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PURPOSE: To describe a new drilling system that allows the surgeon to obtain autologous living bone that, when associated with a plasma rich in growth factors (PRGF), can be used in bone grafting. MATERIALS AND METHODS: Bone particles collected using both conventional and new drilling systems were analyzed by means of optic and electronic microscopy in 10 patients. Blood was collected from 43 volunteers and used to prepare PRGF. Quantitative aspects of the PRGF, including number of platelets and concentration of growth factors (insulin growth factor [IGF-I], transforming growth factor [TGF-beta1], platelet-derived growth factor [PDGF-AB], vascular endothelial growth factor [VEGF], hepatocyte growth factor [HGF], and epidermal growth factor [EGF]) were assessed. A demonstrative case study was presented. RESULTS: Microscopic examination showed that the bone structure and the presence of living cells in the bone chips were conserved in all samples obtained from drilling at low speed, whereas material obtained by conventional drilling did not maintain these qualities. Mean counts for TGF-beta1 (55.27 +/- 16.23 ng/mL), PDGF-AB (27.96 +/- 12.13 ng/mL), VEGF (421.09 +/- 399.0 pg/mL), EGF (455.49 +/- 210.04 pg/mL), and HGF (605.70 +/- 269.20 pg/mL) were significantly correlated with the number of platelets (590,000 +/- 197,000 platelets/microL; P < .05). DISCUSSION AND CONCLUSION: The new drilling procedure was developed based on biologic criteria. The method may reduce damage to the host tissue and can be used to obtain a mass of living bone for subsequent grafting in association with autologous growth factors. This new procedure may present new possibilities for enhanced bone healing and needs to be evaluated in a clinical trial. [ABSTRACT FROM AUTHOR]

Dental implantology is still an expanding field of scientific study because of the number of people that receive dental therapies throughout their lives worldwide. Recovery times associated to dental surgery are still long and demand strategies to improve integration of metallic devices with hard tissues. In this work, an in vitro ceramic coating is proposed to improve and accelerate osseointegration of titanium surfaces conceived to be used as dental implants or hip or knee prosthesis, shaped either as dishes or screws. Such coating consists of hydroxyapatite microdomains on the implant surfaces obtained in vitro by immersion of titanium alloy samples (Ti6Al4V) in a simulated body fluid. This titanium alloy is highly used in implant dentistry and trauma surgery, among other fields. Once the immersion times under physiological conditions yielding to different ceramic topographies on this alloy were set, the acellular coating time of major interest so as to optimize its biological development was determined. For this purpose, dental pulp mesenchymal cells were cultured on titanium coated surfaces with different hydroxyapatite outline, and cell adhesion, proliferation and morphology were followed through histological techniques and scanning electron microscopy. It was found that 4 days of acellular hydroxyapatite coating led to a significant cell adhesion on the titanium alloys at an early stage (6 h). Cells tended although to detach from the surface of the coating over time, but those adhered on domains of intricated topography or hydroxyapatite cauliflowers proliferated on them, leading to isolated large cell clusters. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2723-2729, 2016. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop a biocompatible monolayer substrate based on fibrin and chitosan for in vitro culture of chondrocytes. Fibrin-chitosan composite substrates combined the proved cell adhesion properties of fibrin with the hydrophilicity and poor adhesion capacity of chitosan. Chitosan microspheres were produced by coacervation method, agglomerated within a fibrin network and subsequently crosslinked with genipin. The composite substrate was stable for 28 days of culture due to the high crosslinking density. Human chondrocytes cultured on the composite substrate were viable during the culture period. At the end of culture time (28 days) the composite substrate showed low cellular proliferation, 41% more collagen type II and 13% more production of sulfated glycosaminoglycans with respect to the amounts found at 14 days. The study revealed that dedifferentiated chondrocytes cultured in monolayer on the composite substrate can re-acquire characteristics of differentiated cells without using three-dimensional substrates or chondrogenic media. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013. [ABSTRACT FROM AUTHOR]

Llombart B, Monteagudo C, López-Guerrero J A, Carda C, Jorda E, Sanmartín O, Almenar S, Molina I, Martín J M&Llombart-Bosch A(2005)HistopathologyClinicopathological and immunohistochemical analysis of 20 cases of Merkel cell carcinoma in search of prognostic markersTo evaluate the clinicopathological and immunohistochemical characteristics of Merkel cell carcinoma (MCC) in an attempt to find new, potentially significant, prognostic markers.Clinical data and follow-up, histopathological features (pattern, cell size, thickness, mitoses, vascular invasion, lymphocytic infiltration) and immunohistochemical detection[CK20, thyroid transcription factor (TTF-1), chromogranin A, synaptophysin, p53, Ki67, Fli-1, CD99, c-Kit] were evaluated in 20 cases of MCC. Fli-1 and CD99 were detected in 90% and 55% of cases, respectively. Tumour size> 30 mm, stage II,‘absent’ lymphocytic infiltration, and the presence of> 50% of Ki67+ tumour cells, were found to be prognostic indicators of disease-free interval (DFI), but only‘absent’ lymphocytic infiltration constituted an independent prognostic factor of DFI after multivariate analysis. For overall survival, the same variables, together with local recurrence and lymph node involvement, had prognostic significance, with only local recurrence as an independent prognostic factor after multivariate analysis.Absence of lymphocytic infiltration and Ki67 immunoreactivity in more than 50% of tumour cells should be evaluated in conjunction with other well-known prognostic markers in MCC. Furthermore, recognizing that Fli-1 and CD99 expression is commonly found in MCC by immunohistochemistry may avoid misinterpretation in the differential diagnosis of MCC with other small round cell tumours. [ABSTRACT FROM AUTHOR]

Ultrastructural and immunohistochemical characterization of the so-called giant multinucleate cells in cutaneous collagenomas Aims: This study was undertaken to compare the histopathological, immunohistochemical and ultrastructural features of the so-called giant multinucleatecells in cutaneous collagenomas: giant-cell collagenoma and solitary sclerotic fibroma. Methods and results: We studied four collagenomas: one giant-cell collagenoma and three solitary sclerotic fibromas. All the cases showed an indolent clinical presentation and were histologically constituted by a well-demarcated dome-shaped proliferation of coarse collagen bundles with a varying number of interspersed giant multinucleate cells and stellate mononuclear cells. The immunohistochemical study on paraffin sections revealed that neoplastic cells in both collagenomas were vimentin and CD34-positive, whereas FXIIIa was only expressed in solitary sclerotic fibromas. In regard to the so-called giant multinucleate cells, we have ultrastructurally found that these cells were ‘real’ multinucleate cells in giant-cell collagenoma, whereas in solitary sclerotic fibromas they consisted of closely packed aggregates of individual stellate mononuclear cells. Moreover, perinuclear cisternae focally containing finely textured material of moderate density were unexpectedly found in giant cells of giant-cell collagenoma, a finding which was not observed in solitary sclerotic fibromas. Additionally, a characteristic cell–cell interaction between tumour cells and mast cells was encountered in all collagenomas. Conclusions: This study supports a distinctive immunohistochemical and overall ultrastructural profile of giantmultinucleate cells in giant-cell collagenoma and solitary sclerotic fibroma, which suggests a different pattern of differentiation for these two related cutaneouslesions. [ABSTRACT FROM AUTHOR]

TP53 and MDM2 genes and their protein expression were evaluated in frozen and paraffin-embedded tissue from 27 patients with malignant fibrous histiocytoma to elucidate the relationship between them, their implication in tumor progression mechanisms and their possible diagnostic-prognostic value in malignant fibrous histiocytoma. Single-strand conformation polymorphism analysis and direct sequencing of polymerase chain reaction-amplified DNA were used to establish two TP53 mutations (7.4%): a point mutation and a 63-bp duplication. Amplification of the MDM2 gene was observed in two tumors (7.4%) by means of Southern-blot analysis, one of them also carrying the TP53 point mutation. Immunohistochemical and Western-blot techniques were used to study nuclear accumulation of p53 and mdm2 proteins: 11 cases (40.7%) with p53 protein expression and thirteen cases (48.1%) with mdm2 protein expression were detected. We confirmed overexpression of mdm2 protein in eight of ten cases (80%) with p53 protein expression without TP53 gene mutation. Statistical analysis shows that simultaneous co-expression of p53 and mdm2 in malignant fibrous histiocytoma is significantly correlated with survival in absence of gene alteration in contrast to the lack of statistical correlation with survival of p53 protein expression alone. [ABSTRACT FROM AUTHOR]

Uterine sarcomas are approximately 3% of all malignant uterine corpus tumours. Of these, the tumours that originate solely in the stromal elements of the uterine wall are infrequent and have not been well characterized cytogenetically. We report two cases of endometrial stromal sarcomas (ESS), one low grade and one high grade, diagnosed by conventional histology, immunocytochemistry, electron microscopy and cytogenetics. Morphologically clear-cut differential structures were seen at optical, immunohistochemical, and electron microscopic levels, permitting a clear differential diagnosis. The low-grade ESS expressed hormonal receptors and vimentin, whereas the high-grade ESS showed no hormone receptors, high Ki-67 activity, and occasional cytokeratin-positive cells. Ultrastructurally, no malignant epithelial differentiation was seen in the tumour cells, but cilia were found in both cases. Cytogenetic study of the low-grade ESS showed pseudodiploid karyotype with chromosomes 6 and 20 rearranged. The high-grade ESS showed a complex karyotype with clonal numerical and structural anomalies. The chromosomes involved in the structural rearrangements were 1, 3, 6, 7, 13, 14, 15, 17, 19, and 21. [ABSTRACT FROM AUTHOR]

A new case of giant-cell tumour (GCT) of bone with benign histological features, clinical stage II, has been reviewed with immunohistochemistry and electron microscopy. After short-term tissue culture the karyotype, using G-banding techniques, presented a consistent translocation t(12;19)(q13;q13). Nude mice xenografts of the tumour were unsuccessful after 6 months of follow-up. Presence of such chromosomal rearrangement may be related to locally aggressive, histologically benign giant-cell tumors of bone. [ABSTRACT FROM AUTHOR]

Esthesioneuroblastoma (ESTH) is a neuroepithelial-cell-derived neoplasm of the olfactory mucosa composed of homogeneous small round cells which contain neurosecretory granules. Melanin has been detected in such tumours only occasionally. Here we describe a new case of ESTH with divergent differentiation. The primary neoplasm was found in a 67 year-old female, involving the left nasal and maxillary sinus; she died of cerebral metastasis ten months after diagnosis. Histologically only small round cells were seen, with S-100 and NSE positivity. Electron microscopy revealed neurosecretory granules and filaments, as well as the occasional presence of melanosomes. A nude mice xenograft line has been established, and is presently in its ninth transfer. Two cell types are present: small round-to-spindle shaped cells with neural features, and large epithelial-like ones. Both immunohistochemistry and electron microscopy confirm this dual differentiation, with the presence of membrane-bound dense-core neural secretion, as well as melanosomes of neuroectodermal origin. Additionally, an in vitro cell line has been established. Cytogenetic analysis confirmed the presence of both malignant human melanoma patterns; non-random abnormalities in chromosomes 1 and 6, extra copies of chromosome 7. Duplication of the long arm of chromosome 14, as seen in olfactory neuroblastoma, is also seen. [ABSTRACT FROM AUTHOR]

During hyposensitization therapy with aluminium-precipitated antigen solutions, a small % of patients develop persistent subcutaneous nodules at the injection site; the existence of delayed sensitivity to aluminium has been implicated in the pathogenesis of these nodules. We studied the prevalence of aluminium sensitivity (using patch, prick and intradermal tests) and common contact allergens (TRUE Test™) in 20 healthy subjects, and in 40 patients treated with aluminium- containing extracts, 20 of whom had persistent subcutaneous nodules that remained for more than 2 months, the other half having no nodular reactions or nodules that remained for less than 2 months. Aluminium sensitivity was found only in those patients of the treated group who had persistent nodular reactions, 4 cases of positivity to an aluminium chloride patch test being found. All 4 cases were women, nodules remained for more than 6 months, and intracutaneous tests were negative. 3 of them also had contact sensitivity to nickel. In 2 cases, nodules were removed for histological and histochemical examination, showing non-specific inflammatory granulomas, and aluminium crystals being found in only 1 case. It is concluded that delayed sensitivity to aluminium appears to be implicated in the pathogenesis of persistent nodular reactions, but sensitivity to aluminium was not found in patients treated with aluminium-precipitated extracts without persistent nodular reactions. [ABSTRACT FROM AUTHOR]

Small cell sarcomas of bone are difficult to classify and diagnose. The present case deals with such a tumor in which the original biopsy and the resected specimen, studied by histology before chemotherapy, provided no final information about its real nature. Thus several techniques were applied to discern its histogenesis and biology. Myogenin proved positive in isolated cells of the primary neoplasm but was extensively expressed in nude mice xenografts. Electron microscopy confirmed the existence of myofilaments. The cytogenetic analysis revealed a large number of chromosomal abnormalities, but not those found in the Ewing's/PNET (peripheral neuroectodermal tumor) family of tumors. This was confirmed by polymerase chain reaction (RT-PCR) wherein no EWS/Fli 1 or PAX3/FKHR gene rearrangements were detected. Based upon these studies, a rhabdomyosarcoma of bone was diagnosed. Clinically the neoplasm showed a highly aggressive behavior, causing death of the patient within 2 years after diagnosis. [ABSTRACT FROM AUTHOR]

Presents photographs of diffuse hypersecretory microcapillary invasive carcinoma, an unusual ductal carcinoma. Appearance of a lumpectomy specimen with numerous small cysts with colloid material; Epithelium lining cysts covered by pluristratified layers.