Endothelial Cell Migration Regulated by Surface Topography of Poly(ε-caprolactone) Nanofibers.

The study of cell migration on biomaterials is of great significance in tissue engineering and regenerative medicine. In recent years, there has been increasing evidence that the physical properties of the extracellular matrix (ECM), such as surface topography, affect various cellular behaviors such as proliferation, adhesion, and migration. However, the biological mechanism of surface topography influencing cellular behavior is still unclear. In this study, we prepared polycaprolactone (PCL) fibrous materials with different surface microstructures by solvent casting, electrospinning, and self-induced crystallization. The corresponding topographical structure obtained is a two-dimensional (2D) flat surface, 2.5-dimensional (2.5D) fibers, and three-dimensional (3D) fibers with a multilevel microstructure. We then investigated the effects of the complex topographical structure on endothelial cell migration. Our study demonstrates that cells can sense the changes of micro- and nanomorphology on the surface of materials, adapt to the physical environment through biochemical reactions, and regulate actin polymerization and directional migration through Rac1 and Cdc42. The cells on the nanofibers are elongated spindles, and the positive feedback of cell adhesion and actin polymerization along the fiber direction makes the plasma membrane continue to protrude, promoting cell polarization and directional migration. This study might provide new insights into the biomaterial design, especially those used for artificial vascular grafts.

Stretchable Strain Sensor for Human Motion Monitoring Based on an Intertwined-Coil Configuration.

Wearable electronics, such as sensors, actuators, and supercapacitors, have attracted broad interest owing to their promising applications. Nevertheless, practical problems involving their sensitivity and stretchability remain as challenges. In this work, efforts were devoted to fabricating a highly stretchable and sensitive strain sensor based on dip-coating of graphene onto an electrospun thermoplastic polyurethane (TPU) nanofibrous membrane, followed by spinning of the TPU/graphene nanomembrane into an intertwined-coil configuration. Owing to the intertwined-coil configuration and the synergy of the two structures (nanoscale fiber gap and microscale twisting of the fiber gap), the conductive strain sensor showed a stretchability of 1100%. The self-inter-locking of the sensor prevents the coils from uncoiling. Thanks to the intertwined-coil configuration, most of the fibers were wrapped into the coils in the configuration, thus avoiding the falling off of graphene. This special configuration also endowed our strain sensor with an ability of recovery under a strain of 400%, which is higher than the stretching limit of knees and elbows in human motion. The strain sensor detected not only subtle movements (such as perceiving a pulse and identifying spoken words), but also large movements (such as recognizing the motion of fingers, wrists, knees, etc.), showing promising application potential to perform as flexible strain sensors.

Endothelial Cell Migration on Poly(ε-caprolactone) Nanofibers Coated with a Nanohybrid Shish-Kebab Structure Mimicking Collagen Fibrils.

Regulating cell migration dynamics is of significance in tissue engineering and regenerative medicine. A 3D scaffold was created to provide various topographies based on a poly(ε-caprolactone) (PCL) self-induced nanohybrid shish-kebab structure, which consisted of aligned PCL nanofibers and spaced PCL crystal lamellae grown on the fibers. Electrospinning was applied followed by self-induced crystallization. The results resembled natural collagen fibrils in an extracellular matrix. This variable microstructure enabled control of cell adhesion and migration. The kebab size was controlled by initial PCL concentrations. The geometry of cells seeded on the fibers was less elongated, but the adhesion was more polarized with a higher nuclear shape index and faster migration speed. These results could aid in rapid endothelialization in tissue engineering.

Polycaprolactone Nanofibers Containing Vascular Endothelial Growth Factor-Encapsulated Gelatin Particles Enhance Mesenchymal Stem Cell Differentiation and Angiogenesis of Endothelial Cells.

During the regeneration of tissues and organs, growth factors (GFs) play a vital role by affecting cell behavior. However, because of the low half-life time and quick degradation of GFs, their stimulations on cells are relatively short and discontinuous. In this study, a releasing scaffold platform, consisting of polycaprolactone (PCL) nanofibers and vascular endothelial growth factor (VEGF)-encapsulated gelatin particles, was developed to extend the influence of GFs on mesenchymal stem cells (MSCs) and endothelial cells (ECs). The results showed that this kind of scaffold can direct the differentiation of MSCs to ECs and maintain the stability of the tubular structure, an indicator of the angiogenesis ability of ECs, for an extended period of time. Therefore, the results suggest the potential application of PCL/VEGF-encapsulated gelatin particles (PCL/VGPs) as a growth factor (GF)-releasing scaffold platform in vascular tissue engineering.

Endogenous biological factors modulated by substrate stiffness regulate endothelial differentiation of mesenchymal stem cells.

During the process of tissue regeneration facilitated by stem cells, physical properties of a scaffold affect behavior and activities of the cell. To enhance differentiation of human mesenchymal stem cells (MSCs) into endothelial-like cells (ELCs), we used electrospun fibrous substrates with different stiffness to enhance the differentiation. A simple method of annealing with different lengths of treatment time was employed to modulate stiffness of electrospun fibrous substrates without changing their chemistry. We seeded MSCs on substrates with different stiffness to study how stiffness of a culture substrate affects differentiation of MSCs into ELCs. Results of RT-PCR and western blotting revealed that stiffer substrates with the average surface modulus of 7.82 MPa induced differentiated MSCs to express more VEGF, CD31, and vWF mRNA transcripts and proteins than softer ones with that of 3.8 or 1.44 MPa. We also found that the production of macrophage migration inhibitory factor (MIF) in ELCs was increased with substrate stiffness. After silencing MIF mRNA, MSCs during differentiation showed lower expression levels of VEGF, CD31, and vWF than control cells whereas VEGF-silenced and control cells expressed comparable levels of MIF, indicating that MIF is an upstream molecule regulating VEGF in the mechanism. Our findings provide new insight into how stiffness of a culture substrate regulates differentiation of MSCs into ELCs. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1595-1603, 2018.

Electrospun polycaprolactone/gelatin composites with enhanced cell-matrix interactions as blood vessel endothelial layer scaffolds.

During the fabrication of tissue engineering scaffolds and subsequent tissue regeneration, surface bioactivity is vital for cell adhesion, spreading, and proliferation, especially for endothelium dysfunction repair. In this paper, synthetic polymer polycaprolactone (PCL) was blended with natural polymer gelatin at four different weight ratios followed by crosslinking (i.e., 100:0, 70:30, 50:50, 30:70, labeled as PCL-C, P7G3-C, P5G5-C, and P3G7-C) to impart enhanced bioactivity and tunable mechanical properties. The PCL/gelatin blends were first dissolved in 2,2,2-trifluroethanol (TFE) and supplementary acetic acid (1% relative to TFE) solvent, electrospun, and then cross-linked to produce PBS-proof fibrous scaffolds. Scanning electron micrographs (SEM) indicated that fibers of each sample were smooth and homogeneous, with the fiber diameters increasing from 1.01±0.51µm to 1.61±0.46µm as the content of gelatin increased. While thermal resistance and crystallization of the blends were affected by the presence of gelatin, as reflected by differential scanning calorimetry (DSC) results, water contact angle (WCA) tests confirmed that the scaffold surfaces became more hydrophilic. Tensile tests showed that PCL-C and P7G3-C scaffolds had mechanical properties comparable to those of human coronary arteries. As for cytocompatibility, skeleton staining images showed that human mesenchymal stem cells (hMSCs) had more favorable binding sites on PCL/gelatin scaffolds than those on PCL scaffolds. Cell proliferation assays revealed that P7G3-C scaffolds could support the most number of hMSCs. The results of this study demonstrated the enhanced cell-matrix interactions and potential use of electrospun PCL/gelatin scaffolds in the tissue engineering field, especially in wound dressings and endothelium regeneration.

[The regulation of T3 on maldeveloped gonocyte (Go) and expression of NCAM in flutamide-induced cryptorchidism rat].

OBJECTIVE: To investigate the regulation of T3 on the maldeveloped gonocytes (Go) and the expression of NCAM in Flutamide-induced cryptorchidism SD rat. METHODS: Postnatal day (PD) 1 and PD20 cryptorchidism SD rat induced by Flu were treated with 15 microg/(kg x d) T3 by subcutaneous injections. Incidence of the cryptorchidism, histology were evaluated. And the expression of NCAM was detected by immunohistochemisty and RT-PCR. RESULTS: The incidence of cryptorchidism in the group of Flu-induced offsprings treated by 15 microg/(kg x d) T3 from PD1 was 25% (8/32), which was lower than that of Flu group 40.9% (9/22). No Go and NCAM expression were observed in the seminiferous tubules. NCAM mRNA expression in Flu-induced PD13, PD20 testes was up-regulated when compared with control (P < 0.01, P < 0.05), while after treated with 15 microg/ (kg x d) T3 the NCAM expression was significantly down-regulated compared to that of non T3 treated group (P < 0.01), and was not significantly different from that of control (P > 0.05). CONCLUSION: The incidence of Flu-induced cryptorchidism can be decreased by T3 treatment after birth. T3 may contribute to the down-regulation of the NCAM expression, and reduction or elimination of the remainded gonocytes in crytorcidism seminiferous tubules.