In Vitro Evaluation of Acellular Dermal Matrix as a Three-Dimensional Scaffold for Gingival Fibroblasts Seeding

Background: Tissue engineering principles could improve the incorporation of acellular dermal matrix (ADM). The aim of this study is to verify if ADM is a suitable three-dimensional matrix for gingival fibroblasts and cancerous cells ingrowth, and also if cultured medium conditioned in ADM affect cellular behavior. Methods: Canine gingival fibroblasts (CGF), human gingival fibroblasts (HGF), and murine melanoma cell line (B16F10) were seeded on ADM for up to 14 days. The following parameters were assessed: morphology and distribution of CGF, HGF, and B16F10; CGF and HGF viability; and the effect of ADM conditioned medium (CM) on CGF viability. Results: Epifluorescence revealed that CGF were unevenly distributed on the ADM surface, showing no increase in cell number over the periods of study; HGF formed a monolayer on the ADM surface in a higher number at 14 days (P<0.05); B16F10 exhibited an increase in cell number within 7 days (P<0.05), and were mainly arranged in cell aggregates on the ADM, forming a continuous layer at 14 days. A higher percentage of cells on the ADM surface (P<0.05) compared to inside was observed for all cell types. 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MU) values indicated higher cell viability in samples cultured with HGF compared to CGF (P=0.024). A significantly lower cell viability for CGF grown in CM compared to cells grown in non-CM was observed at 48 and 72 hours (P<0.05). Conclusions: ADM is not suitable as a three-dimensional matrix for gingival fibroblasts ingrowth. Gingival fibroblasts and highly proliferative cells as B16F10 can only be superficially located on ADM, and CGF are negatively affected by culture medium conditioned in ADM, reducing its viability. J Periodontol 2011;82:293-301.

An alternative technique to the harvesting of a connective tissue graft from a thin palate: Enhanced wound healing

This article describes an alternative surgical approach to the harvesting of subepithelial connective tissue grafts from thin palates. A partial-thickness flap is raised, and a graft composed of epithelium and connective tissue is removed from the palate. The superficial layer ( epithelium and a thin zone of connective tissue) is then dissected from the graft and replaced at the donor site to facilitate faster healing. The subjacent layer of connective tissue is placed as needed to obtain root coverage. The clinical application of this technique is described in two patients with multiple gingival recessions.

Clinical and histological evaluation of subepithelial connective tissue after collagen sponge implantation in the human palate

Rocha AL, Shirasu BK, Hayacibara RM, Magro-Filho O, Zanoni JN, Araujo MG. Clinical and histological evaluation of subepithelial connective tissue after collagen sponge implantation in the human palate. J Periodont Res 2012; 47: 758765. (c) 2012 John Wiley & Sons A/S Background and Objective: Successful root-coverage treatment depends on the thickness of the donor tissue. This study aimed to evaluate the thickness of donor tissue after augmentation of the connective tissue in the palatal area by implantation of lyophilized collagen sponge (Hemospon (R)). Material and Methods: Ten patients with an indication for root coverage, whose palate was deficient in adequate connective tissue, were recruited. The procedure was carried out in two stages. In the first stage, the palatal thickness in the donor site was measured at three standardized points (points 1, 2 and 3), from the distal of the canine to the distal of the first molar, and the lyophilized collagen sponge was inserted. In the second stage, the palatal thickness over the implant was measured (at points 1, 2 and 3), two biopsies of the palatal mucosa were collected one over the implant (experimental sample) and the other on the contralateral side (control sample) and then root-coverage treatment was performed. Analyses consisted of clinical assessment of the palatal measurements before and after sponge implantation, and histological assessment of the experimental and control biopsy samples. Data were analyzed using the Wilcoxon test. Results: Both analyses showed a significant increase in mean thickness, of 1.08 mm of neoformed tissue in the clinical analysis (the tissue at point 2 was the thickest of the three points) and of 0.53 mm in the histological analysis. Conclusion: The insertion of lyophilized collagen sponge induced a significant increase in the thickness of palatal connective tissue.

Use of the connective tissue graft for the coverage of composite resin-restored root surfaces in maxillary central incisors

The purpose of this article is to report the use of the subepithelial connective tissue graft technique combined with the coronally positioned flap on a composite resin-restored root surface to treat Miller Class I gingival recessions associated with deep cervical abrasions in maxillary central incisors. Clinical measurements, including gingival recession height, probing depth, and bleeding on probing (BoP), were recorded during the preoperative clinical examination and at 2, 6, 12, and 24 months postoperatively. During the follow-up periods, no periodontal pockets or BoP were observed. The periodontal tissue of the teeth presented normal color, texture, and contouring. In addition, it was observed that creeping attachment had occurred on the restoration. This case report shows that this form of treatment can be highly effective and predictable in resolving gingival recession associated with a deep cervical abrasion. (Quintessence Int 2012;43:597-602)

Treatment of gingival fibromatosis associated with Zimmermann-Laband syndrome

Background: Gingival fibromatosis is a rare condition characterized by a generalized enlargement of the buccal and lingual aspects of the attached and marginal gingiva.Methods: This case report describes the periodontal management of a 13-year-old female patient with gingival fibromatosis associated with Zimmermann-Laband syndrome. The patient presented with gingival enlargement involving the maxillary and the mandibular arches, anterior open bite, and non-erupted teeth. Periodontal treatment included gingivectomy in all four quadrants.Results: Histopathologic evaluation of the excised tissue supported the diagnosis of gingival flbromatosis. A significant improvement in esthetic appearance and eruption of the non-erupted teeth were obtained. The patient was referred for appropriate orthodontic treatment and has been closely followed for the earliest signs of recurrence of gingival enlargement.Conclusions: the successful therapy for gingival fibromatosis depends on correctly identifying the etiological factors and improving the impaired function and esthetic appearance through surgical intervention and adjunctive orthodontics. Maintaining treatment results depends on preservation of periodontal health.

Prostaglandin production by human gingival fibroblasts inhibited by triclosan in the presence of cetylpyridinium chloride

Background: the effect of triclosan plus the cationic detergent cetylpyridinium chloride (CPC) was evaluated for prostaglandin inhibition in human gingival fibroblasts. Since triclosan has previously been shown to inhibit proinflammatory cytokine induced prostaglandin E-2 (PGE(2)) production, we wanted to determine if triclosan, in the presence of CPC, could enhance these effects.Methods: Initial studies determined that both triclosan and CPC were cytotoxic to human gingival fibroblasts in concentrations exceeding 1.0 mu g/ml for either agent longer than 24 hours in a tissue culture. Therefore, subsequent studies measuring prostaglandin biosynthesis and cyclooxygenase (COX)-1 and COX-2 mRNA expression were performed in concentrations and times that did not significantly affect cell viability.Results: PGE2 biosynthesis was dose dependently inhibited by both triclosan and triclosan and CPC when challenged by tumor necrosis factor (TNF)-alpha or interleukin (IL)-1 beta. At pharmacologically relevant concentrations, triclosan and CPC inhibited ILAP-induced PGE(2) production to a greater extent than triclosan alone (P = 0.02). Moreover, enhanced COX-2 mRNA repression was observed with triclosan and CPC in comparison to triclosan alone in IL-1 beta and TNF-alpha stimulated cells. No effect on COX-I gene expression was observed. Further analysis of cell signaling mechanisms of triclosan and CPC indicates that nuclear factor-kappa B (NF-kappa B) and not p38 mitogen-activated protein kinase (MAPK) signaling may be impaired in the presence of triclosan and CPC.Conclusion: This study indicates that triclosan and CPC are more effective at inhibiting PGE(2) at the level of COX-2 gene regulation, and this combination may offer a potentially better anti -inflammatory agent in the treatment of inflammatory lesions in the oral cavity.

Quantitation of malondialdehyde in gingival crevicular fluid by a high-performance liquid chromatography-based method

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Clinical and histomorphometric characteristics of three different families with hereditary gingival fibromatosis

Objective. To examine the histomorphologic and histomorphometric features of tissue from 3 unrelated families with hereditary gingival fibromatosis (HGF).Study design. Twelve affected individuals from 3 HGF families and 3 control subjects were evaluated. Gingival samples were fixed in formalin and embedded in paraffin for hematoxylin and eosin stain to count the number of fibroblast and inflammatory cells. Sirius red staining was performed to quantitate the amount of collagen present.Results. Histomorphologic analysis of HGF showed extension of epithelial rete ridges into the underlying lamina propria and the presence of collagen bundles in the connective tissue. Analysis of the mean area fraction of collagen showed that there were significant increases in the collagen fraction for all HGF types compared with control subjects (P < .05). There were significant increases in the number of fibroblasts for HGFa and HGFb compared with control subjects (P < .05). The number of fibroblasts for HGFc were similar to that for control subjects.Conclusions. The collagen fraction was significantly greater in all HGF types compared with controls. The number of fibroblasts was significantly increased in 2 of the 3 HGF types compared with controls. These data indicate that different mechanisms may be responsible for tissue enlargement in different forms of HGF.

New attachment achieved by guided tissue regeneration using a bioresorbable polylactic acid membrane in dogs

Created periodontal defects in dogs were randomly assigned for experimental (Guidor bioresorbable membranes) or control (conventional therapy) treatment the results showed that the new connective issue attachment was significantly greater in test sites than in controls. This new attachment averaged 2.79 +/- 0.74 mm and 1.47 +/- 0.20 mm at test and control sites, respectively (P < 0.05). Epithelial downgrowth was also reduced in the test sites (P < 0.05). No differences in bone response were found. The bioresorbable barrier was effective in blocking gingival epithelial downgrowth and connective tissue proliferation, promoting new attachment according to the principles of guided tissue regeneration.

Gingival metastasis from salivary duct carcinoma of the parotid gland

Background: This article reports a rare case of metastasis of salivary duct carcinoma of the parotid gland to the gingiva and reviews the occurrence of metastatic processes to the oral mucosa.Methods: A 67-year-old white male presented with a chief complaint of a painless nodular tissue growth on the gingiva with reportedly 5 months of evolution. The intraoral examination revealed a reddish, superflcially ulcerated nodular lesion (similar to 2 cm in diameter) on the right mandibular buccal attached gingiva, and the clinical aspect was that of a benign reactive lesion. The patient had undergone a parotidectomy for removal of a salivary duct carcinoma of the parotid gland almost 1 year before. A biopsy of the gingival lesion was performed, and the biopsied tissue was forwarded for histopathologic examination.Results: The analysis of the histopathologic sections of the gingival lesion revealed histomorphologic characteristics very similar to those of the primary parotid gland tumor. The definitive diagnosis was gingival metastasis from a salivary duct carcinoma of the parotid gland. The patient died of complications of a pulmonary metastasis I month after the diagnosis of the oral metastatic lesion.Conclusions: Gingival lesions that mimic reactive and hyperplastic lesions may be metastases from malignant neoplasias of diverse origins. An accurate and timely diagnosis is crucial to establish proper and immediate treatment of the metastatic tumor and possibly identify an occult primary malignant neoplasia.

Interference of the blood clot on granulation tissue formation after tooth extraction. Histomorphological study in rats.

The interference of a blood clot in the first postoperative hours of dental extraction wounds was studied in rats. Sixty male albino rats were divided into two groups: Group I, immediately after extraction of right maxillary incisor the gingival mucosa was approximated and sutured; Group II, after 6 to 8 minutes postoperatively the blood clot was removed with saline irrigation and absorbent paper cones. The mucosa was then approximated and sutured. Six animals in each group were sacrificed after 12 hours, 1, 4, 7 and 10 days. There was a profound delay in healing in Group II since, although a new blood clot was later formed, it was not organized. The quality and the constitution, maintenance and retraction of the clot are the regulating factors in connective tissue formation during alveolar healing.

Morphometric evaluation of gingival overgrowth and regression caused by cyclosporin in rats

Cyclosporin A is a selective immunosuppressant, used in organ transplants to prevent graft rejection. Cyclosporin A can cause various side effects including gingival overgrowth. The aim of this work was to evaluate gingival overgrowth of rats treated daily with 10 mg/kg body weight of Cyclosporin A for 60 days, as well as the regression after the interruption of treatment. All rats treated with Cyclosporin A developed gingival overgrowth, with increased thickness of the epithelium, height and width of the connective tissue. The density of fibroblasts and collagen fibers also increased. Five to 90 days after the interruption of treatment with Cyclosporin A, there was a progressive reduction of the gingival volume and of collagen fibers and fibroblast densities. The reduction was more pronounced in the initial periods and after 90 days did not return to the normal values.

Combined effects of cyclosporin and nifedipine on gingival overgrowth in rats is not age dependent

Background: Cyclosporin A and nifedipine cause gingival overgrowth in rat, and the combined use of these drugs increases the overgrowth severity. Objective: The purpose of this study was to compare gingival overgrowth of rats of differents ages treated with cyclosporin A and nifedipine alone or given concurrently. Materials and methods: Rats 15, 30, 60 and 90 d old were treated with 10 mg/kg body weight of cyclosporin A and/or 50 mg/kg body weight of nifedipine in the chow. Results: Young rats showed evident gingival overgrowth with nifedipine, cyclosporin A, and cyclosporin A and nifedipine given concurrently. Adult rats did not show significant gingival alterations when treated with cyclosporin A and nifedipine alone. Nevertheless evident gingival overgrowth with alterations of the epithelium and connective tissue were observed when treated simultaneously with cyclosporin A and nifedipine. Conclusion: These results suggest that the combined effects of cyclosporin A and nifedipine on gingival overgrowth in rat is not age dependent.

Naso-orbicular tissue necrosis by Streptococcus parasanguis in a patient with Fanconi anemia: Clinical and laboratory aspects

Fanconi anemia (FA) is a rare autosomal recessive disorder, characterized by pancytopenia and progressive hypoplasia of the bone marrow. A 23-year-old woman with FA showed severe pancytopenia and developed an abscess on the infraorbicular region on the right side of the face that progressed to phlegmon and caused tissue necrosis of the nostrils, nasal septum, nasal fossa, and posterior orbital region. Laboratory examination showed Streptococcus parasanguis as the etiologic agent of the phlegmon. Supportive treatment was recommended due to donor incompatibility for bone marrow transplant. The intraoral examination showed spontaneous gingival bleeding, edema of the interdental papillae, hematomas on the superior and inferior lips, bacterial and fungal infections, and adequate oral hygiene. The patient was treated with the administration of an antibiotic (imipenem), an antifungal (amphotericin B), and mouth washing with antiseptic solutions. Periodontal prophylaxis and orientation to and control of oral hygiene and diet were also used during the remission period. For functional and esthetic rehabilitation of the alar regions and nasal dorsum, an acrylic resin nasal prosthesis was made, supported by a spectacle frame.

Periodontal plastic surgery associated with treatment for the removal of gingival overgrowth

Background: Excisional biopsies of gingival overgrowths, performed with safety margins, frequently result in mucogingival defects. These defects may produce esthetic problems and increase the chances of dentin hyperesthesia and its possibility of hindering oral hygiene. Methods: Two clinical cases are reported in which gingival overgrowths were removed by excisional biopsy, resulting in unsightly defects. The first clinical case presents an invasive approach for the treatment of a recurrent pyogenic granuloma in the anterior maxilla, and the second depicts a complete removal of a peripheral odontogenic fibroma in the posterior maxilla. In both situations, the soft-tissue defects were repaired by periodontal plastic surgery, including a laterally positioned flap and a coronally positioned flap, respectively. Results: Periodontal plastic surgery successfully restored the defects that resulted from biopsies, and no recurrence has been noticed in the 5-year postoperative follow-up period. Conclusions: The combination of biopsy and periodontal plastic surgery in a one-step procedure seems to be suitable to remove gingival overgrowths in most areas of the mouth, regardless of esthetic significance. Such procedures seem to restore gingival health, encourage healing, and create both esthetics and function in the excised area.