Really Close Up! Surveying Surfaces at Sub‐micrometre Resolution: The Measurement of Osteoclastic Resorption Lacunae

Abstract

The authors discuss systems for the three dimensional measurement of microscopic features, in particular the pits made by isolated bone resorbing cells (osteoclasts) in test substrates with flat surfaces; such measurements may be used to assay the work done by these cells. Confocal light microscopy, with its very shallow depth of jield, seems to be replacing scanning electron microscopic stereophotogrammetry as the preferred method for such measurement.BOYDE, A,, 1968. Height measurements from stereopair scanning electron micrographs. Beitruege zur Elektronenmikroskopische Direktabbrldung yon Oberjlaechen. Munster, I: 97-105. BOYDE, A., 1970a. Practical methods, application and problems of 3-D analysis of scanning electron microscope (SEM) images. Scanning Electron Microscopy 1970 (Ed. 0. Johari). Illinois Institute of Technology Research Institute, Chicago, Illinois. Pages 193-200. BOYDE, A,, 1970b. Calibration of the specimen stage of the Stereoscan. Beitruege zur Elektronenrnikroskopische Direktabbildung van Oberjlaechen, Munster, 3: 403-4 10. BOYDE, A,, 1971. The tooth surface. In Theprevenlion ofperiodontaldisease(J. E. Eastoe, D. C. A. Picton and A. G. Alexander, Editors). Henry Kimpton, London. 244 pages: 46-63. BOYDE, A., 1973. Quantitative photogrammetric analyses and qualitative stereoscopic analysis of SEM images. Journal of Microscopy, 98(3): 452-47 I. BOYDE, A,, I974a. A stereo-plotting device for SEM micrographs: and a real-time 3-D system for the SEM. Scanning Electron Microscopy I974 (Ed. 0. Johari and 1. Corvin). Illinois Institute of Technology Research Institute, Chicago, Illinois. Page 93-100. BOYDE, A,, 1974b. Photogrammetry of stereopair SEM images using separate measurements from the two images. /bid.: 101-108. BOYDE, A,, 1 9 7 4~. Real-time stereo TV speed SEM. Beitraege zur Elektronenmikroskopische Direktabbildung yon Oberjaechen. Munster. 7: 22 1-230. BOYDE, A,, 1975. Measurement of specimen height difference and beam tilt angle in anaglyph real-time

Modeled gravitational unloading triggers differentiation and apoptosis in preosteoclastic cells

“…FLG 29.1 cells cultured in standard conditions at 1 Â g are immature elements unable to adhere to or resorb bone. In fact, as expected, no resorption activity was found in bone slices from the control samples at 1 Â g. On the contrary, a mean value of resorbed bone volume equal to 2 AE 0.5 mm 3 per mm 2 was found in the bone slices from samples exposed to modeled low g. This value is in accordance with data reported by Boyde and Jones [1992], who reviewed the more reliable methods used in the study of shape and dimension of Howship's lacunae in order to extrapolate the volume of resorbed bone. Surface analysis by SEM and Digital wide-field microscopy with grazing lighting gave visual evidence that FLG 29.1 cells cultured in the RPM were able to adhere to the bone surface and resorb bone.…”

Targeting Drug Delivery in the Elderly: Are Nanoparticles an Option for Treating Osteoporosis?

“…The sequence described above is a summary of the findings reported in an extensive series of papers over a 20-year period (about 1954-1985), resulting largely from work by the research team associated with Prof. Alan Boyde. Prominent amongst the series of papers were these: [46][47][48][49][50][51][52][53][54][55][56][57][58][59][60]. A smaller number of authors have considered the distribution of resorption areas throughout the skeleton: for example, in rabbits [61] or human [62].…”

“…Narrow and deep pits-for example, 6 µm wide and 20 µm deep-are unlikely to have strong flows of blood circulating in them and might be best targeted by use of the smallest size nanoparticles. The measurement of Howship's pit depths is technically difficult and has required ingenuity and specialized instrumentation to achieve reliable measurements [58][59][60]. The pits often reach volumes of 1000 µm 3 , and volumes of 2000 µm 3 are not rare; much more rarely, volumes as large as 10,000 µm 3 can be found.…”

“…The pits often reach volumes of 1000 µm 3 , and volumes of 2000 µm 3 are not rare; much more rarely, volumes as large as 10,000 µm 3 can be found. A pit of width 10-40 µm and depth 4-6 µm can be considered "normal" [60]. As is often the case in nanomedicine, the biological conditions within the target tissue must be considered equally as carefully as the design of the nanoparticles to be employed.…”