Abstract
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.
Highlights
Periodontitis is a chronic inflammatory bacterial infection, where the oral flora organizes a biofilm subgingivally, which constitutes a continuous challenge to the host immune system that responds by continuous inflammatory cytokine shower that affects the body homeostasis; with time, deregulated immune response eventually results, and a hyperresponsive immune reaction causes the destruction of the tooth-supporting apparatus, leading to tooth loss.Stem Cells InternationalPeriodontitis is considered an irreversible degenerative disease of the odontogenic supporting tissue; this throws light on the immune-mediated nature of periodontal disease [1].A historical debate did exist about the periosteum’s role in bone growth, repair, and regeneration
Seeded gingival mesenchymal stem cells (GMSC) in P10 dish showed typical distinctive fibroblast-like colonies of 50 to cells/colony after on average 14 days of culturing in vitro; all experiments were done in triplicate (n = 3) for each group; no significant difference was noted between the heterogeneous GMSC group and CD146-positive homogeneous GMSCs in shape, form, or number of cells in colonies (p > 0:05; Mann–Whitney U test); the only difference noted was that the homogeneous CD146-positive group reached 100 cell colonies 1 day earlier on average compared to the heterogeneous group (Figure 4(a))
In 2018, Al Bahrawy et al [4] proved the concept that GMSCs can migrate through microperforated membranes of Guided tissue regeneration (GTR) in the presence of a suitable chemoattractant, while in the absence of the chemoattractant, the membranes were totally occlusive for cell migration; this can be a basis for a new generation of the GTR technique, where a selective barrier membrane was employed, which was occlusive for undesired cells in the gingival tissue, namely, the epithelium and connective tissue cells, and allowed the homing of GMSCs from the gingival tissue to the periodontal wound by utilizing a suitable chemoattractant in the wound area [4]
Summary
Periodontitis is a chronic inflammatory bacterial infection, where the oral flora organizes a biofilm subgingivally, which constitutes a continuous challenge to the host immune system that responds by continuous inflammatory cytokine shower that affects the body homeostasis; with time, deregulated immune response eventually results, and a hyperresponsive immune reaction causes the destruction of the tooth-supporting apparatus, leading to tooth loss.Stem Cells InternationalPeriodontitis is considered an irreversible degenerative disease of the odontogenic supporting tissue; this throws light on the immune-mediated nature of periodontal disease [1].A historical debate did exist about the periosteum’s role in bone growth, repair, and regeneration. 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. 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. 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. 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
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