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PubMed1996

The effects of acetone, ethanol, HEMA, and air on the stiffness of human decalcified dentin matrix.

During resin-bonding procedures, dentin surfaces are treated with acidic conditioners to remove the smear layer and decalcify the surface to expose the collagen fibrils of the underlying matrix. These decalcified surfaces are then either air-dried or treated with dehydrating solvents, procedures which may modify the physical properties of the dentin matrix. The purpose of this study was to evaluate the effects of dehydration on the stiffness of the decalcified dentin matrix. Small (8 x 1.7 x 0.9 mm) beams of dentin were prepared from mid-coronal dentin of extracted human molars. The ends were covered with varnish for protection, and the specimens were placed in 0.5 M EDTA for 5 days to decalcify. The stiffness was measured by both the cantilever technique and by conventional stress-strain testing. Specimens tested by the cantilever technique were sequentially exposed to water, acetone, alcohol, HEMA, and glutaraldehyde. Specimens tested by conventional stress-strain testing were exposed either to water, acetone, or HEMA, or were allowed to air-dry. The results indicate that the stiffness of decalcified human dentin matrix is very low (ca. 7 MPa), if the specimens are wet with water. As they are dehydrated, either chemically in water-miscible organic solvents or physically in air, the stiffness increases 20- to 38-fold at low strains or three- to six-fold at high strains. These increases in modulus were rapidly reversed by rehydration in water. Exposure to glutaraldehyde also produced an increase in stiffness that was not reversible when the specimens were placed back in water.

Acetone↗
PubMed1982

The determination of bone viability: a histochemical method for identification of lactate dehydrogenase activity in osteocytes in fresh calcified and decalcified sections of human bone.

We have developed a technique for assessing bone viability by the histochemical demonstration of lactate dehydrogenase (LDH) activity in osteocytes. Fresh sawn and ground (75 microns) sections were prepared from femoral heads removed at operation from patients with osteoarthritis of the hip. The sections were decalcified overnight in cold 10% EDTA, pH 7.0. LDH activity was shown by the tetrazolium-formazan reaction with nitroblue tetrazolium as indicator and lithium lactate as substrate. Osteocytes were regarded as viable if their cytoplasm stained dark blue, indicating LDH activity; lacunae containing non-viable osteocytes could be identified by interference contrast illumination. Nearly all osteocytes were viable in the samples studied. Small trabecular fragments, such as could be obtained by needle biopsy, were also suitable for staining after grinding to approximately 50 microns. The method should have application both in research and in diagnosis of ischemic bone disease.

Bone and Bones↗
PubMed1993

A technique for the tensile testing of demineralised bone.

The purpose of this study was to devise a method for the tensile testing of the organic component of demineralised bone. It was hoped that the method could be applied to determine whether an observed reduction in the strength of bone in copper-deficient rats could be attributed to a reduction in the cross-linking of collagen. Tensile testing of a fully demineralised specimen proved impossible to perform because of the difficulty in gripping the material, which tended to slide out of its holding clamps. The technique devised involved the demineralisation of only the diaphysis of the bone, leaving the bone ends intact. This enabled the bone to be gripped firmly at both ends in an Instron tensile testing machine. Elongation under loading of the demineralised diaphysis was measured by the Instron and a video recording was made of the elongation of the central portion. The strain was found to be uniform. Tensile strength was found to correlate with copper levels in the liver (r = 0.54, p < 0.05), although no significant differences were found between the strength, the stiffness, or the strain to failure for bone from the copper-deficient animals and bone from the normal controls. The test method itself should be useful in other studies in which the tensile properties of the organic phase of bone need to be measured.

Animals↗
PubMedDate not supplied

Autogenous demineralized dentin matrix for tissue engineering applications: radiographic and histomorphometric studies.

PURPOSE: This work evaluated the osteoconductive properties of autogenous demineralized dentin matrix (ADDM) on surgical bone defects in the parietal bone of rabbits, using the guided bone regeneration technique and polytetrafluoroethylene (PTFE) membrane. MATERIALS AND METHODS: Surgical bone defects were created in 24 adult rabbits and repaired with either ADDM and PTFE (experimental group) or PTFE alone (control group). The ADDM had been obtained from the central incisors of the experimental rabbits. The rabbits were sacrificed after 15, 30, 60, and 90 days and the defects examined radiographically and histologically. RESULTS: Radiographically, the defects in the experimental animals achieved radiopacity more quickly than the defects in the control group. DISCUSSION: After 15, 30, 60, and 90 days of observation following surgery, the ADDM slices appeared to stimulate new bone formation. The dentin slices were completely incorporated into the new bone tissue and were resorbed during the bone repair. CONCLUSIONS: Bone repair was accelerated on the bone defects treated with ADDM when compared to the control group.

Animals↗
PubMed2006

Root canal morphology of mandibular incisors in a Jordanian population.

AIM: To investigate the root canal morphology of mandibular incisors in a Jordanian population using a canal staining and tooth-clearing technique. METHODOLOGY: Four hundred and fifty extracted mandibular incisors were collected from dental clinics within north Jordan. Following pulp tissue removal and staining of the canal systems, the teeth were decalcified with 5% nitric acid, dehydrated with ascending concentrations of alcohol and rendered clear by immersion in methyl salicylate. Cleared teeth were examined by eye and the following features were evaluated: (i) number and type of root canals; (ii) presence and location of lateral canals and intercanal communications; (iii) location of apical foramina; and (iv) frequency of apical deltas. RESULTS: The majority of mandibular incisors had a single canal (73.8% of teeth possessed a Type I canal system). Although 26.2% of the roots possessed two canals, only 8.7% had two separate apical foramina. CONCLUSIONS: The prevalence of two canals in this group of mandibular incisors was 26.2% and is within the range of previous studies performed on populations of different racial origin.

Coloring Agents↗
PubMed1998

Two canals in mesiobuccal roots of maxillary molars.

The purpose of this study was to examine extracted root canal treated maxillary molars cleared for: (i) the presence of a mesiopalatal (MP) canal in both first and second molars, (ii) the extension of MP canal from the pulp to the apical area, and (iii) the incidence of two foramina in the MB root. Seventy-two extracted maxillary molars (42 first and 30 second molars) were root canal treated by graduate students. The mesiobuccal (MB) roots of these samples were rendered transparent using a clearing technique. The results demonstrated that 52.3% of first and 40% of second molars had two canals obturated in the MB root. After clearing the same roots, the presence of MP canals rose to 80.9% and 66.6%, respectively. The MP canals were root canal treated as far as the foramen in 35.2% of first and 35% of second molars. However, after making them transparent, 91.1% and 90% showed the presence of this canal to the anatomical apex. The MB roots of the root canal treated first molars showed the presence of two foramina in 47% of cases but in 88.2% after clearing. The second molars showed 50% and 70%, respectively. The differences between root canal treated teeth before and after clearing were significantly different in almost all comparisons in both first and second molars. The only exception was when the presence of two foramina was compared before and after the clearing process in second molars (McNemar test P < 0.05).

Coloring Agents↗
PubMed2002

Can application of exogenous fibronectin enhance periodontal regeneration?

BACKGROUND: Although fibronectin (FN) is an important extracellular glycoprotein involved in periodontal wound healing, it is not clear whether the application of exogenous fibronectin (ExoFN) offers any clinical benefit. The purpose of this preliminary in vitro study was to determine the binding of FN from three different sources, viz. endogenous EDTA-plasma, endogenous serum and exogenous commercial purified human fibronectin in PBS buffer, to demineralized and non-demineralized root powder. METHOD: The binding of FN to a known quantity of mineralized and non-demineralized root powder by overnight incubation at 15 degrees C was studied by enzyme immunoassay (EIA) technique. The criteria for optimal performance of EIA procedure for the determination of FN was established. Particle size of powdered root structure was standardized using a Vibratory Sieve Shaker. RESULTS: The EDTA-plasma and the serum FN exhibited binding of (17.8 +/- 2.1 microg) and (6.5 +/- 4.5 microg), respectively, to the non-demineralized root powder. However, the binding was only significant for the EDTA-plasma FN (p < 0.01) when compared to controls. In the demineralized group there was no ascertainable binding of FN from either endogenous or exogenous sources. ExoFN in buffer exhibited no binding at all to the non-demineralized or demineralized root powder. CONCLUSION: The preliminary data suggest that additional plasma and serum factors may facilitate the binding of FN to root powder. High levels of FN in blood do not necessarily indicate that FN is available for binding to the root surface during periodontal surgery.

Analysis of Variance↗
PubMed1999

Discrimination of morphological findings in dentine from osteogenesis imperfecta patients using combinations of polarized light microscopy, microradiography and scanning electron microscopy.

OBJECTIVES: The aim of this study was to investigate the morphological appearance of dentine in teeth from individuals with osteogenesis imperfecta type I, III and IV using different histological techniques, and to correlate morphological findings to different types of osteogenesis imperfecta. SAMPLE AND METHODS: Extracted or exfoliated primary and permanent teeth were collected from 15 patients with the osteogenesis imperfecta diagnoses I, III or IV, with or without the additional diagnosis dentinogenesis imperfecta. Ground and decalcified sections were prepared from the teeth. Histo-morphological studies of the dentine were performed utilizing light and polarized light microscopy, microradiography and scanning electron microscopy. RESULTS: Characteristic findings were irregular tubules, remnants of capillary inclusions and obliterated pulps. All types of osteogenesis imperfecta exhibited similar types of dentine aberrations, but patients with type III or IV had a higher frequency of aberrations when compared to type I. CONCLUSIONS: The combination of either polarized light microscopy or micro-radiography, together with scanning electron microscopy, gave the most amount of morphological information from dentine samples. In addition, aberrations in dentine structure were more clearly observable. Light microscopy was not critical for the analyses.

Adolescent↗
PubMed1983

Intracapsular and intraosseous vascular supply to the mature dog's coxofemoral joint.

The intracapsular and intraosseous arterial supply to the mature dog's coxofemoral joint was investigated using a vascular injection, tissue-clearing technique in 6 mixed-breed dogs. Ascending arteries of the femoral neck, arising from an extracapsular vascular ring at the base of the femoral neck, penetrated the articular capsule and traveled subsynovially over the surface of the femoral neck. These intracapsular arteries formed an incomplete intracapsular vascular ring at the junction of the femoral head and neck. Bony components of the acetabulum, ie, ilium, ischium, and acetabular bones, received intraosseous vascular supply from the nutrient artery of the ilium and minute vessels entering nutrient foramina around the rim of the acetabulum. The femoral head received branches (epiphyseal arteries) from the ascending arteries of the femoral neck at the intracapsular vascular ring. Epiphyseal arteries formed anastomosing vascular arches of decreasing caliber up to the subchondral bone of the femoral head. The proximal femoral metaphysis, particularly the femoral neck, received vascular supply from 4 sources: (i) branches from the epiphyseal arteries on the surface of the physeal scar which descended, penetrating the physeal scar, (ii) distally directed branches from the intracapsular vascular ring, (iii) ascending nutrient vessels from the trochanteric fossa, and (iv) terminal branches of the ascending nutrient artery of the femur.

Acetabulum↗
PubMed1998

Bone maintenance 5 to 10 years after sinus grafting.

PURPOSE: This radiographic study determined the amount of bone around hydroxyapatite (HA)-coated dental implants that were placed into bone-grafted maxillary sinuses. PATIENTS AND METHODS: Postoperative complex motion tomograms using the Grossman technique were taken on 16 patients who had 27 maxillary sinus grafts performed using particulate autogenous iliac bone with and without demineralized bone, autogenous iliac corticocancellous block with and without demineralized bone, and autogenous jaw bone with demineralized bone. Bone levels were measured from the new floor of the grafted sinus to the apex of the implant and to the alveolar crest. The resulting bone level measures were compared with the type of graft used. All patients had been restored for 5 to 10 years after simultaneous graft and implant placement. RESULTS: For all patients summed together, the average amount of bone from the top of the graft to the apex of the implant was 3.3 +/- 3.1 mm, and the average amount of bone from the top of the graft to the alveolar crest was 17.6 +/- 3.1 mm. The average level of bone in the sinuses of patients grafted with autogenous iliac bone was greater than the average level of bone in those grafted with autogenous bone combined with demineralized bone. CONCLUSION: The results of this study indicate that autogenous bone grafts are maintained in the maxillary sinus, but the results with autogenous bone alone are better than when demineralized bone is added. However, this difference may not be clinically significant.

Adolescent↗
PubMed1995

Material and compositional properties of selectively demineralized cortical bone.

Timed immersion in buffered ethylenediamine-tetraacetic acid (EDTA) was used to selectively alter the mineral content at each level in the cortical bone structural hierarchy. The effects on the mechanical behavior were investigated using a combination of experimental techniques which provide collectively a wide range of resolution (5 microns to 3 mm). Optical microscopy and histological analysis demonstrated a heterogeneous structure consisting of a mineralized tissue core surrounded by a layer of demineralized tissue (collagen) whose thickness varied depending on the immersion time. The mechanical behaviors of treated samples with (intact) and without (core) the surrounding demineralized layer were evaluated using three-point flexure. Overall, the intact specimens became significantly less brittle with increased immersion time in buffered-EDTA. For the core specimens, there was a systematic decrease in the elastic flexural properties (E, sigma e, epsilon e). The site-specific properties of the specimens were determined using microhardness testing, scanning acoustic microscopy, and wavelength dispersive analysis. The mineralization and site-specific properties of the mineralized cores were not significantly affected by buffered-EDTA immersion; however, histomorphometric analysis showed a decrease in the mineralized volume fraction via widening of the pre-existing vascular channels. The experimental hierarchy was effective in discerning site-specific property changes and the localized heterogeneities resulting from the buffered-EDTA treatment. Based on the results of this study, buffered-EDTA treatment can be used to facilitate the determination of material and physical properties of intact and demineralized tissues within a single cortical bone sample.

Animals↗
PubMed2000

Quality of apical seal in curved canals using three types of spreaders.

The aim of the present study was to evaluate the influence of spreader shape on the quality of obturation. Forty-five single canal teeth with varying degrees of apical curvature were used throughout. The canals were prepared using the step-back technique and obturated with lateral condensation. The teeth were classified into three groups (A, B, and C) and treated as follows. Group A was treated with markedly conical, flat-ended spreaders (A to D; Dentsply-Maillefer, Ballaigues, Switzerland). Group B was treated with slightly conical (15 to 40) sharp-pointed CC-cord spreaders. Group C was treated with CC-cord, slightly conical but flat-ended spreaders. The obturated teeth were decalcified in 7.5% nitric acid to obtain four cross-sections: the first at 2 mm from the apex and then the following three sections at 1-mm intervals. Section thickness was approximately 1 mm. Each section was photographed under a stereoscopic microscope. The following areas were measured on each of the microphotographs using a planimeter: (a) gutta-percha mass, (b) mass of sealer, (c) foreign bodies, and (d) empty spaces. These four areas were added to determine total cross-sectional canal area and then the percent of that area attributable to a and to b + c + d was calculated. The statistical analysis of the data (parametric Student's t test for independent groups) revealed that only in group B was the area occupied by a, the gutta-percha mass, significantly greater at all levels than the area occupied by b + c + d. Next in obturation effectiveness was group C, followed by group A.

Decalcification Technique↗
PubMed1996

The periodontal tissues in healed degree III furcation defects. An experimental study in dogs.

The aim of the present study was (i) to describe the periodontal tissue that formed in degree III furcation defects in mandibular molars of dogs following GTR therapy and (ii) to compare this healed periodontium to the corresponding tissue of pristine furcations. The study was performed in 10 mongrels dogs. In 6 of the dogs (group A), the 2nd and 4th premolars in both sides of the mandible were extracted 2 months prior to the start of the study. The 4 remaining dogs (group B) were used as normal untreated controls. In the dogs of group A, a furcation defect was produced in the 3rd mandibular premolars and reconstructive surgery was later performed in accordance with the GTR technique. 5 months after reconstructive surgery, all 10 animals (groups A+B) were sacrificed and perfused with a solution of 10% neutral, buffered formalin through the carotid arteries. Tissue blocks containing the experimental teeth were excised, demineralized in EDTA, embedded in paraffin. Serial sections were cut in the mesiodistal plane and parallel with the long axis of the roots. The microtome was set at 7 microns. The sections were stained in hematoxyline and eosin or Van Gieson's connective tissue staining. From each biopsy, 3 sections, 14 microns apart, and representing the central part of the furcation, were selected for light microscopic examination. In the healed furcation sites, histometric and morphometric measurements were performed at different levels (zones), either at the mesial or distal root surfaces; (zone 1) immediately apical of the notch; (zone 2) coronal to the notch where the newly formed alveolar bone was in continuity with the reduced bone crest; (zone 3) coronal to the notch; representing the most coronally positioned area of new bone formation; (zone 4) coronal to the notch; representing areas with no alveolar bone present. In the pristine furcation sites, the measurements were made at zones which corresponded to the location of the zones in the healed furcations. The present data demonstrated that all furcation sites in group A after 5 months of healing exhibited comprehensive de novo cementum formation in the previously exposed parts of the intraradicular root surfaces, and that collagen fibers invested in the newly formed cementum. Comparisons between the pristine and the healed furcations disclosed that the periodontal ligament of the healed furcations was poorly organised, and that bone formation was frequently incomplete.

Alveolar Process↗
PubMed1992

HIV inactivation in a bone allograft.

The use of exclusionary techniques in the procurement of donors for bone allografts greatly reduces chances for disease transmission. Furthermore, treatment of HIV with either chemical agents or strong acids will effectively inactivate the AIDS virus. These data are taken as indirect proof that the risk of obtaining AIDS from a freeze-dried bone allograft is highly remote. The purpose of this study is to obtain direct evidence that the processing of a demineralized freeze-dried bone allograft would render the allograft safe for human use. In Part I, human cortical bone was obtained from a cadaveric source and tested to be free of HIV contamination. The bone was spiked with 5.26 x 10(9) viral particles. This corresponded to 148 micrograms of total viral protein. In Part II, cortical bone was procured from a donor who died of AIDS. In both Parts I and II, the cortical bone was ground to yield particle sizes of 90 to 500 microns. Test samples were treated with a virucidal agent and demineralized in HCl. Control samples were left untreated. All samples were cocultivated with stimulated peripheral blood lymphocytes and assayed for p24 core protein, reverse transcriptase, and viral gag gene by polymerase chain reaction (PCR). In Part I, the HIV spiking experiment, untreated virus infected particulate bone was positive for HIV replication. Treated samples were negative when assayed for HIV. Bone samples in Part II, HIV infected bone, were positive by PCR. Replication of viable HIV could not be demonstrated after treatment. It was concluded that demineralization and treatment with a virucidal agent inactivates HIV in spiked and infected bone.

Acquired Immunodeficiency Syndrome↗
PubMed2003

Immunohistochemistry of matrix markers in Technovit 9100 New-embedded undecalcified bone sections.

Trabecular bone is routinely analysed by histomorphological-histometrical and immunohistochemical techniques as means of assessing the differentiation status of bone deposition and growth. Currently few embedding resins exist for which both morphological and immunohistochemical analyses can be performed on mineralised tissue. Paraffin, the standard embedding medium for bone enzyme and immunohistochemistry, can only be used on demineralised tissue, but then trabecular structure may be badly preserved. Methyl methacrylate (MMA), the resin of choice for undecalcified bone histology can only be used for bone immunohistochemistry if the usual, highly exothermic polymerisation procedure is avoided which destroys tissue antigenicity. Consequently, most current practices involve cutting samples in half to be processed in separate resins when more than one type of analysis is required. Technovit 9100 New is a low temperature MMA embedding system that is purported to significantly improve tissue antigenicity preservation allowing polymerisation at -20 degrees C. In this study, Technovit 9100 New-embedded undecalcified trabecular bone samples (adult human, young bovine and ovine) yielded immunolabelling with several bone matrix markers and preserved morphological features in 7 microm sections when stained with Masson-Goldner, von Kossa, or toluidine blue. Bone samples from all resins used were immunolabelled with antibodies against osteocalcin, alkaline phosphatase, osteopontin, osteonectin, bone sialoprotein and procollagen type I amino-terminal propeptide. Technovit-embedded bone yielded more reliable immunolabelling of the matrix proteins when compared with heat or cold-cured LR White or standard embedded MMA samples. Technovit 9100 New provided better routine histology than LR White, and was comparable to MMA. Results demonstrated that Technovit 9100 New can be used as a low-temperature acrylic resin embedding method for routine undecalcified bone histology, as well as for immunohistochemistry.

Acrylic Resins↗
PubMed2003

Effect of 830-nm laser light on the repair of bone defects grafted with inorganic bovine bone and decalcified cortical osseous membrane.

OBJECTIVE: The aim of this study was to assess histologically the effect of LLLT (lambda830 nm) on the repair of standardized bone defects on the femur of Wistar albinus rats grafted with inorganic bovine bone and associated or not to decalcified bovine cortical bone membrane. BACKGROUND DATA: Bone loss may be a result of several pathologies, trauma or a consequence of surgical procedures. This led to extensive studies on the process of bone repair and development of techniques for the correction of bone defects, including the use of several types of grafts, membranes and the association of both techniques. There is evidence in the literature of the positive effect of LLLT on the healing of soft tissue wounds. However, its effect on bone is not completely understood. MATERIALS AND METHODS: Five randomized groups were studied: Group I (Control); Group IIA (Gen-ox); Group IIB (Gen-ox + LLLT); Group IIIA (Gen-ox + Gen-derm) and Group IIIB (Gen-ox + Gen-derm + LLLT). Bone defects were created at the femur of the animals and were treated according to the group. The animals of the irradiated groups were irradiated every 48 h during 15 days; the first irradiation was performed immediately after the surgical procedure. The animals were irradiated transcutaneously in four points around the defect. At each point a dose of 4 J/cm2 was given (phi approximately 0.6 mm, 40 mW) and the total dose per session was 16 J/cm2. The animals were humanely killed 15, 21, and 30 days after surgery. The specimens were routinely processed to wax, serially cut, and stained with H&E and Picrosirius stains and analyzed under light microscopy. RESULTS: The results showed evidence of a more advanced repair on the irradiated groups when compared to non-irradiated ones. The repair of irradiated groups was characterized by both increased bone formation and amount of collagen fibers around the graft within the cavity since the 15th day after surgery, through analysis of the osteoconductive capacity of the Gen-ox and the increment of the cortical repair in specimens with Gen-derm membrane. CONCLUSION: It is concluded that LLLT had a positive effect on the repair of bone defect submitted the implantation of graft.

Animals↗
PubMed1997

The extraosseous and intraosseous arterial anatomy of the adult elbow.

UNLABELLED: We investigated the extraosseous and intraosseous arterial anatomy of the human adult elbow. Twenty-two fresh adult cadaveric upper extremities were studied with a technique of combined India-ink and latex injection followed by chemical débridement. The intraosseous vascularity of twelve extremities was then evaluated with a rapid Spalteholz clearing technique. Our findings demonstrated consistent patterns of extraosseous and intraosseous vascular anatomy, which were organized into three vascular arcades: medial, lateral, and posterior. The medial arcade was formed by the superior and inferior ulnar collateral arteries and the posterior ulnar recurrent artery. The lateral arcade was formed by the radial and middle collateral, radial recurrent, and interosseous recurrent arteries. The posterior arcade was formed by the medial and lateral arcades and the middle collateral artery. The intraosseous circulation of the elbow, which was segmental in organization, appeared to be dependent on the local blood supply. The capitellum and the lateral aspect of the trochlea were supplied by posterior perforating vessels arising from the radial recurrent, radial collateral, and interosseous recurrent arteries. The medial aspect of the trochlea was supplied by a circumferential vascular ring originating from the inferior ulnar collateral artery. Watershed areas were apparent between the blood supplies to the medial and lateral aspects of the distal end of the humerus. The olecranon was richly supplied by vessels coursing along its medial aspect from the posterior ulnar recurrent artery and along its lateral aspect from the interosseous recurrent artery. The radial head had a dual extraosseous blood supply from a single branch of the radial recurrent artery, which supplied the head directly, and from additional vessels from both the radial and the interosseous recurrent artery, which penetrated the capsular insertion at the neck of the radius. CLINICAL RELEVANCE: Our findings demonstrate that arterial contributions to the intraosseous circulation of the elbow are more specific than previously appreciated. The intraosseous circulation of the elbow is derived mainly from perforating vessels that arise from neighboring extraosseous arteries. These perforating arteries may be damaged by trauma or by extensile dissection during reconstruction of the elbow. An understanding of the extraosseous and intraosseous circulation of the elbow may help to avoid iatrogenic injury to the intraosseous circulation.

Adult↗