Calcification during bone healing in a standardised rat calvarial defect assessed by micro‐<scp>CT</scp>and<scp>SEM</scp>‐<scp>EDX</scp>

Okata, Hiroshi; Nakamura, Megumi; Henmi, Akiko; Yamaguchi, Satoko; Mikami, Yasuto; Shimauchi, Hidetoshi; Sasano, Yasuyuki

doi:10.1111/odi.12212

Cited by 18 publications

(17 citation statements)

References 43 publications

(61 reference statements)

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“…Ca and P have accumulated in the bone matrix (＊ in d-i) at E16 and the area expands through week 1, matching the region of developing bone (＊ in a-c), whereas C becomes localized around bone (＊ in j-l). The area with higher concentrations of Ca and P shows a lower concentration of C. ratios decrease during bone development and healing in calvaria (5,12). Fourier transform infrared spectroscopy (FTIR) analysis indicated that the proportion of protein to mineral decreases during calvarial bone development (5).…”

Section: Discussionmentioning

confidence: 99%

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Calcification in rat developing mandibular bone demonstrated by whole mount staining, microcomputed tomography and scanning electron microscopy with energy dispersive X-ray spectroscopy

et al. 2017

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The study was designed to investigate calcification in developing rat mandibular bone using whole mount staining, micro-computed tomography (micro-CT) and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). Wistar rats at embryonic days 16, 18, and 20 and postnatal weeks 1 and 6 were used. Rats were fixed with 4% paraformaldehyde and heads were resected, frozen and sectioned for histology, then analysed with SEM-EDX. Some of the specimens were observed with micro-CT. Other rats were fixed and stained with alcian blue and alizarin red for whole mount staining. Histology and whole mount staining showed that osteoid was deposited around Meckel's cartilage at day 15 and developed into bone at day 16. Accumulation of Ca and P was identified in the bone matrix with SEM-EDX. The area of bone expanded until week 6. The Ca/P ratio increased, whereas the C/Ca and C/P ratios decreased during development. Micro-CT demonstrated an increase in radio-opacity with bone development. The results suggest that rat mandibular bone formation is initiated around Meckel's cartilage at day 15. Deposition and maturation of the calcium phosphate mineral increase gradually with decrease in the organic component as the rat mandible develops.Rat mandibular bone formation begins with deposition of uncalcified bone matrix in the middle of the mandibular process, lateral to Meckel's cartilage, at embryonic day 15 (E15). The uncalcified bone matrix calcifies at E16 (2,3,13,14). Various regions of the mandibular bone differentiate and expand from the ossification site with time. The mandible has been shown to be one of the first bones to calcify in pigs (6, 10) and rabbits (4). Calcification of the bone matrix in the mandible may proceed by deposition, and then by increase and maturation of calcium phosphate minerals, as reported in calvaria (5). However, little information is available about the process of mandibular bone calcification during development. The study was designed to investigate the process of calcification in developing rat mandibular bone. We examined the developing mandible with histology and whole mount staining to demonstrate the spatiotemporal three-dimensional architecture of developing bone. The process of calcification related to mandibular bone development is shown using micro-computed tomography (micro-CT). Analysis of the concentrations and distributions of elements that constitute the bone matrix, that is, calcium (Ca), phosphorus (P) and carbon (C) is performed by scanning electron microscope equipped with an energy dispersive X-ray spectroscope (SEM-EDX).

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Section: Discussionmentioning

confidence: 99%

“…The samples were reconstructed by using a software program (Tri3DBon64; Ratoc, Tokyo, Japan). The threshold for standardising micro-CT images for separating bone from soft tissue was determined, as previously reported (12).…”

Section: Micro-ctmentioning

confidence: 99%

Calcification in rat developing mandibular bone demonstrated by whole mount staining, microcomputed tomography and scanning electron microscopy with energy dispersive X-ray spectroscopy

et al. 2017

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“…Our data of R Ca/P (around 1.55) in AIS was in agreement with previous report of lower R Ca/P value in younger bone tissue and indicated adequate homeostasis of calcium and phosphorous deposition in AIS. The ratio of calcium to phosphorus (R Ca/P ) was widely accepted as an indicator for assessing the status of bone mineralization2526. Carbon (C) was determined as the reference to assess the calcium content in the bone tissue because of its abundant and relatively stable content and being the major component of bone tissue262728.…”

Section: Resultsmentioning

confidence: 99%

“…The ratio of calcium to phosphorus (R Ca/P ) was widely accepted as an indicator for assessing the status of bone mineralization2526. Carbon (C) was determined as the reference to assess the calcium content in the bone tissue because of its abundant and relatively stable content and being the major component of bone tissue262728. EDX scanning showed significantly lower R Ca/C in AIS, but there was absence of significant difference in R Ca/P between AIS and Con, suggesting decreased mineralization which was in agreement with more osteoid as indicated by histomorphometric analyses (Table 6).…”

Section: Resultsmentioning

confidence: 99%

Unique local bone tissue characteristics in iliac crest bone biopsy from adolescent idiopathic scoliosis with severe spinal deformity

Wang

Chen

et al. 2017

Sci Rep

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Adolescent idiopathic scoliosis is a complex disease with unclear etiopathogenesis. Systemic and persistent low bone mineral density is an independent prognostic factor for curve progression. The fundamental question of how bone quality is affected in AIS remains controversy because there is lack of site-matched control for detailed analysis on bone-related parameters. In this case-control study, trabecular bone biopsies from iliac crest were collected intra-operatively from 28 severe AIS patients and 10 matched controls with similar skeletal and sexual maturity, anthropometry and femoral neck BMD Z-score to control confounding effects. In addition to static histomorphometry, micro-computed tomography (μCT) and real time-PCR (qPCR) analyses, individual trabecula segmentation (ITS)-based analysis, finite element analysis (FEA), energy dispersive X-ray spectroscopy (EDX) were conducted to provide advanced analysis of structural, mechanical and mineralization features. μCT and histomorphometry showed consistently reduced trabecular number and connectivity. ITS revealed predominant change in trabecular rods, and EDX confirmed less mineralization. The structural and mineralization abnormality led to slight reduction in apparent modulus, which could be attributed to differential down-regulation of Runx2, and up-regulation of Spp1 and TRAP. In conclusion, this is the first comprehensive study providing direct evidence of undefined unique pathological changes at different bone hierarchical levels in AIS.

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“…However, aging‐related changes in the structure and function of osteocyte LCN might alter the ability of osteocytes to sense mechanical loading, orchestrate bone remodelling and detect microdamage by fluid flow resulting in delayed insufficient bone repair and fragile bone. Emerging evidence showed that osteocytes could release secretory factors at the fracture site and facilitate bone regeneration by robust cellular recruitment during fracture healing . Osteocytes expressed proteins known to play profound roles during the healing process.…”

Section: Introductionmentioning

confidence: 99%

Can we enhance osteoporotic metaphyseal fracture healing through enhancing ultrastructural and functional changes of osteocytes in cortical bone with low‐magnitude high‐frequency vibration?

Choy

Wong

et al. 2020

FASEB j.

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Fragility fractures are related to the loss of bone integrity and deteriorated morphology of osteocytes. Our previous studies have reported that low‐magnitude high‐frequency vibration (LMHFV) promoted osteoporotic fracture healing. As osteocytes are known for mechanosensing and initiating bone repair, we hypothesized that LMHFV could enhance osteoporotic fracture healing through enhancing morphological changes in the osteocyte lacuna‐canalicular network (LCN) and mineralization. A metaphyseal fracture model was established in female Sprague‐Dawley rats to investigate changes in osteocytes and healing outcomes from early to late phase post‐fracture. Our results showed that the LCN exhibited an exuberant outgrowth of canaliculi in the osteoporotic fractured bone at day 14 after LMHFV. LMHFV upregulated the E11, dentin matrix protein 1 (DMP1), and fibroblast growth factor 23 (FGF23), but downregulated sclerostin (Sost) in osteocytes. Moreover, LMHFV promoted mineralization with significant enhancements of Ca/P ratio, mineral apposition rate (MAR), mineralizing surface (MS/BS), and bone mineral density (BMD) in the osteoporotic group. Consistently, better healing was confirmed by microarchitecture and mechanical properties, whereas the enhancement in osteoporotic group was comparable or even greater than the normal group. This is the first report to reveal the enhancement effect of LMHFV on the osteocytes’ morphology and functions in osteoporotic fracture healing.

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Calcification during bone healing in a standardised rat calvarial defect assessed by micro‐CTandSEM‐EDX

Abstract: Healing bone is immaturely calcified initially and proceeds calcification gradually, that is, as the bone volume increases, mineral increases in density and matures in quality, while organic components decrease.

Cited by 18 publications

References 43 publications

<b>Calcification in rat developing mandibular bone demonstrated by whole mount staining, microcomputed tomography and scanning electron microscopy with energy dispersive X-ray spectr</b><b>oscopy </b>

<b>Calcification in rat developing mandibular bone demonstrated by whole mount staining, microcomputed tomography and scanning electron microscopy with energy dispersive X-ray spectr</b><b>oscopy </b>

Unique local bone tissue characteristics in iliac crest bone biopsy from adolescent idiopathic scoliosis with severe spinal deformity

Can we enhance osteoporotic metaphyseal fracture healing through enhancing ultrastructural and functional changes of osteocytes in cortical bone with low‐magnitude high‐frequency vibration?

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