Background. In the field of periodontal guided tissue regeneration, microperforated membranes have recently proved to be very promising periodontal regenerative tissue engineering tools. Regenerative periodontal approaches, employing gingival mesenchymal stem/progenitor cells in combination with these novel membranes, would occur mostly in inflamed microenvironmental conditions intraorally. This in turn entails the investigation into how inflammation would affect the proliferation as well as the migration dynamics of gingival mesenchymal stem/progenitor cells. Materials and Methods. Clones of human gingival mesenchymal stem/progenitor cells (GMSCs) from inflamed gingival tissues were characterized for stem/progenitor cells’ characteristics and compared to clones of healthy human GMSCs (n=3), to be subsequently seeded on perforated collagen-coated poly-tetra-floro-ethylene (PTFE) membranes with a pore size 0.4 and 3 microns and polycarbonic acid membranes of 8 microns pore size in Transwell systems. The population doubling time and the MTT test of both populations were determined. Fetal bovine serum (FBS) was used as a chemoattractant in the culturing systems, and both groups were compared to their negative controls without FBS. Following 24 hours of incubation period, migrating cells were determined on the undersurface of microperforated membranes and the membrane-seeded cells were examined by scanning electron microscopy. Results. GMSCs demonstrated all predefined stem/progenitor cell characteristics. GMSCs from inflamed gingival tissues showed significantly shorter population doubling times. GMSCs of inflamed and healthy tissues did not show significant differences in their migration abilities towards the chemoattractant, with no cellular migration observed in the absence of FBS. GMSCs from healthy gingival tissue migrated significantly better through larger micropores (8 microns). Scanning electron microscopic images proved the migratory activity of the cells through the membrane pores. Conclusions. Inflammation appears to boost the proliferative abilities of GMSCs. In terms of migration through membrane pores, GMSCs from healthy as well as inflamed gingival tissues do not demonstrate a difference in their migration abilities through smaller pore sizes, whereas GMSCs from healthy gingival tissues appear to migrate significantly better through larger micropores.
Background. Guided tissue regeneration (GTR) is a powerful modality for periodontal regeneration, but it blocks the periosteum and gingival stem cells (GMSCs), from supporting periodontal wound by the nutrients, growth factors, and regenerative cells. The microperforated membrane considered a rewarding solution for this major drawback; GMSCs can migrate through a GTR microperforated membrane toward a chemoattractant, with the blocking of other unfavorable epithelial cells and fibroblasts. In the absence of a sole marker for MSC, a homogeneous population of GMSC is difficult to isolate; using CD146 as confirmatory markers for MSC identification, testing the behaviour of such homogeneous population in migration dynamics was the question to answer in this study. Materials and Methods. GMSCs from healthy crown lengthening tissue was isolated ( n = 3 ), its stem cell nature was confirmed, CD146 and CD271 markers were confirmatory markers to confirm homogenous stem cell population, and magnetic sorting was used to isolate GMSC with CD146 markers. A homogenous CD146 population was compared to heterogeneous GMSCs of origin; the population doubling time and MTT test of the two populations were compared. Migration dynamics were examined in a transwell migration chamber through 8 μm perforated polycarbonic acid membrane, and 0.4 μm and 3 μm perforated collagen-coated polytetrafluoroethylene membrane (PTFE) and 10% fetal bovine serum (FBS) were the chemoattractants used in the lower compartment to induce cell migration, were incubated in a humidified environment for 24 hours, then migrated the cell in the lower compartment examined by a light and electron microscope. Results. GMSCs fulfilled all the minimal criteria of stem cells and showed low signal 10% for CD146 on average and extremely low signal 2% for CD271 on average. Magnetic sorting optimized the signal of CD146 marker to 55%. GMSC CD146 population showed nonstatistically significant shorter population doubling time. CD146 homogeneous population migrated cell numbers were statistically significant compared to the heterogeneous population, through 0.4 μm and 3 μm perforated collagen membrane and 8 μm perforated polycarbonate membrane. Scanning electron microscopy proved the migration of the cells. Conclusions. A subset of the isolated GMSC showed a CD146 marker, which is considered a dependable confirmatory marker for the stem cells. In terms of GMSC migration through the microperforated membrane, a homogeneous CD146 population migrates more statistically significant than a heterogeneous GMSC population.
Objectives: Periodontitis is characterized by an increased alveolar bone turnover with dominance of bone resorption over bone formation leading to alveolar bone loss and loss of attachment. Osteocalcin is the major noncollagenous calcium-binding single chain protein of bone and dentin matrix and it is a bone marker whose concentration in circulation has been used as a reflector of bone turnover.The present study was conducted to evaluate osteocalcin levels in healthy and diseased periodontium and to correlate the changes in its levels with the changes in the clinical and radiographic parameters before and after treatment. Materials & Methods: Double blind clinical randomized trial, 40 patients divided into 3 groups; 15 patients diagnosed as periodontitis stage III grade B, 15 patients diagnosed as periodontitis stage III grade C, 10 healthy cases as control. Clinical parameters namely plaque index (PI), Gingival index (GI), Pocket depth (PD), Clinical attachment loss (CAL), radiographic examination, beside gingival crevicular fluid (GCF) samples collection for Osteocalcin level, tested at 3 time points 1 before and 2 after periodontal treatment, which included phase I therapy, and respective periodontal surgery accompanied by systemic antibiotics protocol. Results: revealed a positive correlation in the changes of osteocalcin levels in relation to probing depth, clinical attachment level, plaque index and gingival index and a negative correlation with the changes in bone density although these correlations were insignificant. Conclusions: Osteocalcin in gingival crevicular fluid as marker for diagnosis , prognosis, and follow up of the periodontitis infection activity, As a second conclusion cumulative non surgical, surgical and systemic antibiotic medication have high ability in the control of periodontal infection which could be measured clinically and radiographically. Clinical relevance: Osteocalcin would be suitable for diagnosis and follow up of the disease, but not suitable for the prognosis of periodontal disease infection.
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