Regenerative potential of cultured gingival fibroblasts in treatment of periodontal intrabony defects (randomized clinical and biochemical trial) Mahetab Abdal‐Wahab, Khaled A. Abdel Ghaffar, Ola M. Ezzatt, Ahmed Abdel Aziz Hassan, Mervat Mohamed S. El Ansary, et al. Journal of Periodontal Research, 2020 BACKGROUND Defective cellular elements constitute an important challenge to achieve predictable periodontal regeneration. In an attempt to improve the cellularity of periodontal defects, gingival fibroblasts were implanted without their associated extracellular elements in periodontal defects to expose them to periodontal tissue mediators. In order to investigate the regenerative potential of gingival fibroblasts translocated into periodontal defects, the present study was designed to clinically and biochemically investigate the use of gingival fibroblasts (GF) and their associated mesenchymal stem cells (GMSC) in the treatment of intrabony periodontal defects. METHODS A total of 20 subjects were randomly divided into two groups (n = 20). Group I: ten patients were included with ten intrabony periodontal defects that received β-calcium triphosphate (β-TCP) followed by collagen membrane defect coverage, while group II: (10 patients) ten periodontal defects received cultured gingival fibroblasts (GF) on the β-TCP scaffold and covered by a collagen membrane. The clinical evaluation was carried out at the beginning and at 6 months. Gingival crevicular fluid (GCF) samples were collected directly from the test sites for the quantitative measurement of PDGF-BB and BMP-2 using the ELISA kit at 1, 7, 14, and 21 days after surgery. RESULTS Group II reported a significantly greater reduction in vertical pocket depth (VPD) and CAL gain compared with group I after 6 months. Radiographic bone gain was statistically higher in group II compared with group I. A significantly higher concentration of PDGF-BB was observed in group II on days 1, 3, and 7 compared with group I. CONCLUSIONS Translocation of gingival fibroblasts from gingival tissue to periodontal defects could be a promising option that increases cellular elements with regeneration potential. The concept of total isolation of gingival fibroblasts using occlusive membranes must be re-evaluated.
Guided tissue regeneration of intrabony defects with perforated barrier membranes, simvastatin, and EDTA root surface modification: A clinical and biochemical study Dalia Rasheed Issa, Khaled A. Abdel‐Ghaffar, Mohamed A. Al‐Shahat, Ahmed Abdel Aziz Hassan, Vincent J. Iacono, et al. Journal of Periodontal Research, 2020 BACKGROUND Perforated barrier membranes (PBM) were suggested to enhance periodontal regeneration by allowing positive charity of wanted elements from the gingival tissue side. The present study was designed to evaluate clinically and biochemically the use of PBM combined with simvastatin (SMV) gel with and without an associated EDTA gel root surface etching as a suggested option that could improve SMV availability and clinical outcomes of PBM. METHODS Forty patients having moderate-to-severe chronic periodontitis with 40 intrabony defects were randomly divided into four treatment groups (10 sites each). Patients in group 1 received 1.2% SMV gel and covering the defect with occlusive membrane (OM). Patients in group 2 received 1.2% SMV gel and covering the defect with PBM. Group 3 received 24% EDTA root surface etching, 1.2% SMV gel, and defect coverage with OM (eOM). Patients in group 4 were treated as in group 3 but the defect was covered with PBM (ePBM). Clinical parameters were recorded at baseline before surgical procedures and were reassessed at 6 and 9 months after therapy. The mean concentration of SMV in gingival crevicular fluid (GCF) was estimated by reverse-phase high-performance liquid chromatography at days 1, 7, 14, 21, and 30. RESULTS At 6- and 9-month observation periods, groups 3 and 4 showed a statistically significant improvement in PD reduction and CAL gain compared with groups 1 and 2. Group 4 showed a statistically significant more defect fill compared with groups 1, 2, and 3 (P ≤ .05). Group 2 showed statistically significant higher defect fill compared with group 1 and group 3 (P < .05). Bone density was significantly increased with no significant difference between the four groups at 6- and 9-month observation periods. SMV-GCF concentration in group 4 showed the highest mean concentration with no significant difference than that of group 3. CONCLUSION The use of perforated barrier membranes in association with SMV enhances the clinical hard tissue parameters compared with occlusive ones in treating intrabony periodontal defects. Moreover, EDTA root surface treatment could enhance SMV availability in the defect area.
In vitro evaluation of the human gingival fibroblast/gingival mesenchymal stem cell dynamics through perforated guided tissue membranes: cell migration, proliferation and membrane stiffness assay A. Y. Gamal, N. N. Al-Berry, A. A. Hassan, L. A. Rashed, V. J. Iacono Journal of Periodontal Research, 2017 BACKGROUND Migration of gingival fibroblasts/gingival mesenchymal stem cells through macro-perforated barrier membranes may allow them to participate positively in periodontal regeneration. The optimal guided tissue membrane perforation diameter that could favor maximum cell migration into the defect area and at the same time act as an occlusive barrier for gingival epithelium and its associated gingival extracellular matrix component is not yet identified. MATERIAL AND METHODS Cultured human gingival fibroblasts/gingival mesenchymal stem cells were placed in the upper chambers of 12-well collagen-coated polytetrafluoroethylene transwells, which were manually perforated with 0.2, 0.4 and 0.7 mm sized pores. The lower chambers of the transwells received blood clot as an attraction medium. The number of cells that have migrated to the lower chambers was calculated. Proliferation of these cells was evaluated using MTT assay. Scanning electron microscopy images were obtained for the lower surfaces of the transwell membranes. Perforated bovine collagen membranes (Tutopatch® ) were subjected to mechanical testing to determine the tensile strength and modulus of elasticity. RESULTS Group 3 (0.7 mm) showed significantly higher values for cell migration and proliferation. All groups showed a small degree of extracellular matrix migration through membrane perforations. Scanning electron microscopy evaluation revealed variable numbers of cells in fibrin matrices located mainly around the pore edges. There were non-significant differences between groups regarding mechanical properties. CONCLUSIONS The present study demonstrated that macro-membrane perforations of 0.2, 0.4 and 0.7 mm are suitable pore diameters that could maintain membrane stiffness and allow for cellular migration. However, these membrane perforation diameters did not allow for total gingival connective tissue isolation.