ArticleJournal of materials science. Materials in medicine2026
Multimodal analysis of biomineralization within a collagen scaffolding in a rat calvarial defect model by using decalcified and undecalcified specimens.
Article in Journal of materials science. Materials in medicine, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
This study histologically evaluated the biomineralization process during bone regeneration and the in vivo behavior of a collagen sheet used as scaffolding in a rat 5-mm calvarial defect model. Two experimental groups were established: a group using collagen sheet and bone substitute (BC group), and a group using bone substitute alone (BO group). Bone regeneration was assessed by computed tomography (CT) and both decalcified and undecalcified sections were analyzed using histological staining (hematoxylin and eosin, Villanueva-Goldner [VG], von Kossa, and Join of the Five dyes Revealing CoLlagenous tissue [JFRL]), immunohistochemistry, polarized light microscopy, and low-vacuum scanning electron microscopy (LV-SEM) combined with energy-dispersive X-ray spectroscopy (EDX). CT revealed time-dependent defect reductions, progressing significantly faster in the BC group. In undecalcified specimens, VG staining demonstrated a thick, red, osteoid layer, and serial sections stained with von Kossa showed granular blackish-brown deposits within this layer. LV-SEM/EDX confirmed localized Ca/P accumulation in these deposits, indicating initial biomineralization foci. In decalcified JFRL-stained sections, JFRL color profiles corresponded to gray-scale contrast in LV-SEM images, reflecting collagen fibril organization and the degree of biomineralization. Polarized observation of undecalcified, VG-stained, polished sections revealed the emergence and temporal expansion of orange birefringence within the transplanted collagen sheet and surrounding connective tissue. Immunohistochemistry demonstrated BrdU-, Runx2-, and osterix-positive cells, and osteopontin localization within newly formed matrix in the defect, indicating active osteoblastogenesis. Collagen sheets appear to function not only as physical scaffolding, but also as a bioactive matrix promoting biomineralization by modulating cellular activity and matrix remodeling.
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