Root coverage with cultured gingival dermal substitute composed of gingival fibroblasts and matrix: a case series.

Cultured gingival dermal substitute (CGDS), composed of gingival fibroblasts and matrix and fabricated using tissue-engineering techniques, has been used for root coverage procedures. Fourteen sites from four patients with > or = 2 mm of Miller Class I or II facial gingival tissue recession were treated. The autologous CGDS sheet, prepared prior to surgical treatment, was grafted over the teeth with gingival recession and then covered with a coronally positioned flap. Vertical and horizontal recession was measured at baseline (prior to the surgical procedure) and 13 to 40 weeks (average: 30.7 +/- 9.6 weeks) after surgery. The average vertical and horizontal root coverage after surgery was 79.1% +/- 25.7% and 75.2% +/- 31.4%, respectively. Moreover, there was a significant increase of keratinized and attached gingival tissue at the final clinical evaluation compared with preoperative measurements (P < .05). These results demonstrate CGDS as a promising grafting material for use with root coverage procedures in periodontal therapy.

Immunohistochemical localization of cytokeratin 19, involucrin and proliferating cell nuclear antigen (PCNA) in cultured human gingival epithelial sheets.

It has been suggested that human cultured gingival epithelial sheets may serve as a possible grafting material. The purpose of this study was to examine the biological characteristics of human cultured gingival epithelial sheets by epithelial differentiation and proliferation markers. Immunohistochemical localization of cytokeratin 19, involucrin and proliferating cell nuclear antigen (PCNA) were examined in human cultured gingival epithelial sheets samples from twenty patients. Cytokeratin 19-immunopositive cells were scattered mainly in the suprabasal layer. Immunoreactivity for involucrin was observed in all layers except for the basal layer. The majority of proliferating cell nuclear antigen-immunopositive cells was found in the basal layer. These results suggested that the cultured human gingival epithelial sheets were biologically active and in proliferative condition, which implies that this biological product may be a potential grafting material.

Immunohistochemical localization of cytokeratin 19, involucrin and proliferating cell nuclear antigen (PCNA) in cultured human gingival epithelial sheets.

It has been suggested that human cultured gingival epithelial sheets may serve as a possible grafting material. The purpose of this study was to examine the biological characteristics of human cultured gingival epithelial sheets by epithelial differentiation and proliferation markers. Immunohistochemical localization of cytokeratin 19, involucrin and proliferating cell nuclear antigen (PCNA) were examined in human cultured gingival epithelial sheets samples from twenty patients. Cytokeratin 19-immunopositive cells were scattered mainly in the suprabasal layer. Immunoreactivity for involucrin was observed in all layers except for the basal layer. The majority of proliferating cell nuclear antigen-immunopositive cells was found in the basal layer. These results suggested that the cultured human gingival epithelial sheets were biologically active and in proliferative condition, which implies that this biological product may be a potential grafting material.

Vascular endothelial growth factor and transforming growth factor-alpha and -beta1 are released from human cultured gingival epithelial sheets.

BACKGROUND: It has been demonstrated that human cultured epithelial sheets prepared by tissue engineering techniques provide useful graft material for wound healing and tissue regeneration. However, limited information is available with regard to biological effects such as release of growth factors from human cultured gingival epithelial sheets (HCGES). The purpose of this study was to measure the levels of growth factors released from HCGES into culture medium. METHODS: Twenty patients aged 44 to 71 years with generalized chronic periodontitis were recruited, and their gingival tissues obtained during periodontal flap surgery. The levels of vascular endothelial growth factor (VEGF), transforming growth factor-alpha and -beta1 (TGF-alpha and -beta1), and epidermal growth factor (EGF) released into the culture medium were determined using enzyme-linked immunosorbent assay at the just confluent (T1) and the adequate stratification (T2) culture stages. The medium without cells was collected as a control (T0). Statistical tests were performed by analysis of variance and Sheffé multiple range test among T0, T1, and T2. RESULTS: Significantly higher levels of VEGF and TGF-alpha were observed at T1 and T2 compared to T0 (P<0.001). In addition, there was a significant difference in the TGF-alpha levels between T2 and T1 (P<0.001). TGF-beta1 at T1 was significantly higher in comparison to T0 (P <0.01). EGF had been released only in a small amount at T2. CONCLUSION: This study indicates that meaningful amounts of VEGF and TGF-alpha and -beta1 are released from HCGES, which suggests potential for promoting wound healing and tissue regeneration after grafting.

Platelet-rich plasma contains high levels of platelet-derived growth factor and transforming growth factor-beta and modulates the proliferation of periodontally related cells in vitro.

BACKGROUND: Platelet-rich plasma (PRP) is a fraction of plasma, in which platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta) are thought to be concentrated. It is plausible that topically-applied PRP up-regulates cellular activity and subsequently promotes periodontal regeneration in vivo. However, the concentrations of these growth factors in PRP have not been specifically determined and the biological effects of PRP at the cellular and molecular levels have not been determined. METHODS: PRP obtained from 20 healthy subjects was prepared from plasma by centrifugation. These PRP preparations were immediately subjected to an evaluation for PDGF-AB and TGF-beta1 using enzyme-linked immunosorbent assay (ELISA) kits. The biological effects of the PRP preparations were evaluated on osteoblastic, epithelial, fibroblastic, and periodontal ligament cells. Cellular mitogenic activity was evaluated by counting cell numbers or evaluating 5-bromodeoxyuridine (BrdU) incorporation. Expression of alkaline phosphatase (ALP) was immunocytochemically evaluated. RESULTS: In the PRP preparations, platelets were concentrated up to 70.9 x 10(4) cells/microl (283.4% of the unconcentrated plasma). The levels of PDGF-AB and TGF-beta1 were also concentrated up to 182.0 ng/ml (440.6%) and 140.9 ng/ml (346.6%), respectively. Scatter plots revealed significant correlations between platelet counts and levels of these growth factors. PRP stimulated osteoblastic DNA synthesis and cell division (138% of control), with simultaneous down-regulation of ALP, but suppressed epithelial cell division (80% of control). PRP also stimulated DNA synthesis in gingival fibroblasts and periodontal ligament cells. CONCLUSIONS: These data demonstrated that both PDGF-AB and TGF-beta1 were highly concentrated in the PRP preparations. It is suggested PRP modulates cell proliferation in a cell type-specific manner similar to what has been observed with TGF-beta1. Since synchronized behavior of related cell types is thought to be required for successful periodontal regeneration, it is further suggested these cell type-specific actions may be beneficial for periodontal regenerative therapy.

Treatment of chronic desquamative gingivitis using tissue-engineered human cultured gingival epithelial sheets: a case report.

Human cultured gingival epithelial sheets were used as an autologous grafting material for regenerating gingival tissue in the maxillary left and mandibular right quadrants of a patient with chronic desquamative gingivitis. Six months post-surgery in both treated areas, there were gains in keratinized gingiva and no signs of gingival inflammation compared to presurgery. In the maxillary left quadrant, preoperative histopathologic findings revealed the epithelium was separated from the connective tissue and inflammatory cells were extensive. After grafting with the gingival epithelial sheets, inflammatory cells were decreased and separation between epithelium and connective tissue was not observed. The human cultured gingival epithelial sheets fabricated using tissue engineering technology showed significant promise for gingival augmentation in periodontal therapy.

Vascular Endothelial Growth Factor and Transforming Growth Factor-α and -ß1 Are Released From Human Cultured Gingival Epithelial Sheets.

BACKGROUND: It has been demonstrated that human cultured epithelial sheets prepared by tissue engineering techniques provide useful graft material for wound healing and tissue regeneration. However, limited information is available with regard to biological effects such as release of growth factors from human cultured gingival epithelial sheets (HCGES). The purpose of this study was to measure the levels of growth factors released from HCGES into culture medium Methods: Twenty patients aged 44 to 71 years with generalized chronic periodontitis were recruited, and their gingival tissues obtained during periodontal flap surgery. The levels of vascular endothelial growth factor (VEGF), transforming growth factor-α and -ßl (TGF-α and -ßl), and epidermal growth factor (EGF) released into the culture medium were determined using enzyme-linked immunosorbent assay at the just confluent (Tl) and the adequate stratification (T2) culture stages. The medium without cells was collected as a control (TO) statistical tests were performed by analysis of variance and sheffé multiple range test among TO, T1, and T2. RESULTS: Significantly higher levels of VEGF and TGF-α were observed at T1 and T2 compared to To (P<0.001). In addition, there was a significant difference in the TGF-α levels between T2 and T1 (P<0.001). TGF-ßl at T1 was significantly higher in comparison to T0 (P<0.0l). EGF had been released only in a small amount at T2. CONCLUSION: This study indicates that meaningful amounts of VEGF and TGF-α and -ßl are released from HCGES, which suggests potential for promoting wound healing and tissue regeneration after grafting. J Periodontol 2002;73:748-753.

Osteocalcin, deoxypyridinoline and interleukin-1beta in peri-implant crevicular fluid of patients with peri-implantitis.

The aim of the present study was to analyze the levels of osteocalcin, deoxypyridinoline (Dpd) and interleukin-1beta as markers of bone metabolism in peri-implant crevicular fluid (PICF) from peri-implantitis patients. PICF was sampled from a total of 34 endosseous titanium implants from 16 patients; nine females (mean age 52.8, range 40-62 years) and seven males (mean age 56.0, range 36-66 years). The implants had been in place for a period of 9-112 months (mean; 35.8 months) since the loading. These sites were categorized as six peri-implantitis, eight peri-implant mucositis and 20 healthy implant. PICF volume from peri-implantitis sites was significantly higher than mucositis and healthy implant sites (P < 0.01). Osteocalcin levels in PICF from mucositis sites were significantly higher than healthy implants (P < 0.05), whereas peri-implantitis sites were not significantly different from either mucositis or healthy implant sites. Dpd could not be detected in any of the samples examined. IL-1beta levels in PICF from peri-implantitis sites were significantly higher than levels from peri-implant mucositis (P < 0.05) and healthy implant sites (P < 0.01). In conclusion, osteocalcin in PICF may reflect increased local bone turnover around implants. Further, IL-1beta should be a useful marker for peri-implant inflammation.