Comparison of the Migration Potential through Microperforated Membranes of CD146+ GMSC Population versus Heterogeneous GMSC Population

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.

Unveiling heterogeneity in MSCs: exploring marker-based strategies for defining MSC subpopulations

“…Moreover, functional diversity is evident among MSC subpopulations. For instance, CD146 + MSCs exhibit strong chemotactic attraction and immune suppression, while CD362 + MSCs display enhanced colony formation and immune suppression, validated in COVID-19 clinical trials [27,[94][95][96][97][98][99][105][106][107][108]. Additionally, CD106 + MSCs from placenta and umbilical cord tissues demonstrate potent immunomodulation and pro-angiogenic activities [28,112,114,115].…”

“…And it has been proposed that CD146 is an MSC marker of multipotency [90][91][92][93]. CD146 + MSCs have a much stronger chemotactic attraction [94][95][96][97], and enhanced immune suppression activities in vitro and in vivo [27,[97][98][99]. Higher levels of CD146 expression correlate with a faster proliferation rate, enhanced multilineage differentiation potentials, stronger stemness characteristics, and less senescent phenotypes [98][99][100].…”

“…Within the CD271 + MSCs, both CD146 + and CD146 − share similar genotypes and phenotypes [ 101 ]. Furthermore, other studies have also demonstrated that CD146 + and CD146 − share similar levels of MSC marker expression, colony-forming, proliferation and differentiation capabilities [ 94 , 96 , 102 , 103 ]. And the CD146 − MSCs even proliferate significantly faster than the CD146 + population [ 103 ].…”

Dental Pulp Fibroblast: A Star Cell

“…Although it has been stated that fibroblasts and MSCs express the same surface markers 36,37 , some markers and genes have been found to allow their correct differentiation. CD 106 39,[57][58][59] , CD 146 57,[59][60][61][62][63] , ITGA11 57,64 , SSEA-4 57,65 , GD-2 57 , Stro-1 66 , CD 271 66 , CD 166 57 , and IGF-2 57 are specific to MSCs, and CD 9 57,58 , CD 10 52,57,67 , and CD 26 52,57 to fibroblasts. Therefore, although some surface markers have been used to identify fibroblasts in distinct tissues, when used alone or in combination, they do not reliably identify fibroblast subpopulations in all tissues 68 .…”

Managing the Heterogeneity of Mesenchymal Stem Cells for Cartilage Regenerative Therapy: A Review

“…In a three-dimensional scaffold, CD146 + MSCs demonstrate spontaneous chondrogenesis [ 119 ], while intra-articular implantation of CD146 + MSCs has led to chondroprotective effects in inflammatory environments [ 120 ]. A magnetically-sorted CD146 + MSC population has also shown improved cell migration compared to a heterogenous cell population [ 121 ]. Compared to unsorted AD-MSCs, CD146 + MSCs promote long-term cartilage repair in vivo and produce less inflammation [ 122 ].…”