Decellularized human amniotic membrane associated with adipose derived mesenchymal stromal cells as a bioscaffold: Physical, histological and molecular analysis

Abstract

The human amniotic membrane (hAM) has a great potential as a wound dressing and its combination with mesenchymal stromal cells (MSCs) represents an alternative therapy for wound healing. The scaffold should maintain the stemness of cells, so that they can present their immunomodulatory and chemoattractant properties when transplanted into the injured area. In this work we analyzed the histological and physical characteristics of amniotic membrane preserved in glycerol, in the presence and absence of human adipose derived mesenchymal stromal cells (AD-MSCs). No statistical difference was observed when comparing collagen fibers types I and III in intact (I-hAM) and amniotic membrane (D-hAM). We compared the degradation temperature with the thermogravimetric analysis in I-hAM, D-hAM, and D-hAM with AD-MSCs and characterized the structure and composition of the biological tissue material with Fourier Transform Infrared (FTIR). The intact hAM showed to be more thermally stable; however, it was observed that the hAM, intact and decellularized, is stable at high temperatures. The AD-MSCs were able to grow and expand on D-hAM without showing alterations on cell biology parameters as adhesion, viability, nuclei morphologyand actin cytoskeleton. Importantly, the AD-MSCs did not lose their differentiation potential and their mesenchymal cell markers expression after growth on D-hAM. These results constitute a main step for the use of this bioscaffold in pre-clinical models.