Abstract
Potent signaling agents stimulate and guide pulp tissue regeneration, especially in endodontic treatment of teeth with incomplete root formation. Objective This study evaluated the bioactive properties of low concentrations of extracellular matrix proteins on human apical papilla cells (hAPCs).Methodology Different concentrations (1, 5, and 10 µg/mL) of fibronectin (FN), laminin (LM), and type I collagen (COL) were applied to the bottom of non-treated wells of sterilized 96-well plates. Non-treated and pre-treated wells were used as negative (NC) and positive (PC) controls. After seeding the hAPCs (5×103 cells/well) on the different substrates, we assessed the following parameters: adhesion, proliferation, spreading, total collagen/type I collagen synthesis and gene expression (ITGA5, ITGAV, COL1A1, COL3A1) (ANOVA/Tukey; α=0.05).Results We observed greater attachment potential for cells on the FN substrate, with the effect depending on concentration. Concentrations of 5 and 10 µg/mL of FN yielded the highest cell proliferation, spreading and collagen synthesis values with 10 µg/mL concentration increasing the ITGA5, ITGAV, and COL1A1 expression compared with PC. LM (5 and 10 µg/mL) showed higher bioactivity values than NC, but those were lower than PC, and COL showed no bioactivity at all.Conclusion We conclude that FN at 10 µg/mL concentration exerted the most intense bioactive effects on hAPCs.
Highlights
The concepts of tissue engineering were introduced to Dentistry in the last decade giving special attention to inducting pulp regeneration.1-4 The principles of regenerative endodontics are four: 1. disinfection and detoxification of root canals; 2. use of biomaterials as scaffold for cell adhesion, proliferation, and differentiation; 3. presence of cells with potential to regenerate a new tissue similar to the original; and 4. use of signaling agents to induce cell migration and enhance the bioactive action of the biomaterial.5,6Two strategies emerged as the most promising therapies for tissue regeneration: cell approaches and cell-free approaches.7,8 The first involves implanting pre-cultured cells associated with a biomaterial at the site of injury
Cell proliferation: Figure 3 shows increased cell proliferation occurred over time-points for all groups (p0.05)
In the present study, we assessed the biological activities of low concentrations of extracellular matrix proteins (ECMp) on human apical papilla cells (hAPCs)
Summary
The concepts of tissue engineering were introduced to Dentistry in the last decade giving special attention to inducting pulp regeneration. The principles of regenerative endodontics are four: 1. disinfection and detoxification of root canals; 2. use of biomaterials as scaffold for cell adhesion, proliferation, and differentiation; 3. presence of cells with potential to regenerate a new tissue similar to the original; and 4. use of signaling agents to induce cell migration and enhance the bioactive action of the biomaterial.5,6Two strategies emerged as the most promising therapies for tissue regeneration: cell approaches and cell-free approaches. The first involves implanting pre-cultured cells associated with a biomaterial at the site of injury. Use of biomaterials as scaffold for cell adhesion, proliferation, and differentiation; 3. Presence of cells with potential to regenerate a new tissue similar to the original; and 4. Use of signaling agents to induce cell migration and enhance the bioactive action of the biomaterial.. Two strategies emerged as the most promising therapies for tissue regeneration: cell approaches and cell-free approaches.. The first involves implanting pre-cultured cells associated with a biomaterial at the site of injury. The bioactive materials provide a porous three-dimensional structure and act as a temporary extracellular matrix where attached cells can grow to regenerate the tissue. The second, cellfree approaches, involve the use of biomaterials along potent signaling agents. When put into the injury site, these agents stimulate cell migration towards the site of interest, as well as their proliferation and differentiation, favoring tissue healing.
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