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Results for “Decalcification Technique”

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At least 19 recordsLinked to original sources
PubMed2002

Routine acid decalcification of bone marrow samples can preserve DNA for FISH and CGH studies in metastatic prostate cancer.

Production of paraffin-section material from tissue samples that contain bone requires decalcification. Techniques such as acidic decalcification or EDTA chelation are suitable methods. Acid decalcification is generally quicker than EDTA chelation but studies have suggested that it may result in hydrolysis of DNA. Here we show that limited acid decalcification (less than 24 hr) in 5% formic acid can preserve DNA sufficient for fluorescent in situ hybridization (FISH) or comparative genomic hybridization (CGH) and that prolonged 10% formic acid decalcification results in failure of FISH and only limited retrieval of DNA for CGH studies.

Bone Marrow↗
PubMed1986

A technique for decalcification and demonstration of substance P-like immunoreactivity.

Substance P-like immunoreactivity (SPLI) was demonstrated in mouse spinal cord by an indirect immunofluorescence method after decalcification of the vertebra with a mixture of EDTA and Zamboni's fixative. SPLI was observed mainly in the gray matter of the spinal cord, especially the superficial layers of the dorsal horn; the distribution was the same as in the control spinal cord. No diffusion and depletion of SPLI were recognized after decalcification and no specific fluorescence was observed. The findings reported here indicate that decalcification with a mixture of EDTA and Zamboni's fixative is a useful method for examining SPLI in nervous tissue surrounded in situ by calcified tissues.

Animals↗
PubMed1991

Decalcification by perfusion. A new method for rapid softening of temporal bones.

We describe a new technique, decalcification by perfusion, for the softening of bony tissue. The blood circulatory system was perfused in 16 rats via a cannula through the left heart ventricle with a fixative followed by New Decalc (an acidic demineralizer) for 30-240 minutes. Perfusion decalcification for 120 minutes softened all heads and middle ear specimens could be easily sampled and prepared for studies by both light and electron microscope. For comparison, a conventional immersion technique required 72 hours of decalcification to accomplish softening. The perfusion technique considerably reduced the time needed to decalcify the tissue and preserved the morphology better than did the immersion procedure.

Animals↗
PubMed2001

Immunohistological demonstration of erythroid cells in canine bone marrow.

In an immunohistological/cytological study of canine bone marrow, the aim was to demonstrate canine erythroid cells with the help of various commercially available antibodies against human antigens (monoclonal antibody against glycophorin A, polyclonal antibodies against haemoglobin and spectrin). In order to preserve possible cross-reacting epitopes various fixation methods (cross-linking, precipitating and dehydrating fixing agents, partly in combination with unmasking measures), decalcification techniques [acid or ethylenediaminetetraacetic acid (EDTA) decalcification] and tissue-embedding methods (paraffin embedding, cryostat sectioning technique) were used. Alternative methods, such as the preparation of cell smears and immunoblotting, were also employed. The only result that was of use for routine diagnostic procedures (paraffin sections) was that obtained by using polyclonal antibodies against haemoglobin. Best results were achieved when tissue was fixed in a formaldehyde-glutaraldehyde mixture, decalcified in EDTA and treated with microwave irradiation. The primary antibody was used in a dilution of 1:500 and incubated for 16 h. With the exception of mature red blood cells and proerythroblasts, different stages of erythrocytopoietic cells in canine bone marrow were shown to be arranged in erythrons. The polyclonal antibody against spectrin also showed clear cross-reactivity, but was only employable in other systems (immunoblotting). The monoclonal antibody against glycophorin A reacted only when used on human tissue or cells.

Animals↗
PubMed2001

Intraoperative microwave processing of bone margins during resection of head and neck cancer.

OBJECTIVE: To develop an accurate method for performing histopathologic analysis for a full cross-section of cortical bone within 2 to 3 hours. METHODS: Microwave technology was used to augment and to more rapidly perform fixation and decalcification of cortical bone. RESULTS: Using the methods described, slides suitable for histopathologic analyses regarding the presence or absence of malignant tumor were prepared in 2 to 3 hours and, in 10 patients studied, had a 100% correlation with slides prepared in 7 days using the standard decalcification technique. CONCLUSION: Microwave technology allows accurate assessment of a full cross-section of cortical bone regarding the presence or absence of malignant tumor within the time limits required for resection of a malignant head and neck tumor and reconstruction of the surgical defect.

Adult↗
PubMedDate not supplied

Ultrastructural morphology and protein content of the internal organic material of rat otoconia.

Otoconia taken from the maculae of the saccule and utricle of the rat inner ear were investigated using transmission electron microscopy (TEM). Their ultrastructure was compared using various decalcification techniques. Similar preparations were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to assess their protein constituents. TEM results show loss of organic material during prolonged decalcification. This is not seen in samples decalcified overnight. The SDS-PAGE results show that rat otoconia contain a major broad band of 90-100 kDa, a major thin band of 56 kDa, and three other bands of 33, 45, and 50 kDa. The band of 56 kDa may be associated with the surface of the otoconia.

Animals↗
PubMed1998

Abrasion-debridement of aortic valve calcific stenosis. Immediate and mid-term clinical results.

BACKGROUND: Ultrasonic debridement of aortic valve calcific stenosis has been abandoned because of high incidence of late insufficiency, due to retraction of cusps damaged by ultrasonic energy. The feasibility of alternative decalcification technique has been tested at the Department of Thoracic and Cardiovascular Surgery of the University of Siena, Italy, using the abrasive power of a diamond burr, actioned by electrical high speed motor. METHODS: Between January 1995 and March 1996, fifteen patients underwent aortic decalcification. Mean age was 69.6 years. RESULTS: All were symptomatic and 46% had associated coronary or mitral valve disease. Mean preoperative peak systolic echo-cardiographic gradient was 76+/-24.2 mmHs and valve area 0.87+/-0.31 at echoplanimetry. No moderate to severe aortic insufficiency was allowed. Valve debridement was achieved without mortality and morbidity procedure-related. Transvalvular hemodynamic gradient disappeared in 8 patients and fell in 7 from a mean value of 81.5 to 29.7 mmHg. The valvular area increased to a mean value of 2.4 cm2. At mid-term follow-up (mean 16.9 months, range 12 to 25) all patients remained clinically improved and free of complications. No case of new aortic incontinence nor worsening has been observed at echocardiographic control. The valve area was calculated 1.54+/-0.47 by continuity equation. CONCLUSIONS: Although these results are encouraging, more experience and longer follow-up are necessary, before giving affordable guide-lines regarding the indication for abrasion-debridement technique in a clinical setting.

Aged↗
PubMed1985

Mode of growth of bacterial pathogens in chronic polymicrobial human osteomyelitis.

Direct examination of material from two cases of persistent (2 and 60 years) osteomyelitis by morphological and culture techniques showed that the pathogens comprised several bacterial species whose cells grew predominantly in discrete exopolysaccharide-enclosed microcolonies made up of a single bacterial morphotype. Bacterial microcolonies were seen between tissue elements in infected connective tissue, and the microcolonies adherent to bone surfaces coalesced to form extensive biofilms that occluded the surfaces of dead bone in sequestrae. Decalcification techniques were required to examine the interior of infected bone, but recognizable remnants were associated with very large amounts of fibrous, ruthenium red-stained material. All bacterial growth in these persistent infections occurred within an intercellular matrix, and some elements of this matrix, which was fibrous in transmission electron microscopy and amorphous in scanning electron microscopy, were associated with the surfaces of bacterial cells in a manner that suggested their production by these organisms. All of the implications of this microcolony mode of bacterial growth in osteomyelitis, and in other chronic bacterial diseases, have yet to be determined.

Adult↗
PubMed2002

[Comparison of three neuro-tracing techniques for identification of the sciatic spinal nerve origin in mice].

To identify sensory and motor neurons associated with the sciatic nerve in adult mice, three methods for applying fluorescent tracers (Fluorogold and Dil) were investigated: direct application, intraneural injection and impregnation of a sectioned nerve in a silicone chamber. Most accurate localization of the neurons on the dorsal root ganglia and spinal cord was accomplished by introducing the proximal stump of a transected sciatic nerve into a silicone chamber, filled with tracers and then decalcifying the tissue. Fluorogold was an effective tracing agent, in contrast to Dil, which was not. In addition to associations with cephalic ganglia L4, L5 and L6, as seen in rats, contributory neurons to the sciatic nerve were located in other ganglia in the mouse. These findings show that the silicone chamber-tissue decalcification technique is a viable tool for obtaining comparative neuroanatomical information in the mouse model.

Animals↗
PubMed2004

Experimental study on allogenic decalcified bone matrix as carrier for bone tissue engineering.

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rabbits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good biocompatibility and ectopic induced osteogenic activity.

Animals↗
PubMed1996

Change in trabecular architecture as measured by fractal dimension.

The objective of this work was to expose dried trabecular bone material to a decalcifying environment and to quantify the change in the spatial distribution of the bone with a fractal measure. Digitized radiographic images were produced from four separate slices of human vertebral bone as they dissolved within a solution of nitric acid. Pixel data from a region of interest (ROI) within the trabecular bone were used to estimate the time-dependent change in fractal dimension of the ROI as the bone dissolved. Results demonstrated that a change in the spatial distribution of trabecular material may be expressed in terms of a concurrently changing estimate of the fractal dimension.

Algorithms↗
PubMed1994

Intercavernous venous communications in the human skull base.

The intercavernous communications of the skull base were studied in 32 sphenoid blocks using electrolytic decalcification techniques, vascular filling, x-rays, and serial anatomical sections. In this study four intercavernous connections were found: anterior intercavernous sinus (AIS), posterior intercavernous sinus (PIS), inferior intercavernous sinus (IIS), and basilar plexus (BP). The AIS was present in 100% of the cases, with diameters ranging from 0.57 mm to 5.43 mm; in 17 cases (53.12%) it took up the whole anterior wall of the hypophyseal fossa. The PIS was also detected in 100% of the cases, and its diameters ranged from 0.71 mm to 4.14 mm. The IIS was identified in 31 cases (96.9%), assuming three different forms: plexuslike, venous lake, and mixed. The BP was found in 100% of the material analyzed; in 23 cases (71.9%) it proved to be the widest intercavernous communication.In 12 cases of this series the hypophysis was completely enveloped by venous structures, except at the level of the sellar diaphragm.

Journal Article↗
PubMed2000

Effect of alendronate on immature human dental root explants.

This study investigated the use of alendronate in the formation of new dentin in vitro. Extracted human premolar and molar teeth with immature apices were grown in tissue culture medium for 60 days. Six control specimens were grown without alendronate in the medium, and 22 experimental specimens were grown with alendronate at [10(-9) M] in the medium. Newly formed dentin was stained with tetracycline and procion brilliant red at days 1 to 3, 30 to 33, and 60. Specimens were decalcified and 5-micron sections were prepared for examination using fluorescent microscopy. New dentin formation was measured in microns at the most apical region, at 125-micron from the apical measurement and at 250-micron from the apical measurement. The alendronate group had 57.15% more growth than the control group at the most apical region, and this difference was significant (p = 0.0001). The results indicate that alendronate at [10(-9) M] is effective in accelerating dentin formation in vitro.

Alendronate↗