About of the possibility of using of Adobe Photoshop software for the morphometric analysis of digital images of micropreparations during the immunohistochemical study of the bone matrix
The article proposes a method of objective evaluation of the content of bone matrix proteins (collagen I, collagen II and osteocalcin) using Adobe Photoshop software. The study was conducted on 70 adult male rats “Wistar” line, to which formed an open transverse fracture of the middle third of the tibial shaft. In the experimental group (EG) the animals to fracture on day 1 and 2 injected in 0,5 ml “Vinfar” preparation containing metabolites Bacillus subtillis 804, in the control group (CG) - 0,5 ml 0,9% NaCl. On days 3, 21, 44 and 61 osteoreparative process used digital analysis of photomicrographs of immunohistochemical preparations using Adobe Photoshop Extended software for calculate the area sites of structures containing studied proteins, followed objective calculation of the relative content in the tissues by the formula: RVD (%) = (Sa / St) x 100, where the RVD - the relative volume density, Sa - the total area of all the the selected sites in the analyzed image, St - total area of the digital image. It was established that on day 44 in animals CG periosteal fracture zone RVD of the osteocalcin is 1,156 ± 0,034%, and in EG RVD of the osteocalcin already at this period reaches values (density 1,582 ± 0,026%) close to normal bone tissue (about 1,6%). At day 61 the expression of type I collagen in EG (RVD 69,25 ± 2,23%) more than a third higher than that of CG (RVD 46,09 ± 1,74%), reaching a value close to RVD of the collagen i of normal bone tissue (about 70.0%). Thus, the determination of RVD of the collagen l, collagen il and osteocalcin using Adobe Photoshop software allows to objectively evaluate their content in the bone matrix, which gives an indication of the character of the healing of bone fractures.