Angiogenesis induction as a key step in cardiac tissue Regeneration: From angiogenic agents to biomaterials.

Cardiovascular diseases are the leading cause of death worldwide. After myocardial infarction, the vascular supply of the heart is damaged or blocked, leading to the formation of scar tissue, followed by several cardiac dysfunctions or even death. In this regard, induction of angiogenesis is considered as a vital process for supplying nutrients and oxygen to the cells in cardiac tissue engineering. The current review aims to summarize different approaches of angiogenesis induction for effective cardiac tissue repair. Accordingly, a comprehensive classification of induction of pro-angiogenic signaling pathways through using engineered biomaterials, drugs, angiogenic factors, as well as combinatorial approaches is introduced as a potential platform for cardiac regeneration application. The angiogenic induction for cardiac repair can enhance patient treatment outcomes and generate economic prospects for the biomedical industry. The development and commercialization of angiogenesis methods often involves collaboration between academic institutions, research organizations, and biomedical companies.

Synthesis and Characterization of Thymol-Loaded Niosomal Film for the Prevention of Implant-Related Infection.

Background: Infection is one of the significant challenges in medical implant-related surgeries. Despite systemic antibiotic therapies, bacterial growth after implantation may cause implant failure. Nowadays, unlike the systemic therapy, local controlled release of antibiotic agents is considered an effective approach for the prevention of implant-related infections. The present study aimed to develop a niosomal nanocarrier incorporated into fibroin films for local and continuous delivery of thymol, a natural plant-derived antimicrobial agent for preventing infections caused by implant-related. Methods: Niosomes containing thymol were prepared by thin-film hydration technique. Thymol sustained release from the prepared films was assessed for 14 days. Antibacterial activities of the synthesized films were also evaluated by the agar diffusion technique against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Results: The release behavior from the niosomal thymol films showed a sustained manner, in which the amount of the released thymol reached 40% after 14 days. The films containing thymol with and without niosome showed a significant viability against L929 fibroblast cells compared to other groups after 24 and 48 h, using MTT assay. Also, samples exhibited potent antibacterial activity against Gram-negative and Gram-positive bacteria. Conclusion: The results of this study demonstrate that the niosomal thymol-loaded fibroin film is a promising candidate for the controlled release of thymol and prevention of implant-related infection.

Lawsonella clevelandensis, a case series of vascular graft infections caused by a rare pathogen.

Lawsonella clevelandensis is a fastidious Gram-positive, partially acid-fast, anaerobic, catalase positive bacterium that has been reported to be a rare cause of abdominal, breast, spinal, and liver abscesses. Here, three L. clevelandensis vascular graft infections (VGIs) and cardiac infections are reported.

A review on in vitro/in vivo response of additively manufactured Ti-6Al-4V alloy.

Bone replacement using porous and solid metallic implants, such as Ti-alloy implants, is regarded as one of the most practical therapeutic approaches in biomedical engineering. The bone is a complex tissue with various mechanical properties based on the site of action. Patient-specific Ti-6Al-4V constructs may address the key needs in bone treatment for having customized implants that mimic the complex structure of the natural tissue and diminish the risk of implant failure. This review focuses on the most promising methods of fabricating such patient-specific Ti-6Al-4V implants using additive manufacturing (AM) with a specific emphasis on the popular subcategory, which is powder bed fusion (PBF). Characteristics of the ideal implant to promote optimized tissue-implant interactions, as well as physical, mechanical/chemical treatments and modifications will be discussed. Accordingly, such investigations will be classified into 3B-based approaches (Biofunctionality, Bioactivity, and Biostability), which mainly govern native body response and ultimately the success in implantation.

Angiogenic Effect of a Nanoniosomal Deferoxamine-Loaded Poly(vinyl alcohol)-Egg White Film as a Promising Wound Dressing.

Owing to the noticeable increase in the number of patients with impaired wound healing capabilities, developing bioactive wound dressings with supportive physicomechanical and biological properties for clinical wound management has attracted much more attention nowadays. In this regard, engineered dressings with angiogenesis potential are vital for accelerated tissue regeneration. In the current study, nanoniosomal deferoxamine (DFO)-loaded transparent films of egg white-poly(vinyl alcohol) (PVA/EW/ND) were successfully fabricated at three different PVA/EW ratios (1:0, 1:1, and 1:1.5 wt/wt %) through the thin film hydration and solvent casting methods. The developed films' characterizations were carried out using scanning electron microscopy, Fourier transform infrared spectroscopy analysis, uniaxial tensile strength, water uptake, water vapor transmission rate, in vitro degradation, and drug release. The results demonstrated that the various weight ratios of PVA/EW have a significant effect on the microscopic morphology, equilibrium swelling, degradation, and mechanical properties of the films. The drug release profile exhibited a sustained release of DFO with controlled burst-lag phases resembling the Korsmeyer-Peppas pattern. The cytotoxicity and adhesion analysis using human dermal fibroblasts displays the biocompatibility of the developed PVA/EW/ND films and the formation of cellular colonies on the surface. The in vitro angiogenic capability of the developed films evaluated by the scratch wound assay and microbead-assisted tube formation study showed a significant increase in the rate of migration of human umbilical vein endothelial cells and in the number of tube-like structures. Therefore, the achieved results suggest that the presented PVA/EW/ND film has promising potential for effective wound healing applications.

Skin wound healing assisted by angiogenic targeted tissue engineering: A comprehensive review of bioengineered approaches.

Skin injuries and in particular, chronic wounds, are one of the major prevalent medical problems, worldwide. Due to the pivotal role of angiogenesis in tissue regeneration, impaired angiogenesis can cause several complications during the wound healing process and skin regeneration. Therefore, induction or promotion of angiogenesis can be considered as a promising approach to accelerate wound healing. This article presents a comprehensive overview of current and emerging angiogenesis induction methods applied in several studies for skin regeneration, which are classified into the cell, growth factor, scaffold, and biological/chemical compound-based strategies. In addition, the advantages and disadvantages of these angiogenic strategies along with related research examples are discussed in order to demonstrate their potential in the treatment of wounds.

Overexpression of the chloroplastic 2-oxoglutarate/malate transporter disturbs carbon and nitrogen homeostasis in rice.

The chloroplastic 2-oxaloacetate (OAA)/malate transporter (OMT1 or DiT1) takes part in the malate valve that protects chloroplasts from excessive redox poise through export of malate and import of OAA. Together with the glutamate/malate transporter (DCT1 or DiT2), it connects carbon with nitrogen assimilation, by providing 2-oxoglutarate for the GS/GOGAT (glutamine synthetase/glutamate synthase) reaction and exporting glutamate to the cytoplasm. OMT1 further plays a prominent role in C4 photosynthesis: OAA resulting from phosphoenolpyruvate carboxylation is imported into the chloroplast, reduced to malate by plastidic NADP-malate dehydrogenase, and then exported for transport to bundle sheath cells. Both transport steps are catalyzed by OMT1, at the rate of net carbon assimilation. To engineer C4 photosynthesis into C3 crops, OMT1 must be expressed in high amounts on top of core C4 metabolic enzymes. We report here high-level expression of ZmOMT1 from maize in rice (Oryza sativa ssp. indica IR64). Increased activity of the transporter in transgenic rice was confirmed by reconstitution of transporter activity into proteoliposomes. Unexpectedly, overexpression of ZmOMT1 in rice negatively affected growth, CO2 assimilation rate, total free amino acid content, tricarboxylic acid cycle metabolites, as well as sucrose and starch contents. Accumulation of high amounts of aspartate and the impaired growth phenotype of OMT1 rice lines could be suppressed by simultaneous overexpression of ZmDiT2. Implications for engineering C4 rice are discussed.

A review of accelerated wound healing approaches: biomaterial- assisted tissue remodeling.

Nowadays, due to a growing number of tissue injuries, in particular, skin wounds, induction and promotion of tissue healing responses can be considered as a crucial step towards a complete regeneration. Recently, biomaterial design has been oriented towards promoting a powerful, effective, and successful healing. Biomaterials with wound management abilities have been developed for different applications such as providing a native microenvironment and supportive matrices that induce the growth of tissue, creating physical obstacles against microbial contamination, and to be used as delivery systems for therapeutic reagents. Until now, numerous strategies aiming to accelerate the wound healing process have been utilized and studied with their own pros and cons. In this review, tissue remodeling phenomena, wound healing mechanisms, and their related factors will be discussed. In addition, different methods for induction and acceleration of healing via cell therapy, bioactive therapeutic delivery, and/or biomaterial-based approaches will be reviewed.

In Vitro Analysis of Metabolite Transport Proteins.

The photorespiratory cycle is distributed over four cellular compartments, the chloroplast, peroxisomes, cytoplasm, and mitochondria. Shuttling of photorespiratory intermediates between these compartments is essential to maintain the function of photorespiration. Specific transport proteins mediate the transport across biological membranes and represent important components of the cellular metabolism. Although significant progress was made in the last years on identifying and characterizing new transport proteins, the overall picture of intracellular metabolite transporters is still rather incomplete. The photorespiratory cycle requires at least 25 transmembrane transport steps; however to date only plastidic glycolate/glycerate transporter and the accessory 2-oxoglutarate/malate and glutamate/malate transporters as well as the mitochondrial transporter BOU1 have been identified. The characterization of transport proteins and defining their substrates and kinetics are still major challenges.Here we present a detailed set of protocols for the in vitro characterization of transport proteins. We provide protocols for the isolation of recombinant transport protein expressed in E. coli or Saccharomyces cerevisiae and the extraction of total leaf membrane protein for in vitro analysis of transporter proteins. Further we explain the process of reconstituting transport proteins in artificial lipid vesicles and elucidate the details of transport assays.

Loss of cytosolic phosphoglucose isomerase affects carbohydrate metabolism in leaves and is essential for fertility of Arabidopsis.

Carbohydrate metabolism in plants is tightly linked to photosynthesis and is essential for energy and carbon skeleton supply of the entire organism. Thus, the hexose phosphate pools of the cytosol and the chloroplast represent important metabolic resources that are maintained through action of phosphoglucose isomerase (PGI) and phosphoglucose mutase interconverting glucose 6-phosphate, fructose 6-phosphate, and glucose 1-phosphate. Here, we investigated the impact of disrupted cytosolic PGI (cPGI) function on plant viability and metabolism. Overexpressing an artificial microRNA targeted against cPGI (amiR-cpgi) resulted in adult plants with vegetative tissue essentially free of cPGI activity. These plants displayed diminished growth compared with the wild type and accumulated excess starch in chloroplasts but maintained low sucrose content in leaves at the end of the night. Moreover, amiR-cpgi plants exhibited increased nonphotochemical chlorophyll a quenching during photosynthesis. In contrast to amiR-cpgi plants, viable transfer DNA insertion mutants disrupted in cPGI function could only be identified as heterozygous individuals. However, homozygous transfer DNA insertion mutants could be isolated among plants ectopically expressing cPGI. Intriguingly, these plants were only fertile when expression was driven by the ubiquitin10 promoter but sterile when the seed-specific unknown seed protein promoter or the Cauliflower mosaic virus 35S promoter were employed. These data show that metabolism is apparently able to compensate for missing cPGI activity in adult amiR-cpgi plants and indicate an essential function for cPGI in plant reproduction. Moreover, our data suggest a feedback regulation in amiR-cpgi plants that fine-tunes cytosolic sucrose metabolism with plastidic starch turnover.

Shifts in the vascular endothelial growth factor isoforms result in transcriptome changes correlated with early neural stem cell proliferation and differentiation in mouse forebrain.

Regulation of neural stem cell (NSC) fate decisions is critical during the transition from a multicellular mammalian forebrain neuroepithelium to the multilayered neocortex. Forebrain development requires coordinated vascular investment alongside NSC differentiation. Vascular endothelial growth factor A (Vegf) has proven to be a pleiotrophic gene whose multiple protein isoforms regulate a broad range of effects in neurovascular systems. To test the hypothesis that the Vegf isoforms (120, 164, and 188) are required for normal forebrain development, we analyzed the forebrain transcriptome of mice expressing specific Vegf isoforms, Vegf120, VegfF188, or a combination of Vegf120/188. Transcriptome analysis identified differentially expressed genes in embryonic day (E) 9.5 forebrain, a time point preceding dramatic neuroepithelial expansion and vascular investment in the telencephalon. Meta-analysis identified gene pathways linked to chromosome-level modifications, cell fate regulation, and neurogenesis that were altered in Vegf isoform mice. Based on these gene network shifts, we predicted that NSC populations would be affected in later stages of forebrain development. In the E11.5 telencephalon, we quantified mitotic cells [Phospho-Histone H3 (pHH3)-positive] and intermediate progenitor cells (Tbr2/Eomes-positive), observing quantitative and qualitative shifts in these populations. We observed qualitative shifts in cortical layering at P0, particularly with Ctip2-positive cells in layer V. The results identify a suite of genes and functional gene networks that can be used to further dissect the role of Vegf in regulating NSC differentiation and downstream consequences for NSC fate decisions.