Journal articles: 'Bone decalcification'

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Salih, Magdi Mansour. "Comparison between Conventional Decalcification and a Microwave-Assisted Method in Bone Tissue Affected with Mycetoma." Biochemistry Research International 2020 (August 1, 2020): 1–6. http://dx.doi.org/10.1155/2020/6561980.

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Mycetoma is a lifelong granulomatous disease of subcutaneous tissues and bones. Histopathology is a substantiated indicative method based on the assumption of a definitive diagnosis of mycetoma. It requires efficient processing of tissues including bone decalcification. The decalcification process must ensure complete removal of calcium and also a proper preservation of tissue and microorganisms’ staining ability. Objectives. To compare the conventional method used in decalcification with the microwave method using different decalcification solutions. Different characteristics were tested, including the speed of decalcification and morphological and fungal preservation in bone tissue affected with mycetoma. Materials and Methods. Three decalcification solutions were employed to remove calcium from 50 bone tissue samples affected with mycetoma, including 10% neutral buffered EDTA (pH 7.4), 5% nitric acid, and 5% hydrochloric acid. Conventional and microwave methods were used. Haematoxylin-eosin (HE) stain, Gridley’s stain, and Grocott hexamine-silver stain were employed to evaluate the bone and fungi morphologies. Results. The decalcification time of the conventional method compared with the microwave method with 10% EDTA (pH 7.4) took 120 hours and 29 hours, while 5% hydrochloric acid and 5% nitric acid took 8 hours and 3 hours, separately. Also, 10% EDTA is the best decalcifying agent for HE staining and fungal stains. 5% hydrochloric acid and 5% nitric acid can be used for fungal staining. Conclusion. The current study investigated the effects of different decalcifying agents as well as two decalcification procedures on the preservation of the bone structure and fungal staining, which will help to develop suitable protocols for the analyses of the bone tissue affected with mycetoma infection.

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El Khassawna, Thaqif, Diaa Eldin S. Daghma, Sabine Stoetzel, Seemun Ray, Stefanie Kern, Deeksha Malhan, Volker Alt, et al. "Postembedding Decalcification of Mineralized Tissue Sections Preserves the Integrity of Implanted Biomaterials and Minimizes Number of Experimental Animals." BioMed Research International 2017 (2017): 1–10. http://dx.doi.org/10.1155/2017/2023853.

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Bone histology of decalcified or undecalcified samples depends on the investigation. However, in research each method provides different information to answer the scientific question. Decalcification is the first step after sample fixation and governs what analysis is later feasible on the sections. Besides, decalcification is favored for immunostaining and in situ hybridization. Otherwise, sample decalcification can be damaging to bone biomaterials implants that contains calcium or strontium. On the other hand, after decalcification mineralization cannot be assessed using histology or imaging mass spectrometry. The current study provides a solution to the hardship caused by material presence within the bone tissue. The protocol presents a possibility of gaining sequential and alternating decalcified and undecalcified sections from the same bone sample. In this manner, investigations using histology, protein signaling, in situ hybridization, and mass spectrometry on the same sample can better answer the intended research question. Indeed, decalcification of sections and grindings resulted in well-preserved sample and biomaterials integrity. Immunostaining was comparable to that of classically decalcified samples. The study offers a novel approach that incites correlative analysis on the same sample and reduces the number of processed samples whether clinical biopsies or experimental animals.

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Bogoevski, Kristofor, Anna Woloszyk, Keith Blackwood, Maria A. Woodruff, and Vaida Glatt. "Tissue Morphology and Antigenicity in Mouse and Rat Tibia: Comparing 12 Different Decalcification Conditions." Journal of Histochemistry & Cytochemistry 67, no. 8 (May 15, 2019): 545–61. http://dx.doi.org/10.1369/0022155419850099.

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Conventional bone decalcification is a time-consuming process and is therefore unsuitable for clinical applications and time-limited research projects. Consequently, we compared the effect of four different decalcification solutions applied at three different temperatures, and assessed the rate of decalcification and the implications on tissue morphology and antigenicity of mouse and rat tibiae. Bones were decalcified with 10% ethylenediaminetetraacetic acid (EDTA), 10% formic acid, 5% hydrochloric acid, and 5% nitric acid at 4C, 25C, and 37C. Decalcification in both species was fastest in nitric acid at 37C and slowest in EDTA at 4C. Histological and immunohistochemical staining confirmed that the conventional protocols of EDTA at 4C and 25C remain the best option regarding the quality of tissue preservation. Whereas formic acid at 4C is a good alternative saving about 90% of the decalcification time, hydrochloric and nitric acids should be avoided particularly in case of rat tibia. By contrast, due to their smaller size, mouse tibiae had shorter decalcification times and tolerated higher temperatures and exposure to acids much better. In conclusion, this study demonstrated that depending on the specific research question and sample size, alternative decalcification methods could be used to decrease the time of decalcification while maintaining histological accuracy.

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Rodgers, Griffin, Guido R. Sigron, Christine Tanner, Simone E. Hieber, Felix Beckmann, Georg Schulz, Arnaud Scherberich, Claude Jaquiéry, Christoph Kunz, and Bert Müller. "Combining High-Resolution Hard X-ray Tomography and Histology for Stem Cell-Mediated Distraction Osteogenesis." Applied Sciences 12, no. 12 (June 20, 2022): 6286. http://dx.doi.org/10.3390/app12126286.

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Distraction osteogenesis is a clinically established technique for lengthening, molding and shaping bone by new bone formation. The experimental evaluation of this expensive and time-consuming treatment is of high impact for better understanding of tissue engineering but mainly relies on a limited number of histological slices. These tissue slices contain two-dimensional information comprising only about one percent of the volume of interest. In order to analyze the soft and hard tissues of the entire jaw of a single rat in a multimodal assessment, we combined micro computed tomography (µCT) with histology. The µCT data acquired before and after decalcification were registered to determine the impact of decalcification on local tissue shrinkage. Identification of the location of the H&E-stained specimen within the synchrotron radiation-based µCT data collected after decalcification was achieved via non-rigid slice-to-volume registration. The resulting bi- and tri-variate histograms were divided into clusters related to anatomical features from bone and soft tissues, which allowed for a comparison of the approaches and resulted in the hypothesis that the combination of laboratory-based µCT before decalcification, synchrotron radiation-based µCT after decalcification and histology with hematoxylin-and-eosin staining could be used to discriminate between different types of collagen, key components of new bone formation.

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Uma, K., Vidya Chandavarkar, and R. Sangeetha. "Comparison of routine decalcification methods with microwave decalcification of bone and teeth." Journal of Oral and Maxillofacial Pathology 17, no. 3 (2013): 386. http://dx.doi.org/10.4103/0973-029x.125204.

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Bychkov, Aleksey, Vyacheslav Koptev, Varvara Zaharova, Polina Reshetnikova, Elena Trofimova, Elena Bychkova, Ekaterina Podgorbunskikh, and Oleg Lomovsky. "Experimental Testing of the Action of Vitamin D and Silicon Chelates in Bone Fracture Healing and Bone Turnover in Mice and Rats." Nutrients 14, no. 10 (May 10, 2022): 1992. http://dx.doi.org/10.3390/nu14101992.

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This study presents findings on the biological action of an integrated supplement containing the following components involved in osteogenesis and mineralization: vitamin D and silicon in the bioavailable and soluble form. A hypothesis that these components potentiate one another’s action and make calcium absorption by the body more efficient was tested. Biological tests of the effect of vitamin D and silicon chelates on bone fracture healing and bone turnover were conducted using ICR mice and albino Wistar rats. Radiographic and biochemical studies show that the supplement simultaneously containing silicon chelates and vitamin D stimulates bone tissue regeneration upon mechanical defects and accelerates differentiation of osteogenic cells, regeneration of spongy and compact bones, and restoration of bone structure due to activation of osteoblast performance. Bone structure restoration was accompanied by less damage to skeletal bones, apparently due to better absorption of calcium from food. The studied supplement has a similar effect when used to manage physiologically induced decalcification, thus holding potential for the treatment of osteomalacia during pregnancy or occupational diseases (e.g., for managing bone decalcification in astronauts).

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Duncan, Ian, Natalie Danziger, Daniel Duncan, Amanda Hemmerich, Claire Edgerly, Richard Huang, Jo-Anne Vergilio, et al. "Acid-Based Decalcification Methods Compromise Genomic Profiling from DNA and RNA." Blood 134, Supplement_1 (November 13, 2019): 4659. http://dx.doi.org/10.1182/blood-2019-131362.

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BACKGROUND: Comprehensive genomic profiling (CGP) performed by next-generation sequencing of DNA detects genomic alterations including point mutations, insertions/deletions, copy number variations, and select gene rearrangements. When RNA sequencing is included in CGP, it allows for expanded detection of gene fusions, which are common in hematologic malignancies and sarcomas. When such tumors involve bone, a decalcification step is frequently employed to soften tissues prior to processing and sectioning. While commonly used acid-based decalcification methods work quickly, the resulting nucleic acid damage can be profound. In this study, we examine the effects of decalcification on DNA and RNA sequencing in the clinical setting. DESIGN: 1711 consecutive formalin-fixed paraffin embedded samples were evaluated by CGP during routine clinical care via DNA and RNA sequencing, using a hybrid-capture next-generation sequencing assay (FoundationOne®Heme). Specimen site [e.g. bone/ bone marrow or soft tissue] and decalcification status were extracted from pathology reports and H&E review. Samples were considered decalcified if reported as such in the pathology report or if visible decalcified bone was present on the H&E. Samples documented to be processed with fixatives other than formalin were excluded. Sequencing failures were defined as samples that failed DNA extraction (DNAx), RNA extraction (RNAx), or library construction (LC) due to insufficient nucleic acid to advance into sequencing. Samples were only evaluated for RNA if DNAx was successful (1594 cases). RESULTS: Specimen site was a strong predictor of sequencing failure, with a significant increase in failure rate from bone/bone marrow samples (n=619) compared to samples from soft tissue sites (n=1092) for both DNA (13.4% vs 4.6%, p=4.7E-9) and RNA (42.5% vs 13.5%, p<2.2E-16). Of the bone/bone marrow samples, 237 of 619 samples were decalcified. Decalcification was associated with significantly higher failure rates than non-decalcified samples for both DNA (29.1% vs 3.7%) and RNA (67.4% vs 30.8%) (Table 2). One method of avoiding decalcification for bone marrow samples is utilization of clot preparations, where aspirates are processed as an FFPE block. Clot preparations fail sequencing significantly less often than decalcified core biopsies (DNA: 3.3% vs 18.8%, p=9.2E-06; RNA: 39.2% vs 70.4%, p=2.5E-03) (Table 3). CONCLUSIONS: CGP of samples acquired from bone and bone marrow sites is challenging, with a lower success rate for DNA and RNA sequencing than soft tissue sites. The higher overall failure rate correlates with use of decalcification agents leading to degradation of nucleic acids and impacts RNA sequencing significantly more than DNA (67.4% vs 30.8% failed). Clot preparations of bone marrow samples performed better than core biopsies for both DNA and RNA. The higher overall RNA sequencing failure rates still observed in in non-decalcified bone/bone marrow are predominantly due to RNA failure of non-decalcified clot preparations. These samples likely have increased failure rates secondary the use of non-standard fixatives (e.g. B+, Bouin's, AZF, etc.) not documented in the pathology report and the frequency of hypocellular clot preparations in conjunction with higher requirements for RNA yield compared to DNA yield. To increase CGP success rates, decalcification should be avoided when possible. Peripheral blood and bone marrow aspirate samples rarely fail sequencing (<1%, data not shown) and are preferable to decalcified samples if adequate tumor is present. Bone marrow clot preparations perform better than bone marrow core biopsies and clot preparations should be fixed with 10% neutral buffered formalin. If decalcification is required for processing, EDTA based decalcification methods and/or minimizing decalcification times is recommended. Disclosures Duncan: Foundation Medicine, Inc.: Employment. Danziger:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. Duncan:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. Hemmerich:F. Hoffman La Roche, Ltd.: Equity Ownership; Foundation Medicine, Inc.: Employment. Edgerly:F. Hoffman La Roche, Ltd.: Equity Ownership; Foundation Medicine, Inc: Employment. Huang:F. Hoffman La Roche, Ltd.: Equity Ownership; Foundation Medicine, Inc.: Employment. Vergilio:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. Elvin:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. He:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. Britt:Foundation Medicine, Inc: Employment. Reddy:F. Hoffman La Roche, Ltd.: Equity Ownership; Foundation Medicine, Inc: Employment. Sathyan:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. Alexander:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. Ross:F. Hoffman La Roche, Ltd.: Equity Ownership; Foundation Medicine, Inc.: Employment. Brown:Foundation Medicine, Inc.: Employment; F. Hoffman La Roche, Ltd.: Equity Ownership. Ramkissoon:F. Hoffman La Roche, Ltd.: Equity Ownership; Foundation Medicine, Inc.: Employment. Severson:F. Hoffman La Roche, Ltd.: Equity Ownership; Foundation Medicine, Inc.: Employment.

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Pitol, Dimitrius Leonardo, Flavio Henrique Caetano, and Laurelúcia Orive Lunardi. "Microwave-induced fast decalcification of rat bone for electron microscopic analysis: an ultrastructural and cytochemical study." Brazilian Dental Journal 18, no. 2 (2007): 153–57. http://dx.doi.org/10.1590/s0103-64402007000200013.

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Bone decalcification is a time-consuming process. It takes weeks and preservation of the tissue structure depends on the quality and velocity of the demineralization process. In the present study, a decalcification methodology was adapted using microwaving to accelerate the decalcification of rat bone for electron microscopic analysis. The ultrastructure of the bone decalcified by microwave energy was observed. Wistar rats were perfused with paraformaldehyde and maxillary segments were removed and fixed in glutaraldehyde. Half of specimens were decalcified by conventional treatment with immersion in Warshawsky solution at 4ºC during 45 days, and the other half of specimens were placed into the beaker with 20 mL of the Warshawsky solution in ice bath and thereafter submitted to irradiation in a domestic microwave oven (700 maximum power) during 20 s/350 W/±37ºC. In the first day, the specimens were irradiated 9 times and stored at 40ºC overnight. In the second day, the specimens were irradiated 20 times changing the solution and the ice after each bath. After decalcification, some specimens were postfixed in osmium tetroxide and others in osmium tetroxide and potassium pyroantimonate. The specimens were observed under transmission electron microscopy. The results showed an increase in the decalcification rate in the specimens activated by microwaving and a reduction of total experiment time from 45 days in the conventional method to 48 hours in the microwave-aided method.

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Kristensen, Harald K. "STAINING OF HUMAN BONE MARROW AFTER DECALCIFICATION." Acta Pathologica Microbiologica Scandinavica 26, no. 5 (August 18, 2009): 715–18. http://dx.doi.org/10.1111/j.1699-0463.1949.tb00773.x.

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Hiratai, Rumi, Miho Nakamura, Akiko Nagai, and Kimihiro Yamashita. "The Storing Properties of Electric Energy in Bone." Key Engineering Materials 493-494 (October 2011): 170–74. http://dx.doi.org/10.4028/www.scientific.net/kem.493-494.170.

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We have shown that hydroxyapatite (HA), which characteristics were similar to those of bone’s inorganic components, had polarization capability and was possible to accumulate electricity under high temperature and pressure. Then, we presumed that bones had polarization capability which enabled electrical storage and conducted the experiment to measure the polarization capability of bones using rabbit’s femurs. After preparing and polarizing bone samples using KOH treatment (koh), KOH and baking treatment (koh+bake) and decalcification treatment (decalcification) as well as the bone without any treatment (untreat), quantitative amounts of stored charge in samples were determined by thermally stimulated depolarization current (TSDC) measurement of these samples. Under the condition of 400 °C for 1 h with the electric fields of 5kV/cm, samples of koh, koh+bake, and untreat showed polarization capability. In addition, under the polarization condition of 37 °C for 1 hour with the electric fields of 5kV/cm, all samples showed polarization capability. Those findings can be summarized that bones have the polarization capability which enables electrical storage and polarization of bones is possible even under the low temperature condition, which was at 37 °C in our experiment, where polarization is impossible for HA.

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Rakhmawati, Handina, Adrian Situmeang, Nurhidayat Nurhidayat, Andri Maruli Tua Lubis, Harry Murti, and Arief Boediono. "Efektivitas Larutan Dekalsifikasi pada Os tibia Domba Garut (Ovis aries) (THE EFFECTIVENESS OF DECALCIFYING SOLUTIONS ON THE TIBIAL OF GARUT SHEEP (OVIS ARIES))." Jurnal Veteriner 20, no. 3 (November 27, 2019): 403. http://dx.doi.org/10.19087/jveteriner.2019.20.3.403.

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Bone is a tissue that has a density of extracellular matrix structures and composed by organic and inorganic components. Decalcification is a stage that plays an important role in bone histology using various types of solutions. The sample used in this study was lateral condyle from tibia of three garut sheeps (Ovis aries) which had been fixed with 10% Neutral Buffered Formalin (NBF) for 24 hours. Sample were cut into pieces ranging from 1 cm x 1 cm x 1 cm in size, the decalcification using three solutions; 10% nitric acid, 10% EDTA (pH 7.4) and 10% EDTA (pH 7.4) + TBD-1®. The aim of this study was to evaluate the effectiveness of three solutions for the decalcification process of lateral tibial condyle of garut sheep. Observation parameter in this study includes: duration of decalcification, sectioning process of ribboning formation, structural integrity and absorption of hematoxylin-eosin (HE) staining. The results shows that 10% EDTA (pH 7.4) solution provides a long duration of decalcification which is ease sectioning process ribboning, the best structural integrity of lateral tibial condyle. In hematoxylin-eosin (HE) staining shows that 10% nitric acid solution does not absorb the optimum color, opposite in 10% EDTA and 10% EDTA + TBD-1® solution, the color intensity between hematoxylin and eosin in the tissue shows the best results. Based on these results, it can be concluded that the 10% EDTA (pH 7.4) is the best decalcification solution for lateral condyle from tibial of garut sheeps.

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Liu, Haixia, Ruyuan Zhu, Chenyue Liu, Rufeng Ma, Lili Wang, Beibei Chen, Lin Li, et al. "Evaluation of Decalcification Techniques for Rat Femurs Using HE and Immunohistochemical Staining." BioMed Research International 2017 (2017): 1–6. http://dx.doi.org/10.1155/2017/9050754.

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Aim. In routine histopathology, decalcification is an essential step for mineralized tissues. The purpose of this study is to evaluate the effects of different decalcification solutions on the morphological and antigenicity preservation in Sprague Dawley (SD) rat femurs. Materials and Methods. Four different decalcification solutions were employed to remove the mineral substances from rat femurs, including 10% neutral buffered EDTA, 3% nitric acid, 5% nitric acid, and 8% hydrochloric acid/formic acid. Shaking and low temperature were used to process the samples. The stainings of hematoxylin-eosin (HE) and immunohistochemical (IHC) were employed to evaluate the bone morphology and antigenicity. Key Findings. Different decalcification solutions may affect the quality of morphology and the staining of paraffin-embedded sections in pathological examinations. Among four decalcifying solutions, 3% nitric acid is the best decalcifying agent for HE staining. 10% neutral buffered EDTA and 5% nitric acid are the preferred decalcifying agents for IHC staining. Significance. The current study investigated the effects of different decalcifying agents on the preservation of the bone structure and antigenicity, which will help to develop suitable protocols for the analyses of the bony tissue.

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Nageris, Benny, and Dan Gazit. "Method for Embedding Temporal Bones of Rats in Methyl-Methacrylate." Annals of Otology, Rhinology & Laryngology 104, no. 10 (October 1995): 783–85. http://dx.doi.org/10.1177/000348949510401006.

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The commonly used method of preparing the temporal bone for light microscopy is a refinement of a basic formula that has been employed for a century. This process includes fixation, decalcification, neutralization, dehydration, embedding in celloidin, and hardening. The main disadvantage of this process is that decalcification is performed. This article describes a new method for preparing the temporal bone of rats for light microscopy. The main advantage of this new method is that no decalcification is involved, so that all bony elements are retained in their normal shape and location, and even retain some enzymatic activity. Other advantages are that the fixation is reversible and the process is short (approximately 2 weeks) and therefore relatively inexpensive. Our vast and positive experience with this technique has led us to report this method not in a specific experiment, but rather as a specific laboratory technique.

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Nath, Shriram Vaidia, Michelle Tamblyn, Susan Telfer, Tony Henwood, Peter Gilham, Colin Story, Greg Hodge, et al. "Ethylene Diamine Tetra Acetic Acid (EDTA) Decalcification of Paediatric Bone Marrow Trephines In a Diagnostic Laboratory." Blood 116, no. 21 (November 19, 2010): 2566. http://dx.doi.org/10.1182/blood.v116.21.2566.2566.

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Abstract Abstract 2566 Background: In paediatric patients with haematological disorders such as acute lymphoblastic leukaemia (ALL), bone marrow aspiration is sometimes difficult to obtain and bone marrow trephine biopsy (BMTB) is a valuable source of material. In a diagnostic laboratory, the turnaround time is critical for a bone marrow trephine to be decalcified, processed and embedded. In our laboratory, 48 hours was routinely required from the time the bone marrow was performed until the sections were ready for reporting. A hydrochloric acid-EDTA (Ethylene-Diamine-Tetra-Acetic acid) decalcifying solution was used for 4 hours but rendered the trephines unsuitable for special studies such as Fluorescence in Situ Hybridisation (FISH). Aim: To evaluate an alternative decalcification method which preserved the ability to perform FISH on formalin-fixed paraffin-embedded (FFPE) tissue without compromising the turnaround time as a laboratory quality improvement measure. Method (EDTA decalcification): Following overnight formalin fixation, the BMTB was decalcified in a solution containing 20% EDTA with continuous stirring for 7.5 hours. The 20% EDTA, pH 7.1 stock solution was prepared by adding Ammonium Hydroxide (25%, concentrated ammonia) (Merck) to distilled water. EDTA disodium salt (372.24; Ajax Finechem) was added and the pH adjusted to pH 7.1 using concentrated ammonia. BM trephines were then processed routinely, embedded in paraffin and 4μm sections were mounted on Super Frost Plus slides. Haematoxylin-Eosin (H&E) staining, Silver Nitrate staining for Reticulin was performed on all slides and Immunocytochemistry, Immunofluorescence and FISH on selected slides. Patient Characteristics: 20 trephines from 15 patients underwent 7.5 hour EDTA decalcification. The diagnosis in 9 patients was Precursor-B ALL while one each had T-Cell ALL, Acute Myeloid Leukaemia, Hodgkin's disease, Refractory Anaemia, Drug Induced Anaemia and Idiopathic Thrombocytopenic Purpura. The mean age was 9.3 years (range 1.9–16.7years) and the mean trephine length was 12.8mm (range 6–21mm). Results: 100% decalcification was achieved in 18 trephines while in 2 trephines 95% decalcification was achieved on morphological examination. The turnaround time was 48 hours. The quality of H&E, reticulin stain, immunohistochemistry and immunofluorescence was maintained and FISH was successful on these FFPE BMTB tissues. This has lead to incorporation of this method for routine use in our laboratory. Conclusion: 20% EDTA decalcification of paediatric BMTB specimens is feasible without affecting the quality of histological preparations or turnaround time. The main advantage of the EDTA decalcification process is that the tissue is amenable to FISH analysis should it be required. Disclosures: No relevant conflicts of interest to declare.

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Inaas Mahamad Apandi, Nurul, Nor Hasilah Mokhtar, Anitha Krishnan Pandarathodiyil, and Anand Ramanathan. "Bone Decalcification for Histological Examination of Biopsy Specimen." Acta Scientific Dental Scienecs 5, no. 5 (April 30, 2021): 132–35. http://dx.doi.org/10.31080/asds.2021.05.1107.

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Jeong, Daewon, Hyun-Sook Lim, and Young-Hee Kang. "Mineral Imbalance: Bone Decalcification and Soft Tissue Calcification." Journal of the Korean Society of Food Science and Nutrition 38, no. 12 (December 31, 2009): 1815–19. http://dx.doi.org/10.3746/jkfn.2009.38.12.1815.

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HIRONAKA, Ryouji, Kazuo SANO, and Tsugio INOKUCHI. "Experimental studies on ultrasonic decalcification of bone implants." Japanese Journal of Oral & Maxillofacial Surgery 37, no. 1 (1991): 1–7. http://dx.doi.org/10.5794/jjoms.37.1.

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Thorat, Rekha, Ujjwala Joshi, Naveen Krishnamoorthy, and Nirmala Jambhekar. "Simultaneous fixation and decalcification protocol for bone specimens." Journal of Histotechnology 34, no. 4 (December 2011): 162–64. http://dx.doi.org/10.1179/204602311x13214380951905.

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Hao, Zhengling, Vicki L. Kalscheur, and Peter Muir. "Decalcification of Bone for Histochemistry and Immunohistochemistry Procedures." Journal of Histotechnology 25, no. 1 (March 2002): 33–37. http://dx.doi.org/10.1179/his.2002.25.1.33.

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Kacena, Melissa A., Nancy W. Troiano, Christiane E. Coady, and Mark C. Horowitz. "Decalcification of Mounted Bone Sections Enhances Immunohistochemical Staining." Journal of Histotechnology 26, no. 2 (June 2003): 105–9. http://dx.doi.org/10.1179/his.2003.26.2.105.

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Mishra, V., N. Singh, D. V. Rai, U. Tiwari, G. C. Poddar, S. C. Jain, S. K. Mondal, and P. Kapur. "Fiber Bragg grating sensor for monitoring bone decalcification." Orthopaedics & Traumatology: Surgery & Research 96, no. 6 (October 2010): 646–51. http://dx.doi.org/10.1016/j.otsr.2010.04.010.

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Jiang, Qian-li, Shan Jiang, Fei-li Chen, Li-qiong Zhu, Pei-ran Zhao, Hao-jia Huang, Xiao-hong He, Chang-xin Yin, and Fan Yi Meng. "Semi-Solid Decalcification and Research System: a Novel Method to Study Fluorescence Protein Gene Modified Stem Cells In Bone." Blood 116, no. 21 (November 19, 2010): 2625. http://dx.doi.org/10.1182/blood.v116.21.2625.2625.

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Abstract Abstract 2625 Background: It remains a huge challenge to observe fluorescent protein (GFP, RFP, etc.) gene-marked cells in bone, since bone is compact, poor lucency and porous, with different tissues such as vessels, nerve, cells, matrix and blood interlacing inside. However, it is very important to study the location, growth, migration and interaction of different stem cells and their offspring in bone. Aim: To better study different fluorescent protein gene-marked stem cells and microenvironment in bone, we establish a novel semi-solid decalcification (SSD) and research system. Methods: 1)Transplantation: Male BABL/C-GFP(H-2d) transgenic mice as donors and female FVB-RFP(H-2q) transgenic mice as recipients. Each RFP recipient mice were injected i.v. 5×10e6 allogeneic GFP bone marrow cells after 8 Gy TBI (n=10). Routinely, mice survival, weight, hemogram, GVHD manifestation were observed, with the fluorescence cells in peripheral blood and organs being traced. 2)Sections preparation: Total body perfusion fixation was performed with paraformaldehyde 21d after transplantation, and then different samples were collected for pathological examinations. The femurs were made frozen sections after semi-solid decalcification (SSD) system, while GMA plastic-embedding sections without decalcification, paraffin sections after EDTA decalcification, frozen sections after EDTA decalcification were also prepared as controls. Sections were observed by confocal microscopy. 3)Other researches: After SSD, observation and three-dimensional reconstruction were done by confocal microscopy; target tissue and cells were picked up for real-time quantified PCR for fluorescent protein genes and cell proliferation cytokines. Results: 1)Recipients RFP mice gained WBC recovery on (18.0±1.2)d, 90.0%±2.3% peripheral cells were GFP+ (n=10), 6 of 10 developed GVHD within 3m. 2)During SSD, hard component of the bone disappeared slowly, replaced gradually by semi-solid substance. SSD is even workable when the bone's diameter is large than 10cm. Frozen sections after SSD clearly showed unchanged position, form, and fluorescence of the GFP and RFP cells with repeatable hematoxylin and eosin(HE)and Wright-giemsa (RG) staining and immunohistochemical staining, fluorescence chromosomal in situ hybridization (FISH) after fluorescence observation and information from different tests of the same section can also be synthesized by computer. However, GMA cold embedding section could keep the cells where they are while losing the fluorescence, further more, embedding section only works well when the bone tissue is small (diameter<2mm). Frozen section after EDTA decalcification could keep the fluorescence with changed position and form during the progress. Paraffin sections can't keep neither the fluorescence nor the normal cell position and morphological characteristics. 3)Three-dimensional reconstruction shows the interesting relationships between different cells with different fluorescence and microenvironment by confocal microscopy. Quantified PCR described the cytokine expression profile of different fluorescence gene-marked cells. Conclusion: The SSD system shows great potency for the research of stem cells in vivo in bone while maintaining the morphological characteristics and structures between different cells without losing fluorescence signals. Another fantastic advantage is that a large number of techniques can be combined to our system to help us understand the homing, growth, proliferation, differentiation, migration and interaction of different target stem cells and their offspring. Disclosures: No relevant conflicts of interest to declare.

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Choi, Myung-Sub, Hyunsup Lee, Hyuk-Chul Kwon, Moon-Hwan Bae, Young-Hye Ko, Hee-Jin Kim, Beom-Se Lee, and Bon-Kyung Koo. "Optimal Fixation and Decalcification Methods for Bone Marrow Biopsy." Korean Journal of Clinical Laboratory Science 47, no. 4 (December 31, 2015): 243–50. http://dx.doi.org/10.15324/kjcls.2015.47.4.243.

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Iyavoo, S., S. Hadi, and W. Goodwin. "Evaluation of decalcification for recovery of DNA from bone." Forensic Science International: Genetics Supplement Series 6 (December 2017): e270-e272. http://dx.doi.org/10.1016/j.fsigss.2017.09.087.

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Saraji, Alireza, Anne Offermann, Janine Stegmann-Frehse, Katharina Hempel, Duan Kang, Rosemarie Krupar, Christian Watermann, et al. "Cracking it - successful mRNA extraction for digital gene expression analysis from decalcified, formalin-fixed and paraffin-embedded bone tissue." PLOS ONE 16, no. 9 (September 16, 2021): e0257416. http://dx.doi.org/10.1371/journal.pone.0257416.

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With the advance of precision medicine, the availability of tumor tissue for molecular analysis has become a limiting factor. This is particularly the case for bone metastases which are frequently occurring in cancer types such as prostate cancer. Due to the necessary decalcification process it was long thought that transcriptome analysis will not be feasible from decalcified formalin-fixed, paraffin-embedded (DFFPE) in a large manner. Here we demonstrate that mRNA extraction from DFFPE is feasible, quick, robust and reproducible and that decalcification does not hamper subsequent gene expression analysis. This might assist in implementing transcriptome analysis from DFFPE into every day practice.

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Hoshiyama, Ken, Shintaro Nakao, Satomi Shiose, Hiroshi Yoshikawa, Kumiko Kano, Yoshihiro Kaizu, Toshinori Murata, and Koh-Hei Sonoda. "Optical Coherence Tomography Angiography Detects Choriocapillaris Loss in Decalcification of Choroidal Osteoma." Journal of VitreoRetinal Diseases 3, no. 4 (July 2019): 263–68. http://dx.doi.org/10.1177/2474126419855314.

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Purpose: Choroidal osteoma, which typically affects young women, is a benign intraocular tumor composed of mature bone within the choroid. Tumor decalcification and subfoveal choroidal neovascularization often lead to poor visual acuity although the etiology is unknown. Choriocapillaris characteristics in choroidal osteoma also are unknown. Methods: We report 4 cases of choroidal osteoma with decalcification in which the choriocapillaris could be imaged by optical coherence tomography angiography (OCTA). Results: OCTA showed that the choriocapillaris structure was maintained in the calcified portion, whereas a loss occurred in parts of the decalcified portion in all cases. Conclusions: OCTA may be useful for understanding the pathological states of choroidal osteoma.

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Gaulier, A., C. Fourcade, G. Szekeres, and M. Pulik. "Bone marrow one step fixation-decalcification in Lowy FMA solution." Pathology - Research and Practice 190, no. 12 (December 1994): 1149–61. http://dx.doi.org/10.1016/s0344-0338(11)80441-3.

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Valiya Kambrath, Anuradha, Justin N. Williams, and Uma Sankar. "An Improved Methodology to Evaluate Cell and Molecular Signals in the Reparative Callus During Fracture Healing." Journal of Histochemistry & Cytochemistry 68, no. 3 (January 11, 2020): 199–208. http://dx.doi.org/10.1369/0022155419900915.

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Approximately 5% to 10% of all bone fractures do not heal completely, contributing to significant patient suffering and medical costs. Even in healthy individuals, fracture healing is associated with significant downtime and loss of productivity. However, no pharmacological treatments are currently available to promote efficient bone healing. A better understanding of the underlying molecular mechanisms is crucial for developing novel therapies to hasten healing. The early reparative callus that forms around the site of bone injury is a fragile tissue consisting of shifting cell populations held together by loose connective tissue. The delicate callus is challenging to section and is vulnerable to disintegration during the harsh steps of immunostaining, namely, decalcification, deparaffinization, and antigen retrieval. Here, we describe an improved methodology for processing early-stage fracture calluses and immunofluorescence labeling of the sections to visualize the temporal (timing) and spatial (location) patterns of cellular and molecular events that regulate bone healing. This method has a short turnaround time from sample collection to microscopy as it does not require lengthy decalcification. It preserves the structural integrity of the fragile callus as the method does not entail deparaffinization or harsh methods of antigen retrieval. Our method can be adapted for high-throughput screening of drugs that promote efficacious bone healing:

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Tuross, Noreen. "Comparative Decalcification Methods, Radiocarbon Dates, and Stable Isotopes of the VIRI Bones." Radiocarbon 54, no. 3-4 (2012): 837–44. http://dx.doi.org/10.1017/s0033822200047482.

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The Fifth International Radiocarbon Comparison (VIRI) provided a suite of 5 bone samples with consensus ages ranging from 969 to 39,305 14C yr BP (Scott et al. 2010). These bones were used herein in a comparison of decalcification methods using either HCl or EDTA to produce collagen, and the results demonstrate age concordance between both preparation methods and the VIRI consensus values. Additional isotopic analyses of the collagen (δ13C, δ15N, and δ18O) illustrate the increasing sensitivity of carbon, nitrogen, and oxygen isotopes in assessing gelatin degradation.

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Savadori, Paolo, Sophia Dalfino, Marco Piazzoni, Francesco Inchingolo, Massimo Del Fabbro, Gianluca Martino Tartaglia, and Luciano Giardino. "Arduino Automated Microwave Oven for Tissue Decalcification." Bioengineering 10, no. 1 (January 6, 2023): 79. http://dx.doi.org/10.3390/bioengineering10010079.

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Decalcification of hard tissues such as bone and teeth is a complex process that requires using chemicals such as acids and chelating agents. Acids act faster than chelating agents, but they have a greater risk of damaging biological samples. Increasing the reaction speed of the chelating agent may solve this issue. There are several strategies to speed up this process, and using microwaves seems to be one of the most effective. However, lab-dedicated microwave ovens are expensive, and their purchase may seem unjustified. Therefore, a low-cost modification of a commercial microwave oven, consisting of an Arduino automation device, has been developed. The setup has proven reliable for continuous work, thanks to implementing an electronic safety circuit. In addition, it may reduce the decalcification time using a chelating agent, achieving optimal results regarding tissue preservation and quality of histological sections.

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Sanchis, Pilar, Ángel-Arturo López-González, Antonia Costa-Bauzá, Carla Busquets-Cortés, Pere Riutord, Paula Calvo, and Felix Grases. "Understanding the Protective Effect of Phytate in Bone Decalcification Related-Diseases." Nutrients 13, no. 8 (August 20, 2021): 2859. http://dx.doi.org/10.3390/nu13082859.

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Myo-inositol hexaphosphate (phytate; IP6) is a natural compound that is abundant in cereals, legumes, and nuts, and it can bind to crystal surfaces and disturb crystal development, acting as crystallization inhibitor. The adsorption of such inhibitors to crystal faces can also inhibit crystal dissolution. The binding of phytate to metal cofactors suggests that it could be used for treatment of osteoporosis. Our in-vitro study showed that phytate inhibits dissolution of hydroxyapatite (HAP). The effect of phytate was similar to that of alendronate and greater than that of etidronate. This led us to perform a cross-sectional study to investigate the impact of consumption of IP6 on bone mineral density (BMD) in post-menopausal women. Our data indicate that BMD and t-score of lumbar spine increased with increasing phytate consumption, and a phytate consumption higher than 307 mg/day was associated with a normal BMD (t-score > −1). These data suggest that phytate may have a protective effect in bone decalcification by adsorbing on the surfaces of HAP, and a daily consumption of phytate-rich foods (at least one serving/day of legumes or nuts) may help to prevent or minimize bone-loss disorders, such as osteoporosis. However, further studies are needed to gain a better understanding about the mechanism of inhibition of phytate in bone-related diseases (see graphical abstract).

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Gądek, Aneta, Leszek Wojnar, Maciej Tęsiorowski, and Barbara Jasiewicz. "A NEW METHOD FOR QUANTIFICATION OF REGENERATED BONE TISSUE ON X-RAY IMAGES OF ELONGATED BONES." Image Analysis & Stereology 22, no. 3 (May 3, 2011): 183. http://dx.doi.org/10.5566/ias.v22.p183-191.

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A new method for quantification of bone regenerate on the basis of computer-aided analysis of digitized Xray images is presented and its applicability in bone lengthening using Ilizarov method is demonstrated. In contrary to classical methods the internal part of the bone image is taken into consideration instead of the bone edges. Theoretical background of this concept is presented and experimentally verified. Experimental results show that the method proposed allows us for assessment of the bone regenerate, precise choice of the moment of external fixator removal as well as prediction of abnormalities in the osteogenesis process (excluding overall decalcification). However, the rules of interpretation of the results are not discussed in details.

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Jiang, Qianli, Yan-yan Ye, Le-zhong Yuan, Shan Jiang, Jing Sun, Pei-ran Zhao, Chang-xin Yin, et al. "Study Of Stem Cell Homing and Donor-Recipient Cellular Interaction In Allogenic Hematopoietic Stem Cell Transplantation Mice Model." Blood 122, no. 21 (November 15, 2013): 5424. http://dx.doi.org/10.1182/blood.v122.21.5424.5424.

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Abstract Background Allogeneic hematopoietic stem cell transplantation is an effective method for treatment of hematological malignancies, while GVHD and graft rejection are main complications, which seriously affect patients' survival rates and quality of life. Aim Establishing allo-transplantation mice model with mRFP and GFP transgenic mice, to simulate clinical hematopoietic stem cell transplantation and explore the mechanism of stem cell homing and GVHD. Methods 1) Thirteen C57BL/6 GFP transgenic mice, used as recipients, were irradiated with 7 Gy. Each mouse was injected through caudal vein with 2*106 bone marrow cells isolated from FVB mRFP transgenic mice. 2) Symptoms like weight loss, depilation, diarrhea were observed as GVHD manifestation while survival rates were evaluated. Routine blood test and FACS were performed at different time points to confirm hematopoiesis reconstitution. 3) Mice were perfused with paraformaldehyde under anesthesia to fix the tissue, while pathological examination and real-time PCR were performed for studying donor and recipient cells interactions in different organs. 4) Semi-solid decalcification was used to treat the femora before observing under confocal microscope directly or after making frozen section, three-dimensional reconstruction were made to observe the cellular interaction, especially for cells within the bone marrow. Result 1) Depilation, wrinkled skin, hunchback and sharp decline of weight were observed in 8/13 mice. Routine blood test implicated hematopoietic reconstitution. FACS showed 86.1%±7.8% mRFP+ cells in peripheral blood of recipients. 2) mRFP+ cells were found distributing throughout the body's organs. mRFP+ Lymphocyte infiltration and inflammatory exudate were seen especially in the small intestine, lung, liver and skin (Fig.1). GFP+ cells were found surrounding mRFP+ cells in the bone marrow of the femora decalcified with semi-solid decalcification. Their interactions can be further observed clearly in bone marrow microenvironment in three-dimensional reconstruction by confocal microscope (Fig.2). Discussion Owing to RFP on donors' cells and GFP on recipients' cells, together with our novel protocol named semi-solid decalcification, we can visually observe the donor and recipient cells' location, ratio and cellular interaction, as well as morphological changes. Within various tissues especially for such tissues as bone marrow and lung, the details between cells can be studied lively by fluorescence microscope and confocal microscope. In recipient mice with GVHD, donor cells can be found in various target tissues such as intestine, lung, liver and skin. Gene marked cells with fluorescence protein can benefit morphological, immunological, cytogenetic and molecular studies in recipients after HSCT. Conclusion The allo-transplantation model with mRFP and GFP transgenic mice is powerful in study of Stem Cell Homing and Donor-Recipient Cellular Interaction. The cellular interaction can be easily observed by three-dimensional reconstruction after semi-solid decalcification, especially for bone marrow and lung. Disclosures: No relevant conflicts of interest to declare.

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Fujii, H., D. J. Wood, J. M. Papadimitriou, and M. H. Zheng. "Application of Confocal Laser Scanning Microscopy in Bone." Journal of Musculoskeletal Research 02, no. 01 (March 1998): 65–71. http://dx.doi.org/10.1142/s0218957798000093.

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The optical sectioning method of confocal laser scanning microscopy provides higher resolution than standard light microscope techniques. The use of optical rather than physical sections for detailed histological analyses of bone obviates the need for either decalcification or complex plastic embedding processes which are required as a routine for the preparation of thin microtome sections. In this study we have used confocal laser scanning microscopy for the morphological analyses of fresh unembedding human cortical bone, bone allograft and bone cement interfaces. Our results have indicated that such an approach has provided a relatively easy and rapid means for the assessment of the histology of normal and pathological bone.

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Harms, John F., Lynn R. Budgeon, Neil D. Christensen, and Danny R. Welch. "Maintaining GFP Tissue Fluorescence through Bone Decalcification and Long-Term Storage." BioTechniques 33, no. 6 (December 2002): 1197–200. http://dx.doi.org/10.2144/02336bm02.

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Cheng, Guoping, Liang Chen, Huanhuan Feng, Bo Jiang, and Yi Ding. "Preliminary Study on Fish Scale Collagen Lamellar Matrix as Artificial Cornea." Membranes 11, no. 10 (September 28, 2021): 737. http://dx.doi.org/10.3390/membranes11100737.

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To construct a novel artificial cornea biomaterial, a method to prepare collagen lamellar matrix was developed in this study using grass carp scales as raw materials. The relationship between the structure of fish scale collagen lamellar matrix and the optical and mechanical properties was analyzed, and co-culture of it and rat bone marrow mesenchymal stem cells (BMSCs) was performed to preliminarily analyze the cellular compatibility of fish scale collagen lamellar matrix. The results show that the grass carp scales could be divided into base region, lateral region and parietal region according to the surface morphology. The inorganic calcium in the surface layer could be effectively removed by decalcification, and the decalcification rate could reach 99%. After etching treatment, homogeneous collagen lamellar matrix could be obtained. With the decalcification and etching treatment, the water content of the sample increased gradually, but the cross-linking treatment had no obvious effect on the water content of fish scale collagen lamellar matrix. Fish scale collagen lamellar matrix has good transparency, refractive index, mechanical properties and cellular compatibility, which may represent a prospect for the construction of cornea tissue engineering products.

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Mullink, H., S. C. Henzen-Logmans, T. M. Tadema, J. J. Mol, and C. J. Meijer. "Influence of fixation and decalcification on the immunohistochemical staining of cell-specific markers in paraffin-embedded human bone biopsies." Journal of Histochemistry & Cytochemistry 33, no. 11 (November 1985): 1103–9. http://dx.doi.org/10.1177/33.11.2414361.

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A number of fixation and decalcification procedures were evaluated to determine their suitability for immunohistochemistry on trephine samples of bone marrow after paraffin embedding. In particular, the immunoreactivity of antigens characteristic for various hematopoietic cell lines (immunoglobulin heavy and light chains for plasmacytoid cells; elastase for neutrophil myeloid cells; lysozyme, alpha-1-antitrypsin and alpha-1-antichymotrypsin for hystiocytic cells; leukocyte common antigen for lymphocytes; hemoglobin and glycophorin A for erythroid cells; Factor VIII-related antigen for thrombocytoid cells) as well as some antigens specific for epithelial tumors (CEA, 115D8, and keratin) were investigated. Fixation in a mercuric chloride-formaldehyde mixture followed by decalcification in acetic acid-formaldehyde-saline proved to be the best procedure for antigen preservation and retention of morphologic detail. Moreover, there is no need of trypsinization when using this procedure. The only exception was Factor VIII-related antigen in megakaryocytes, which was best demonstrated in trypsin-digested sections of formalin-fixed and acetic acid-decalcified biopsies.

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Neat, Michael J., Mufaddal T. Moonim, Robert G. Dunn, Helen Geoghegan, and Nicola J. Foot. "Fluorescence in situ hybridisation analysis of bone marrow trephine biopsy specimens; an additional tool in the diagnostic armoury." Journal of Clinical Pathology 66, no. 1 (October 4, 2012): 54–57. http://dx.doi.org/10.1136/jclinpath-2012-201131.

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Fluorescence in situ hybridisation (FISH) analysis is now widely employed in the diagnosis and risk stratification of a wide range of malignant diseases. While this technique is used successfully with formalin-fixed paraffin-embedded (FFPE) sections from numerous tissue types, FISH analysis of FFPE tissue sections from trephine biopsy specimens has been less widely reported, possibly due to technical limitations relating to the decalcification protocols employed. During the last 4 years FISH analysis has been carried out successfully in 42 out of 55 (76%) consecutive trephine biopsy specimens received as part of the standard diagnostic service at our institution. Samples decalcified using EDTA-based protocols were analysed successfully in 31/31 cases (100%), whereas only 11/24 samples (46%) decalcified using formic acid-based protocols were successful. In our experience, FISH analysis of trephine biopsy specimens is a highly reproducible technique and a very useful adjunctive tool in the diagnostic armoury; however, its use in a standard diagnostic setting relies on the use of EDTA-based decalcification protocols.

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Yamazaki, Y., M. Yuguchi, S. Kubota, and K. Isokawa. "Whole-mount bone and cartilage staining of chick embryos with minimal decalcification." Biotechnic & Histochemistry 86, no. 5 (August 12, 2010): 351–58. http://dx.doi.org/10.3109/10520295.2010.506158.

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Miquelestorena-Standley, Elodie, Marie-Lise Jourdan, Christine Collin, Corinne Bouvier, Frédérique Larousserie, Sébastien Aubert, Anne Gomez-Brouchet, et al. "Effect of decalcification protocols on immunohistochemistry and molecular analyses of bone samples." Modern Pathology 33, no. 8 (February 24, 2020): 1505–17. http://dx.doi.org/10.1038/s41379-020-0503-6.

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Chow, Dick Ho Kiu, Li-Zhen Zheng, Li Tian, Kam-Sing Ho, Ling Qin, and Xia Guo. "Application of ultrasound shortened the decalcification duration of human cortical bone sample." Journal of Orthopaedic Translation 7 (October 2016): 119. http://dx.doi.org/10.1016/j.jot.2016.06.143.

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Fukui, Kenji, Ryo Tsubota, and Mami Matsukawa. "Dependence of local wave velocity in bovine cortical bone on the decalcification." Journal of the Acoustical Society of America 131, no. 4 (April 2012): 3426. http://dx.doi.org/10.1121/1.4708851.

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Mailhac, N., and G. Daculsi. "Bone Ingrowth for Sinus Lift Augmentation with Micro Macroporous Biphasic Calcium Human Cases Evaluation Using MicroCT and Histomorphometry." Key Engineering Materials 361-363 (November 2007): 1347–50. http://dx.doi.org/10.4028/www.scientific.net/kem.361-363.1347.

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The development of implantology requires enough bone support, sufficient bone architecture. The use of autograft remains the gold standard; however the surgeons use cortical bone coming from mandibular part or craniofacial site, involving severe anaesthetic bone loss. The strategy of bone substitutes in place of autograft can be an efficient method. Several patients having a sinus lift augmentation using MBCP, and BioOss have been performed in human, and bone biopsies were realized during the preparation of the site for dental implantation. Biopsies were analyzed in classical histology without decalcification and by 3D reconstruction using micro CT. Both techniques revealed bone ingrowth and MBCP resorption. For BioOss, no bone ingrtowth and resorption process were observed in spite of stability of the implant and clinical efficiency. These case reports confirm the performance of bone substitutes for Sinus Lift augmentation.

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Wang, Jingbo, Xue Zhao, Huimin Shi, Ling Zhu, and Xiaofeng Tao. "Radiological diagnostic features of uremic leontiasis ossea: a case report." Dentomaxillofacial Radiology 49, no. 1 (January 2020): 20190253. http://dx.doi.org/10.1259/dmfr.20190253.

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Uremic leontiasis ossea (ULO), which occurs in the craniomaxillofacial region, is a sign of terminal stage osteitis fibrosa cystica or brown tumors and primarily caused by secondary hyperparathyroidism induced by renal failure. Pathophysiological changes include osteoclasts or osteoblasts proliferation, bone resorption, bone decalcification, and connective tissue proliferation. In this paper, we report a case of a 24-year-old female patient, who was diagnosed with ULO and presented with multiple facial swellings. Imaging features included zonal patterns with alternating rings of hypo- and hyperattenuated craniomaxillofacial bones, and diffused mixed sclerotic tissues with lytic changes in CT imaging. T1 weighted image and T2 weighted image in MRI were characterized by alternating rings of low and intermediate signal intensity patterns. To the best of our knowledge, this case is the first example of pathologically proved ULO with maxillofacial MRI.

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Vermeulen, A. H., C. Vermeer, and F. T. Bosman. "Histochemical detection of osteocalcin in normal and pathological human bone." Journal of Histochemistry & Cytochemistry 37, no. 10 (October 1989): 1503–8. http://dx.doi.org/10.1177/37.10.2789247.

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We investigated the immunohistochemical localization of osteocalcin in demineralized, paraffin-embedded normal and pathological human bone. Acid decalcification protocols appeared to be more suitable for osteocalcin detection than mild chelating agents. In normal lamellar bone, osteocalcin was detected in osteocytes and along the lamellar bone matrix in fine granular deposits. Under pathological conditions (osteomyelitis, neoplasia), appositional bone showed immunoreactivity in osteoblasts and osteocytes but not in the provisory woven bone matrix. Intense immunoreactivity could be seen at the cell borders of osteoclasts and the bone margins of Howship lacunae. In primary bone-forming tumors, osteocalcin immunoreactivity was detected in osteoblasts and their malignant counterparts. On the basis of these results, we conclude that optimal preservation of osteocalcin is obtained through mild acid decalcifiers. Osteocalcin is deposited in bone matrix, especially that of metabolically inactive bone. In neoplasms, osteocalcin could be a marker of osteoblastic differentiation.

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Tomoto, Takashi, and Akihiro Moriyoshi. "Decalcification mechanism of concrete by organic matters in atmosphere." Canadian Journal of Civil Engineering 35, no. 7 (July 2008): 744–50. http://dx.doi.org/10.1139/l08-038.

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This study confirmed that concrete structures and historical structures made of limestone absorb environmental endocrine disrupters such as anion surfactant and phthalic acid esters contained in windshield washer fluid by using 1H nuclear magnetic resonance (NMR), high-performance liquid chromatography (HPLC), gas chromatograph mass spectrometer (GC–MS), and other analytical techniques. Furthermore, it was confirmed that environmental endocrine disrupting organic matter, including phthalic acid esters, penetrated into concrete and accelerated its deterioration and that calcium salt of phthalic acid esters existed in decalcified concrete by means of solid-state cross polarization magic angle spinning carbon-13 (CPMAS 13C) NMR and electron probe microanalysis (EPMA). This phenomenon was demonstrated in a laboratory experiment. Bone mineral content (BMC) and a specific surface area of decalcified concrete slabs were measured in each layer in the depth direction, and it was revealed that deterioration was more severe and cement paste was more porous in the layers closer to the slab surface.

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Cantadori, Lucas Oliveira, Rafael Dezen Gaiolla, Ligia Niero-Melo, and Cristiano Claudino Oliveira. "Bone Marrow Aspirate Clot: A Useful Technique in Diagnosis and Follow-Up of Hematological Disorders." Case Reports in Hematology 2019 (March 10, 2019): 1–5. http://dx.doi.org/10.1155/2019/7590948.

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Bone marrow biopsy is a diagnostic tool largely used in the evaluation of a broad number of disorders that could affect the hematopoietic system. Differently, bone marrow aspirate clot technique is rarely performed even though it has been described in literature. Here, we highlight the utility of the bone marrow aspirate clot, exemplifying through the discussion of three clinical cases in which this technique was used for diagnosis and follow-up purposes: megaloblastic hemopathy, multiple myeloma, and chronic lymphocytic leukemia. Bone marrow clot analysis increases sensitivity to diagnose hemopathies and offers the possibility of morphological evaluation and anatomopathological study, with the advantage of not needing decalcification processes, hence improving antigenic expression in immunohistochemical and FISH techniques. It is an easy-to-perform technique, offering a quick, reliable, and more comfortable procedure for patients.

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Fernandes, Marilene Issa, Eduardo José Gaio, Cassiano Kuchenbecker Rosing, Rui Vicente Oppermann, and Pantelis Varvaki Rado. "Microscopic qualitative evaluation of fixation time and decalcification media in rat maxillary periodontium." Brazilian Oral Research 21, no. 2 (June 2007): 134–39. http://dx.doi.org/10.1590/s1806-83242007000200007.

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The rat model is widely used in periodontal research and the quality of histological sections is essential. The purpose of this study was to evaluate the histological characteristics of periodontal tissues in Wistar rat maxillae, with different times of fixation and decalcified by nitric acid or formic acid (Anna Morse Solution). Fifteen rats were used. Fixation was performed for 24, 48 and 72 hours. The maxillae were hemi-sectioned and each part was decalcified either in nitric acid for 7 days or in Anna Morse solution for 35 days. Two trained and blinded examiners performed the evaluation. Fourty eight hours of fixation and decalcification with Anna Morse solution showed more clear characteristics of the epithelium-connective tissue interface and of the periodontal structures. Mean measurements between the cementum-enamel junction and the bone crest varied in the different experimental times from 176.5 (± 60.45) to 210.94 (± 39.33) pixels on the buccal aspect, and from 199.69 (± 38.33) to 298.55 (± 70.81) pixels on the palatal aspect, with no statistically significant differences (ANOVA, p > 0.05). In the same fixation period, decalcification with nitric acid or Anna Morse solution did not display any statistically significant differences. It may be concluded that for a qualitative histological analysis, fixation should preferably be for 48 hours and the demineralization should be made by Anna Morse solution. For a histomorphometric analysis, the decalcification solution does not interfere in the results.

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Park, Ji Young, and Kyung Hee Han. "Analysis of the Effects of Bone Marrow Biopsy Decalcification Methods on Histopathological Examination." Korean Journal of Clinical Laboratory Science 48, no. 4 (December 31, 2016): 371–77. http://dx.doi.org/10.15324/kjcls.2016.48.4.371.

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Shah, K. M., J. C. H. Goh, R. Karunanithy, S. L. Low, S. De Das, and K. Bose. "Effect of decalcification on bone mineral content and bending strength of feline femur." Calcified Tissue International 56, no. 1 (January 1995): 78–82. http://dx.doi.org/10.1007/bf00298748.

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Journal articles on the topic 'Bone decalcification'

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