ABSTRACT
Several in vitro investigations have indicated that the particulate phase of cigarette smoke, such as nicotine, affects many cell types, including gingival fibroblasts. However, few studies have been performed on the effects of the volatile fraction on the cellular structures that are involved in cell functions, such as adhesion and proliferation. Since the survival and reproduction of gingival fibroblasts are fundamental in maintaining the integrity of the oral connective tissue, as well as in wound healing, the effects on the cytoskeleton of acrolein and acetaldehyde, which are the volatile fractions of cigarette smoke, were examined in vitro for human gingival fibroblasts (HGFs). HGF strains that were taken from healthy subjects with non-inflamed-gingiva were utilized in this investigation. The cells were incubated in the presence of different concentrations of acrolein and acetaldehyde. Cell adhesion and viability were evaluated after incubation for 3 h and 5 days, respectively. The influence on cytoskeletal structures (tubulin, actin and vimentin intermediate filaments) was investigated with the indirect immunofluorescence technique. The results show that both substances produced similar effects, which resulted in a dose-dependent inhibition of HGF adhesion and viability. Disturbance of the HGF cytoskeleton consisted of disruption of the microtubules, actin filaments and vimentin microfilaments, which was accompanied by alterations to cell shape. Our experimental findings suggest that the volatile fractions of cigarette smoke, such as acrolein and acetaldehyde, have a cytotoxic effect on HGFs, with the result that they lose their capacity for adhesion and proliferation. The consequences of this could be impairment of the maintenance, integrity and remodelling of the oral connective tissue. According to our morphological evidence, these findings show that cigarette smoke can lead to the development and progression of periodontal disease, and indicate the need for appropriate therapy.
Subject(s)
Acetaldehyde/toxicity , Acrolein/toxicity , Cytoskeleton/drug effects , Gingiva/drug effects , Smoking/adverse effects , Actin Cytoskeleton/drug effects , Cell Adhesion/drug effects , Cell Survival/drug effects , Cells, Cultured , Fibroblasts/drug effects , Fluorescent Antibody Technique, Indirect , Gingiva/cytology , Humans , Intermediate Filaments/drug effects , Microscopy, Fluorescence , Microtubules/drug effectsABSTRACT
The aim of this study is to evaluate the possibility to recognize a condylar-mandibular asymmetry through a panoramic radiograph. Results from a previous work, in which 100 skulls from the Museum of the Institute of Anatomy of the University of Pavia were studied and measured, showed the presence of asymmetry. Using the same skulls we examined the possible correlation between morphological and radiological data. We did not find out correlation's between the condylar asymmetry evaluated at the anatomical level and the radiological asymmetry which was indeed found. This is probably due to the different positioning of the jaws during the two different measuring processes. Nevertheless our results confirm the daily experience of dentistry: asymmetry of mandibular condyle can be one of the mayor causes for the asymmetry of the stomatognatic apparatus.
Subject(s)
Facial Asymmetry/diagnostic imaging , Mandibular Condyle/diagnostic imaging , Mandibular Diseases/diagnostic imaging , Radiography, Panoramic , Algorithms , Cephalometry , Facial Asymmetry/pathology , Humans , Mandible/diagnostic imaging , Mandible/pathology , Mandibular Condyle/pathology , Mandibular Diseases/pathology , Radiography, Panoramic/instrumentation , Radiography, Panoramic/methodsABSTRACT
BACKGROUND: Several in vitro investigations have indicated that the particulate phase of cigarette smoke as nicotine affects many cell types including gingival fibroblasts, but few studies have examined the effect of volatile fraction on cellular structures involved in cell functions such as adhesion and proliferation. Since gingival fibroblast survival and reproduction are fundamental to maintaining the oral connective tissue as well as to wound healing, the effects of acrolein and acetaldehyde, volatile fractions of cigarette smoke, on cytoskeleton were examined in human gingival fibroblasts (HGFs) in vitro. METHODS: Human gingival fibroblast (HGF) strains from healthy subjects with non-inflamed gingiva were utilized. The cells were incubated in different concentrations of acrolein and acetaldehyde. Cell adhesion was evaluated after 3 hours. The influence of both substances on cytoskeletal structures, tubulin and vimentin intermediate filaments (VIF), was investigated using indirect immunofluorescence technique. RESULTS: The results show that both substances produced similar effects, resulting in a dose-dependent inhibition of HGF adhesion. Disturbance of HGF cytoskeleton consisted of a disruption of microtubules and vimentin microfilaments with alterations in cell shape. CONCLUSIONS: Our experimental findings suggest that volatile fractions of cigarette smoke such as acrolein and acetaldehyde, because their ability to bind and interact with the cytoskeleton, prevent HGF adhesion. Consequently the maintenance of the oral connective tissue and integrity and remodeling could be impaired. According to our morphological evidence, these findings confirm other clinical and epidemiological investigations reporting that volatile components of cigarette smoke could lead to the initiation and progression of periodontal disease.
Subject(s)
Acetaldehyde/adverse effects , Acrolein/adverse effects , Cytoskeleton/ultrastructure , Fibroblasts/ultrastructure , Gingiva/ultrastructure , Nicotiana/adverse effects , Plants, Toxic , Smoke/adverse effects , Acetaldehyde/administration & dosage , Acrolein/administration & dosage , Actin Cytoskeleton/drug effects , Actin Cytoskeleton/ultrastructure , Cell Adhesion/drug effects , Cell Division/drug effects , Cell Size/drug effects , Cell Survival/drug effects , Cells, Cultured , Connective Tissue Cells/drug effects , Connective Tissue Cells/ultrastructure , Cytoskeleton/drug effects , Dose-Response Relationship, Drug , Fibroblasts/drug effects , Fluorescent Antibody Technique, Indirect , Gingiva/drug effects , Humans , Intermediate Filaments/drug effects , Intermediate Filaments/ultrastructure , Microtubules/drug effects , Microtubules/ultrastructure , Statistics as Topic , Time Factors , Tubulin/drug effects , Vimentin/drug effects , Wound Healing/drug effectsABSTRACT
Tobacco smoke, particularly its non-volatile fraction e. g. nicotine, is considered to be a major risk factor for the development and progression of periodontal disease. The purpose of this study has been to determine the effects of acrolein and acetaldehyde of the volatile fraction of tobacco smoking, on human gingival fibroblasts (HGFs) cultured in vitro with particular attention to cytoskeletal structures. A human gingival fibroblast strain derived from healthy gingiva was utilized in this study. The cells were exposed to acrolein and acetaldehyde at various concentrations. Control and treated cells were compared as regards their adhesion on cell culture dishes. Their cytoskeletal structures [tubulin and vimentin intermediate filaments (VIFs)] were examined by fluorescence microscopy. The results revealed that both substances produced similar effects resulting in a dose dependent decrease in cell adhesion and alterations of HGF cytoskeleton consisting of rearrangement and/or disruption of microtubules and vimentin associated filaments. Changes in cell shape and decrease in cell size were also seen. On the basis of this in vitro study, it appears that tobacco, through its volatile components, may directly affect the main functions of HGFs.
Subject(s)
Acetaldehyde/pharmacology , Acrolein/pharmacology , Fibroblasts/drug effects , Gingiva/cytology , Intermediate Filaments/drug effects , Microtubules/drug effects , Vimentin/drug effects , Cell Adhesion/drug effects , Cell Size , Cells, Cultured , Fibroblasts/cytology , Fibroblasts/physiology , Gingiva/physiology , Humans , Intermediate Filaments/physiology , Intermediate Filaments/ultrastructure , Microtubules/physiology , Microtubules/ultrastructure , Reference Values , Tubulin/drug effects , Tubulin/physiology , Vimentin/physiologyABSTRACT
Tobacco and some of its volatile and non-volatile components have been found to affect many types of cells including gingival fibroblasts. Because normal gingival fibroblast functioning is fundamental to the maintenance of the oral connective tissue as well as to wound healing, we examined the effect of two vapour phase smoke components (acrolein and acetaldehyde) on proliferation and ultrastructure of human gingival fibroblasts (HGFs) in culture. A human gingival fibroblast strain derived from healthy individuals was used in this study. The cells were incubated in the presence of different concentrations of acrolein and acetaldehyde and cell proliferation and fine morphology were evaluated. The results show that acrolein and acetaldehyde produced dose dependent inhibition of HGF viability and alteration of cytoplasmic organelles. The main ultrastructural finding for the HGF cytoplasm was the presence of vacuoles and lysosomal structures which became prominent with increasing concentration of acrolein and acetaldehyde. Our results suggest that the ultrastructural alterations we observed in HGFs may be due to the uptake and storage of acrolein and acetaldehyde by the cells.
Subject(s)
Acetaldehyde/pharmacology , Acrolein/pharmacology , Fibroblasts/ultrastructure , Gingiva/cytology , Smoke , Actin Cytoskeleton/drug effects , Actin Cytoskeleton/ultrastructure , Fibroblasts/drug effects , Gingiva/drug effects , Gingiva/ultrastructure , Humans , Polyribosomes/drug effects , Polyribosomes/ultrastructure , Reference Values , Vacuoles/drug effects , Vacuoles/ultrastructureABSTRACT
BACKGROUND: Tobacco and some of its volatile and non-volatile components have been found to affect many types of cells including gingival fibroblasts. Since normal gingival fibroblast functioning is fundamental to the maintenance of the periodontal connective tissue, as well as to wound healing, we examined the effect of acrolein and acetaldehyde, volatile components of cigarette smoke, on proliferation, attachment, and ultrastructure of human gingival fibroblasts (HGFs) in culture. METHODS: Human gingival fibroblast (HGF) strains derived from healthy individuals with non-inflamed gingiva were used in this study. The cells were incubated in the presence of different concentrations of acrolein and acetaldehyde. Cell attachment and proliferation were evaluated after incubation for 3 hours and 5 days, respectively. In addition, the cells were examined with a transmission electron microscope in order to evaluate their morphology. RESULTS: The results show that acrolein and acetaldehyde produced dose-dependent inhibition of HGF attachment and proliferation. The cytotoxic effect was, however, reversible when both substances were removed, after 3 days, from the medium. The main ultrastructural finding for the HGF cytoplasm was the presence of vacuoles and lysosomal structures that became prominent with increasing concentration of acrolein and acetaldehyde. CONCLUSIONS: Our experimental data suggest that acrolein and acetaldehyde, volatile components of tobacco smoke, are detrimental to HGF survival and consequently to the oral connective tissue. According to our morpho-functional evidence, these findings corroborate clinical and epidemiological investigations demonstrating smoke as a risk factor in the development of periodontal disease.
Subject(s)
Acetaldehyde/adverse effects , Acrolein/adverse effects , Fibroblasts/drug effects , Gingiva/drug effects , Nicotiana , Plants, Toxic , Smoke/analysis , Acetaldehyde/administration & dosage , Acrolein/administration & dosage , Adult , Cell Adhesion/drug effects , Cell Division/drug effects , Cell Survival/drug effects , Cells, Cultured , Connective Tissue/drug effects , Cytoplasm/drug effects , Cytotoxins/adverse effects , Dose-Response Relationship, Drug , Fibroblasts/physiology , Fibroblasts/ultrastructure , Gingiva/cytology , Gingiva/ultrastructure , Humans , Lysosomes/drug effects , Microscopy, Electron , Periodontal Diseases/etiology , Risk Factors , Time Factors , Vacuoles/drug effectsABSTRACT
In recent years, epidemiological studies have pointed to a significant correlation between cigarette smoke and poor periodontal status. Cigarette smoking is a significant risk factor for the onset and development of periodontal disease, and an association between reduced healing response subsequent to periodontal therapies and cigarette smoking has been found. The epidemiological studies reported here are also supported by the results of an in vitro study on the cytotoxicity of two of the volatile components of cigarette smoke that we ourselves conducted, in which the investigated compounds were found to damage human gingival fibroblasts. We concluded that this damage would be reflected in periodontal health and could slow down wound healing. Patients should thus be alerted by clinicians to the risks smoking poses to oral and dental health.
Subject(s)
Periodontal Diseases/etiology , Smoking/adverse effects , Acetaldehyde/adverse effects , Acrolein/adverse effects , Adult , Age Factors , Aged , Fibroblasts/drug effects , Gingiva/cytology , Gingiva/drug effects , Humans , Middle Aged , Nicotine/adverse effects , Periodontal Diseases/physiopathology , Periodontal Diseases/therapy , Periodontitis/etiology , Periodontitis/physiopathology , Recurrence , Risk Factors , Wound HealingABSTRACT
PTFE membranes are used for guided tissue regeneration in order to treat angular bone defects or forcation involvements in surgical treatment. Ultrastructural investigations have been performed by means of electron transmission and scanning microscopy. In agreement with previous reports, fibroblast cells adhering to the reticular structure of PTFE membrane were observed; these were interposed among coagulated clusters of fibrinous material and blood cells round shaped. Elongated bacterial cells were always present in the microscope fields analysed. These observations were confirmed by means of transmission microscopy; moreover specific techniques enabled us to demonstrate that fibroblast cells were synthetizing collagen, which was present in the form of extracellular fibers mixed to fibrin clusters. Roundish and elongate bacterial cells were always observed both in the extracellular matrix and into macrophages.