Isolation and functional characterization of cold-induced gene (AmCIP) promoter from Ammopiptanthus mongolicus.

AmCIP is a dehydrin-like protein which involved in abiotic stress tolerance in xerophytes evergreen woody plant A. mongolicus. AmCIP could be induced in the cotyledon and radicle during cold acclimation. To further elucidate the regulation of the upstream region of the gene, we isolated and characterized the promoter of AmCIP. Herein, a 1115 bp 5'-flanking region of AmCIP genomic DNA was isolated and cloned by genome walking from A. mongolicus and the segment sequence was identified as "PrAmCIP" promoter. Analysis of the promoter sequence revealed the presences of some basic cis-acting elements, which were related to various environmental stresses and plant hormones. GUS histochemical staining of transgene tobacco showed that PrAmCIP was induced by 4℃, 55℃, NaCl, mannitol and ABA, whereas it could hardly drive GUS gene expression under normal conditions. Furthermore, we constructed three deletion fragments and genetically transformed them into Arabidopsis thaliana. GUS histochemical staining showed that the MYCATERD1 element of the CP7 fragment (-189 âˆ¼ -1) may be a key element in response to drought. In conclusion, we provide an inducible promoter, PrAmCIP, which can be applied to the development of transgenic plants for abiotic stresse tolerance.

Genome-wide identification and functional analysis of silicon transporter family genes in moso bamboo (Phyllostachys edulis).

Silicon (Si) has crucial effects on plant development and stress resistance. Silicon transporters regulate Si absorption, transport, and distribution in plants. In this study, we identified and characterized the Si transporter gene family of moso bamboo (Phyllostachys edulis) and cloned seven putative Si transporter genes. Moso bamboo Si transporters contain conserved functional domains that mediate the accumulation of considerable amounts of Si. The analysis of gene duplication patterns and divergence times suggested that the expansion of the moso bamboo Si transporter family was mainly due to segmental duplications. The expression of moso bamboo Si transporter genes, which varied among organs, was significantly modulated by Si treatments. The subcellular localization analysis showed that Si transporters are plasma membrane proteins. The Si content increased in transgenic Arabidopsis overexpressing PeLsi1-1 or PeLsi1-2, which affected vegetative and reproductive growth. Our single-particle tracking analysis revealed the four diffusion modes of PeLsi1-1 on the plasma membrane. Moreover, the particle velocity, dwell time, and motion range of PeLsi1-1 decreased in response to Si treatments. The results of this study will further clarify the molecular mechanisms underlying Si absorption and accumulation in bamboo plants.

Bilayer silk fibroin/sodium alginate scaffold promotes vascularization and advances inflammation stage in full-thickness wound.

An ideal wound dressing for full-thickness wound regeneration should offer desirable biocompatibility, adequate mechanical properties, barrier function, and cellular regulation. Here, a bilayer scaffold resembling the hierarchical structure of human skin was developed using silk fibroin and sodium alginate. The upper membrane was prepared through casting and functioned as the epidermis, whereas the lower porous scaffold was prepared by freeze-drying and mimicked extracellular matrix structures. The membrane had nonporous structure, desirable mechanical properties, moderate hydrophilic surface, and suitable water vapor transmission rate, whereas the porous scaffold revealed 157.61 ± 41.67µm pore size, 86.10 ± 3.60% porosity, and capability of stimulating fibroblast proliferation. The combination of the two structures reinforced the tensile strength by five-fold and provided protection from wound dehydration. A suitable degradation rate reduced potential administration frequency. Furthermore, anin vivorabbit full-thickness wound healing test demonstrated that the bilayer scaffold facilitated wound closure, granulation tissue formation, re-epithelialization and skin component transition towards normal skin by providing a moist wound environment, advancing the inflammation stage, and stimulating angiogenesis. Collectively, as an off-the-shelf and cell-free wound dressing with single topical administration, the bilayer scaffold is a promising wound dressing for full-thickness wound regeneration.

Development of silk fibroin­sodium alginate scaffold loaded silk fibroin nanoparticles for hemostasis and cell adhesion.

During wound healing process, it is essential to promote hemostasis and cell adhesion. Herein, we incorporated a scaffold with nanoparticles to improve the hemostatic properties and stimulate cell adhesion. The nanoparticles were prepared by self-assembling of silk fibroin, and the scaffold loaded nanoparticles were synthesized by crosslinking and freeze-drying. Macroscopical images showed that the nanoparticles distributed uniformly and increased the surface roughness of scaffold pore wall. The addition of nanoparticles decreased the pore size, enhanced the compression strength, lowered the degradation rate, and maintained the resilience and water uptake capacity. Compared with pure scaffold, the scaffold loaded nanoparticles revealed higher blood clotting index and promoted platelets adhesion. Furthermore, in vitro tests showed that scaffold loaded nanoparticles exhibited excellent biocompatibility, and stimulation effects on cell proliferation, migration, and adhesion for both L929 cells and HUVECs. Therefore, the scaffold loaded nanoparticles possessed great potential as a wound dressing for efficient hemostasis and subsequent wound healing.

Research progress, models and simulation of electrospinning technology: a review.

In recent years, nanomaterials have aroused extensive research interest in the world's material science community. Electrospinning has the advantages of wide range of available raw materials, simple process, small fiber diameter and high porosity. Electrospinning as a nanomaterial preparation technology with obvious advantages has been studied, such as its influencing parameters, physical models and computer simulation. In this review, the influencing parameters, simulation and models of electrospinning technology are summarized. In addition, the progresses in applications of the technology in biomedicine, energy and catalysis are reported. This technology has many applications in many fields, such as electrospun polymers in various aspects of biomedical engineering. The latest achievements in recent years are summarized, and the existing problems and development trends are analyzed and discussed.

Fabrication of Hydroxyapatite/Tantalum Composites by Pressureless Sintering in Different Atmosphere.

The effect of sintering atmosphere (air and Ar) and temperature (1100, 1200, 1300 °C) on the microstructure, mechanical properties, and bioactivity of hydroxyapatite/tantalum (HA/Ta) composites were systematically investigated by pressureless sintering of the mixture of HA and Ta powders. It shows that the sintering atmosphere greatly impacts the phase composition and microstructure of the HA/Ta composites. The higher diffusion of atoms promotes shrinkage and causes deeper reaction fusion between the HA matrix and Ta, which improved the interfacial binding of the HA/Ta composites. The refined grain structure and improved interfacial binding obtained within the Ar atmosphere compared to the air atmosphere benefit the mechanical properties. The maximum bending strength and shrinkage observed for the composites sintered at 1300 °C in the Ar atmosphere are 27.24 MPa and 6.65%, respectively. The cell counting kit-8 (CCK-8) method was used to investigate the in vitro cytocompatibility of HA/Ta composites. The results revealed that the HA/Ta composites sintered with different conditions have no cytotoxicity. The simulated body fluid (SBF) soaking results showed that all of the studied composites possess desirable bioactivity, as demonstrated by their ability to form calcium-deficient carbonate apatite layer on the surfaces. For composites sintered at 1300 °C, the surface apatite layer coverage of the composites obtained in the Ar atmosphere was increased by 139.7% than the ones obtained in air, which confirmed an enhanced bioactive mineralization ability. The results indicated that the HA/Ta composites sintered at 1300 °C in Ar possess desirable mechanical properties and bioactivity. This work opens up the new possibility for preparing HA-based composites and is of great value in biomedical applications.

Maintaining Inducibility of Dermal Follicle Cells on Silk Fibroin/Sodium Alginate Scaffold for Enhanced Hair Follicle Regeneration.

The extracellular matrix (ECM) is important for maintaining cell phenotype and promoting cell proliferation and differentiation. In order to better solve the problem of skin appendage regeneration, a combination of mechanical/enzymatic digestion methods was used to self-extract dermal papilla cells (DPCs), which were seeded on silk fibroin/sodium alginate scaffolds as seed cells to evaluate the possibility of skin regeneration/regeneration of accessory organs. Scanning electron microscopy (SEM) graphs showed that the interconnected pores inside the scaffold had a pore diameter in the range of 153-311 µm and a porosity of 41-82%. Immunofluorescence (IF) staining and cell morphological staining proved that the extracted cells were DPCs. The results of a Cell Counting Kit-8 (CCK-8) and Calcein-AM/PI live-dead cell staining showed that the DPCs grew well in the composite scaffold extract. Normal cell morphology and characteristics of aggregation growth were maintained during the 3-day culture, which showed that the silk fibroin/sodium alginate (SF/SA) composite scaffold had good cell-compatibility. Hematoxylin-eosin (H&E) staining of tissue sections further proved that the cells adhered closely and aggregated to the pore wall of the scaffold, and retained the ability to induce differentiation of hair follicles. All these results indicate that, compared with a pure scaffold, the composite scaffold promotes the adhesion and growth of DPCs. We transplanted the SF/SA scaffolds into the back wounds of SD rats, and evaluated the damage model constructed in vivo. The results showed that the scaffold inoculated with DPCs could accelerate the repair of the skin and promote the regeneration of the hair follicle structure.