Selective extraction of myoglobin from human serum with antibody-biomimetic magnetic nanoparticles.

Myoglobin (Mb) is an ideal biochemical marker for the diagnosis of certain diseases caused by damage to heart muscle or skeletal muscle. Nevertheless, serum myoglobin levels are usually very low while the interference components in real sample are extremely abundent. Hence, it is of great clinical significance to establish an effective method for Mb targeting. To obtain desired selectivity, targeting biomolecules like antibody and aptamer are essential to 'the state of the art'. However, such biomolecules suffer from many disadvantages, such as hard to prepare, susceptible to protease degradation, and high cost. Thus, novel alternatives that can overcome these issues are highly desirable. Herein, we pioneered a template-anchored controllable surface imprinting strategy for selective extraction of Mb from human serum via combining with facile magnetic separation of magnetic nanoparticles (MNPs). Mb-imprinted MNPs, as antibody-biomimetic materials, were prepared using amino group-modified MNPs as substrates and water-soluble self-polymerizable dopamine as imprinting monomer. The optimized imprinting time was 70 min, giving an optimal performance with high practical imprinting efficiency (up to 41%), high imprinting factor (4.2), high binding affinity (Kd=(2.05 ± 0.09) × 10-5 M), as well as excellent recognition selectivity. Moreover, compared to bare MNPs, Mb-imprinted MNPs possessed markedly better pH tolerance. Finally, the selective extraction of Mb from human serum sample by Mb-imprinted MNPs was experimentally confirmed and the recoveries of Mb in spiked serum ranged from (91.12 ± 6.81)% to (107.99 ± 7.76)%, indicating that the Mb-imprinted MNPs could be competent for the selective analysis of Mb in real bio-samples like human serum with high precision and reliability.

Inhibition of profibrotic signalling enhances the 5-azacytidine-induced reprogramming of fibroblasts into cardiomyocytes.

Recent studies have shown that cardiac fibroblasts (CFs) can be transformed into induced cardiomyocytes (iCMs). This phenomenon represents a potential method for rescuing damaged myocardia after myocardial infarction. The mechanism underlying cardiac reprogramming regulation must be clarified to improve the induction efficiency of iCMs. In this study, we treated CFs with 5-aza for 24 h and added TGF-ß inhibitor A83-01 for 2 weeks in vitro to investigate the effect of inhibiting fibrosis on myocardial differentiation. Inhibition of TGF-ß1 activity with A83-01 significantly decreased the expressions of collagen III and α-SMA and increased the expression of myocardial specific marker cTnT and gap junction protein Cx43 in CFs, enhanced cardiac reprogramming as opposed to 2 weeks with 5-aza alone. Transcriptome and quantitative real-time reverse transcription-polymerase chain reaction analysis at the 14th day postinduction of A83-01 revealed that the expression of genes involved in cardiac development increased in the presence of 5-aza. These findings suggest that the addition of A83-01 remarkably inhibits profibrotic signalling and improved the efficiency of iCMs, provide new insights into the molecular mechanisms of cardiac reprogramming and promote the use of iCMs in clinical applications.

Sulfated polysaccharides interact with fibroblast growth factors and protect from denaturation.

Fibroblast growth factors (FGFs) regulate embryonic development and homeostasis, including tissue and organ repair and specific aspects of metabolism. The basic FGF and acidic FGF, now known as FGF2 and FGF1, are widely used protein drugs for tissue repair. However, they are susceptible to denaturation at ambient temperatures and during long-time storage, which will reduce their biological activity. The interaction of FGFs with the sulfated domains of heparan sulfate and heparin is essential for their cellular signaling and stability. Therefore, we analyzed the interactions of FGF1 and FGF2 with four sulfated polysaccharides: heparin, dextran sulfate (DXS), λ-carrageenan, and chondroitin sulfate. The results of thermal stability and cell proliferation assays demonstrate that heparin, DXS, and λ-carrageenan bound to both FGFs and protected them from denaturation. Our results suggest heparin, DXS, and λ-carrageenan are potential formulation materials that bind and stabilize FGFs, and which may also potentiate their activity and control their delivery.