Abstract
The aim of this study was to develop a three-dimensional in vitro model of periodontium to investigate the osteogenic and cementogenic differentiation potential of the periodontal ligament fibroblast (PDLF) spheroids within a dentin-membrane complex. PDLFs were cultured in both spheroid forms and monolayers and were seeded onto two biological collagen-based and synthetic membranes. Cell-membrane composites were then transferred onto dentin slices with fibroblasts facing the dentin surface and further cultured for 20 days. The composites were then processed for histology and immunohistochemical analyses for osteocalcin, Runx2, periostin, and cementum attachment protein (CAP). Both membranes seeded with PDLF-derived cells adhered to dentin and fibroblasts were present at the dentin interface and spread within both membranes. All membrane-cell-dentine composites showed positive staining for osteocalcin, Runx2, and periostin. However, CAP was not expressed by any of the tissue composites. It can be concluded that PDLFs exhibited some osteogenic potential when cultured in a 3D matrix in the presence of dentin as shown by the expression of osteocalcin. However the interaction of cells and dentin in this study was unable to stimulate cementum formation. The type of membrane did not have a significant effect upon differentiation, but fibroblast seeded-PGA membrane demonstrated better attachment to dentin than the collagen membrane.
Highlights
Autologous transplantation of periodontal fibroblasts and stem cells may be a promising technique to induce tissue regeneration in the treatment of periodontal disease [1,2,3,4]
At 20 days, polyglycolic acid (PGA) cultured with either periodontal spheroids or monolayers showed a better attachment on dentin compared to collagen membrane
PGA showed a well-blended appearance and complete attachment onto the dentin surface; collagen membrane showed some nonadherent areas to the dentin surface mainly on the margins of the membrane (Figure 1)
Summary
Autologous transplantation of periodontal fibroblasts and stem cells may be a promising technique to induce tissue regeneration in the treatment of periodontal disease [1,2,3,4]. 3D spheroidal cultures of periodontal fibroblasts were developed and characterized in vitro with respect to their potential use in conjunction with the biological membranes for periodontal tissue repair and regeneration [8]. The interaction of periodontal fibroblast spheroids with dentin has not been thoroughly investigated and it is not clear how the periodontal fibroblasts would behave in a 3D matrix in the presence of both dentin and biological membranes. Experimental studies have accentuated the interactions between dentin, cells from the tooth, and periodontal tissues and reveal dentin to be an adhesive, signaling, and migratory
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