Instrument separation during endodontic therapy is a frequent accident with rotary instruments being more likely to separate than manual ones. The treatment of cases with a separated instrument can be either conservative or surgical. A conservative approach involves the following treatment choices: a) bypass of the fragment, b) removal of the fragment, c) instrumentation and obturation coronally to the fragment. Concerning the removal of a separated instrument, a variety of techniques and systems have been developed. Ultrasonics, in combination with the operative microscope constitute the most effective and reliable tools for removing a separated endodontic instrument from a root canal. The likelihood of successful removal depends on: the level of separation (coronal, middle or apical third); location in relation to the root canal curvature; the type of separated instrument; its length; the degree of canal curvature and the tooth type. Several complications may occur during the management of a separated instrument: separation of the ultrasonic tip or file used for bypassing or removing the instrument; further separation of the fragment; perforation; ledge; extrusion of the file into periapical tissues; tooth weakening due to dentin removal, as well as excessive temperature rise in periodontal tissues. Prognosis for a tooth retaining a separated instrument depends on the presence of a periapical lesion, the microbial load of the root canal during the time of separation and the quality of the obturation.
The purpose of this study was the evaluation of 3 different histological methods for studying pulpal blood vessels in combination with 2 types of confocal microscope and computer assisted 3-dimensional reconstruction. 10 human, healthy, free of restorations or caries teeth that were extracted for orthodontic reasons were used. From these teeth, the pulp tissues of 5 were removed, fixed in formalin solution, dehydrated and embedded in paraffin. Serial cross sections 5μm thick were taken from 3 of the above mentioned pulpal tissues and stained with CD34 according to the immunohistochemical ABC technique, while the rest 2 were stained with CD34 and Cy5 by means of immunofluorescence after serial cross sectioning of 10μm. 5 of the 10 teeth were fixed, decalcified, serial cross sectioned (30μm thickness) and stained with eosin. The physical sections were examined under 2 types of confocal laser microscope. Serial images were taken for each section, alignment of the images was followed and finally 3-dimensional reconstructions of the pulpal vessels were achieved.
The combined use of immunofluorescence, confocal microscope and automatic segmentation proved to be a useful method for the detailed study of pulpal vasculature. The above method provides deep knowledge of the form and spatial relationship even of the smallest pulpal blood vessels with neighbouring structures like odontoblasts, which are essential for the fully understanding of their role and function within the dental pulp.