title: Quantitative assessment of the regenerative and mineralogenic performances of the zebrafish caudal fin.
abstract: The ability of zebrafish to fully regenerate its caudal fin has been explored to better understand the mechanisms underlying de novo bone formation and to develop screening methods towards the discovery of compounds with therapeutic potential. Quantifying caudal fin regeneration largely depends on successfully measuring new tissue formation through methods that require optimization and standardization. Here, we present an improved methodology to characterize and analyse overall caudal fin and bone regeneration in adult zebrafish. First, regenerated and mineralized areas are evaluated through broad, rapid and specific chronological and morphometric analysis in alizarin red stained fins. Then, following a more refined strategy, the intensity of the staining within a 2D longitudinal plane is determined through pixel intensity analysis, as an indicator of density or thickness/volume. The applicability of this methodology on live specimens, to reduce animal experimentation and provide a tool for in vivo tracking of the regenerative process, was successfully demonstrated. Finally, the methodology was validated on retinoic acid- and warfarin-treated specimens, and further confirmed by micro-computed tomography. Because it is easily implementable, accurate and does not require sophisticated equipment, the present methodology will certainly provide valuable technical standardization for research in tissue engineering, regenerative medicine and skeletal biology.
authors.affiliation: ProRegeM PhD Programme, Department of Biomedical Sciences and Medicine, University of Algarve, Campus de Gambelas, Faro, Portugal.|Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, Faro, Portugal.
authors.forename: João
authors.initials: J
authors.lastname: Cardeira
authors.affiliation: Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, Faro, Portugal.|Department of Biomedical Sciences and Medicine, University of Algarve, Campus de Gambelas, Faro, Portugal.
authors.forename: Paulo J
authors.initials: PJ
authors.lastname: Gavaia
authors.affiliation: Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, Faro, Portugal.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Fernández
authors.affiliation: 3B's Research Group, Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Avepark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco GMR, Portugal.|ICVS/3B's, PT Government Associated Laboratory, Portugal.
authors.forename: Ibrahim Fatih
authors.initials: IF
authors.lastname: Cengiz
authors.affiliation: ICVS/3B's, PT Government Associated Laboratory, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Moreira-Silva
authors.affiliation: 3B's Research Group, Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Avepark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco GMR, Portugal.|ICVS/3B's, PT Government Associated Laboratory, Portugal.
authors.forename: Joaquim Miguel
authors.initials: JM
authors.lastname: Oliveira
authors.affiliation: 3B's Research Group, Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Avepark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco GMR, Portugal.|ICVS/3B's, PT Government Associated Laboratory, Portugal.
authors.forename: Rui L
authors.initials: RL
authors.lastname: Reis
authors.affiliation: Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, Faro, Portugal.|Department of Biomedical Sciences and Medicine, University of Algarve, Campus de Gambelas, Faro, Portugal.
authors.forename: M Leonor
authors.initials: ML
authors.lastname: Cancela
authors.affiliation: Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, Faro, Portugal.
authors.forename: Vincent
authors.initials: V
authors.lastname: Laizé
mesh_terms: D058500:Animal Fins / Q000473:pathology / Q000502:physiology*; D000818:Animals; D001861:Bone Regeneration / Q000187:drug effects / Q000502:physiology; D001842:Bone and Bones / Q000502:physiology; D002113:Calcification, Physiologic / Q000187:drug effects; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D014212:Tretinoin / Q000494:pharmacology; D014859:Warfarin / Q000494:pharmacology; D055114:X-Ray Microtomography; D015027:Zebrafish / Q000502:physiology*
chemical_list: D014212:Tretinoin; D014859:Warfarin