Growth Factor Liberation and DPSC Response Following Dentine Conditioning.

Liberation of the sequestrated bioactive molecules from dentine by the action of applied dental materials has been proposed as an important mechanism in inducing a dentinogenic response in teeth with viable pulps. Although adhesive restorations and dentine-bonding procedures are routinely practiced, clinical protocols to improve pulp protection and dentine regeneration are not currently driven by biological knowledge. This study investigated the effect of dentine (powder and slice) conditioning by etchants/conditioners relevant to adhesive restorative systems on growth factor solubilization and odontoblast-like cell differentiation of human dental pulp progenitor cells (DPSCs). The agents included ethylenediaminetetraacetic acid (EDTA; 10%, pH 7.2), phosphoric acid (37%, pH <1), citric acid (10%, pH 1.5), and polyacrylic acid (25%, pH 3.9). Growth factors were detected in dentine matrix extracts drawn by EDTA, phosphoric acid, and citric acid from powdered dentine. The dentine matrix extracts were shown to be bioactive, capable of stimulating odontogenic/osteogenic differentiation as observed by gene expression and phenotypic changes in DPSCs cultured in monolayer on plastic. Polyacrylic acid failed to solubilize proteins from powdered dentine and was therefore considered ineffective in triggering a growth factor-mediated response in cells. The study went on to investigate the effect of conditioning dentine slices on growth factor liberation and DPSC behavior. Conditioning by EDTA, phosphoric acid, and citric acid exposed growth factors on dentine and triggered an upregulation in genes associated with mineralized differentiation, osteopontin, and alkaline phosphatase in DPSCs cultured on dentine. The cells demonstrated odontoblast-like appearances with elongated bodies and long extracellular processes extending on dentine surface. However, phosphoric acid-treated dentine appeared strikingly less populated with cells, suggesting a detrimental impact on cell attachment and growth when conditioning by this agent. These findings take crucial steps in informing clinical practice on dentine-conditioning protocols as far as treatment of operatively exposed dentine in teeth with vital pulps is concerned.

Cytocompatibility of a tissue conditioner containing vinyl ester as a plasticizer.

In the current study, we examined the cytocompatibility of eight vinyl esters as candidate plasticizers for producing phthalate- and ethanol-free tissue conditioners. We measured the estrogenic activity and cytotoxicity of vinyl esters in human fibroblasts and keratinocytes using an E-screen assay and a mitochondrial dye conversion assay, respectively. We also assessed the cytotoxicity of three prototype materials and commercially available tissue conditioners on human fibroblasts grown in collagen gels. Finally, we measured the effects of these materials on the expression of cytokines in three-dimensional cultures by reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assays. None of the tested vinyl esters had estrogenic activity. Vinyl octanoate and vinyl pivalate were the least cytotoxic of the eight tested vinyl esters. In the same vein, a prototype tissue conditioner containing vinyl octanoate had equivalent or weaker cytotoxicity and induction of cytokine expression than conventional materials.

Cytocompatibility and viscoelastic properties of phthalate ester-free tissue conditioners.

We prepared prototype phthalate ester-free tissue conditioners (PFT) from a powder of poly(ethyl methacrylate) and a mixture of several liquids, including di-n-butyl sebacate, benzyl benzoate (BB), and ethanol. The estrogenic activities of the liquids in the PFT were measured by an E-screen assay. We also assessed the cytotoxicity of the prototype and commercial tissue conditioners against a living skin equivalent. Finally, the viscoelastic properties were determined by measuring the rubber hardness and initial flow, while the effect of the mixing liquid on the mechanical properties of a denture resin was assessed by three-point bending test. PFT did not show any estrogenic activity and displayed a low or a similar level of cytotoxicity as the conventional materials. The viscoelastic properties and the effect of the mixing liquid on mechanical properties were influenced by the quantity of BB present in the mixing liquid. The prototype is therefore a reasonable PFT candidate.

Estrogenic activity of tissue conditioners in vitro.

OBJECTIVES: In order to assess the estrogenic activities of plasticizers used in tissue conditioners and four commercial tissue conditioners, we carried out in vitro tests. METHODS: Seven plasticizers and two metabolites were diluted to concentrations ranging from 10(-9) to 10(-4)M. Four commercial tissue conditioners were also diluted to concentrations ranging from 2 x 10(-8) to 2 x 10(-3)g/ml. Estrogenic activities were tested by the E-screen test using MCF-7 cells. When estrogen is present, the cells proliferate. Instead of counting the cells or nuclei directly, cell numbers were assessed by measurement of the total protein content using the sulforhodamine B assay. The liquid compositions of the four commercial tissue conditioners were examined by high-performance liquid chromatography. RESULTS: n-Butyl benzyl phthalate, dibutyl phthalate, n-butyl phthalyl n-butyl glycolate, di-2-ethylhexyl phthalate and benzyl salicylate significantly increased proliferation of MCF-7 cells. The remaining two plasticizers, di-2-ethylhexyl adipate and benzyl benzoate, as well as two metabolites of dibutyl phthalate and di-2-ethylhexyl phthalate, i.e. monobutyl phthalate and mono-2-ethylhexyl phthalate, respectively, did not increase proliferation of MCF-7 cells at the concentrations tested. Four commercial tissue conditioners, Coe comfort (CC), Tissue Conditioner (TC), Hydro Cast (HC) and Denture Soft (DS) II, significantly increased proliferation of MCF-7 cells. HPLC data revealed the commercial products contained plasticizers: benzyl benzoate and dibutyl phthalate in CC, dibutyl phthalate in TC, n-butyl benzyl phthalate in HC and n-butyl phthalyl n-butyl glycolate in DS II. SIGNIFICANCE: Except for benzyl benzoate and di-2-ethylhexyl adipate, the plasticizers tested showed estrogenic activity. The four commercial tissue conditioners tested also showed estrogenic activity, and HC showed especially strong estrogenicity.

Pressure distribution using tissue conditioners on simplified edentulous ridge models. Part 1: The influence of the height of the residual ridge.

This study was conducted to measure the changes of pressure distribution over time on the denture-bearing mucosal surface resulting from relining the intaglio surface of dentures with tissue conditioners. Pressure sensors of electrostatic capacitance variation were used to detect changes on three models of different residual mandibular ridge height. It is suggested that the shrinkage of tissue conditioners influences the dimensional accuracy of a dynamic impression technique.

Dental iatrogenesis, Part 2.

To substantiate biocompatibility of dental materials and techniques, and thereby reduce iatrogenesis, tests for pulp and dentinal responses have been developed. These have shown that high-speed tooth cutting techniques are superior to low-speed techniques even when both incorporate air-water coolant sprays; pressure when placing restorative materials intensifies pulp responses induced by the cutting procedure, and that chemically self-cured resin composites requiring the application of a matrix to enhance adaptation intensify pulp response as compared to visible light-cured equivalents applied incrementally, fully cured throughout, and not requiring the pressure of a matrix. Most cements are irritating to the pulp when used as luting agents and when used less than 0.5 mm from it. Conditioning agents using weak acids for short periods induce little irritation while bonding agents appear to be beneficial by providing protection from subsequently placed restorative material. Clinicians should make every effort to minimize the development of pulp responses and reduce iatrogenic efforts.

An ultrastructural study comparing new gingival tissue attachment on chemically exposed fibrils and retained periodontal ligament.

Successful periodontal healing has been demonstrated in animals following application of citric acid to exposed root surface. However, we have been unable to achieve comparable results on clinical patients. Accordingly, the present study, using monkeys, was designed to re-evaluate the degree of interdigitation of newly-formed gingival fibrils with ones exposed to citric acid compared to normal attachment tissues of the retained periodontal ligament. We found that superficial collagen fibrils in both cases suffer degradation following normal inflammation and healing secondary to flap surgery. On retained periodontal ligament tissues, this change was limited to superficial levels and newly-formed collagen fibrils were linked immediately with old ones. In contrast, when exposed collagen fibrils were conditioned by exposure to citric acid, the degradative change occurred more deeply, and healing was delayed. Thus, the citric acid may not be the best choice of root treatments in terms of promoting better gingival tissue attachment. Furthermore, the results indicated that fibrils preserved on the root should not be curetted as possible during flap surgery as they offer an improved chance for optimal reattachment.