An immunohistochemical, histological, and electron-microscopic study of the human periodontal ligament during orthodontic treatment

Abstract The periodontal ligament lies between the hard tissues of alveolar bone and cementum of teeth and serves to anchor the tooth to the alveolus and functions as a cushion between these hard tissues to migrate occlusal force during mastication. This tissue is always exposed to mechanical stress during mastication. When occlusal forces exceed the adaptive capacity of the periodontal ligament, the periodontal ligament tissue will be injured and then occlusal trauma will occur. The different modifications of periodontal ligament during load deformation can be monitored by analysis of the expression of different collagen types and fibronectin, with immunohistochemical techniques, and by morphological study of ligament, with light- and transmission electron-microscopic techniques. The use of continued and light orthodontic force generates a pressure of ligament with ejection of parodontal fluid externally and partial closing of vessels. On these basis we performed a study in order to evaluate periodontal ligament collagen types I and IV and the fibronectin modifications induced by application of a precalibrated orthodontic strength. We integrated these results, with light and transmission electron-microscopic observations, in order to evaluate the morphological modifications of periodontal tissue. Our observations showed that the type I collagen immunofluorescence staining is increased in the pressure side; in the tension side, it shows prior to treatment an increase, and after 72 h of treatment, a diminution of the staining pattern. Type IV collagen staining is reduced in both sides, but increased gradually after 7 days from treatment; finally, fibronectin staining pattern is gradually increased in the pressure side and reduced in the tension side. In light and transmission electron-microscopic observations it is possible to show a reduction of vessels at 72 h from treatment, and an increase of vessels after 7 days from treatment. The Malassez's epithelial residues are decreased at 72 h, while they are increased after 7 days from treatment. The modifications of immunofluorescence staining patterns of tested proteins revealed angiogenesis and reparative processes, and a thickening of fibrillar matrix as a defensive reply to mechanical stress. The modification of normal staining patterns of tested protein in our observations, could be determined by variation of scaffold geometry of periodontal ligament. The reduced movements of contraction and relaxation of periodontal ligament, due to orthodontic treatment, provoke a loss of mechanical stresses transmitted over ligament surface. Mechanical signals, therefore, could be integrated with other environmental signals and transduced into biochemical signals through force-dependent changes in scaffold geometry. Physical forces of gravity, hemodynamic stresses, and movement play a critical role in tissues, since the cells use tensegrity architecture for their structural organization.