Development of an ultrathin sheet plastination technique in rat humeral joints with osteoarthritis induced by monosodium iodoacetate for neovascularization study
Injection with monosodium iodoacetate (MIA) is widely used to produce osteoarthritis (OA). Ultrathin sheet plastination has been used to study the morphology of structures, with strong application in anatomical education and research. Our aim was to carry out, for the first time, ultrathin sheet plastination of rat humeral joints to observe the neovascularization provoked by OA. We injected 0.1 mL of MIA into the left humeral joints of ten Sprague–Dawley rats. The right shoulders of the same rats were used as control. Sixteen weeks after the injection, the animals were euthanized and were given an immediate red epoxy resin injection through the thoracic aorta. The samples were fixed in 10% formalin, prior to the plastination process, without decalcification. Samples were dehydrated with acetone (100%) at − 25 °C, for 10 days. Later, for degreasing, samples were immersed in methylene chloride at room temperature during 1 week. Forced impregnation was performed inside a stove within a vacuum chamber. The plastinated blocks obtained were cut with a slow velocity diamond blade saw. Slices were placed in curing chambers to achieve curing and final tissue transparentation. 230 μm thickness slices were obtained. The slices were analyzed under magnifying glass and microscope, achieving visualization of OA neovascularization. The cartilage affected by OA loses its ability to remain avascular, and blood vessels invade it from the subchondral bone to the calcified and uncalcified cartilage. Ultra-thin sheet plastination is useful to observe articular cartilage neovascularization, caused by OA induced with MIA in humeral rat joint.
KeywordsPlastination Ultrathin Osteoarthritis Animal model Epoxy
This work was supported by a grant from the Dental School of Universidad de La Frontera: “Research and Productivity Internationalization Project 2018” (Proyecto Internacionalización para Investigación y Productividad 2018, Facultad de Odontología, Universidad de La Frontera) and a grant from Dirección de Investigación, Universidad de La Frontera, Project DIUFRO FONDOS PROPIOS; Grant number: DFP16-0012.
CAV: data acquisition (osteoarthritis protocol, ultrathin sheet plastination protocol; histological analysis), conceptualization, analysis, interpretation and drafting of the manuscript. CACB: data acquisition (ultrathin sheet plastination protocol), conceptualization and critical revision of manuscript. MDS: conceptualization, critical revision of manuscript. CSV: data acquisition (histological analysis) and conceptualization of the manuscript. BV: data acquisition (histological analysis) and conceptualization of the manuscript. CV: data acquisition (osteoarthritis protocol) and conceptualization of the manuscript. NEO: data acquisition (osteoarthritis protocol, ultrathin sheet plastination protocol; histological analysis), conceptualization, analysis, interpretation and drafting of the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This study was approved by the Scientific Ethics Committee of Universidad de La Frontera (Decision N°084/17 of 9 August 2017).
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