title: Human periodontal ligament fibroblasts stimulated by nanocrystalline hydroxyapatite paste or enamel matrix derivative. An in vitro assessment of PDL attachment, migration, and proliferation.
abstract: We determined the effects of soluble or coated nanocrystalline hydroxyapatite paste (nano-HA) and enamel matrix derivative (EMD) on proliferation, adhesion, and migration of periodontal ligament fibroblasts (PDLs). Cultured PDLs were stimulated with nano-HA paste or EMD in a soluble form or were coated to the surface of cell culture dishes. Proliferation of PDLs on coated nano-HA and EMD was quantified by various methods including bromodeoxyuridine (BrdU) incorporation and Western blot. Cell migration was investigated in a modified Boyden chamber. The surface integrin profile of PDLs was determined using an integrin-specific ELISA, and integrin-specific signaling was measured by immunoblotting of phosphorylated focal adhesion kinase (FAK). Coated nano-HA stimulated PDL proliferation to a larger extent as compared with coated EMD. PDL migration towards a nano-HA or EMD gradient was more efficiently mediated by soluble EMD as compared with nano-HA but vice versa, adhesion of PDLs to compound-coated dishes was more effectively mediated by nano-HA as compared with EMD. Mechanistically, majorly integrin α5β1-mediated adhesion of PDL and both coated compounds mediated a significant increase in FAK activation though to a different extent. Current findings offer two different modes of action for EMD and nano-HA paste. EMD efficiently acts as a chemoattractant in its soluble form, while nano-HA paste effectively serves as a synthetic extracellular matrix component in its coated form. Our findings suggest that EMD and nano-HA paste display different molecular characteristics and apply alternative routes to mediate their beneficial effects on periodontal tissues.
authors.affiliation: Department of Operative Dentistry and Periodontology, Johannes Gutenberg-University, Augustusplatz 2, 55131 Mainz, Germany. Kasaj@gmx.de
authors.forename: Adrian
authors.initials: A
authors.lastname: Kasaj
authors.forename: Brita
authors.initials: B
authors.lastname: Willershausen
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Junker
authors.forename: Stefan-Ioan
authors.initials: SI
authors.lastname: Stratul
authors.forename: Mirko
authors.initials: M
authors.lastname: Schmidt
mesh_terms: D000293:Adolescent; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000494:pharmacology*; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D017886:Durapatite / Q000494:pharmacology*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism; D008297:Male; D053758:Nanoparticles; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D012038:Regeneration / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D013268:Stimulation, Chemical
chemical_list: D020099:Coated Materials, Biocompatible; D003746:Dental Enamel Proteins; D039081:Integrin alpha5beta1; C109069:enamel matrix proteins; D017886:Durapatite; D051416:Focal Adhesion Protein-Tyrosine Kinases