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Histologic changes after semicircular canal occlusion in guinea pigs.

HYPOTHESIS: Histologic changes occurring after varying degrees of surgical trauma to the inner ear in guinea pigs can reveal the mechanism of hearing preservation/loss. BACKGROUND: Surgical approaches to the inner ear that allow for hearing preservation have gained increasing acceptance in neurotologic surgery. The mechanisms responsible for hearing preservation and hearing loss after partial labyrinthectomy are as yet poorly understood. METHODS: Ten animals underwent semicircular canal occlusion, suctioning of perilymph, ampullectomy, or wide vestibulotomy. Tone-burst auditory brain stem response (ABR) thresholds were performed at weekly intervals after surgery. After 4 weeks, temporal bone specimens were processed to obtain 10-jim sections from plastic-embedded ears. The histologic findings were correlated with the initial and final ABR thresholds. RESULTS: After surgical occlusion of one or more semicircular canals, ABR thresholds were preserved, as the authors reported previously. Suctioning of inner ear fluid led to transient loss of thresholds with recovery. Ampullectomy produced dichotomous results, with some subjects preserving auditory function and others losing auditory function. Wide vestibulotomy resulted in permanent loss of auditory function in most cases. Histologically, there was intraluminal fibrosis and inflammation near the site of surgical entry. Most specimens showed normal cochlear architecture and hair cell counts, irrespective of the degree of hearing loss. Vestibular hair cells were also well preserved, even when they were close to the site of surgical injury. CONCLUSIONS: These findings suggest that electromechanical changes, rather than cell death, are responsible for changes in auditory and vestibular function after partial labyrinthectomy.

Animals↗

mRNA in situ hybridization of TIGR/MYOC in human trabecular meshwork.

PURPOSE: To determine the distribution of mRNA expression of the trabecular meshwork-induced glucocorticoid response protein/myocilin (TIGR/MYOC) in human trabecular meshwork. METHODS: In situ hybridization using a 1.25-kb probe obtained from reverse transcription-polymerase chain reaction of TIGR/MYOC cDNA was performed to determine the location of cell labeling within the different regions of the meshwork. The effect of dexamethasone on the pattern of labeling was studied in organ cultured meshwork. Trabecular meshwork from three sources was studied: enucleated eyes obtained at autopsy, trabeculectomy specimens obtained during filtration surgery, and meshworks from anterior segments in perfusion organ culture. Hybridization was performed on frozen sections, paraffin sections, and sections from JB-4 plastic-embedded tissue. RESULTS: Labeling for TIGR/MYOC mRNA was present in most trabecular cells of the uveal, corneoscleral, and juxtacanalicular regions but only variably present in the endothelial cells of Schlemm's canal. A similar pattern was found in the trabeculectomy specimens from eyes with primary open-angle or pseudoexfoliative glaucoma. Dexamethasone treatment increased the labeling intensity and number of labeled cells in meshwork, and also the number of labeled endothelial cells of Schlemm's canal. Fresh tissue processed within 12 hours postmortem gave more consistent labeling than older tissue, although some label was found up to 48 hours postmortem. Labeling was found in tissue from all three sources, and with all three embedding techniques; JB-4 sections provided the best morphologic resolution. CONCLUSIONS: In situ hybridization reveals that mRNA expression for TIGR/MYOC is present in most cells in all regions of the meshwork but only variably present in the endothelial cells of Schlemm's canal. Dexamethasone treatment increased the number and intensity of labeled cells, and also increased the number of labeled cells in the endothelial lining of Schlemm's canal.

Aged↗

Degeneration in the cochlea after noise damage: primary versus secondary events.

PURPOSE: To determine if noise damage in the organ of Corti is different in the low- and high-frequency regions of the cochlea. MATERIALS AND METHODS: Chinchillas were exposed for 2 to 432 days to a 0.5 (low-frequency) or 4 kHz (high-frequency) octave band of noise at 47 to 95 dB sound pressure level. Auditory thresholds were determined before, during, and after the noise exposure. The cochleas were examined microscopically as plastic-embedded flat preparations. Missing cells were counted, and the sequence of degeneration was determined as a function of recovery time (0-30 days). RESULTS: With high-frequency noise, primary damage began as small focal losses of outer hair cells in the 4-8 kHz region. With continued exposure, damage progressed to involve loss of an entire segment of the organ of Corti, along with adjacent myelinated nerve fibers. Much of the latter loss is secondary to the intermixing of cochlear fluids through the damaged reticular lamina. With low-frequency noise, primary damage appeared as outer hair cell loss scattered over a broad area in the apex. With continued exposure, additional apical outer hair cells degenerated, while supporting cells, inner hair cells, and nerve fibers remained intact. Continued exposure to low-frequency noise also resulted in focal lesions in the basal cochlea that were indistinguishable from those resulting from exposure to high-frequency noise. CONCLUSIONS: The patterns of cochlear damage and their relation to functional measures of hearing in noise-exposed chinchillas are similar to those seen in noise-exposed humans. Thus, the chinchilla is an excellent model for studying noise effects, with the long-term goal of identifying ways to limit noise-induced hearing loss in humans.

Animals↗

Behavioral disturbances and vestibular pathology following crotonitrile exposure in rats.

The dinitrile compound 3,3'-iminodipropionitrile causes a number of toxic effects in sensory systems, including degeneration of the vestibular sensory hair cells, as well as a neurofilamentous pathology in motor and sensory neurons. The chemical also causes permanent changes in behavior. These were initially attributed to the effect on neurofilaments, but have been recently linked to the vestibular toxicity. The present work studied the behavioral and pathological effects of the mononitrile compound crotonitrile. Adult male Long-Evans rats (n = 8/group) were exposed to crotonitrile (0, 100, 125, 150 mg/kg/day, for 3 days, i.p., in 1 ml/kg corn oil) and assessed for changes in rearing activity, locomotor activity, and rating scores in tests of vestibular function. Surface preparations of the vestibular sensory epithelia were observed for hair cell loss by scanning electron microscopy (n = 3/group). Control (n = 2) and 3 x 150 mg/kg crotonitrile (n = 3) rats were assessed for neurofilament accumulation in vestibular and dorsal root ganglion neurons by light microscopy observation of semi-thin sections from plastic-embedded ganglia. Crotonitrile dose-dependently increased locomotor activity and rating scores for vestibular dysfunction. A dose-dependent loss of vestibular hair cells was observed in the same animals. In contrast, no obvious neurofilament accumulations were observed in vestibular and dorsal root neurons. We conclude that vestibular toxicity is a property of the nitrile group, and that behavioral disturbances ensue from this toxic action.

Animals↗

Oxidation causes melanin fluorescence.

PURPOSE: The goal of this study is the characterization of the strong yellow fluorescence of oxidized melanin in the retinal pigment epithelium (RPE) and the choroid. METHODS: Naturally occurring melanin in the human retina and choroid was oxidized by exposing fixed and plastic-embedded sections of a human eye to light and hydrogen peroxide. Synthetic melanin was also oxidized in vitro by exposure to light and hydrogen peroxide. The fluorescence of oxidized melanin was examined by absorption spectroscopy, fluorescence spectroscopy, and fluorescence microscopy. RESULTS: Naturally occurring melanin oxidized in situ exhibited a lipofuscin-like yellow fluorescence. Oxidation of melanin in vitro degraded the melanin polymer, resulting in a fluorescent solution. Fluorescence spectroscopy gave an excitation maximum at approximately 470 nm and an emission maximum at approximately 540 nm for both natural and synthetic melanin. Increasing the time of exposure to light or hydrogen peroxide increased melanin fluorescence. CONCLUSIONS: The results indicate that the strong yellow fluorescence of melanin in the RPE and choroid in situ is a property of oxidized melanin and is not due to contamination of the melanin by proteinaceous or lipid materials. The data presented allow a reinterpretation of the results obtained from fluorescence investigations of melanin-containing tissue and suggest a link between melanin degradation and lipofuscin formation.

Choroid↗

[Clinicopathological studies on bone marrow involvement of non-Hodgkin's lymphoma].

OBJECTIVE: To investigate the relationship between pathomorphological features and clinical manifestations of non-Hodgkin's lymphoma (NHL) with bone marrow involvement (BMI). METHODS: Plastic-embedded section of bone marrow biopsy was stained with H-Giemsa-E. Immunotyping of NHL was performed immunohistochemically. RESULTS: A total of 70 patients with NHLBMI(male: 52, female: 18; median age: 49 years) was studied. There were 20 patients with T cell-lymphoma and 50 patients with B cell-lymphoma. The extent of bone marrow involvement was minimal in 15 cases, moderate in 16 cases and severe in 39 cases. Bone marrow involvement was of interstitial type in 23 cases, nodular type in 7 cases, and mixed type in 18 cases and diffuse type in 22 cases. The frequency of splenomegaly in nodular type NHLBMI was significantly higher than that in any other type. Nodular type NHLBMI occurred mainly in B cell-lymphoma. Lymphoma cell leukemia (LCL) developed in 14 of 39 (35.9%) cases of NHL with severe bone marrow involvement which was significantly more frequent than that in NHL with mild and moderate bone marrow involvement. CONCLUSION: Difference in the extent and pattern of bone marrow involvement in NHL is related to clinical manifestations. Bone marrow biopsy helps evaluate response to treatment.

Adolescent↗

Distribution of actin and the actin-associated proteins myosin, tropomyosin, alpha-actinin, vinculin, and villin in rat and bovine exocrine glands.

Actin, myosin, and the actin-associated proteins tropomyosin, alpha-actinin, vinculin, and villin were localized in acinar cells of rat and bovine pancreas, parotid, and prostate glands by means of immunofluorescent staining of both frozen tissue sections and semithin sections of quick-frozen, freeze-dried, and plastic-embedded tissues. Antibodies to actin, myosin, tropomyosin, alpha-actinin, and villin reacted strongly with a narrow cytoplasmic band extending beneath the luminal border of acinar cells. The presence of villin, which has so far been demonstrated only in intestinal and kidney brush border, was further confirmed by antibody staining of blotted electrophoresis gels of whole acinar cell extracts. Fluorescently labelled phalloidin, which reacts specifically with F-actin, gave similar staining, within the cell apex to that obtained with antibodies to actin, myosin, tropomyosin, alpha-actinin, and villin. In contrast, immunostaining with antibodies to vinculin was restricted to the area of the junctional complex. Ultrastructurally, the apical immunoreactive band corresponded to a dense web composed of interwoven microfilaments, which could be decorated with heavy meromyosin. Outside this apical terminal web, antibodies to myosin and tropomyosin gave only a weak immunostaining (confined to the lateral cell borders) whereas antibodies to actin and alpha-actinin led to a rather strong bead-like staining along the lateral and basal cell membrane most probably marking microfilament-associated desmosomes. Anti-villin immunofluorescence was confined to the apical terminal web. It is suggested that the apical terminal web is important for the control of transport and access of secretory granules to the luminal plasma membrane and that villin, which is known to bundle or sever actin filaments in a Ca(++)-dependent manner, might participate in the regulation of actin polymerization within this strategically located network of contractile proteins.

Actinin↗

[Changes in peritoneal mesothelial cells in patients on peritoneal dialysis].

INTRODUCTION: Some thirty years ago peritoneal dialysis (PD) became a respectable modality of renal replacement therapy. That is why peritoneal membrane attracted interest of investigators. Certain changes, known as uremic serositis, appear in morphology of serous membranes in end stage kidney disease (ESKD). The aim of our investigation was to examine the morphology of peritoneal lining cells in control group of healthy persons and morphology of peritoneal lining cells in patients on PD. MATERIAL AND METHODS: Peritoneal biopsies were taken in 10 healthy volunteers during the kidney donation and in 15 patients on PD during clinically indicated extirpation. Biopsy samples were prepared for standard routine HE staining and for plastic embedded fine sections studying. Sections were mounted in an ultramicrotome, stained with Toluidine blue (TB) and studied by light microscope (SM), while fine sections were mounted in an ultramicrotome and studied by transmission electron microscope (TEM). RESULTS: One layer mesothelium of the cuboidal or flattened lining cells were present over the lamina propria connective tissue. Mesothelial cells were overlapped like tiles on the roof. These cells were interconnected with different types of cell junctions (unpermeable, adhesion and communication junctions) positioned on lateral parts of the interdigitated cell membranes. A great number of microvilli were often present on the appical surface, as well as a kinocilia and lamellar bodies. Nuclei were euchromatic with well developed nucleoli. Many ribosomes, mitochondria, cisternae of rough endoplasmic reticulum (RER) and Golgi apparatus, lamellar bodies and lipid inclusions were present in the cytoplasm. Using TEM in analyzing fine sections of biopsies of patients on PD, characteristic ultrastructural changes including epithelial defects with only remaining parts of destroyed cells were established, as well as significantly greater number of rough endoplasmic reticulum (RER) cisternae and immature mesothelial cells in lamina propria indicating intensive regeneration of this epithelium. The cytoplasm of new mesothelial cells were of less electron density on TEM photomicrographs, whereas the nuclei of mesothelial cells in these patients were euchromatic with prominent nucleoli and numerous perichromatic granules and fibrogranular nuclear bodies, indicating cells of great activity. Cytoplasmic protrusions of different shape and content were often recognized on the apical surface of cells. Lamellar bodies were also present in this group of patients within the mesothelial cells, as well as between two mesothelial cells or on their apical surface. Mitochondria were picnotic in many of the mesothelial cells of peritoneum in this patient group. In these mesothelial cells intracytoplasmic paracrystaline inclusions were established. TEM photomicrographs showed basal lamina multiplication in this epithelium. CONCLUSION: Our findings comply with reports of other authors. It should be stressed that TEM examination detects characteristic ultrastructural changes in mesothelial lining cells of peritoneum in patients on PD, which could compromise the function of peritoneum as a membrane for dialysis.

Epithelium↗

[Ultrastructure of peritoneal mesothelial cells].

The introduction of peritoneal dialysis (PD) as a respectable modality of renal replacement therapy some three decades ago, suddenly drew attention of many authors to peritoneal membrane as insufficiently investigated structure. In order to explain the pathological changes in peritoneum due to renal diseases, it became necessary to explore the normal peritoneal structure. The aim of this study was to examine the morphology of peritoneal lining cells in healthy persons. Biopsies of the peritoneum were performed on 20 volunteer kidney donors. Tissue samples were taken during renal transplantation. Special care was taken in getting appropriate samples without artificial damage because of the extreme fragility of the peritoneal tissue. The preparing procedure was standard for routine HE staining and for plastic embedded semifine and fine sections studies. Semifine sections were made on ultramicrotome, stained with Toluidin blue and studied by light microscope, while fine sections were made by ultramicrotome and studied by transmission electron microscope. One layer of cuboidal or flattened lining cells present over the lamina propria connective tissue presented mesothelium. The cells were overlapped like tiles on the roof. Lateral parts of their interdigitated membranes were interconnected with different types of cell junctions: unpermeable, adhesion and communication junctions; inhibiting intercellular transport. Cell surface was often covered with great number of microvilli and lamellar bodies. A single kinocilia was also often present on apical cell surface. Nuclei were euchromatic with well developed nucleoli. Cytoplasm was filled with a great number of ribosomes, mitochondria, cisterns of rough endoplasmatic reticulum and Golgi apparatus, lamellar bodies and lipid inclusions. Numerous pinocytic vesicles on all parts of the membrane as well as in the cytoplasm indicating active endocytosis, egsocytosis and transcytosys in the process of secretion and reabsorption of serous liquid in peritoneal cavity, were visible. Euchromatic nuclei with prominent nucleoli and numerous mitochondria indicate cells of great metabolic activity.

Epithelial Cells↗

[The value of combined use of bone marrow smear and section in lymphoma for diagnosis and staging].

OBJECTIVE: To explore the value of combined use of bone marrow smear and section in lymphoma for diagnosis and staging. METHODS: Smears of peripheral blood and bone marrow from 114 cases of lymphoma were stained according to Wright. Plastic embedded sections of the 114 cases were stained according to HE, HGF and Gomori. RESULTS: None of the 7 patients with Hodgkin's lymphoma (HD) presented bone marrow involvement, while 32 (29.9%) of the 107 patients with Non-Hodgkin's lymphoma (NHL) presented bone marrow involvement, 13 (40.6%) of the 32 cases in stage I, II, III were promoted to stage IV after bone marrow examination. In 5 of the 32 patients without lymphadenopathy the diagnosis of lymphoma was established through bone marrow study. Lymphoma cells in bone marrow and peripheral blood varied in morphology. Bone marrow involvement might be focal (81.3%) or diffused (18.7%). There were mainly five type: (1) Small lymphocyte type. (2) blast-like lymphocyte type. (3) multimorphology type. (4) lymphoplasmacytoid cell type. (5) granuloma type. CONCLUSION: Bone marrow examination is of importance in patients with known or suspected lymphomas and it may be diagnostic in patients without lymphadenopathy or in cases with no conclusive histological evidence from other sites.

Adolescent↗

BS-SEM evaluation of the tissular interactions between cortical bone and calcium-phosphate covered titanium implants.

The improvement of the reliability of the contact between the osseous tissues and the implant materials has been tested by recovering the metallic implants with ceramic materials, usually calcium phosphates. In our study, the calcium phosphate recovering layers were deposited by means of a pulsed-laser deposition technique. Our aim was to to evaluate the tissue interactions established between cortical bone and titanium implants covered by five different layers, ranging from amorphous calcium phosphate to crystalline hydroxyapatite, obtained by altering the parameters of the laser ablation process. The surgical protocol of the study consisted in the simultaneous implantation of the five types of implants in both the tibial dyaphisis of three Beagle dogs, sacrificed respectively one, two and three months after the last surgical procedures. After the sacrifice, the samples were submitted to a scheduled procedure of embedding in plastic polymers without prior decalcification, in order to perform the ultrastructural studies: scanning microscopy with secondary and backscattered electrons (BS-SEM). Our observations show that both in terms of the calcified tissues appearing as a response to the presence of the different coatings and of time of recovering, the implants coated with crystalline calcium phosphate layers by laser ablation present a better result than the amorphous-calcium-phosphate-coated implants. Moreover, the constant presence of chondroid tissue, related with the mechanical induction by forces applied on the recovering area, strongly suggests that the mechanisms implied in osteointegration are related to endomembranous, rather than endochondral ossification processes.

Alloys↗

Multilating granuloma inguinale.

Granuloma inguinale is an uncommon infectious granulomatous disease of the inguinale area that can insidiously engulf and mutilate tissue. The reported case of granuloma inguinale demonstrated the painless progression of the disease to destroy most penile, scrotal, and inguinal tissue and also showed the subsequent satisfactory therapeutic result with antibiotic treatment. Light microscopy of plastic-embedded thin-sectioned tissue specimens allowed clear demonstration of the Donovan bodies and may be valuable in diagnosis.

Granuloma Inguinale↗

The porcine ear skin as a model system for the human integument: influence of storage conditions on basic features of epidermis structure and function--a histological and histochemical study.

Based on careful tissue processing, detailed structural analysis, and histochemical as well as cytophotometrical evaluation of the epidermis, the study presents data with respect to changes of tissue integrity during two storing modes (room temperature and 4 degrees C) and various storage times of the porcine auricle. Structural degeneration was first noted in the barrier region of the epidermis from where such changes spread, independent of storage conditions, from small horizontal necrotic islands and continuously with increasing storage time. The histochemical results corroborated these observations, emphasizing, however, that the lower epidermal layers seemed intact for a longer time period than the upper layers. Cytophotometrical evaluation of histochemical stainings showed, with regard to the enzyme succinate dehydrogenase, that oxidative metabolism was negatively affected in the early stages of storage, whereas epidermal lipids (neutral fats, glycolipids) remained relatively stable, even during storage at room temperature. In conclusion, it was obvious that the barrier region is the most sensitive element of the porcine ear epidermis. Taking into consideration that this part of the epidermis is most important for permeation studies, it seems reasonable to avoid any storage of porcine auricles at room temperature, and to use only auricles that have been stored at 4 degrees C for not more than 4 to 6 hours, immediately after delivery from the slaughter-house. In this way better tissue preservation can be achieved, whereby the use of shinkage-free water-soluble plastic embedding would generally improve the histological control of structural integrity, and the application of an easy to handle enzyme histochemical procedure (e.g. succinate dehydrogenase demonstration) to unfixed fresh-frozen sections would help to control basic aspects of tissue functions. The results are discussed in relation to the use of porcine integument as a model in human dermatological research.

Animals↗

[Relationship between macrophages and apoptosis in patients with myelodysplastic syndromes].

OBJECTIVE: To observe the relationship between macrophage proliferation and cell apoptosis in patients with myelodysplastic syndromes (MDS). METHODS: A double labelling method of immunohistochemistry (alkaline phosphatase anti-alkaline phosphatase, APAAP) and ISEL (DNA in situ end labelling) was used to detect the positive CD68 expression (macrophages) and apoptosis on cold plastic embedded bone marrow biopsy sections in 30 MDS cases. 12 cases of iron deficient diseases (IDA) were used as the control. RESULTS: (1) The number of CD68 positive cells in MDS were higher than that in controls (29.2 +/- 33.0/mm(2) bone marrow tissue vs 21.2 +/- 16.7/mm(2)) (P > 0.05); (2) The number of apoptotic cells in MDS group was much higher than that in the controls (71.5 +/- 70.9/mm(2) vs 37.3 +/- 23.0/mm(2), P < 0.05); (3) The number of CD68 expression (35.5 +/- 37.0/mm(2)) and apoptosis (90.7 +/- 74.6/mm(2)) in less advanced MDS were much higher than that in advanced MDS group (14.6 +/- 11.7/mm(2) and 26.8 +/- 33.1/mm(2), P < 0.05 and < 0.01 respectively); (4) CD68 expression showed an obvious positive correlation to apoptosis in MDS cases (r = 0.83, P < 0.001); (5) CD68 positive cells did not show location correlation to apoptotic cells; (6) CD 68 positive cells in MDS showed simultaneous apoptosis. CONCLUSIONS: Over-apoptosis existed in MDS. Less advanced group has a higher ratio of apoptosis than in advanced group. The correlation between macrophages and apoptosis indicates the participation of TNFalpha in apoptosis-induction during MDS development.

Adolescent↗

[Stereological study of the fat cells in bone marrows with a heterogeneous distribution of adipose tissue].

PURPOSE: 1) To analyze to what extent the fat tissue fraction of the human bone marrow with heterogeneous distribution depends on size and number of adipocytes. 2) To infer the influence of local factors on the two aforementioned parameters. MATERIAL AND METHODS: The material was made up of 15 specimens of bone marrow biopsy with markedly heterogeneous distribution of fat tissue, alternating normal or hyperplastic zones (area I) with aplastic ones (area II). The method of study was the stereological technique on plastic-embedded specimen sections. RESULTS: In the area II, with a fat tissue fraction markedly higher than in area I, both adipocyte number and size were significatively increased. The fat tissue fraction difference (Dif FRGR) between both areas was significatively correlated with the difference of the adipocytes number (Dif Nv) but not of the size (Dif D). However, in multiple regression both Dif Nv and Dif D contributed significatively to Dif FRGR. CONCLUSIONS: Since the histopathological pattern investigated in this work represents a model caused by intervention of local factors, it can be concluded that these can modify both the size and number of adipocytes.

Adipose Tissue↗

LIGHT AND ELECTRON MICROSCOPIC STUDIES OF "MYOGRANULES" IN A CHILD WITH HYPOTONIA AND MUSCLE WEAKNESS.

Examination by light and electron microscopy of more than 100 muscle biopsies revealed one very unusual case. A 4-year-old boy with non-progressive muscle weakness and hypotonia was found to have small particles, termed "myogranules", in many muscle fibres from two gastrocnemius biopsies. Paraffin sections and thin sections of plastic-embedded muscle showed that the rod-shaped myogranules measured between 0.1 and 5 microns in length, and were usually orientated in the long axis of the fibre. Normal cross-striations could not be seen in areas occupied by myogranules, although adjacent parts of the same fibre were normal. Electron micrographs showed myofilaments running through the myogranules and a periodicity similar to sections of recrystallized muscle protein paramyosin. It is possible that this child has a disturbance of muscle proteins.

Actin Cytoskeleton↗

Lumbar intervertebral body fusion cages: histological evaluation of clinically failed cages retrieved from humans.

BACKGROUND: Although interbody cages are widely used, there is little histological documentation of the tissue within cages in the human spine. The purpose of this study was to describe the contents of retrieved, clinically failed, interbody cages from human patients, with special reference to the influence of graft type on the viability of bone in the cages. METHODS: Seventy-eight cages that had been retrieved from forty-eight patients were analyzed. There were eight carbon-fiber cages and seventy threaded metal cages. Of the sixty-seven cages for which information about grafting was available, fifty-six had been packed with autograft only, six had local autograft mixed with demineralized bone matrix, four had allograft, and one had demineralized bone matrix only. The indications for cage retrieval included a failed fusion, malposition or migration of the cage, trauma (a compression fracture at the fusion site), low-back pain, progressive spondylosis, nerve-root impingement, and/or infection. The cages had been in situ for an average of twenty-two months. Undecalcified sections through the center of each plastic embedded cage were reviewed, and the approximate areas occupied by viable bone, necrotic bone, fibrocartilage, hyaline cartilage, fibrous tissue, and graft substitute were visually estimated. Debris particles were estimated by a semiquantitative scoring system. RESULTS: Seventy-one of the seventy-eight cages showed evidence of vascular ingrowth and areas of histologically viable bone, representing incorporating bone graft. The average area occupied by viable bone was 44% (range, 0% to 80%). In some cages, relatively large fragments of cortical bone graft were associated with only minimal new-bone formation. Fibrocartilage occupied up to 50% of the available area in these failed cages. Some cages also contained small fibrocartilage seams connecting segments of bone in a pattern that suggested motion in vivo. In thirty-one of the seventy-eight cages, > or = 5% of the available area was occupied by hyaline cartilage, probably from vertebral end plates or facet joints. CONCLUSIONS: While this study was not designed to test the efficacy of cages or of bone graft, the prevalence of hyaline and fibrocartilage in these failed cages illustrates the importance of graft and graft-site preparation to maximize bone-graft incorporation. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See Instructions to Authors for a complete description of levels of evidence.

Adult↗

Uptake of 111In-Z2D3 on SPECT imaging in a swine model of coronary stent restenosis correlated with cell proliferation.

UNLABELLED: Small targets such as cell proliferation in the coronary arteries may potentially be detected with single-photon imaging using high-radiotracer-specific activity. We hypothesized that an antibody linked to polymers to increase specific radioactivity can be visualized on SPECT images and that counts in the target will correlate with the strength of the biologic signal. METHODS: Twenty-four stents were placed using the balloon overexpansion technique in the coronary arteries of 14 juvenile domestic swine. One week later, the animals received 74 MBq of (111)In-diethylenetriaminepentaacetic acid-polylysine Z2D3-F(ab')(2), and SPECT imaging was performed at 24 h. The coronary vessels were removed, and the stented vessels were processed with plastic embedding and sectioning. Medial and neointimal areas, percentage of vessel stenosis, and cell proliferation indices were quantified using a 5-bromo-2-deoxyuridine (BrdU) labeling index. Reconstructed SPECT images were interpreted for tracer uptake in coronary vessels. RESULTS: Sixteen of the vessels were positive on SPECT imaging and 10 were negative. The percentage injected dose was 0.85 +/- 0.28 x 10(-3) in scan-positive vessels and 0.34 +/- 0.11 x 10(-3) in scan-negative vessels (P < 0.001). The medial-plus-neointimal proliferative index was 42 +/- 11 in scan-positive vessels and 11 +/- 11 in scan-negative vessels (P < 0.0001). The percentage stenoses were 21% +/- 22% versus 19% +/- 15% (not statistically significant). When individual values for the stented-to-control vessel counts were plotted against BrdU labeling index, a significant relationship was found (r(2) = 0.441; P = 0.0014). CONCLUSION: These data indicate that small targets relevant to human coronary vascular disease may be detected using polymer-modified radiolabeled antibodies.

Animals↗