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Poly(aspartic acid) (PASP) is an environmentally friendly biopolymer used as a fertilizer synergist and known to increase agricultural yields. The mechanism of PASP enhancement has, however, not been established, although the general hypothesis is that the polymer functions to hold nutrients closer to the root zone. The objective of this study was to determine the physiological and molecular changes that occur when plants are exposed to PASP, with future directions leading to a proposed mode of action. A whole genome transcriptome study was conducted. Arabidopsis thaliana seeds were germinated and grown in sterile plates treated with 250 ppm PASP under continuous light. Total RNA was extracted from whole seedlings and sequenced. The results revealed 462 differentially expressed genes (DEGs), 245 of which were upregulated and 217 downregulated. Gene Ontology, KEGG and MAPman analyses revealed DEGs involved in photosynthesis with 11 light harvesting complexes upregulated (e.g. LHCB1.1, LHCB2.2, LHCA1, LHCB4.2, LHCB2.1, LHCA4, LHCB1.1, LHCB3, LHCA3); the peroxisome pathway had 6 DEGs (CAT1, KAT1 and XDH2) upregulated (CSD1, CSD2 and FSD2) downregulated, the phenylpropanoid biosynthesis pathway had 7 DEGs upregulated. Other key DEGs were associated with the amino acid (e.g. ASN1) and nitrogen metabolism pathways. Physiology assessment results showed significant differences between control and treated plants with a 33 % increase in leaf area, 25 % increase chlorophyll content (p ≤ 0.05) and a 4-fold increase in photosynthetic rate (p ≤ 0.001). This information helps to increase our understanding of the key genes and metabolic pathways associated with plant response to PASP.

Poly(aspartic acid) (pAsp)-based fiber networks were developed via electrospinning and photo-crosslinking. Fiber networks hold the advantage that they exhibit a high surface-to-volume ratio, giving rise to a high water uptake and retention capacity, along with the ability to release moisture under desired circumstances (e.g. reduced relative humidity, mechanical pressure, etc.). Herein, polysuccinimide (PSI), the precursor of pAsp, was modified with 5-norbornene-2-methylamine to obtain crosslinkable norbornene-modified PSI (PSI-NB) with two different degrees of substitution, i.e. 19% and 46%. These derivatives were electrospun into thin uniform fibers after optimization of the processing parameters. The fiber sheets were crosslinked via a thiol-ene step-growth mechanism with three different thiol crosslinkers exploiting UV-A irradiation in the presence of TPO-L as photo-initiator. Using this strategy, fiber networks with diameters ranging between 1.27 ± 0.29 and 2.20 ± 1.05 µm were obtained, as visualized with scanning electron microscopy (SEM). Successful crosslinking was evidenced by a dissolution test in dimethylformamide and through X-ray photoelectron spectroscopy. Finally, the PSI-NB fiber networks were hydrolyzed to obtain pAsp-NB fiber networks by alkaline hydrolysis in a carbonate buffer solution, as confirmed by Fourier-transform infrared spectroscopy. The morphology of the fibers following hydrolysis was visualized by SEM and the average fiber diameters were calculated and compared to the diameters before hydrolysis, generally showing a diameter increase due to swelling of the fibers in aqueous solution. In conclusion, pAsp-based fiber networks were successfully developed and stabilized via photo-crosslinking.

Abstract The onset of osteoporosis leads to a gradual decrease in bone density due to an imbalance between bone formation and resorption. To achieve optimal drug efficacy with minimal side effects, targeted drug delivery to the bone is necessary. Previous studies have utilized peptides that bind to hydroxyapatite, a mineral component of bone, for bone‐targeted drug delivery. In this study, a hydroxyapatite binding (HAB) tag is fused to 30Kc19α‐Runt‐related transcription factor 2 (RUNX2) for bone‐targeting. This recombinant protein can penetrate the nucleus of human mesenchymal stem cells (hMSCs) and act as a master transcription factor for osteogenesis. The HAB tag increases the binding affinity of 30Kc19α‐RUNX2 to mineral deposition in mature osteoblasts and bone tissue, without affecting its osteogenic induction capability. In the osteoporosis mouse model, intravenous injection of HAB‐30Kc19α‐RUNX2 results in preferential accumulation in the femur and promotes bone formation while reducing toxicity in the spleen. These findings suggest that HAB‐30Kc19α‐RUNX2 may be a promising candidate for bone‐targeted therapy in osteoporosis.

The adsorption mechanism of different polymers containing ionic polyamino acids monomers in the chain structure at the solid–liquid interface was investigated. Initially, the influence of molecular weight and solution pH on simple polyamino acids (poly(L-aspartic acid) and poly(L-lysine) binding was determined. Considering the obtained dependencies, the polymer adsorption layer conformation was proposed in the systems containing block copolymers (both diblock and symmetrical triblock) consisting of polypeptide as well as poly(ethylene glycol) fragments. The presented studies focused on the application of two experimental methods. The polymer adsorption was carried out using the batch method and the adsorbate concentration was determined spectrophotometrically. Then, the turbidimetric measurements were taken. The analysis of the obtained results showed that the adsorption process of block copolymers depends on two factors. Firstly, the solution pH determines both the nature of the interactions of the copolymer structural units with the solid surface and the conformation of the polypeptide chains. The second parameter influencing the adsorption layer structure is the ratio of the lengths of both blocks. Introducing a short PEG fragment into the polymer main chain may improve the polymer adsorption properties by increasing the number of interactions with the adsorbent surface.

The innovation of N fertilizer and N management practices is essential to maximize crop yield with fewer N inputs. A long-term field fertilization experiment was established in 2015 on the North China Plain (NCP) to determine the effects of a control treatment (CN) and the eco-friendly material poly(aspartic acid)-coated urea (PN), applied as a one-time basal application method, on winter wheat yield and N use efficiency at four N application rates: 0 (N0), 63 (N63), 125 (N125), and 188 (N188) kg N ha–1. The results indicated that compared to CN, PN resulted in a significant increase in wheat yield by 9.6% and 9.2% at N63 and N125, respectively, across the three experimental years, whereas no significant (p < 0.05) difference was detected at N188. Leaf area duration (LAD), crop growth rate (CGR), and dry matter accumulation (DMA) increased with increasing N rates, while PN significantly increased LAD and CGR by 5.1%–16.4% and 5.4%–64.3%, respectively, during the anthesis-ripening growth stage and DMA by 13.7% and 10.1% at N63 and N125, respectively, after the anthesis stage compared to CN. During the grain-filling stage, PN significantly increased the kernel maximum grain-filling rate (Gmax) by 21.7% and the kernel weight at the maximum grain-filling rate (Wmax) by 6.7% at N125 compared to CN. Additionally, compared to CN, PN significantly improved the stover and grain N content at harvest and increased NUT, NPFP, and NAE by 5.7%–40.1%, 2.5%–23.3%, and 3.9%–42.8%, respectively, at N63–N125. Therefore, PN applied using a single basal nitrogen fertilizer application method showed promising potential in maintaining a stable wheat yield and increasing N use efficiency with a 33% urea cut (approximately 63 kg N ha–1) compared to CN at the current wheat yield level on the NCP.

Polymer hydrogels are ideal scaffolds for both tissue engineering and drug delivery. A great advantage of poly(amino acid)-based hydrogels is their high similarity to natural proteins. However, their expensive and complicated synthesis often limits their application. The use of poly(aspartic acid) (PASP) seems an appropriate solution for this problem due to the relatively cheap and simple synthesis of PASP. Using amino acids not only as building blocks in the polymer backbone but also as cross-linkers can improve the biocompatibility and the biodegradability of the hydrogel. In this paper, PASP cross-linked with cystamine (CYS) and lysine-methylester (LYS) was introduced as fully amino acid-based polymer hydrogel. Gels were synthesized employing six different ratios of CYS and LYS. The pH dependent swelling degree and the concentration of the elastically active chain were determined. After reduction of the disulfide bonds of CYS, the presence of thiol side groups was also detected. To determine the concentration of the reactive cross-linkers in the hydrogels, a new method based on the examination of the swelling behavior was established. Using metoprolol as a model drug, cell proliferation and drug release kinetics were studied at different LYS contents and in the presence of thiol groups. The optimal ratio of cross-linkers for the proliferation of periodontal ligament cells was found to be 60−80% LYS and 20−40% CYS. The reductive conditions resulted in an increased drug release due to the cleavage of disulfide bridges in the hydrogels. Consequently, these hydrogels provide new possibilities in the fields of both tissue engineering and controlled drug delivery.

This review presents an overview on the recent progress in the synthesis, crosslinking, interpenetrating networks, and applications of poly(aspartic acid) (PASP)-based hydrogels. PASP is a synthetic acidic polypeptide that has drawn a great deal of attention in diverse applications due particularly to its biocompatibility and biodegradability. Facile modification of its precursor, poly(succinimide) (PSI), by primary amines has opened a wide window for the design of state-of-the-art hydrogels. Apart from pH-sensitivity, PASP hydrogels can be modified with suitable species in order to respond to the other desired stimuli such as temperature and reducing/oxidizing media as well. Strategies for fabrication of nanostructured PASP-based hydrogels in the form of particle and fiber are also discussed. Different cross-linking agents for PSI/PASP such as diamines, dopamine, cysteamine, and aminosilanes are also introduced. Finally, applications of PASP-based hydrogels in diverse areas particularly in biomedical are reviewed.

Bone infection is a feared complication for patients with surgically fixed bone fractures and local antibiotic delivery is important in prophylaxis and treatment of these infections. Recent studies indicated that Staphylococcus aureus can penetrate bone tissue through micron-sized canaliculi and evade systemic and currently available local antibiotic treatments. Targeting bacteria within the bone requires highly efficient delivery of antimicrobials to the infected bone tissue. In this work, a biodegradable microsphere carrier loaded with antibiotics and with specific affinity to bone mineral was developed. Two widely used antibiotics, i.e., Gentamicin-dioctyl sulfosuccinate (GM-AOT) and Ciprofloxacin (CF) were embedded in poly(ϵ-caprolactone) (PCL) microspheres fabricated by oil-in-water emulsion techniques with carboxylated poly(vinyl alcohol) (cPVA) as surfactant. The carboxylic acid groups present at the Poly(ϵ-caprolactone)/cPVA (PCL-cPVA) microsphere surface were functionalized with aspartic acid oligomers (ASP) granting bone targeting properties. We report on cPVA synthesis, microsphere formulation, and antibiotic loading of PCL/cPVA-ASP microspheres. Antibiotic loaded PCL/cPVA-ASP microspheres show sustained release of its antibiotic load and can inhibit bacterial growth in vitro for up to 6 days. PCL/cPVA-ASP microspheres show enhanced affinity to mineralized substrates compared to non-functionalized PCL/cPVA microspheres. These findings support further development of these bone targeting antibiotic carriers for potential treatment of persistent bone infections.

A 2,4,6-trinitrobenzenesulphonic acid (TNBS)-based assay is developed to determine the degree of chemical cross-linking in aspartic acid-based polymer gels. The conventional colourimetric method for the quantitative determination of amine groups is difficult to use in polymer networks; thus, an improved method is developed to analyse polymer gels swollen in dimethyl sulfoxide (DMSO). Reaction products of the derivatizing reaction are examined by NMR. The chemical stability of the reagent is increased in DMSO, and the method shows satisfactory linearity and accuracy. The degree of chemical cross-linking in the investigated gels is close to its theoretical maximum, but the conversion of the pendant amine groups to cross-linking points is strongly dependent on the feed composition of the gels.