Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia

Abstract Chondrogenesis and angiogenesis drive endochondral ossification. Using the atmospheric scanning electron microscopy (ASEM) without decalcification and dehydration, we directly imaged angiogenesis-driven ossification at different developmental stages shortly after aldehyde fixation, using aq...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Eiko Sakai, Mari Sato, Nassirhadjy Memtily, Takayuki Tsukuba, Chikara Sato
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6dec791a1f8f4d63bd70a4f74e81274e
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:6dec791a1f8f4d63bd70a4f74e81274e
record_format	dspace
spelling	oai:doaj.org-article:6dec791a1f8f4d63bd70a4f74e81274e2021-12-02T13:20:03ZLiquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia10.1038/s41598-021-84202-z2045-2322https://doaj.org/article/6dec791a1f8f4d63bd70a4f74e81274e2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-84202-zhttps://doaj.org/toc/2045-2322Abstract Chondrogenesis and angiogenesis drive endochondral ossification. Using the atmospheric scanning electron microscopy (ASEM) without decalcification and dehydration, we directly imaged angiogenesis-driven ossification at different developmental stages shortly after aldehyde fixation, using aqueous radical scavenger glucose solution to preserve water-rich structures. An embryonic day 15.5 mouse femur was fixed and stained with phosphotungstic acid (PTA), and blood vessel penetration into the hypertrophic chondrocyte zone was visualised. We observed a novel envelope between the perichondrium and proliferating chondrocytes, which was lined with spindle-shaped cells that could be borderline chondrocytes. At postnatal day (P)1, trabecular and cortical bone mineralisation was imaged without staining. Additional PTA staining visualised surrounding soft tissues; filamentous connections between osteoblast-like cells and osteocytes in cortical bone were interpreted as the osteocytic lacunar-canalicular system. By P10, resorption pits had formed on the tibial trabecular bone surface. The applicability of ASEM for pathological analysis was addressed using knockout mice of Keap1, an oxidative-stress sensor. In Keap1 −/− femurs, we observed impaired calcification and angiogenesis of epiphyseal cartilage, suggesting impaired bone development. Overall, the quick ASEM method we developed revealed mineralisation and new structures in wet bone tissue at EM resolution and can be used to study mineralisation-associated phenomena of any hydrated tissue.Eiko SakaiMari SatoNassirhadjy MemtilyTakayuki TsukubaChikara SatoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eiko Sakai Mari Sato Nassirhadjy Memtily Takayuki Tsukuba Chikara Sato Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia
description	Abstract Chondrogenesis and angiogenesis drive endochondral ossification. Using the atmospheric scanning electron microscopy (ASEM) without decalcification and dehydration, we directly imaged angiogenesis-driven ossification at different developmental stages shortly after aldehyde fixation, using aqueous radical scavenger glucose solution to preserve water-rich structures. An embryonic day 15.5 mouse femur was fixed and stained with phosphotungstic acid (PTA), and blood vessel penetration into the hypertrophic chondrocyte zone was visualised. We observed a novel envelope between the perichondrium and proliferating chondrocytes, which was lined with spindle-shaped cells that could be borderline chondrocytes. At postnatal day (P)1, trabecular and cortical bone mineralisation was imaged without staining. Additional PTA staining visualised surrounding soft tissues; filamentous connections between osteoblast-like cells and osteocytes in cortical bone were interpreted as the osteocytic lacunar-canalicular system. By P10, resorption pits had formed on the tibial trabecular bone surface. The applicability of ASEM for pathological analysis was addressed using knockout mice of Keap1, an oxidative-stress sensor. In Keap1 −/− femurs, we observed impaired calcification and angiogenesis of epiphyseal cartilage, suggesting impaired bone development. Overall, the quick ASEM method we developed revealed mineralisation and new structures in wet bone tissue at EM resolution and can be used to study mineralisation-associated phenomena of any hydrated tissue.
format	article
author	Eiko Sakai Mari Sato Nassirhadjy Memtily Takayuki Tsukuba Chikara Sato
author_facet	Eiko Sakai Mari Sato Nassirhadjy Memtily Takayuki Tsukuba Chikara Sato
author_sort	Eiko Sakai
title	Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia
title_short	Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia
title_full	Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia
title_fullStr	Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia
title_full_unstemmed	Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia
title_sort	liquid-phase asem imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: keap1-deficient osteomalacia
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/6dec791a1f8f4d63bd70a4f74e81274e
work_keys_str_mv	AT eikosakai liquidphaseasemimagingofcellularandstructuraldetailsincartilageandboneformedduringendochondralossificationkeap1deficientosteomalacia AT marisato liquidphaseasemimagingofcellularandstructuraldetailsincartilageandboneformedduringendochondralossificationkeap1deficientosteomalacia AT nassirhadjymemtily liquidphaseasemimagingofcellularandstructuraldetailsincartilageandboneformedduringendochondralossificationkeap1deficientosteomalacia AT takayukitsukuba liquidphaseasemimagingofcellularandstructuraldetailsincartilageandboneformedduringendochondralossificationkeap1deficientosteomalacia AT chikarasato liquidphaseasemimagingofcellularandstructuraldetailsincartilageandboneformedduringendochondralossificationkeap1deficientosteomalacia
_version_	1718393217549860864

Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia

Ejemplares similares