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Titel: Scratch Migration Assay and Dorsal Skinfold Chamber for In Vitro and In Vivo Analysis of Wound Healing
VerfasserIn: Flockerzi, Veit
Menger, Michael D.
Laschke, Matthias W.
Belkacemi, Anouar
Sprache: Englisch
Titel: Journal of Visualized Experiments
Heft: 151
Verlag/Plattform: JoVE
Erscheinungsjahr: 2019
DDC-Sachgruppe: 610 Medizin, Gesundheit
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: Impaired cutaneous wound healing is a major concern for patients suffering from diabetes and for elderly people, and there is a need for an effective treatment. Appropriate in vitro and in vivo approaches are essential for the identification of new target molecules for drug treatments to improve the skin wound healing process. We identified the β3 subunit of voltage-gated calcium channels (Cavβ3) as a potential target molecule to influence the wound healing in two independent assays, i.e., the in vitro scratch migration assay and the in vivo dorsal skinfold chamber model. Primary mouse embryonic fibroblasts (MEFs) acutely isolated from wild-type (WT) and Cavβ3-deficient mice (Cavβ3 KO) or fibroblasts acutely isolated from WT mice treated with siRNA to down-regulate the expression of the Cacnb3 gene, encoding Cavβ3, were used. A scratch was applied on a confluent cell monolayer and the gap closure was followed by taking microscopic images at defined time points until complete repopulation of the gap by the migrating cells. These images were analyzed, and the cell migration rate was determined for each condition. In an in vivo assay, we implanted a dorsal skinfold chamber on WT and Cavβ3 KO mice, applied a defined circular wound of 2 mm diameter, covered the wound with a glass coverslip to protect it from infections and desiccation, and monitored the macroscopic wound closure over time. Wound closure was significantly faster in Cacnb3-gene-deficient mice. Because the results of the in vivo and the in vitro assays correlate well, the in vitro assay may be useful for the high-throughput screening before validating the in vitro hits by the in vivo wound healing model. What we have shown here for wild-type and Cavβ3-deficient mice or cells might also be applicable for specific molecules other than Cavβ3.
DOI der Erstveröffentlichung: 10.3791/59608-v
URL der Erstveröffentlichung: https://dx.doi.org/10.3791/59608
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-441574
hdl:20.500.11880/39494
http://dx.doi.org/10.22028/D291-44157
ISSN: 1940-087X
Datum des Eintrags: 27-Jan-2025
Fakultät: M - Medizinische Fakultät
Fachrichtung: M - Chirurgie
M - Experimentelle und Klinische Pharmakologie und Toxikologie
Professur: M - Prof. Dr. Veit Flockerzi
M - Prof. Dr. Michael D. Menger
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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