Orientation of Gingival Fibroblasts in Simulated Periodontal Spaces in Vitro

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Abstract
The present study examined the orientation of gingival fibroblasts in simulated periodontal spaces in vitro. Extracted human teeth were root planed followed by root resection and root canal instrumentation. The middle and cervical thirds of each root were cut transversely to create 600‐μm thick sections. Cortical bovine bone was cut, sectioned and contoured to create bone rings 600 μm thick with an internal diameter large enough to accommodate a root slice leaving a circumferential space varying from approximately 0.1 to 1.0 mm. Root slices and bone rings were incubated in a solution of collagenase and hyaluronidase to remove all remaining soft tissue and partially demineralized in EDTA (18%) for 30 minutes. Human gingival fibroblasts (HGF) were plated to confluency in tissue culture dishes. The dentin slices were then gently placed over the HGF monolayer along with bone rings around them to create simulated periodontal spaces. Control root slices were placed without bone rings around them. Cultures were maintained under standard tissue culture conditions. Representative specimens were obtained after 2, 3 and 4 weeks of culture and processed for scanning electron microscopy (SEM).At 2 weeks, the HGF had formed sheets of cells attached to the periphery of the root slices at one end and to the inner surface of bone rings at the other end. The orientation of cell sheets varied from being perpendicular to the periphery of the slice to oblique. At 3 and 4 weeks, the density and size of cell sheets increased and the orientation was maintained. The control root slices at the different observation periods showed cell attachment around the periphery of root slices but the cell sheets were seen only on isolated spots and were smaller in size compared to the experimentais.The present findings suggest that the spatial relationship of substrata (root and bone) can influence the orientation of cells. The present in vitro model appears to be suitable to study the development of oriented fibers in simulated periodontal spaces.
Funding Information
  • National Institutes of Health (DE‐06746)