COMPARISON OF ADHESION AND PROLIFERATION OF MESENCHYMAL STEM CELLS ON AMNIOTIC MEMBRANE AND COLLAGEN PORCINE-DERIVED SCAFFOLD

Abstract

Background Cellular scaffolds are 3D structures that support cell adhesion and proliferation, promoting an environment favorable to cell culture. The amniotic membrane (AM) is a scaffold that has suitable mechanical properties, low immune rejection and low cost of obtaining and processing. The mesenchymal stem cells (MSC) are known for their immunomodulatory capacity, maintenance and renewal of cells and tissues, and its association with AM may have a great potential in tissue engineering. The aim of this study was to compare the capacity of adhesion and proliferation of dental pulp-derived MSC (DPSC) after cultivation on decellularized AM with the commercial porcine collagen membrane (CPCM) already used clinically. Methods The AM was collected, isolated and decellularized. The DPSC were obtained from permanent tooth pulp by mechanical dissociation followed by enzymatic digestion and expanded in vitro. The DPSC were genetically modified to express the luciferase enzyme. These transfected cells were plated at different concentrations: 10,000, 30,000, 50,000 and 100,000 per cm2 on AM and CPCM, and used in bioluminescence analysis to assess adherence and proliferation capacity on both scaffolds over 24 hours, 7 days, 14 days, 21 days and 28 days. Results The concentration of 100,000 cells per cm2 showed better adhesion in all tested samples. There was a significant difference (p\u202f=\u202f0.004) 24 hours after cell culture, with CPCM presenting five times more cells than AM. The difference was also observed on the 7th day (p\u202f=\u202f0.011), with CPCM presenting eight times more cells than AM. On the 14th and 21st days (p\u202f=\u202f0.006 and p\u202f=\u202f0.004, respectively), CPCM showed a considerable decrease in cell proliferation. At the same time, AM presented an increase in cell proliferation, with two times more cells than CPCM. Finally, there was a significant difference on the 28th day (p\u202f=\u202f0.010), where AM presented three times more cells than CPCM. Conclusion The results showed AM is a viable source to be used as a scaffold for cell adhesion and proliferation. When extrapolated to an in vivo environment, it is possible that CPCM s showed more difficulty in maintaining cell adhesion for more than a week which could affect clinical outcomes. In this scenario, AM it seems to be a more stable scaffold, enabling cell adhesion for up to three weeks. The presented technology proved to be accessible, reproducible, and can be an alternative to commercial membranes.