Summary
The question of the initial mineralization in the epiphyseal plate has been investigated to date in specimens prepared by conventional electron microscopical techniques. As conventional techniques can produce artifacts, either a loss of mineral substance or a secondary nucleation, the mineralization process was investigated using freeze dried, vacuum embedded growth cartilage which was neither contrasted nor stained and which had a very short contact with water.
The prevailing theory that the first mineralization begins within extracellular matrix vesicles and that the mineralization outside these vesicles is a secondary process was confirmed. Mineralized matrix vesicles were found in the fully mineralized long septa down to the opening zone. In several cases a mineralization could be observed in those transverse septa in which organic substance was present between the cells. The typical radial arrangement of the apatitic needles and platelets in the matrix vesicles could be explained by the formation of a mineralization in an ionotropic gel, the orientation of the matrix macromolecules to be produced by a vectorial influx of calcium ions and phosphate groups coming from different directions. Thin strands of mineral substance with low contrast, which follow the direction of the longitudinal septum, were assumed to be the mineralized collagen fibrils. In several needles dot-like formations were seen and the distance between the middle of neighbouring dots was found to lie mainly in the range 30–56 Å, while the lateral separation distance between the middle of closely packed parallel chains and needles was found to lie mainly in the range 30-42 Å. Parallel periodic structures which could be visualized in apatitic chains and needles 20–40 Å in diameter were assumed to be the 8.2 Å-(100)-lattice planes of apatite, being an indication that these formations already possess criteria of the apatite lattice.
Zusammenfassung
Die Frage nach der Art und dem Ort der ersten Minerali-sierung in der Epiphysenplatte wurde bis jetzt an Proben untersucht, die nach den herkömmlichen Methoden der elektronenmikroskopischen Präparation vorbereitet worden waren. Da durch solche konventionellen Präparationsmethoden Artefakte erzeugt werden können, die in einem Herauslösen und/oder einer ungewollten Wiederausfällung von Mineralsubstanz im Gewebe bestehen, untersuchten wir den Mineralisierungsvorgang in der Epiphysenfuge an Substanz, die schnell tiefgefroren, gefriergetrocknet, ohne Kontrastierung eingebettet, und von der Ultradünnschnitte angefertigt wurden, die nur einen kurzen Kontakt mit H2O hatten.
Wir konnten die von den meisten Autoren vertretene Theorie erhärten, daß die erste Mineralisierung — vollkommen bzw. vorherrschend — in extrazellulären Matrixvesikeln beginnt, und daß erst sekundär die umliegenden Bezirke mit mineralisiert werden. Mineralisierte Matrixvesikeln wurden in den vollmineralisierten langen Septen bis herab zur Eröffnungszone gefunden. Wir konnten auch Mineralisierungen in transversalen Septen beobachten, in denen bereits organische Substanz angelegt war. Die typische radiale Ausrichtung der apatitischen Nadeln und länglichen Blättchen in den Matrixvesikeln erklärten wir durch die Ausbildung eines ionotropen Gels infolge vektoriellen Einstroms der Ca- und Phosphat-Ionen und durch eine Mineralisierung an den so geordneten Makromolekülen. Durch Vermessungen stellten wir fest, daß die Abstände zwischen den Mitten von benachbarten punktartigen Mineralgebilden innerhalb der Nadeln und Ketten vorwiegend im Bereich von 30–56 Å liegen und die Seitenabstände zwischen dicht zusammenliegenden, parallel orientierten Nadeln vorwiegend im Bereich von 30–42 Å. Parallele periodische Streifen innerhalb solcher apatitischen Nadeln deuteten wir als Abbildungen der 8.2 Å-(100)-Netzebenen des Apatits, ein Zeichen dafür, daß diese Gebilde bereits Kriterien des Apatitgitters besitzen.
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We express our thanks to the Deutsche Forschungsgemeinschaft for financial support and to Dr. A. Boyde, London, for valuable discussions.
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Höhling, H.J., Steffens, H., Stamm, G. et al. Transmission microscopy of freeze dried, unstained epiphyseal cartilage of the guinea pig. Cell Tissue Res. 167, 243–263 (1976). https://doi.org/10.1007/BF00224331
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DOI: https://doi.org/10.1007/BF00224331