize.

X 線マイクロ断層レントゲン写真撮影に基づいてトウモロコシの維管束のミクロン スケール表現型解析技術

It is necessary to accurately quantify the anatomical structures of maize materials based on high-throughput image analysis techniques. Here, we provide a ...

A Murine Model of a Burn Wound Reconstructed with an Allogeneic Skin Graft

Trivial superficial wounds heal without complications by primary intention. Deep wounds, such as full thickness burns, heal by secondary intention and require surgical debridement and skin grafting. Successful integration of the donor graft into a recipient wound bed depends on timely recruitment of immune cells, robust angiogenic response and new extracellular matrix formation. The development of novel therapeutic agents, which target some key processes involved in wound healing, are hindered by the lack of reliable preclinical models with optimized objective assessment of wound closure. Here, we describe an inexpensive and reproducible model of experimental full thickness burn wound reconstructed with an allogeneic skin graft. The wound is induced on the dorsum surface of anaesthetized inbred wild type mice from the BALB/C and SKH1-Hrhr backgrounds. The burn is produced using a brass template measuring 10 mm in diameter, which is preheated to 80 &#176;C and delivered at a constant pressure for 20 s. Burn eschar is excised 24 hours after the injury and replaced with a full thickness graft harvested from the tail of a genetically similar donor mouse. No specialized equipment is required for the procedure and surgical techniques are straightforward to follow. The method may be effortlessly implemented and reproduced in most research settings. Certain limitations are associated with the model. Due to technical difficulties, the harvest of thinner split thickness skin grafts is not possible. The surgical method we describe here allows for the reconstruction of burn wounds using full thickness skin grafts. It may be used to carry out preclinical therapeutic testing.

The aim of this study was to develop a murine model of burn wound healing. A thermal burn was induced on the dorsal skin of mice using a preheated brass template. Burned tissue was debrided and overlaid with a skin graft harvested from the tail of a genetically similar donor mouse.

同種異系皮膚移植片で再建された火傷のマウスモデル

Trivial superficial wounds heal without complications by primary intention. Deep wounds, such as full thickness burns, heal by secondary intention and ...

A Capsule-Based Model for Immature Hard Tick Stages Infestation on Laboratory Mice

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of mouse models in tick research is critical for understanding their biology and tick-host-pathogen interactions. Here we demonstrate a non-laborious technique for the feeding of immature stages of hard ticks on laboratory mice. The benefit of the method is its simplicity, short duration, and the ability to monitor or collect ticks at different time points of an experiment. In addition, the technique allows attachment of two individual capsules on the same mouse, which is beneficial for a variety of experiments where two different groups of ticks are required to feed on the same animal. The non-irritating and flexible capsule is made from easily accessible materials and minimizes the discomfort of the experimental animals. Furthermore, euthanasia is not necessary, mice recover completely after the experiment and are available for re-use.

In this study, a feeding system for nymphal and larval stages of hard tick was developed using a capsule attached to laboratory mouse. The feeding capsule is made from flexible materials and remains firmly attached to the mouse for at least one week and allows comfortable monitoring of tick feeding.

実験室マウスにおける未熟なハードティックステージのカプセルベースモデル

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of ...

Application of Ultrasound and Shear Wave Elastography Imaging in a Rat Model of NAFLD/NASH

Nonalcoholic Steatohepatitis (NASH) is a condition within the spectrum of Non-Alcoholic Fatty Liver Disease (NAFLD), which is characterized by liver fat accumulation (steatosis) and inflammation leading to fibrosis. Preclinical models closely recapitulating human NASH/NAFLD are essential in drug development. While liver biopsy is currently the gold standard for measuring NAFLD/NASH progression and diagnosis in the clinic, in the preclinical space, either collection of whole liver samples at multiple timepoints during a study or biopsy of liver is needed for histological analysis to assess the disease stage.
Conducting a liver biopsy mid-study is an invasive and labor-intensive procedure, and collecting liver samples to assess disease level increases the number of research animals needed for a study. Thus, there is a need for a reliable, translatable, non-invasive imaging biomarker to detect NASH/NAFLD in these preclinical models. Non-invasive ultrasound-based B-mode images and Shear Wave Elastography (SWE) can be used to measure steatosis as well as liver fibrosis. To assess the utility of SWE in preclinical rodent models of NASH, animals were placed on a pro-NASH diet&#160;and underwent non-invasive ultrasound B-mode and shear wave elastography imaging to measure hepatorenal (HR) index and liver elasticity, measuring progression of both liver fat accumulation and tissue stiffness, respectively, at multiple time points over the course of a given NAFLD/NASH study.
The HR index and elasticity numbers were compared to histological markers of steatosis and fibrosis. The results showed strong correlation between the HR index and percentage of Oil Red O (ORO) staining, as well as between elasticity and Picro-Sirius Red (PSR) staining of livers. The strong correlation between classic ex vivo methods and in vivo imaging results provides evidence that shear wave elastography/ultrasound-based imaging can be used to assess disease phenotype and progression in a preclinical model of NAFLD/NASH.

This protocol describes the use of an enhanced ultrasound technique to non-invasively observe and quantify liver tissue changes in rodent models of nonalcoholic fatty liver disease.

NAFLD/NASHラットモデルにおける超音波および剪断波エラストグラフィーイメージングの応用

Nonalcoholic Steatohepatitis (NASH) is a condition within the spectrum of Non-Alcoholic Fatty Liver Disease (NAFLD), which is characterized by liver fat ...

Zebrafish Corneal Wound Healing: From Abrasion to Wound Closure Imaging Analysis

As the transparent surface of the eye, the cornea is instrumental for clear sight. Due to its location, this tissue is prone to environmental insults. Indeed, the eye injuries most frequently encountered clinically are those to the cornea. While corneal wound healing has been extensively studied in small mammals (i.e., mice, rats, and rabbits), corneal physiology studies have neglected other species, including zebrafish, despite zebrafish being a classic research model.
This report describes a method of performing a corneal abrasion on zebrafish. The wound is performed in vivo on anesthetized fish using an ocular burr. This method allows for a reproducible epithelial wound, leaving the rest of the eye intact. After abrasion, wound closure is monitored over the course of 3 h, after which the wound is reepithelialized. By using scanning electron microscopy, followed by image processing, the epithelial cell shape, and apical protrusions can be investigated to study the various steps during corneal epithelial wound closure.
The characteristics of the zebrafish model permit study of the epithelial tissue physiology and the collective behavior of the epithelial cells when the tissue is challenged. Furthermore, the use of a model deprived of the influence of the tear film can produce&#160;new answers regarding corneal response to stress. Finally, this model also allows the delineation of the cellular and molecular events involved in any epithelial tissue subjected to a physical wound. This method can be applied to the evaluation of drug effectiveness in preclinical testing.

This protocol focuses on damaging the ocular surface of zebrafish through abrasion to assess the subsequent wound closure at the cellular level. This approach exploits an ocular burr to partly remove the corneal epithelium and uses scanning electron microscopy to track changes in cell morphology during wound closure.

ゼブラフィッシュ角膜創傷治癒:擦過傷から創傷閉鎖画像分析まで

As the transparent surface of the eye, the cornea is instrumental for clear sight. Due to its location, this tissue is prone to environmental insults. ...

Characterizing Epithelial Wound Healing In Vivo Using the Cnidarian Model Organism Clytia hemisphaerica

All animal organs, from the skin to eyes to intestines, are covered with sheets of epithelial cells that allow them to maintain homeostasis while protecting them from infection. Therefore, it is not surprising that the ability to repair epithelial wounds is critical to all metazoans. Epithelial wound healing in vertebrates involves overlapping processes, including inflammatory responses, vascularization, and re-epithelialization. Regulation of these processes involves complex interactions between epithelial cells, neighboring cells, and the extracellular matrix (ECM); the ECM contains structural proteins, regulatory proteins, and active small molecules. This complexity, together with the fact that most animals have opaque tissues and inaccessible ECMs, makes wound healing difficult to study in live animals. Much work on epithelial wound healing is therefore performed in tissue culture systems, with a single epithelial cell-type plated as a monolayer on an artificial matrix. Clytia hemisphaerica (Clytia) provides a unique and exciting complement to these studies, allowing epithelial wound healing to be studied in an intact animal with an authentic ECM. The ectodermal epithelium of Clytia is a single layer of large squamous epithelial cells, allowing high-resolution imaging using differential interfering contrast (DIC) microscopy in living animals. The absence of migratory fibroblasts, vasculature, or inflammatory responses makes it possible to dissect the critical events in re-epithelialization in vivo. The healing of various types of wounds can be analyzed, including single-cell microwounds, small and large epithelial wounds, and wounds that damage the basement membrane. Lamellipodia formation, purse string contraction, cell stretching, and collective cell migration can all be observed in this system. Furthermore, pharmacological agents can be introduced via the ECM to modify cell:ECM interactions and cellular processes in vivo. This work shows methods for creating wounds in live Clytia, capturing movies of healing, and probing healing mechanisms by microinjecting reagents into the ECM.

Characterizing Epithelial Wound Healing In Vivo Using the Cnidarian Model Organism Clytia hemisphaerica

This paper describes a method to create wounds in the epithelium of a live Clytia hemisphaerica medusa and image wound healing at a high resolution in vivo. Additionally, a technique to introduce dyes and drugs to perturb signaling processes in the epithelial cells and extracellular matrix during wound healing is presented.

刺胞動物モデル生物Clytia hemisphaericaを用いたin vivoでの上皮創傷治癒の特徴付け

All animal organs, from the skin to eyes to intestines, are covered with sheets of epithelial cells that allow them to maintain homeostasis while ...

マウスにおけるアルカリ熱傷誘発角膜血管新生術

A Mouse Model for Corneal Neovascularization by Alkali Burn

Corneal neovascularization (CoNV), a pathological form of angiogenesis, involves the growth of blood and lymph vessels into the avascular cornea from the limbus&#160;and adversely&#160;affects transparency and vision. Alkali burn is one of the most common forms of ocular trauma that leads to CoNV. In this protocol, CoNV is experimentally induced using sodium hydroxide solution in a controlled manner to ensure reproducibility. The alkali burn model is useful for understanding the pathology of CoNV and can be extended to study angiogenesis in general because of the avascularity, transparency, and accessibility of the cornea. In this work, CoNV was analyzed by direct examination under a dissecting microscope and by immunostaining flat-mount corneas using anti-CD31 mAb. Lymphangiogenesis was detected on flat-mount corneas by immunostaining using anti-LYVE-1 mAb. Corneal edema was visualized and quantified using optical coherence tomography (OCT). In summary, this model will help to advance existing neovascularization assays and discover new treatment strategies for pathologic ocular and extraocular angiogenesis.

著者スポットライト:将来の治療戦略のためのアルカリ熱傷モデルによる角膜血管新生術の解読 

This protocol focuses on alkali burn-induced corneal neovascularization in mice. The method generates a reproducible and controllable corneal disease model to study pathological angiogenesis and the associated molecular mechanisms and to test new pharmacological agents to prevent corneal neovascularization.

アルカリ熱傷による角膜血管新生術のマウスモデル

Corneal neovascularization (CoNV), a pathological form of angiogenesis, involves the growth of blood and lymph vessels into the avascular cornea from the ...