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Transmission and scanning electron microscopic analysis of mineralized loci formed by human periodontal ligament cellsin vitro

Transmissions-und rasterelekronenmikroskopische Untersuchung von Mineralisierungsknoten humaner Parodontalligamentzellen: EineIn-vitro-Studie

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Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie Aims and scope Submit manuscript

Abstract

Fibroblasts isolated from human periodontal ligament (PDL) were cultured under a medium supplemented with ascorbic acid, β-glycerophosphate and dexamethasone. The cultures were assessed for their ability to elaborate a mineralized matrix.

Cell cultures stained positive when analyzed for alkaline phosphatase activity throughout the culture period. After about 3 weeks in culture, the cells produced a calcified matrix. Light microscopy showed formation of clusters of different shapes and sizes. Von Kossa staining revealed mineral deposits as amorphous brown-black precipitates. Transmission electron microscopy showed cells in multilayers and mineralized formations in close association with a dense network of collagen fibers. Scanning electron microscopy revealed smooth formations rising over the cultures with an abundant fiber matrix.

We conclude that human PDL fibroblasts can be induced to form a mineralized matrix which shares features with bone mineralized matrix but most likely represents a more immature type ofin vitro mineralization. Moreover, the present study further supports the osteoblastic potential of these cells.

Zusammenfassung

Fibroblasten des humanen Parodontalligaments (PDL) wurden in einem Medium, das mit Vitamin C, β-Glycerophosphat und Dexamethason angereichert war, kultiviert. Diese Kulturen wurden auf ihre Fähigkeit zur Bildung einer mineralisierten Matrix hin untersucht.

In diesen Zellkulturen verlief der Nachweis der alkalischen Phosphatase während des gesamten Kulturzeitraums positiv. Nach einem Kulturzeitraum von etwa drei Wochen produzierten die Zellen eine kalzifizierte Matrix. Lichtmikroskopisch war die Bildung von Zellklustern (Zellaggregaten) unterschiedlicher Gestalt und Größe sichtbar. Die Von-Kossa-Färbung ließ mineralisierte Ablagerungen in Form amorpher braun-schwarzer Präzipitate erkennen. Die Transmissionselektronenmikroskopie zeigte eine mehrschichtige Anordnung der Zellen und bestätigte mineralisierte Ablagerungen, die in enger Verbindung mit einem dichten Netzwerk aus Kollagenfasern standen. Die rasterelektronenmikroskopische Untersuchung verdeutlichte, daß die Kulturen mit Ablagerungen überschichtet waren, die auf die Gegenwart einer faserartigen Matrix hindeuteten.

Die Befunde lassen den Schluß zu, daß in humanen PDL-Fibroblasten die Bildung einer mineralisierten Matrix induziert werden kann, die Charakteristika der mineralisierten Matrix von Knochen aufweist, aber weitestgehend eine eher unreife Form derIn-vitro-Mineralisation darstellt. Darüber hinaus bestätigt die vorliegende Untersuchung das Potential dieser Zellen, Charakteristika von Osteoblasten zu realisieren.

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Authors and Affiliations

  1. Department of Orthodontics and Dentofacial Orthopedics, School of Dental Medicine, University of Heidelberg, Germany

    Gerda Komposch

  2. Poliklinik für Kieferorthopädie der Universität, Im Neuenheimer Feld 400, D-69120, Heidelberg

    Efthimia K. Basdra

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  1. Efthimia K. Basdra
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  2. Gerda Komposch
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Dedicated to Prof. Dr. Emil Witt on the Occasion of his 65th Birthday.

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Basdra, E.K., Komposch, G. Transmission and scanning electron microscopic analysis of mineralized loci formed by human periodontal ligament cellsin vitro . J Orofac Orthop/Fortschr Kieferorthop 60, 77–86 (1999). https://doi.org/10.1007/BF01298958

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  • DOI: https://doi.org/10.1007/BF01298958

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