The effects of three different demineralization agents on osteopontin localization in adult rat bone using immunohistochemistry.

摘要: Immunohistochemical localization of osteopontin, a phosphorylated acidic glycoprotein, was compared in adult rat femur fixed 4% paraformaldehyde at 4° C for 48 h and demineralized ethylenediaminetetraacetic acid (EDTA), modified Jenkin's solution, or 15% formic acid, until radiographs indicated demineralization complete. Formic also evaluated room temperature. EDTA solution (15 days) resulted intense staining osteocytes, periosteal osteoclasts osteoblastic cells osteonal bone. Osteoblasts were negative the periosteum. No megakaryocyte present; however, occasional neutrophils bone marrow non-specifically stained. Demineralization (16 showed osteopontin matrix, hypertrophic articular chondrocytes, osteocytes. In cortical bone, almost all cement lines demarcating osteons very dense labeling. marrow, megakaryocytes immunopositive produced non-specific skeletal muscle connective tissue. (14 days, C) expression osteoblasts, osteoclast precursors, osteoid, lines, chondrocytes; osteoclasts, although present low numbers, positive. More labeled osteoblasts could be identified to demineralization. Also more obtained than with Jenkin's. Harsh, rapid (4 temperature) loss antigenicity demonstrated by reduction intensity not experienced protocol; still localized matrix zone chondrocytes. These results indicate that is compatible retention immunoreactive Both produce excellent immunostaining are preferred over minimize background levels staining.