Tbx5 overexpression in embryoid bodies increases TAK1 expression but does not enhance the differentiation of sinoatrial node cardiomyocytes.

Genetic studies place Tbx5 at the apex of the sinoatrial node (SAN) transcriptional program. To understand its role in SAN differentiation, clonal embryonic stem (ES) cell lines were made that conditionally overexpress Tbx5, Tbx3, Tbx18, Shox2, Islet-1, and MAP3k7/TAK1. Cardiac cells differentiated using embryoid bodies (EBs). EBs overexpressing Tbx5, Islet1, and TAK1 beat faster than cardiac cells differentiated from control ES cell lines, suggesting possible roles in SAN differentiation. Tbx5 overexpressing EBs showed increased expression of TAK1, but cardiomyocytes did not differentiate as SAN cells. EBs showed no change in the expression of the SAN transcription factors Shox2 and Islet1 and decreased expression of the SAN channel protein HCN4. EBs constitutively overexpressing TAK1 direct cardiac differentiation to the SAN fate but have reduced phosphorylation of its targets, p38 and Jnk. This opens the possibility that blocking the phosphorylation of TAK1 targets may have the same impact as forced overexpression. To test this, we treated EBs with 5z-7-Oxozeanol (OXO), an inhibitor of TAK1 phosphorylation. Like TAK1 overexpressing cardiac cells, cardiomyocytes differentiated in the presence of OXO beat faster and showed increased expression of SAN genes (Shox2, HCN4, and Islet1). This suggests that activation of the SAN transcriptional network can be accomplished by blocking the phosphorylation of TAK1.

Egg shell-derived calcium phosphate/carbon dot nanofibrous scaffolds for bone tissue engineering: Fabrication and characterization.

Recent exciting findings of the particular properties of Carbon dot (CDs) have shed light on potential biomedical applications of CDs-containing composites. While CDs so far have been widely used as biosensors and bioimaging agents, in the present study for the first time, we evaluate the osteoconductivity of CDs in poly (ε-caprolactone) (PCL)/polyvinyl alcohol (PVA) [PCL/PVA] nanofibrous scaffolds. Moreover, further studies were performed to evaluate egg shell-derived calcium phosphate (TCP3) and its cellular responses, biocompatibility and in vitro osteogenesis. Scaffolds were fabricated by simultaneous electrospinning of PCL with three different types of calcium phosphate, PVA and CDs. Fabricated scaffolds were characterized by Scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), contact angle measurement and degradation assessment. SEM, the methyl thiazolyl tetrazolium (MTT) assay, and alkaline phosphatase (ALP) activity test were performed to evaluate cell morphology, proliferation and osteogenic differentiation, respectively. The results demonstrated that while the addition of just 1 wt% CDs and TCP3 individually into PCL/PVA nanocomposite enhanced ALP activity and cell proliferation rate (p < 0.05), the synergetic effect of CDs/TCP3 led to highest osteogenic differentiation and proliferation rate compared to other scaffolds (p < 0.05). Hence, CDs and PCL/PVA-TCP3 could serve as a potential candidate for bone tissue regeneration.

Core-shell fibrous scaffold as a vehicle for sustained release of retinal pigmented epithelium-derived factor (PEDF) for photoreceptor differentiation of conjunctiva mesenchymal stem cells.

Coaxial electrospinning technique was introduced as a flexible and promising technique for the fabrication of core-shell fibrous scaffold from poly ethylene glycol/poly caprolactone (PEG/PCL), where retinal pigmented epithelium-derived factor (PEDF) was encapsulated in the core, for photoreceptor differentiation of conjunctiva mesenchymal stem cells (CJMSCs) seed on scaffolds. The morphology and structure of fibers were characterized using SEM and TEM and photoreceptor differentiation was examined by quantitative real time PCR (qPCR). Release study showed that, a sustained release of PEDF from PEG/PCL scaffold was observed over 14 days. qPCR analysis demonstrated that rhodopsin (as a main photoreceptor gene) was significantly expressed in CJMSCs cultured on scaffold loaded with PEDF. According to the result, the core-shell scaffold loaded with PEDF (PEG + PEDF)/PCL) has superior control over factor release profile and has a potential for guiding photoreceptor differentiation of mesenchymal stem cells and promoting retinal regeneration. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3514-3519, 2017.

Effect of parameters on the quality of core-shell fibrous scaffold for retinal differentiation of conjunctiva mesenchymal stem cells.

This article describes the coaxial electrospinning to generate core-shell fibers from polycaprolactone (PCL) and polyethylene glycol (PEG) for differentiation of conjunctiva mesenchymal stem cells (CJMSCs) into photoreceptor-like cells by delivery of taurine. Also, the effects of many parameters such as polymer concentration, nozzle collector distance, applied voltage, and outer solution flow rate on creation of the core-shell structure were examined. The morphology and structure of fibers were characterized using scanning, transmission electron microscopy, and fourier transform infrared spectroscopy and then retinal differentiation was examined by quantitative real time PCR (qPCR). Significant variations between 25% and 35% PEG concentration groups for fiber diameter were documented. As faster flowing rates from the outer nozzle (PCL fluid) were applied, the creation possibility of fibrous scaffold was increased. The lowest diameter and the best quality alignment of core-shell fibrous scaffold were achieved in 22 Kv and 24 Kv. As rising distancing were applied, the fibrous diameter increased and spraying was observed. qPCR analysis demonstrated the differentiation of CJMSCs to photoreceptor like cells on PEG/PCL scaffolds. According to the result, we have proved successful in the creation of core/shell fibrous scaffold of PEG/PCL by coaxial electrospinning for retinal tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 189-197, 2017.