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Analysis of limbal mesenchymal stem cells expanded on human denuded amniotic membrane and porcine small intestine submucosa scaffolds


One of the main limitations in the treatment of corneal injuries is that the number of patients who needs this tissue exceed the availability of donors. These limitations have motivated the development of strategies using the new technological advances in tissue engineering. Traditional tissue engineering strategies involve the combination of cells, growth factors, and scaffolds that can supply cellular biological components allowing to restore the tissue function. The mesenchymal stem cells found in the limbal stroma (L-MSCs) have a self-renewal potential for multilineage differentiation and immunosuppressive properties that aid them to surviving from host immune rejection, which justifies its use for allograft applications and a suitable biocompatible scaffold that does not alter the cellular properties is necessary to transport these cells to the injured area and help to promote the regeneration.

Objetivos - Metodologia - Resultados - Discussão dos Resultados/Objectives - Methodology - Results - Discussion of Results/Objetivos - Metodología - Resultados - Discusión de los resultados

Thus, in this work we isolated the L-MSCs from human sclerocorneal rims and evaluated their potential for multilineage differentiation and the expression of mesenchymal surface markers. These cells were seeded on human amniotic membrane (hAM) and porcine small intestine submucosa (SIS) and we analyzed their viability, actin cytoskeleton, cell density and adhesion, nuclei morphology, and surface markers. Our results showed that cells adhered and integrated on both membranes with a high cell density, an important characteristic for cell therapy. In addition, the analysis of surface markers expression on L-MSCs after two weeks showed a slight increase in the percentages of negative markers for MSCs grown on SIS membrane. Thus, considering a long-term culture, the hAM was considered a better scaffold to maintain the MSCs phenotype. However, regarding the function as scaffolds, SIS was as efficient as the amniotic membrane, considering that these two types of biological matrices maintained the cell viability, actin cytoskeleton, nuclei morphology and mesenchymal phenotype, without causing cell death.

Considerações Finais/Final considerations/Consideraciones finales

Therefore, our data in vitro provide evidences for future pre-clinical studies where these membranes can be used as a support to transport mesenchymal stem cells to the injured area, creating a type of temporary curative, allowing the release of bioactive molecules, such as cytokines and growth factors and then promoting the tissue regeneration, both in human and veterinary medicine.

Palavras-chave/Key words/Palabras clave

amniotic membrane, porcine small intestine submucosa, limbal mesenchymal stem cells, scaffolds, sclerocorneal rims.


Mesenchymal stem cells/adultas


LILIANA IVET SOUS NAASANI, Cristiano Rodrigues, Jéssica G Azevedo, Aline Damo, Silvio Buchner, Marcia R WINK