Anwendungsmöglichkeit der histologischen Aufarbeitung von Gewebe-Implantat-Scaffolds eingebettet in Technovit¾ : (eine methodische Arbeit)
In Germany approximately 17.000 people go blind per year. Mainly because of degenarative retinopathias, while current available therapy strategies are limited and mostly failing to effect a cure. For some of this people, there is a new chance to gain their eyelight again within the Projekt RETINA-IMPLANT. In this project researchers are develloping a retina protheses. One part of this work are long time biocompatibility experiments. Therefore, the silicone coat of the protheses is implanted for different periods in one eye of a cat, pig, minipig or rabbit. Afterwards the animal is killed and both eyes are explanted and histologically examinated. The biological attachment of the silicone coat on the retina should be through a gliosis and not through a fibrosis. The histological examination is not always able to clearly distinguish between gliosis and fibrosis. So immunhistological methods are an optimal supplement to standard histological examinations. In routine procedures, material for such examinations is embedded in paraffin. But with this method, it is not possible to investigate the retina-implant-scaffolds. Paraffin is too smooth. We need a much harder embedding material like Technovit®8100 or 9100. Till now, there is no information about the immunhistological staining method with Technovit®8100 or 9100, except for the product information about Technovit®8100. So the goal of this study is to establish the immunhistological staining method with Technovit®8100 and 9100 on normal eyes without any implantations. Therefore, normal eyes from rabbit, swine and minipig were embedded in paraffin and normal eyes from rabbit and swine were embedded in Technovit®8100 and 9100. Afterwards serial sections of 2 µm were made from all embedded eyes succeded by immunhistological labelling. Additionally, sections of 2 µm thickness from a rabbit eye (RI014/02) embedded in paraffin which has a fibrosis were immunhistological labelled. Staining results were compared, depending on the embedding material and the possibility to clearly distinguish between gliosis and fibrosis. Staining results from the eyes embedded in paraffin served as negative control. The outcome of the experiments is, that with immunhistological labelled sections from eyes embedded in Technovit®8100, we could not clearly identify structural changes like we have within a gliosis or fibrosis. Another disadvantage with this embedding material was, that for positive staining results higher antibody concentrations are needed and it is not possible to get any positive results with the labelling method with DAP. Meanwhile staining results from immunhistological labelled sections from eyes embedded in Technovit®9100 show the same qualitity with similar antibody concentrations like labelling results from eyes embedded in paraffin. Therefore the immunhistochemistry with Technovit®9100 is suitable for the exact characterization between gliosis and fibrosis. The disadvantage of this method is the really costly and time-consumingembedding of the material in Technovit®9100, which takes more than four weeks. So at the time, there is no optimal embedding material for exploring tissue-implantat-scaffolds with immunhistological labelling.
info:eu-repo/classification/ddc/610, Histologie, Schwein, Kaninchen, Miniaturschwein, Gliose, Fibrose, Immuncytochemie, Fluoreszenz, Netzhaut, Retina <Druckschrift>, Medizin, Retina-Implantat
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