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Objectives: We sought to determine whether the technique of celloidin removal influences the results of immunostaining in celloidin-embedded cochleae. Methods: We compared four protocols of celloidin removal, including those using clove oil, acetone, ether-alcohol, and methanol saturated with sodium hydroxide. By optimally fixing our tissue (perfused mice), and keeping constant the fixative type (formalin plus acetic acid), fixation time (25 hours), and decalcification time (ethylenediaminetetraacetic acid for 7 days), we determined whether the technique of celloidin removal influenced the immunostaining results. Six antibodies were used with each removal method: prostaglandin D synthase, sodium, potassium adenosine triphosphatase (Na+,K+-ATPase), aquaporin 1, connective tissue growth factor, tubulin, and 200 kd neurofilament. Results: Clove oil, acetone, and ether-alcohol resulted in incomplete removal of the celloidin, thereby negatively affecting the results of immunostaining. The methanol-sodium hydroxide method was effective in completely removing the celloidin; it produced the cleanest and most reproducible immunostaining for all six antibodies. Conclusions: Freshly prepared methanol saturated with sodium hydroxide and diluted 1:2 with methanol was the best solvent for removing celloidin from mouse temporal bone sections, resulting in consistent and reproducible immunostaining with the six antibodies tested. [ABSTRACT FROM AUTHOR]

The localization of proteins by immunostaining is a powerful method to investigate otologic disorders. However, the use of fixatives and embedding media (necessary for the preservation of morphology) can obscure antigens, making it difficult to perform immunoassays. We performed a systematic investigation of the effects of fixative and embedding medium on morphology and immunostaining of the mouse cochlea. Three different fixative solutions [4% formaldehyde (F), 4% formaldehyde + 1% acetic acid (FA), and 4% formaldehyde + 1% acetic acid + 0.1% glutaraldehyde (FGA)] and 3 different embedding media (paraffin, polyester wax, and celloidin) were used. Morphology was assessed using light microscopy. Immunostaining was studied using a panel of 6 antibodies (to prostaglandin D synthase, aquaporin 1, connective tissue growth factor, 200-kDa neurofilament, tubulin and Na+,K+-ATPase). Preservation of morphology was suboptimal with paraffin, adequate with polyester wax and superb with celloidin. Immunostaining was successful using all 6 antibodies in all 3 fixatives and all 3 embedding media. While there were differences in strength of signal and localization of antigen between the 3 fixatives, overall, FA and FGA gave the most uniform results. For a given fixative and antibody, there was surprisingly little difference in the quality of immunostaining between celloidin and paraffin, while results in polyester wax were not as good in some cases. These results suggest that celloidin may be the embedding medium of choice for both morphological and pathological studies, including immunostaining when morphology must be optimized. Copyright © 2008 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Suggests a method for teaching research and other media center skills by integrating them in the reading and language arts curriculum with the media specialist and classroom teacher team-planning relevant activities. Advantages and disadvantages of this approach are summarized. (MES)

We have investigated the morphometric changes in the cochlear nucleus of patients who had undergone cochlear implantation following profound deafness. The brain stems of 11 adult patients who had undergone implantation and four non-implanted control cases with varying degrees of hearing loss were studied. The volumes of the ventral cochlear nucleus (VCN) and dorsal cochlear nucleus (DCN), and the maximal cross-sectional area and densities of cell bodies in the anterior ventral cochlear nucleus (AVCN) were measured bilaterally by light microscopy assisted by the Neurolucida 2000 image analysis system. In addition, the density of synapses on cells of the AVCN were estimated using immunostaining for the synaptosome-associated protein (SNAP-25) by light microscopy. There was no significant difference in volumes of VCN and DCN, maximal cross-sectional area and density of cell bodies of the AVCN, and SNAP-25 immunostaining between the cochlear nucleus ipsilateral and contralateral to cochlear implantation. In addition, there was no significant correlation between these morphometric parameters and clinical performance. Peripheral deafness seems to reduce the size of neurons in the AVCN in that the maximum diameter of cell bodies was greater in the ear with better hearing preoperatively (chi-square test P<0.05). However, electrical stimulation provided by the cochlear implantation did not alter these morphometric changes in adult cochlear implant patients. [Copyright &y& Elsevier]

A cochlear implant array consists of biomaterials, including metal and polymeric in type which are biocompatible, but not necessarily bio-inert. Histologic evidence of a foreign body reaction has been described in temporal bones in patients who in life had undergone cochlear implantation. In the current study, the cellular immune response was characterized using immunohistochemical stains for B-cell lymphocytes (CD20), T-cell lymphocytes (CD3), and macrophages (CD68). In addition, energy dispersive spectroscopy by scanning electron microscopy (EDS-SEM) was performed to characterize the nature of particulate foreign material seen near the electrode array. Infiltrations of B-cell and Tcell lymphocytes and macrophages were identified immunohistochemically. The track of the electrode array was frequently lined by multi-nucleated foreign body giant cells. Energy dispersive X-ray spectroscopy identified the particulate material found in the fibrous sheeth surrounding the cochlear implant to be consistent with platinum. In conclusion, a cochlear implant generates a vigorous cellular immune response consisting of B and T lymphocytes, foreign body giant cells, and macrophages. Platinum was identified as one of the antigens likely responsible for this cellular response. This foreign body response may in certain cases result in migration or even extrusion of an implant device. [ABSTRACT FROM AUTHOR]

Hypothesis: Silicone as part of a cochlear implant electrode may be responsible for a foreign body response in the human., Background: Clinical evidence of a foreign body response to a cochlear implant has been reported. In a previous study, particulate material found within the fibrous sheath and within macrophages surrounding a cochlear implant has been identified as being consistent with platinum. However, to date, there has been no histologic evidence of a role for silicone in this cellular immune response., Methods: A total of 44 temporal bone specimens from 36 patients were reviewed by light microscopy for evidence of presumed platinum and/or silicone foreign bodies in an extracellular or intracellular location. Identification of cell type involved in phagocytosis of foreign body material was accomplished using CD163 immunostaining. The identity and source of the foreign body material was confirmed using energy-dispersive X-ray spectroscopy and scanning electron microscopy., Results: Evidence for both platinum and silicone was found in all 44 specimens. In three patients, anti-CD 163 immunostaining demonstrated phagocytized platinum and silicone foreign bodies. In five specimens, energy-dispersive X-ray spectroscopy demonstrated that the birefringent foreign bodies were consistent with silicone. Scanning electron microscopy of two electrodes removed from temporal bones demonstrated small cracks, fragmentation, and small circular defects in the silicone carrier., Conclusion: Histologic evidence of a foreign body response to the presence of platinum and silicone in a cochlear implant has been demonstrated and may be responsible for some reported delayed failures or extrusion.