Accidental Modification of Attwood's Stain on Decalcified Preparations, Yielding Improved Differentiation between Woven and Lamellar Bone

2001 ◽  
Vol 4 (1) ◽  
pp. 32-36 ◽  
Author(s):  
S. Lee ◽  
R. Stocks ◽  
D.J. de Sa
Keyword(s):  
Osteogenesis Imperfecta ◽  
Potassium Permanganate ◽  
Periodic Acid ◽  
Previous Method ◽  
Matrix Proteins ◽  
Differential Staining ◽  
Lamellar Bone ◽  
Woven Bone ◽  
Cartilaginous Matrix ◽  
Original Application

In an earlier report, a modification of the Attwood's stain was used to study sections of decalcified bone and the procedure aided in the distinction between lamellar and woven bone. Lamellar bone showed preferential affinity for phloxine (red) yet resisted differentiation with tartrazine (yellow), while woven bone stained well with tartrazine. In addition, osteoid seams were identified by their affinity for tartrazine. Differential staining was abolished by pretreatment with periodic acid. Recently, accidental use of potassium permanganate in an acidified medium of pH 1.94, instead of pH 6.44, yielded unexpected results with a much sharper, consistent distinction between woven and lamellar bone. This probably reflects changes in the noncollagenous, interfibrillary matrix proteins of bone, but the exact reason(s) is not known. In our view, this serendipitous modification represents an advance over the previous method, and has allowed us to study the structure of bone in some of the systemic disorders, e.g., osteogenesis imperfecta, with greater ease than before. This method does, however, suffer the disadvantage of interfering with the intensity of the staining of any cartilaginous matrix, and the original application is still of value.

scholarly journals Is All Bone the Same? Distinctive Distributions and Properties of Non-Collagenous Matrix Proteins in Lamellar Vs. Woven Bone Imply the Existence of Different Underlying Osteogenic Mechanisms

1998 ◽  
Vol 9 (2) ◽  
pp. 201-223 ◽  
Author(s):  
J.P. Gorski
Keyword(s):  
De Novo ◽  
Storage Proteins ◽  
Bone Matrix ◽  
Matrix Gla Protein ◽  
Matrix Proteins ◽  
Lamellar Bone ◽  
Structural Motifs ◽  
Woven Bone ◽  
Collagenous Matrix ◽  
Extracellular Matrix Components

The purpose of this review is to summarize recent functional and structural findings regarding non-collagenous matrix proteins in bone and teeth, to compare gene locations for bone and tooth matrix proteins with loci for hereditary skeletal diseases, and to present several provocative hypotheses which integrate this new information into a physiological context. Hypothesis 1 proposes that the molecular composition of rapidly deposited and mineralized woven bone, as well as the responsiveness of cells synthesizing woven bone to stimuli, is different from that for more slowly synthesized lamellar bone, implying the existence of distinctive osteogenic mechanisms. This review of recent research strongly supports this proposal. Briefly, the protein composition of woven bone matrix is enriched in acidic phosphoproteins BAG-75 and BSP, which are not expressed in lamellar bone, which is itself enriched in osteocalcin. De novo deposition and mineralization of woven bone occurs faster than in lamellar bone by means of a matrix-vesicle-assisted mechanism. Deposition of woven bone occurs at sites experiencing biomechanical strains higher than those experienced by lamellar bone. In addition, woven bone in metaphyseal regions is more susceptible to osteoclastic resorption after space flight, ovariectomy, and loss of weightbearing than is lamellar bone. Finally, osteoprogenitor cells responsive to parathyroid hormone reside in the metaphyseal region of long bones. Taken together, these findings suggest that Hypothesis I represents a useful paradigm for future studies. Specific functions mediated by most individual bone and tooth matrix proteins remain uncertain A review of current literature suggests that the functionality of skeletal matrix proteins is expressed through specific binding sites composed of particular species-conserved structural motifs (Hypothesis 2). Examples include the previously recognized Asp-Ser-Ser motif of dentin phosphophoryns and the gamma-carboxyglutamic acid motif of matrix GLA protein and osteocalcin. A new polyacidic amino acid motif composed of consecutive Asp and Glu residues (n > 7) was defined in extracellular matrix components osteopontin, bone sialoprotein, and bone acidic glycoprotein-75 on the basis of strong functional analogies with similar polyacidic stretches in divalent metal storage proteins of the endoplasmic reticulum and sarcoplasmic reticulum. These structural motifs represent prime targets for future structure-function studies in vivo and in vitro.

scholarly journals Nitric oxide-mediated vasodilation increases blood flow during the early stages of stress fracture healing

2014 ◽  
Vol 116 (4) ◽  
pp. 416-424 ◽  
Author(s):  
Ryan E. Tomlinson ◽  
Kooresh I. Shoghi ◽  
Matthew J. Silva
Keyword(s):  
Nitric Oxide ◽  
Blood Flow ◽  
Bone Formation ◽  
Mechanical Loading ◽  
Stress Fracture ◽  
Flow Rate ◽  
Vascular Function ◽  
Blood Flow Rate ◽  
Lamellar Bone ◽  
Woven Bone

Despite the strong connection between angiogenesis and osteogenesis in skeletal repair conditions such as fracture and distraction osteogenesis, little is known about the vascular requirements for bone formation after repetitive mechanical loading. Here, established protocols of damaging (stress fracture) and nondamaging (physiological) forelimb loading in the adult rat were used to stimulate either woven or lamellar bone formation, respectively. Positron emission tomography was used to evaluate blood flow and fluoride kinetics at the site of bone formation. In the group that received damaging mechanical loading leading to woven bone formation (WBF),15O water (blood) flow rate was significantly increased on day 0 and remained elevated 14 days after loading, whereas18F fluoride uptake peaked 7 days after loading. In the group that received nondamaging mechanical loading leading to lamellar bone formation (LBF),15O water and18F fluoride flow rates in loaded limbs were not significantly different from nonloaded limbs at any time point. The early increase in blood flow rate after WBF loading was associated with local vasodilation. In addition, Nos2 expression in mast cells was increased in WBF-, but not LBF-, loaded limbs. The nitric oxide (NO) synthase inhibitor Nω-nitro-l-arginine methyl ester was used to suppress NO generation, resulting in significant decreases in early blood flow rate and bone formation after WBF loading. These results demonstrate that NO-mediated vasodilation is a key feature of the normal response to stress fracture and precedes woven bone formation. Therefore, patients with impaired vascular function may heal stress fractures more slowly than expected.

scholarly journals In vivo approach on femur bone regeneration of white rat (Rattus norvegicus) with the use of hydroxyapatite from cuttlefish bone (Sepia spp.) as bone filler

2019 ◽  
Vol 12 (6) ◽  
pp. 809-816
Author(s):  
Aminatun Aminatun ◽  
D.E. Fadhilah Handayani ◽  
Prihartini Widiyanti ◽  
Dwi Winarni ◽  
Siswanto Siswanto
Keyword(s):  
Bone Regeneration ◽  
Bone Repair ◽  
Control Group ◽  
Bovine Bone ◽  
Lamellar Bone ◽  
In Vivo Test ◽  
Femur Bone ◽  
Woven Bone ◽  
Bone Filler

Background: Hydroxyapatite (HA) from bovine bone has been widely used as bone filler in many fractures cases. HA can also be made from cuttlefish bone (Sepia spp.) that has abundant availability in Indonesia and contains 84% CaCO3, which is a basic ingredient of HA. However, research on the effects of HA from cuttlefish bone on bone regeneration parameters has not been done yet. Aim: This study aimed to determine femur bone regeneration of white rats (Rattus norvegicus) through the use of HA from cuttlefish bone (Sepia spp.) as bone filler. Materials and Methods: HA was made using the hydrothermal method by mixing 1M aragonite (CaCO3) from cuttlefish bone and 0.6 M NH4H2PO4 at 200°C for 12 h followed by sintering at 900°C for 1 h. In vivo test was carried out in three groups, including control group, bovine bone-derived HA group, and cuttlefish bone-derived HA group. The generation of femur bone was observed through the number of osteoblasts, osteoclasts, woven bone, lamellar bone, havers system, and repair bone through anatomical pathology test for 28 days and 56 days. Results: Anatomical pathology test results are showed that administration of bovine bone-derived HA and cuttlefish bone-derived HA increased the number of osteoblasts, osteoclasts, woven bone, lamellar bone, havers system, and bone repair at recuperation of 56 days. Statistical test using Statistical Package for the Social Sciences with Kruskal–Wallis and Mann–Whitney U-test was resulted in significant differences between the bovine bone-derived HA control group and the cuttlefish-derived HA control group. There was no significant difference toward the indication of bone formation through the growth of osteoblasts, osteoclasts, woven bone, lamellar bone, havers system, and bone repair in the bovine bone-derived HA and cuttlefish bone-derived HA groups. Conclusion: It can be concluded that cuttlefish bone-derived HA has the potential as bone filler based on the characteristics of bone regeneration through in vivo test.

Identification of Glycogen in Thin Sections of Amphibian Embryos

1967 ◽  
Vol 2 (2) ◽  
pp. 257-264
Author(s):  
MARGARET M. PERRY
Keyword(s):  
Staining Method ◽  
Periodic Acid ◽  
Differential Staining ◽  
Small Particles ◽  
Periodic Acid Schiff ◽  
Thin Sections ◽  
Enzymic Digestion ◽  
Cell Structures ◽  
Schiff Reaction ◽  
Glycogen Particles

Embryonic amphibian cells when examined with the electron microscope were observed to contain an abundance of small particles, approximately 325 Å in diameter. The periodic acid/Schiff reaction and enzymic digestion were employed to determine the nature of the particles, and from the results of these tests they were concluded to be glycogen. Treatment of thin sections with periodic acid/lead citrate solutions resulted in a marked increase in contrast of the glycogen particles compared with other cell structures, and in a clearly defined substructure of 40-Å grains appearing within the particles. This differential staining method enabled the particulate glycogen to be distinguished from ribosomes.

scholarly journals Bone Regeneration Induced by Bone Porcine Block with Bone Marrow Stromal Stem Cells in a Minipig Model of Mandibular “Critical Size” Defect

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Antonio Scarano ◽  
Vito Crincoli ◽  
Adriana Di Benedetto ◽  
Valerio Cozzolino ◽  
Felice Lorusso ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Bone Regeneration ◽  
Critical Size ◽  
Stem Cell Differentiation ◽  
Lamellar Bone ◽  
Histomorphometric Analysis ◽  
Osteoblastic Activity ◽  
Woven Bone ◽  
Stromal Stem Cells

Introduction. Adding stem cells to biodegradable scaffolds to enhance bone regeneration is a valuable option. Different kinds of stem cells with osteoblastic activity were tested, such as bone marrow stromal stem cells (BMSSCs). Aim. To assess a correct protocol for osteogenic stem cell differentiation, so BMSSCs were seeded on a bone porcine block (BPB). Materials and Methods. Bone marrow from six minipigs was extracted from tibiae and humeri and treated to isolate BMSSCs. After seeding on BPB, critical-size defects were created on each mandible of the minipigs and implanted with BPB and BPB/BMSSCs. After three months, histomorphometric analysis was performed. Results. Histomorphometric analysis provided percentages of the three groups. Tissues present in control defects were 23 ± 2% lamellar bone, 28 ± 1% woven bone, and 56 ± 4% marrow spaces; in BPB defects were 20 ± 5% BPB, 32 ± 2% lamellar bone, 24 ± 1% woven bone, and 28 ± 2% marrow spaces; in BPB/BMSSCs defects were 17 ± 4% BPB/BMSSCs, 42 ± 2% lamellar bone, 12 ± 1% woven bone, and 22 ± 3% marrow spaces. Conclusion. BPB used as a scaffold to induce bone regeneration may benefit from the addition of BDPSCs in the tissue-engineered constructs.

scholarly journals DIFFERENTIAL STAINING OF ULTRATHN SECTIONS OF EPON-EMBEDDED TISSUES FOR LIGHT MICROSCOPY

1965 ◽  
Vol 13 (7) ◽  
pp. 579-582 ◽  
Author(s):  
BERNARD P. LANE ◽  
DOMINIC L. EUROPA
Keyword(s):  
Fine Structure ◽  
Periodic Acid ◽  
Differential Staining ◽  
Tissue Architecture ◽  
Paraffin Sections ◽  
Periodic Acid Schiff ◽  
Thin Sections ◽  
Hematoxylin Staining ◽  
Ultrathin Sections ◽  
Hematoxylin Eosin

A convenient method is described for the removal of Epon 812 from thin sections, utilizing a saturated solutions of sodium hydroxide. The tissue architecture and ultrastructural details are preserved. Hematoxylin-eosin, periodic acid-Schiff and phosphotungstic-hematoxylin staining modifications are suggested which result in differentiation similar to that seen in paraffin sections. The technique is applicable to ultrathin sections suitable for examination with the electron microscope, allowing comparison of staining characteristics and fine structure of adjacent thin sections.

Osteogenesis Imperfecta of the Temporal Bone and its Relation to Otosclerosis

1988 ◽  
Vol 97 (6) ◽  
pp. 585-593 ◽  
Author(s):  
George T. Nager
Keyword(s):  
Hearing Loss ◽  
Connective Tissue ◽  
Bone Formation ◽  
Osteogenesis Imperfecta ◽  
Temporal Bone ◽  
Bone Dysplasia ◽  
Lamellar Bone ◽  
Current Classification ◽  
Sound Conduction ◽  
Loss Of Hearing

Osteogenesis imperfecta (OI) designates a heterogeneous group of heritable disorders of connective tissue that in addition to bone may affect tendons, ligaments, fascia, skin, sclerae, blood vessels, teeth, and hearing. The current classification identifies at least four major syndrome groups or types. It also recognizes a considerable number of additional syndromes that may represent supplementary types or subgroups. Loss of hearing is the least constant of the prominent features of OI. Its incidence varies between 26% and 60%. In OI, formation and remodelling of bone are variously affected. In the temporal bone the development of the inner ear capsule may be involved severely. In the stapes the disturbance in lamellar bone formation can lead to extreme thinness, dehiscence, and nonunion of the stapedial superstructure with the footplate. Osteogenesis imperfecta can be associated with otosclerosis, another bone dysplasia with a different morphology. Otosclerosis, in turn, may interfere with sound conduction and perception. Thus, the hearing loss encountered in OI may be the result of OI, otosclerosis, or a combination of both.

scholarly journals Immunocytochemical detection of dentin matrix proteins in primary teeth from patients with dentinogenesis imperfecta associated with osteogenesis imperfecta

2014 ◽  
Author(s):  
G. Orsini ◽  
A. Majorana ◽  
A. Mazzoni ◽  
A. Putignano ◽  
M. Falconi ◽  
...  
Keyword(s):  
Osteogenesis Imperfecta ◽  
Matrix Protein ◽  
Primary Teeth ◽  
Type I Collagen ◽  
Immunohistochemical Analysis ◽  
Matrix Proteins ◽  
Type I ◽  
Dentinogenesis Imperfecta ◽  
Dentin Matrix Protein ◽  
Dentin Matrix

Dentinogenesis imperfecta determines structural alterations of the collagen structure still not completely elucidated. Immunohistochemical analysis was used to assay Type I and VI collagen, various non-collagenous proteins distribution in human primary teeth from healthy patients or from patients affected by type I dentinogenesis imperfecta (DGI-I) associated with osteogenesis imperfecta (OI). In sound primary teeth, an organized well-known ordered pattern of the type I collagen fibrils was found, whereas atypical and disorganized fibrillar structures were observed in dentin of DGI-I affected patients. Expression of type I collagen was observed in both normal and affected primary teeth, although normal dentin stained more uniformly than DGI-I affected dentin. Reactivity of type VI collagen was significantly lower in normal teeth than in dentin from DGI-I affected patients (P<0.05). Expressions of dentin matrix protein (DMP)-1 and osteopontin (OPN) were observed in both normal dentin and dentin from DGI-I affected patients, without significant differences, being DMP1 generally more abundantly expressed. Immunolabeling for chondroitin sulfate (CS) and biglycan (BGN) was weaker in dentin from DGI-I-affected patients compared to normal dentin, this decrease being significant only for CS. This study shows ultrastructural alterations in dentin obtained from patients affected by DGI-I, supported by immunocytochemical assays of different collagenous and non-collagenous proteins.

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