Disparity in the Influence of Implant Provisional Materials on Human Gingival Fibroblasts with Different Phases of Cell Settlement: An In Vitro Study.

The development of healthy peri-implant soft tissues is critical to achieving the esthetic and biological success of implant restorations throughout all stages of healing and tissue maturation, starting with provisionalization. The purpose of this study was to investigate the effects of eight different implant provisional materials on human gingival fibroblasts at various stages of cell settlement by examining initial cell attachment, growth, and function. Eight different specimens-bis-acrylic 1 and 2, flowable and bulk-fill composites, self-curing acrylic 1 and 2, milled acrylic, and titanium (Ti) alloy as a control-were fabricated in rectangular plates (n = 3). The condition of human gingival fibroblasts was divided into two groups: those in direct contact with test materials (contact experiment) and those in close proximity to test materials (proximity experiment). The proximity experiment was further divided into three phases: pre-settlement, early settlement, and late settlement. A cell culture insert containing each test plate was placed into a well where the cells were pre-cultured. The number of attached cells, cell proliferation, resistance to detachment, and collagen production were evaluated. In the contact experiment, bis-acrylics and composites showed detrimental effects on cells. The number of cells attached to milled acrylic and self-curing acrylic was relatively high, being approximately 70% and 20-30%, respectively, of that on Ti alloy. There was a significant difference between self-curing acrylic 1 and 2, even with the same curing modality. The cell retention ability also varied considerably among the materials. Although the detrimental effects were mitigated in the proximity experiment compared to the contact experiment, adverse effects on cell growth and collagen production remained significant during all phases of cell settlement for bis-acrylics and flowable composite. Specifically, the early settlement phase was not sufficient to significantly mitigate the material cytotoxicity. The flowable composite was consistently more cytotoxic than the bulk-fill composite. The harmful effects of the provisional materials on gingival fibroblasts vary considerably depending on the curing modality and compositions. Pre-settlement of cells mitigated the harmful effects, implying the susceptibility to material toxicity varies depending on the progress of wound healing and tissue condition. However, cell pre-settlement was not sufficient to fully restore the fibroblastic function to the normal level. Particularly, the adverse effects of bis-acrylics and flowable composite remained significant. Milled and self-curing acrylic exhibited excellent and acceptable biocompatibility, respectively, compared to other materials.

Human Gingival Fibroblast Growth and Function in Response to Laser-Induced Meso-and Microscale Hybrid Topography on Dental Implant Healing Abutments.

PURPOSE: Laser-created titanium surface topographies enhance soft tissue attachment and implant stability. However, knowledge about the underlying mechanisms governing the tissue-level reaction is lacking. The objective of this study was to examine the behavior and function of human gingival fibroblasts growing on healing abutments with or without laser-textured topography. MATERIALS AND METHODS: Human primary gingival connective tissue fibroblasts were cultured on healing abutments with machined or laser-textured (Laser-Lok, BioHorizons) surfaces. Cellular and molecular responses were evaluated by cell density assay (WST-1), fluorescence microscopy, qRT-PCR, and detachment test. RESULTS: The machined surface showed mono-directional traces and scratches from milling, whereas the laser-textured surface showed a distinct morphology consisting of mono-directional meso-scale channels (15 µm pitch) and woven, oblique micro-ridges formed within the channel. There were no differences in initial fibroblast attachment, subsequent fibroblast proliferation, nor collagen production between the machined and laser-textured surfaces. Fibroblasts growing on laser-textured surface spread mono-directionally along the meso-channels, while cells growing on machined surfaces spread randomly. Fibroblasts on laser-textured surfaces were 1.8-times more resistant to detachment than those on machined surfaces. An adhesive glycoprotein (fibronectin) and trans-membrane adhesion linker gene (integrin beta-1) were upregulated on laser-textured surfaces. CONCLUSIONS: The increased fibroblast retention, uniform growth, increased transcription of cell adhesion proteins compellingly explain the enhanced tissue-level response to laser-created, hybrid textured titanium surfaces. These results provide a cellular and molecular rationale for the tissue reaction to this unique surface and support its extended use from implant fixtures and healing abutments to diverse prosthetic components where enhanced soft tissue responses would be desirable.