An Improved Methodology to Evaluate Cell and Molecular Signals in the Reparative Callus During Fracture Healing

8Citations
Citations of this article
13Readers
Mendeley users who have this article in their library.

This article is free to access.

Abstract

Approximately 5% to 10% of all bone fractures do not heal completely, contributing to significant patient suffering and medical costs. Even in healthy individuals, fracture healing is associated with significant downtime and loss of productivity. However, no pharmacological treatments are currently available to promote efficient bone healing. A better understanding of the underlying molecular mechanisms is crucial for developing novel therapies to hasten healing. The early reparative callus that forms around the site of bone injury is a fragile tissue consisting of shifting cell populations held together by loose connective tissue. The delicate callus is challenging to section and is vulnerable to disintegration during the harsh steps of immunostaining, namely, decalcification, deparaffinization, and antigen retrieval. Here, we describe an improved methodology for processing early-stage fracture calluses and immunofluorescence labeling of the sections to visualize the temporal (timing) and spatial (location) patterns of cellular and molecular events that regulate bone healing. This method has a short turnaround time from sample collection to microscopy as it does not require lengthy decalcification. It preserves the structural integrity of the fragile callus as the method does not entail deparaffinization or harsh methods of antigen retrieval. Our method can be adapted for high-throughput screening of drugs that promote efficacious bone healing:.

References Powered by Scopus

Indian hedgehog signaling regulates proliferation and differentiation of chondrocytes and is essential for bone formation

1450Citations
N/AReaders
Get full text

Fracture healing: Mechanisms and interventions

1269Citations
N/AReaders
Get full text

Bone remodeling during fracture repair: The cellular picture

705Citations
N/AReaders
Get full text

Cited by Powered by Scopus

A review of recent developments in the molecular mechanisms of bone healing

26Citations
N/AReaders
Get full text

Porous gelatin microsphere-based scaffolds containing MC3T3-E1 cells and calcitriol for the repair of skull defect

9Citations
N/AReaders
Get full text

CAMKK2 is upregulated in primary human osteoarthritis and its inhibition protects against chondrocyte apoptosis

5Citations
N/AReaders
Get full text

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Cite

CITATION STYLE

APA

Valiya Kambrath, A., Williams, J. N., & Sankar, U. (2020). An Improved Methodology to Evaluate Cell and Molecular Signals in the Reparative Callus During Fracture Healing. Journal of Histochemistry and Cytochemistry, 68(3), 199–208. https://doi.org/10.1369/0022155419900915

Readers' Seniority

Tooltip

PhD / Post grad / Masters / Doc 5

71%

Professor / Associate Prof. 1

14%

Researcher 1

14%

Readers' Discipline

Tooltip

Medicine and Dentistry 4

57%

Agricultural and Biological Sciences 1

14%

Materials Science 1

14%

Engineering 1

14%

Save time finding and organizing research with Mendeley

Sign up for free