Aligned ovine diaphragmatic myoblasts overexpressing human connexin-43 seeded on poly (l-lactic acid) scaffolds for potential use in cardiac regeneration

GIMÉNEZ, CARLOS SEBASTIÁN; LOCATELLI, PAOLA; MONTINI BALLARIN, FLORENCIA; ORLOWSKI, ALEJANDRO; DEWEY, RICARDO A.; PENA, MILAGROS; ABRAHAM, GUSTAVO ABEL; AIELLO, ERNESTO ALEJANDRO; BAUZÁ, MARÍA DEL ROSARIO; CUNIBERTI, LUIS; OLEA, FERNANDA DANIELA; CROTTOGINI, ALBERTO

CYTOTECHNOLOGY (DORDRECHT)

SPRINGER

Lugar: Berlin; Año: 2018 vol. 70 p. 651 - 664

0920-9069

Diaphragmatic myoblasts (DMs) are precursorsof type-1 muscle cells displaying highexhaustion threshold on account that they contractand relax 20 times/min over a lifespan, making thempotentially useful in cardiac regeneration strategies.Besides, it has been shown that biomaterials for stemcell delivery improve cell retention and viability in thetarget organ. In the present study, we aimed atdeveloping a novel approach based on the use of poly(L-lactic acid) (PLLA) scaffolds seeded with DMsoverexpressing connexin-43 (cx43), a gap junctionprotein that promotes inter-cell connectivity. DMsisolated from ovine diaphragm biopsies were characterizedby immunohistochemistry and ability todifferentiate into myotubes (MTs) and transduced witha lentiviral vector encoding cx43. After confirmingcx43 expression (RT-qPCR and Western blot) and itseffect on inter-cell connectivity (fluorescence recoveryafter photobleaching), DMs were grown on fiberalignedor random PLLA scaffolds. DMs were successfullyisolated and characterized. Cx43 mRNA andprotein were overexpressed and favored inter-cellconnectivity. Alignment of the scaffold fibers not onlyaligned but also elongated the cells, increasing thecontact surface between them. This novel approach isfeasible and combines the advantages of bioresorbablescaffolds as delivery method and a cell type that onaccount of its features may be suitable for cardiacregeneration. Future studies on animal models ofmyocardial infarction are needed to establish itsusefulness on scar reduction and cardiac function.