In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen Withdrawal

The bone is a highly dynamic organ that undergoes continuous remodeling through an intricate balance of bone formation and degradation. Hyperactivation of the bone-degrading cells, the osteoclasts (OCs), occurs in disease conditions and hormonal changes in females, resulting in osteoporosis, a disea...

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Main Authors: Cara Fiorino, Safia Omer, Nisha Gandhi, Rene Harrison
Format: Article
Language:English
Published: Bio-protocol LLC 2025-01-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5155&type=0
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author Cara Fiorino
Safia Omer
Nisha Gandhi
Rene Harrison
author_facet Cara Fiorino
Safia Omer
Nisha Gandhi
Rene Harrison
author_sort Cara Fiorino
collection DOAJ
description The bone is a highly dynamic organ that undergoes continuous remodeling through an intricate balance of bone formation and degradation. Hyperactivation of the bone-degrading cells, the osteoclasts (OCs), occurs in disease conditions and hormonal changes in females, resulting in osteoporosis, a disease characterized by altered microarchitecture of the bone tissue, and increased bone fragility. Thus, building robust assays to quantify OC resorptive activity to examine the molecular mechanisms underlying bone degradation is critical. Here, we establish an in vitro model to investigate the effect of estrogen withdrawal on OCs derived from the mouse macrophage RAW 264.7 cell line in a bone biomimetic microenvironment. This simple and robust model can also be adapted to examine the effect of drugs and genetic factors influencing OC resorptive activity in addition to being compatible with fluorescent imaging.
format Article
id doaj-art-134bc9f132f64569b47041438dc76b32
institution Kabale University
issn 2331-8325
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spelling doaj-art-134bc9f132f64569b47041438dc76b322025-02-07T08:16:31ZengBio-protocol LLCBio-Protocol2331-83252025-01-0115110.21769/BioProtoc.5155In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen WithdrawalCara Fiorino0Safia Omer1Nisha Gandhi2Rene Harrison3Department of Biological Sciences, University of Toronto Scarborough, Toronto, ON, CanadaDepartment of Biological Sciences, University of Toronto Scarborough, Toronto, ON, CanadaDepartment of Cell & Systems Biology, University of Toronto Scarborough, Toronto, ON, CanadaDepartment of Biological Sciences, University of Toronto Scarborough, Toronto, ON, CanadaDepartment of Cell & Systems Biology, University of Toronto Scarborough, Toronto, ON, CanadaThe bone is a highly dynamic organ that undergoes continuous remodeling through an intricate balance of bone formation and degradation. Hyperactivation of the bone-degrading cells, the osteoclasts (OCs), occurs in disease conditions and hormonal changes in females, resulting in osteoporosis, a disease characterized by altered microarchitecture of the bone tissue, and increased bone fragility. Thus, building robust assays to quantify OC resorptive activity to examine the molecular mechanisms underlying bone degradation is critical. Here, we establish an in vitro model to investigate the effect of estrogen withdrawal on OCs derived from the mouse macrophage RAW 264.7 cell line in a bone biomimetic microenvironment. This simple and robust model can also be adapted to examine the effect of drugs and genetic factors influencing OC resorptive activity in addition to being compatible with fluorescent imaging.https://bio-protocol.org/en/bpdetail?id=5155&type=0
spellingShingle Cara Fiorino
Safia Omer
Nisha Gandhi
Rene Harrison
In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen Withdrawal
Bio-Protocol
title In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen Withdrawal
title_full In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen Withdrawal
title_fullStr In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen Withdrawal
title_full_unstemmed In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen Withdrawal
title_short In Vitro Assay to Examine Osteoclast Resorptive Activity Under Estrogen Withdrawal
title_sort in vitro assay to examine osteoclast resorptive activity under estrogen withdrawal
url https://bio-protocol.org/en/bpdetail?id=5155&type=0
work_keys_str_mv AT carafiorino invitroassaytoexamineosteoclastresorptiveactivityunderestrogenwithdrawal
AT safiaomer invitroassaytoexamineosteoclastresorptiveactivityunderestrogenwithdrawal
AT nishagandhi invitroassaytoexamineosteoclastresorptiveactivityunderestrogenwithdrawal
AT reneharrison invitroassaytoexamineosteoclastresorptiveactivityunderestrogenwithdrawal