Formation of human cementum following different modalities of regenerative therapy.

The aim of the present study was to compare newly formed cementum following different types of regenerative therapy in humans. Eighteen patients, each displaying one advanced intrabony defect around teeth scheduled for extraction, were included in this study. The defects were treated with either guided tissue regeneration (GTR), enamel matrix protein derivative (EMD), EMD plus bioactive glass, bovine-derived xenograft (BDX), BDX plus GTR, or BDX plus EMD. After healing, the teeth were removed together with their surrounding soft and hard tissues. Cellular content, presence of artifactual splits between the new cementum and the old one or the dentin surface, and thickness of the new cementum were evaluated. Irrespective of treatment, the new cementum was of a reparative, cellular, extrinsic and intrinsic fiber type. There were no differences in cementum thickness among treatments. These findings indicate that in humans, (a) the new cementum formed after different types of regenerative therapy was, irrespective of the treatment, of a reparative, cellular, extrinsic and intrinsic fiber type, and (b) the regenerative modality does not seem to influence the type of newly formed cementum.

Immunohistochemical investigation on the pattern of vimentin expression in regenerated and intact monkey and human periodontal ligament.

The expression of vimentin is well documented in the intact animal and human periodontal ligament (PDL), but there is limited information on the pattern of vimentin expression in the regenerated PDL. The aim of the present study was to investigate the pattern of vimentin expression in the regenerated and intact monkey and human PDL. A total of 12 chronic recession-type defects were created in three monkeys (Macaca fascicularis) and treated either with guided tissue regeneration (GTR), or with an enamel matrix protein derivative (EMD). After 5 months, the animals were sacrificed and specimens containing the defects and surrounding tissues were dissected free, decalcified in EDTA and embedded in paraffin. Sections were labelled immunohistochemically by using monoclonal antibody against vimentin (VIM 3B4). Twelve patients, each of whom displayed one deep intrabony defect scheduled for extraction were treated with GTR, EMD or combination of EMD+natural bone mineral (NBM). Following a healing period of 6 months, the teeth were extracted "en block" and immunohistochemically analysed according to the same protocol as described in monkeys. The results revealed that in both monkeys and humans the newly formed PDL was labelled similarly for vimentin to the intact (non-treated) PDL. In all specimens, the newly formed PDL was in continuation with the intact parts of PDL, thus suggesting that the mesenchymal cells capable of regenerating the attachment apparatus may have their origin in the intact PDL. In conclusion, the present findings indicate that (a) the reformed PDL displayed a similar expression of vimentin to the intact (original) PDL, and (b) the cells capable of regenerating new PDL and new cementum appear to be of mesenchymal origin and their source may be in the intact PDL.

Immunohistochemical evaluation of matrix molecules associated with wound healing following regenerative periodontal treatment in monkeys.

The aim of the study was to investigate immunohistochemically the expression of matrix molecules associated with periodontal tissues reformed after regenerative periodontal treatment. Chronic intrabony defects were treated with guided tissue regeneration, enamel matrix proteins, the combination of both, or access flap surgery. Five months after healing, the animals were killed, and the healed periodontal tissues were evaluated immunohistochemically by means of polyclonal antibodies against osteopontin, collagen I, and collagen III. The intact (nontreated) parts of the periodontium served as controls. As a general observation, the staining for all investigated matrix molecules appeared to be stronger within the regenerated tissues than in the intact ones. The results failed to reveal any differences in terms of staining intensity or distribution pattern of investigated matrix molecules between the four different treatments. Osteopontin expression was most intense at the border near the newly formed cementum and bone. In the regenerated periodontium, collagens I and III were localized throughout the entire periodontal ligament connective tissue. In the regenerated periodontal ligament, collagen III displayed more intense staining than collagen I. The present results suggest that: (1) even after a 5-month period following surgical periodontal therapy, extracellular matrix molecules associated with wound healing and/or remodelling are more strongly expressed in regenerated than in intact tissues and (2) once an environment for periodontal regeneration has been created, the expression of extracellular matrix molecules associated with the healing process seems to display the same pattern, irrespective of treatment modality.

Patterns of cytokeratin expression in monkey and human periodontium following regenerative and conventional periodontal surgery.

The pattern of cytokeratin expression has been extensively described in the normal and inflamed periodontium. However, there is no information regarding the pattern of cytokeratin expression in the periodontium which has been reformed following regenerative periodontal surgery. The aim of the present investigation was to evaluate the pattern of cytokeratin expression in the reformed human and monkey periodontium following regenerative and conventional periodontal surgery. In 3 monkeys, acute fenestration-type and chronic intrabony defects were treated with guided tissue regeneration (GTR), enamel matrix proteins (EMD), or coronally repositioned flap surgery (control). After a healing period of 5 months, the animals were sacrificed and perfused with 10% buffered formalin for fixation. Specimens containing the defects and surrounding tissues were dissected free, decalcified in EDTA and embedded in paraffin. Histological sections were cut with the microtome set at 3 microm. The sections were alternatively stained either with hematoxylin and eosin, or immunohistochemically by using one of the broad range monoclonal antibodies 34betaE 12 (for cytokeratins 1, 5, 10 and 14) or KL 1 (for cytokeratins 1, 2, 5, 6, 7, 8, 10, 11, 16 and 19), or one of the individual monoclonal antibodies LL025 (for cytokeratin 16), DC 10 (for cytokeratin 18), A53-B/A2 (for cytokeratin 19). Twelve patients, each displaying one deep intrabony defect scheduled for extraction due to advanced periodontitis or prosthetic reasons, were treated as described above. Following a healing period of 6 months, the teeth were extracted together with some of their surrounding soft and hard tissues. The histological and immunohistochemical processing of the human biopsies was identical to that described in monkeys. The results revealed that both the normal non-treated (original) monkey and human junctional epithelium stained strongly with all of the monoclonal antibodies used. The reformed junctional epithelium displayed the same cytokeratin expression pattern as the non-treated junctional epithelium. No differences regarding the cytokeratin expression pattern of the junctional epithelium were found between the treatments and types of healing (i.e. regenerative, through a new periodontal ligament, or reparative through a long junctional epithelium). In the intact periodontal ligament, the epithelial rests of Malassez displayed a comparable cytokeratin expression pattern to that of the junctional epithelium. However, no expression of cytokeratins was seen in the newly formed periodontal ligament. In such specimens, cytokeratin expression was observed only until the borderline between the regenerated cementum and the epithelial downgrowth. It was concluded that: a) the reformed junctional epithelium, following any type of surgical procedure, displays a similar pattern of cytokeratin expression to the original junctional epithelium; b) in the newly formed periodontal ligament, no expression of cytokeratins is present; and c) the epithelial rests of Malassez do not seem to reform after regenerative periodontal surgery.