Silk fibroin scaffolds enhance cell commitment of adult rat cardiac progenitor cells

Abstract

The use of three-dimensional (3D) cultures may induce cardiac progenitor cells to synthesize their own extracellular matrix (ECM) and sarcomeric proteins to initiate cardiac differentiation. 3D cultures grown on synthetic scaffolds may favour the implantation and survival of stem cells for cell therapy when pharmacological therapies are not efficient in curing cardiovascular diseases and when organ transplantation remains the only treatment able to rescue the patient's life. Silk fibroin-based scaffolds may be used to increase cell affinity to biomaterials and may be chemically modified to improve cell adhesion. In the present study, porous, partially orientated and electrospun nanometric nets were used. Cardiac progenitor cells isolated from adult rats were seeded by capillarity in the 3D structures and cultured inside inserts for 21 days. Under this condition, the cells expressed a high level of sarcomeric and cardiac proteins and synthesized a great quantity of ECM. In particular, partially orientated scaffolds induced the synthesis of titin, which is a fundamental protein in sarcomere assembly.

Endurance Exercise and Conjugated Linoleic Acid (CLA) Supplementation Up-Regulate CYP17A1 and Stimulate Testosterone Biosynthesis

“…The qPCR technique was previously described in Di Felice et al [22]. Total cellular RNA was isolated from cell cultures and testicle samples using TRIzol® REAGENT (Sigma-Aldrich, USA), according to the manufacturer’s instructions.…”

“…The mRNA levels were normalised to the levels obtained for hypoxanthine phosphoribosyltransferase 1 (HPRT1) and for beta-glucuronidase (GUSB). Changes in the transcript level were calculated using the 2-∆∆Ct method [22,23]. The cDNA was amplified using the primers indicated in Table 1.…”

Cardiac tissue engineering: a reflection after a decade of hurry

“…Most of the investigations showcased the expression of proteins typical of the cardiac tissue, and a spatial distribution or alignment of the cells inside the scaffolds. Some research groups were able to show a striation typical of myofibrils (Engelmayr et al, 2008), others the massive expression of cardiac troponins, formation of intercalated disks, and functional gap junctions (Di Felice et al, 2013; Cristallini et al, 2014). Unfortunately, the extent of differentiation shown is very far from the actual maturation required to rebuild the cardiac tissue complexity.…”

“…Culturing cells in three dimensions or on substrates displaying the mechanical properties of the myocardium has been deemed sufficient to prompt cardiac commitment but not complete maturation per se (Engler et al, 2006; Di Felice et al, 2013). …”

Tissue engineering

“…Bottom-up self-assembly approaches are not rarely inspired by nature itself; nanostructures may be obtained by biologically inspired self-assembly as demonstrated by amphiphilic peptide self-assembly into hydrogels for TE strategies. [100][101][102][103] Top-down fabrication methods involving the use of biodegrad able scaffolds as a longer term substrate for cellular adhesion, pro liferation and new tissue formation may thus be required. However, insufficient ECM production in some cell types is thought to limit cellular migration and formation of cell-cell as well as cell-matrix junctions.…”