Review
Issues in modern bone histomorphometry
☆
R.R. Recker
a,
⁎
, D.B. Kimmel
a
, D. Dempster
b
, R.S. Weinstein
c
, T.J. Wronski
d
, D.B. Burr
e,f
a
Osteoporosis Research Center, Creighton University, 601 N 30th St Omaha, NE, 68131, USA
b
Regional Bone Center, Helen Hayes Hospital, Route 9W, West Haverstraw, NY, 10993, USA
c
Center for Osteoporosis and Metabolic Bone Diseases, Dept. of Medicine, Central Arkansas Veterans Health Care System and University of Arkansas for Medical Sciences, 4301 West
Markham St.- slot 587, Little Rock AR, 72205, USA
d
Dept. of Physiological Sciences, University of Florida, P.O. Box 100144, JHMHC, Gainesville, FL, 32610, USA
e
Dept. of Anatomy and Cell Biology, Indiana University School of Medicine, 635 Barnhill Dr., MS 5035, Indianapolis, IN, 46202, USA
f
Dept. of Biomedical Engineering, Indiana University-Purdue University Indiana (IUPUI), USA
abstractarticle info
Article history:
Received 18 February 2011
Revised 25 June 2011
Accepted 13 July 2011
Available online 23 July 2011
Edited by: R. Baron
Keywords:
Histomorphometry
Bone remodeling
Over-suppression
This review reports on proceedings of a bone histomorphometry session conducted at the Fortieth International
IBMS Sun Valley Skeletal Tissue Biology Workshop held on August 1, 2010. The session was prompted by recent
technical problems encountered in conducting histomorphometry on bone biopsies from humans and animals
treated with anti-remodeling agents such as bisphosphonates and RANKL antibodies. These agents reduce
remodeling substantially, and thus cause problems in calculating bone remodeling dynamics using in vivo
fluorochrome labeling. The tissue specimens often contain few or no fluorochrome labels, and thus create statistical
and other problems in analyzing variables such as mineral apposition rates, mineralizing surface and bone formation
rates. The conference attendees discussed these problems and their resolutions, and the proceedings reported here
summarize their discussions and recommendations.
© 2011 Elsevier Inc. All rights reserved.
Introduction
The understanding of bone physiology accelerated in the 1950s
with the development of plastic-embedding technology for microscopic
examination of undemineralized sections of bone tissue. Prior to
then, bone histology required removal of its principal component, the
mineral. This new technology was refined in several labs, in particular,
the Radiobiology Lab at the University of Utah in studies of the skeletal
effects of plutonium and radium, alpha radiation-emitting, bone-
seeking metals. A benchmark occurred in 1960 with a publication by
Frost [1] entitled, “Measurement of Bone Formation in a 57 Year-old
Man by Means of Tetracyclines.” This opened the door for study of the
complete composite of human bone, including both mineral and organic
matrix. In 1983, a text was produced [2] which summarized the
methodology and stereological theories behind the histomorphometric
calculations. Another benchmark occurred in 1987 when the nomen-
clature, symbols and units for bone histomorphometry were standard-
ized [3]. As a consequence, the use of in-life dual fluorochrome labels in
humans and animals together with the application of bone histomor-
phometric techniques has become a standard method for studying the
tissue level activities of bone modeling and remodeling. Starting in the
late 1960s, Professor Webster Jee began the Sun Valley Workshops on
musculoskeletal biology that continue to the present. In August 2010,
the International Bone and Mineral Society, along with the National
Institutes of Health, sponsored the 40th Annual IBMS/Sun Valley
Workshop on musculoskeletal biology. At that meeting a half-day
session was devoted to discussing issues in modern histomorphometry,
50 years after its introduction. The authors participated in the pre-
sentations and subsequent discussion.
Change in landscape
The success of potent anti-remodeling agents in reducing fracture risk
in osteoporotic patients in recent years has presented new real world
problems for bone histomorphometry, as it is applied to explain their
tissue level mechanisms. Bone biopsies taken from patients, or specimens
taken from animals treated with these agents, may have few or no
fluorochrome labels, creating difficulty in the interpretation of the
dynamic histomorphometric data and evaluation of patients' or animals'
bone remodeling status. This experience has generated a number of
important questions that have been debated in publications and scientific
meetings over the past decade. Other issues involve use of proper
referents for histomorphometric measurements, clarification of the
utility of certain bone histomorphometric measurements and endpoints,
sample size requirements for valid measurements, quantification of
rare events, and the variables that should be reported to describe best
Bone 49 (2011) 955–964
☆
Presented at the Fortieth International IBMS Sun Valley Skeletal Tissue Biology
Workshop; August 1, 2010.
⁎ Corresponding author at: 601 N 30th St, #5766, Omaha, NE, 68131, USA. Fax: +1
402 280 5034.
E-mail address: rrecker@creighton.edu (R.R. Recker).
8756-3282/$ – see front matter © 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.bone.2011.07.017
Contents lists available at ScienceDirect
Bone
journal homepage: www.elsevier.com/locate/bone