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Healing responses to grafting of combined collagen-decalcified bone in periodontal defects in dogs.

The feasibility of a new combined collagen gel-autolysed antigen-extracted allogeneic bone implant was tested. Its effect on gaining new attachment in surgically created defects in four dogs was evaluated over 24 weeks. As controls, sites receiving implants of the bone implant alone, nonimplanted and untreated defects were also evaluated. The collagen gel encouraged ingrowth of regenerative tissue-fibroblasts in the early stages of wound healing while the allogeneic bone induced new bone formation. The graft materials were bicompatible, technically manageable and clinically effective. From the present preliminary study, it was evident that the combined autolysed antigen-extracted allogeneic bone-collagen gel implant may offer advantages over the present implant materials and techniques currently used in the treatment of periodontal intrabony defects.

Alveolar Process↗

[Whole organ plastination in otorhinolaryngology].

Total organ sections have proven to be useful in the evaluation of the clinical growth behavior of malignant tumors. However, histologic techniques using tissues embedded in celloidin or paraffin are difficult to produce and require extensive amounts of time. Total organ plastination is a method superior to the aforementioned techniques. The number of artefacts is also greatly reduced by avoiding the need for decalcification. After removal of fat and dehydration, specimens can be placed in epoxy resin. Following evacuation and hardening of specimens, parallel sections can be separated with a diamond wire saw for precision slicing, with section thicknesses ranging from several centimeters to as small as 40 microns. The loss in material caused by slicing is as little as 0.3 mm per section. A further benefit of this method is the relatively large amount of time saved. The entire procedure yielding complete plastinated specimens requires a time period of approximately 40 days. The plastinated specimens are transparent, odorless and fixed and are ideal for instructional and research purposes. Deplastination for the purpose of further diagnositic study is also achievable with a minimum of technical effort.

Epoxy Resins↗

THE PROBLEM OF DEMINERALISATION IN THIN SECTIONS OF FULLY CALCIFIED BONE.

Thin sections of embryonic avian bone decalcify during preparation for electron microscopy, creating a false impression of mineral distribution. The results of the experiments reported herein show that viscous embedding materials do not penetrate compact formed bone, and so, in thin sections, the calcium apatite crystals may be leeched out by water, both in the collecting trough and in aqueous solutions of stains used to enhance tissue electron opacity. To prevent decalcification, a simple technique is described in which the aqueous fluids that come in contact with thin sections are saturated with respect to calcium and phosphate ions, thereby preventing solution of the bone mineral. The theoretical basis of this technique is briefly discussed.

Animals↗

[Morphological diagnosis of spinal diseases. Current technical possibilities and challenges for the histological preparation of transpedicular biopsies].

The spine is the central component for the mobility of the human body. Both locally limited and diffuse pathologies of the spine are a challenge for the treating physician due to the difficult anatomy. The biopsy of a pathologically altered vertebral body is a fast and reliable basis for further therapy but until now this has not regularly been made use of as a diagnostic standard for spinal diseases, since the tissue gained was often microfractured because of the difficult anatomical position. Our own experience with transpedicular vertebral biopsies of 70 patients with different diseases is reported because of the considerable improvement in the biopsy technique and the methodical possibilities for processing the bony tissue. Methods which have proven particularly valuable are contact radiographs, embedding in plastic, careful decalcifying with EDTA and immunohistological procedures. In this way a definite diagnosis can be made in 97% of the cases. A close cooperation with the clinician carrying out the biopsy and a greater use of methods other than just fast decalcification is recommended.

Biopsy↗

Inner ear proteomics of mouse models for deafness, a discovery strategy.

Inner ear dysfunction is often associated with defective hair cells. Therefore, hair cells are the focus of study in many of the mouse mutants showing auditory and vestibular deficits. However, harvesting sufficient numbers of hair cells from the tiny bony mouse inner ear for proteomic analysis is challenging. New approaches that would take advantage of mouse mutants and avoid processing steps, such as decalcification or microdissetion, would be more suitable for proteomic analysis. Here, we propose a novel approach called SSUMM-Subtractive Strategy Using Mouse Mutants. SSUMM takes advantage of the differences between control and affected or mutant samples. We predict that SSUMM would be a useful method in proteomics, especially in those cases in which the investigator must work with small numbers of diverse cell types from a tiny organ. Here, we discuss the potential utility of SSUMM to unravel the protein expression profiles of hair cells using the Pou4f3 mouse mutant as an example. Pou4f3 mutant mice exhibit a total loss of inner and outer hair cells, but supporting cells remain relatively intact in the cochlea, thus providing an excellent model for identifying proteins and transcripts that are specific to the hair cell at all life stages. SSUMM would maximize the sensitivity of the analyses while obviating the need for tedious sessions of microdissection and collection of hair cells. By comparing the mutant to control ears at specific time points, it is possible to identify direct targets of a gene product of interest. Further, SSUMM could be used to identify and analyze inner ear development markers and other known genes/proteins that are coexpressed in the ear. In this short technical report, we also discuss protein-profiling approaches suitable for SSUMM and briefly discuss other approaches used in the field of proteomics.

Animals↗