Cryopreservation of an artificial human oral mucosa stroma. A viability

Abstract

<font face="AdvGulliv-R" size="1"><font face="AdvGulliv-R" size="1"> <p align="LEFT">The aim of this study was to evaluate the viability and biomechanical properties of artificial human oral</p> <p align="LEFT">mucosa stroma (HOMS) subjected to cryopreservation with different cryoprotectant solutions.</p> <p align="LEFT">Artificial HOMS based on a fibrin?agarose matrix with human gingival fibroblasts cultured 7 days</p> <p align="LEFT">in vitro were cryopreserved with three cryoprotectant solutions: (A) TC-199 Medium, DMSO 15%, albumin;</p> <p align="LEFT">(B) DMEM, FCS, DMSO 10%; (C) QC Medium, glycerol. As controls, artificial HOMS not subjected to</p> <p align="LEFT">cryopreservation (CF) and HOMS cryopreserved without cryoprotectant solution (CS) were used. Histological</p> <p align="LEFT">analysis by light microscopy showed that solutions A and B preserved a pattern of porosity similar</p> <p align="LEFT">to values in CF. Based on the number of intact cells in the fibrin?agarose matrix, substitutes preserved</p> <p align="LEFT">with solution B showed the best results. Cell proliferation detected with PCNA immunochemical methods</p> <p align="LEFT">showed that the cell proliferation index was highest in substitutes cryopreserved with solution B. The</p></font></font> <p align="LEFT"><font face="AdvGulliv-R" size="1"><font face="AdvGulliv-R" size="1">reculture method and cell viability analyses with Live &amp; Dead</font></font><font face="AdvPSSym" size="1"><font face="AdvPSSym" size="1"> </font></font><font face="AdvGulliv-R" size="1"><font face="AdvGulliv-R" size="1">revealed increased number of viable in</font></font></p><font face="AdvGulliv-R" size="1"><font face="AdvGulliv-R" size="1"> <p align="LEFT">cells preserved with solution B. Artificial stroma substitutes in CS control samples showed the greatest</p> <p align="LEFT">alterations in microstructure and cell proliferation. Analysis of the biomechanical properties showed that</p> <p align="LEFT">substitutes cryopreserved with different solutions had adequate rheological parameters (yield stress,</p> <p align="LEFT">elastic modulus and viscous modulus) and were therefore suitable for use in regenerative medicine.</p> <p align="LEFT">These results establish effective methods of cryopreservation for all experimental situations and suggest</p> <p align="LEFT">that solution B (DMEM, FCS, DMSO 10%) was the best cryoprotectant for the cryopreservation of an artificial</p> <p>oral human mucosa substitute based on a fibrin?agarose matrix.</p></font></font>