Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Orthop. Sep 18, 2017; 8(9): 681-687
Published online Sep 18, 2017. doi: 10.5312/wjo.v8.i9.681
Electron probe microanalysis оf experimentally stimulated osteoarthrosis in dogs
Tatyana Stupina, Michael Shchudlo, Michael Stepanov
Tatyana Stupina, Michael Shchudlo, Michael Stepanov, Laboratory of Morphology, FSBI Russian Ilizarov Scientific Center “Restorative Traumatology and Orthopaedics”, 640014 Kurgan, Russia
Author contributions: Stupina T, Shchudlo M and Stepanov M substantially contributed to the conception and design of the study, acquisition, analysis and interpretation of data; all authors drafted the article and made critical revisions related to the intellectual content of the manuscript, and approved the final version of the article to be published.
Supported by The RF Ministry of Health within government-mandated program for FSBI Russian Ilizarov Scientific Center “Restorative Traumatology and Orthopaedics” (RISC “RTO”) for Scientific Research, No. 01201155770.
Institutional review board statement: The article was reviewed by RISC “RTO” Review Board and statement including: (1) the manuscript is not simultaneously being considered by other journals or already published elsewhere; (2) the manuscript has no redundant publication, plagiarism, or data fabrication or falsification; (3) experiments involving animal subjects were designed and performed in compliance with the relevant laws regarding the animal care and use of subjects; and (4) material contained within the manuscript is original, with all information from other sources appropriately referenced.
Institutional animal care and use committee statement: All animal studies are approved by RISC “RTO” Ethical Committee - excerpts from the minutes #4 (50) under date of December 13, 2016.
Conflict-of-interest statement: The authors declare no conflict of interest.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Dr. Tatyana Stupina, Senior Researcher, Laboratory of Morphology, FSBI Russian Ilizarov Scientific Center “Restorative Traumatology and Orthopaedics”, M. Ulianova Street 6, 640014 Kurgan, Russia. stupinasta@mail.ru
Telephone: +7-890-58506789 Fax: +7-352-2454060
Received: January 25, 2017
Peer-review started: February 2, 2017
First decision: May 23, 2017
Revised: May 26, 2017
Accepted: July 14, 2017
Article in press: July 15, 2017
Published online: September 18, 2017
Processing time: 232 Days and 7.8 Hours
Abstract
AIM

To develop methods of articular cartilage preparation for X-ray-electron probe microanalysis and to study its elements content in experimental osteoarthrosis.

METHODS

Twenty dogs aged 2-8 years were divided in research (aged 2 years, induction of osteoarthrosis - IOA) and intact group. Intact group included three subgroups (aged 2, 5 and 8 years). Samples of cartilage after araldite saturation and pouring were partially cut into semithin sections stained with methylene blue and with methylene blue-basic fuchsin. Their smooth surfaces were investigated by X-ray-electron probe microanalysis. Spatial distribution of sulfur, calcium and phosphorus and their concentrations (weight %) were investigated.

RESULTS

X-ray electron probe microanalysis revealed non-uniform sulfur distribution in cartilage of intact animals: Its content increases from superficial zone to deep one, this regularity was preserved in animals with IOA. Differences of IOA with spontaneous chondropathy were revealed. Spontaneous aging was characterized by calcium and phosphorus storage in deep and calcified zones and compensatory increase of sulfated glycosaminoglycans in intermediate and deep cartilage zones as evidenced by the metachromatic reaction and microanalysis data. Unlike spontaneous chondropathy connected with aging in experimentally stimulated osteoarthrosis more intensive storage of calcium but minor phosphorus in intermediate zone were marked. In IOA the calcified cartilage thinning and osteoclastic resorption are apparent with few changes of elements composition; the only difference from control is minority phosphorus content.

CONCLUSION

The obtained results demonstrate specific tricks of X-ray electron probe microanalysis and its possibility in the research of mechanisms of articular cartilage alterations in osteoarthrosis.

Keywords: Osteoarthrosis; Articular cartilage; Dog; X-ray electron probe microanalysis

Core tip: In this basic study we present the development of methods of articular cartilage preparation for X-ray-electron probe microanalysis and elements content in articular cartilage in animal experimentally induced (IOA) osteoarthrosis and during spontaneous animal aging (SA). SA was characterized by calcium and phosphorus storage in deep and calcified articular cartilage zones and compensatory increase of sulfated glycosaminoglycans in intermediate and deep zones. In IOA more intensive storage of calcium but few phosphorus in intermediate zone were marked. As for Sulphur content, all zones of uncalcified cartilage in two-year-old animals with IOA were comparable with cartilage of five-year-old intact animals.