Identification, Selection, and Enrichment of Cardiomyocyte Precursors

The large-scale production of cardiomyocytes is a key step in the development of cell therapy and tissue engineering to treat cardiovascular diseases, particularly those caused by ischemia. the main objective of this study was to establish a procedure for the efficient production of cardiomyocytes by reprogramming mesenchymal stem cells from adipose tissue. First, lentiviral vectors expressing neoR and GFP under the control of promoters expressed specifically during cardiomyogenesis were constructed to monitor cell reprogramming into precardiomyocytes and to select cells for amplification and characterization. Cellular reprogramming was performed using 5'-azacytidine followed by electroporation with plasmid pOKS2a, which expressed Oct4, Sox2, and Klf4. Under these conditions, GFP expression began only after transfection with pOKS2a, and less than 0.015% of cells were GFP(+). These GFP(+) cells were selected for G418 resistance to find molecular markers of cardiomyocytes by RT-PCR and immunocytochemistry. Both genetic and protein markers of cardiomyocytes were present in the selected cells, with some variations among them. Cell doubling time did not change after selection. Together, these results indicate that enrichment with vectors expressing GFP and neoR under cardiomyocyte-specific promoters can produce large numbers of cardiomyocyte precursors (CMPs), which can then be differentiated terminally for cell therapy and tissue engineering.

Low-level laser therapy and light-emitting diode effects in the secretion of neuropeptides SP and CGRP in rat skin

The phototherapy effects in the skin are related to biomodulation, usually to accelerate wound healing. However, there is no direct proof of the interrelation between the effects of low-level laser therapy (LLLT) and light-emitting diode (LED) in neuropeptide secretion, these substances being prematurely involved in the neurogenic inflammation phase of wound healing. This study therefore focused on investigating LLLT and LED in Calcitonin gene-related peptide (CGRP) and substance P (SP) secretion in healthy rat skin. Forty rats were randomly distributed into five groups with eight rats each: Control Group, Blue LED Group (470 nm, 350 mW power), Red LED Group (660 nm, 350 mW power), Red Laser Group (660 nm, 100 mW power), and Infrared Laser Group (808 nm, 100 mW power) (DMCA (R) Equipamentos Ltda., So Carlos, So Paulo, Brazil). the skin of the animals in the experimental groups was irradiated using the punctual contact technique, with a total energy of 40 J, single dose, standardized at one point in the dorsal region. After 14 min of irradiation, the skin samples were collected for CGRP and SP quantification using western blot analysis. SP was released in Infrared Laser Group (p = 0.01); there was no difference in the CGRP secretion among groups. Infrared (808 nm) LLLT enhances neuropeptide SP secretion in healthy rat skin.