GAPDH-Silence Microsphere via Reprogramming Macrophage Metabolism and eradicating Bacteria for Diabetic infection bone regeneration.

Macrophage metabolism dysregulation, which is exacerbated by persistent stimulation in infectious and inflammatory diseases, such as diabetic infectious bone defects (DIBD), eventually leads to the failure of bone repair. Here, we have developed an injectable, macrophage-modulated GAPDH-Silence drug delivery system. This microsphere comprises chondroitin sulfate methacrylate (CM) and methacrylated gelatin (GM), while the dimethyl fumarate (DMF)-loaded liposome (D-lip) is encapsulated within the microsphere (CM@GM), named D-lip/CM@GM. Triggered by the over-expressed collagenase in DIBD, the microspheres degrade and release the encapsulated D-lip. D-lip could modulate metabolism by inhibiting GAPDH, which suppresses the over-activation of glycolysis, thus preventing the inflammatory response of macrophages in vitro. While beneficial for macrophages, D-lip/CM@GM is harmful to bacteria. GAPDH, while crucial for glycolysis of staphylococcal species (S. aureus), can be effectively countered by D-lip/CM@GM. We are utilizing existing drugs in innovative ways to target central metabolism for effective eradication of bacteria. In the DIBD model, our results confirmed that the D-lip/CM@GM enhanced bacteria clearance and reprogrammed dysregulated metabolism, thereby significantly improving bone regeneration. In conclusion, this GAPDH-Silence microsphere system may provide a viable strategy to promote diabetic infection bone regeneration.

Purification and characterization of lysozyme from Chinese Lueyang black-bone Silky fowl egg white.

Lysozyme, an important antibacterial protein, is an enzyme that cleaves the glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine of peptidoglycan in cell walls. The novel lysozyme was purified and characterized from Chinese Lueyang black-bone silky fowl (CBSF) egg white, and its N-terminal amino acid sequence, enzymatic properties, and antibacterial activity were investigated. The CBSF lysozyme was purified using adsorption chromatography, ammonium sulfate precipitation, ion exchange chromatography, and size-exclusion chromatography. The purification fold and yield were 3.28 and 14.69%, respectively. The purified lysozyme was revealed as a single protein band with SDS-PAGE and had a MALDI-TOF/TOF molecular weight of 14305.57 Da and a final specific activity of 3.49 × 105 U/mg protein using Micrococcus lysodeikticus as a substrate. The optimum temperature and pH of the lysozyme were 50 °C and 6.0, respectively. The 20 N-terminal amino acid residues of the purified lysozyme were determined to be KVFGRCELAAAMKRHGLDNY, showing some homology to the N-terminus of the odontophoridae egg white lysozyme. The purified lysozyme exerted a potent antimicrobial activity toward indicator microorganisms, including Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, and Escherichia coli ATCC 25922. However, its inhibition of gram-negative activity was weaker than that of the Gram-positive bacteria.

Biodegradable sizing agents from soy protein via controlled hydrolysis and dis-entanglement for remediation of textile effluents.

Fully biodegradable textile sizes with satisfactory performance properties were developed from soy protein with controlled hydrolysis and dis-entanglement to tackle the intractable environmental issues associated with the non-biodegradable polyvinyl alcohol (PVA) in textile effluents. PVA derived from petroleum is the primary sizing agent due to its excellent sizing performance on polyester-containing yarns, especially in increasingly prevailing high-speed weaving. However, due to the poor biodegradability, PVA causes serious environmental pollution, and thus, should be substituted with more environmentally friendly polymers. Soy protein treated with high amount of triethanolamine was found with acceptable sizing properties. However, triethanolamine is also non-biodegradable and originated from petroleum, therefore, is not an ideal additive. In this research, soy sizes were developed from soy protein treated with glycerol, the biodegradable triol that could also be obtained from soy. The soy sizes had good film properties, adhesion to polyester and abrasion resistance close to PVA, rendering them qualified for sizing applications. Regarding desizing, consumption of water and energy for removal of soy size could be remarkably decreased, comparing to removal of PVA. Moreover, with satisfactory degradability, the wastewater containing soy sizes was readily dischargeable after treated in activated sludge for two days. In summary, the fully biodegradable soy sizes had potential to substitute PVA for sustainable textile processing.

A sustainable slashing industry using biodegradable sizes from modified soy protein to replace petro-based poly(vinyl alcohol).

Biodegradable sizing agents from triethanolamine (TEA) modified soy protein could substitute poly(vinyl alcohol)(PVA) sizes for high-speed weaving of polyester and polyester/cotton yarns to substantially decrease environmental pollution and impel sustainability of textile industry. Nonbiodegradable PVA sizes are widely used and mainly contribute to high chemical oxygen demand (COD) in textile effluents. It has not been possible to effectively degrade, reuse or replace PVA sizes so far. Soy protein with good biodegradability showed potential as warp sizes in our previous studies. However, soy protein sizes lacked film flexibility and adhesion for required high-speed weaving. Additives with multiple hydroxyl groups, nonlinear molecule, and electric charge could physically modify secondary structure of soy protein and lead to about 23.6% and 43.3% improvement in size adhesion and ability of hair coverage comparing to unmodified soy protein. Industrial weaving results showed TEA-soy protein had relative weaving efficiency 3% and 10% higher than PVA and chemically modified starch sizes on polyester/cotton fabrics, and had relative weaving efficiency similar to PVA on polyester fabrics, although with 3- 6% lower add-on. In addition, TEA-soy sizes had a BOD5/COD ratio of 0.44, much higher than 0.03 for PVA, indicating that TEA-soy sizes were easily biodegradable in activated sludge.

Water-stable three-dimensional ultrafine fibrous scaffolds from keratin for cartilage tissue engineering.

Intrinsically water-stable scaffolds composed of ultrafine keratin fibers oriented randomly and evenly in three dimensions were electrospun for cartilage tissue engineering. Keratin has been recognized as a biomaterial that could substantially support the growth and development of multiple cell lines. Besides, three-dimensional (3D) ultrafine fibrous structures were preferred in tissue engineering due to their structural similarity to native extracellular matrices in soft tissues. Recently, we have developed a nontraditional approach to developing 3D fibrous scaffolds from alcohol-soluble corn protein, zein, and verified their structural advantages in tissue engineering. However, keratin with highly cross-linked molecular structures could not be readily dissolved in common solvents for fiber spinning, which required the remarkable drawability of solution. So far, 3D fibrous scaffolds from pure keratin for biomedical applications have not been reported. In this research, the highly cross-linked keratin from chicken feathers was de-cross-linked and disentangled into linear and aligned molecules with preserved molecular weights, forming highly stretchable spinning dope. The solution was readily electrospun into scaffolds with ultrafine keratin fibers oriented randomly in three dimensions. Due to the highly cross-linked molecular structures, keratin scaffolds showed intrinsic water stability. Adipose-derived mesenchymal stem cells could penetrate much deeper, proliferate, and chondrogenically differentiate remarkably better on the 3D keratin scaffolds than on 2D PLA fibrous scaffolds, 3D soy protein fibrous scaffolds, or 3D commercial nonfibrous scaffolds. In summary, the electrospun 3D ultrafine fibrous scaffolds from keratin could be promising candidates for cartilage tissue engineering.

Ultrafine fibrous gelatin scaffolds with deep cell infiltration mimicking 3D ECMs for soft tissue repair.

In this research, ultrafine fibrous scaffolds with deep cell infiltration and sufficient water stability have been developed from gelatin, aiming to mimic the extracellular matrices (ECMs) as three dimensional (3D) stromas for soft tissue repair. The ultrafine fibrous scaffolds produced from the current technologies of electrospinning and phase separation are either lack of 3D oriented fibrous structure or too compact to be penetrated by cells. Whilst electrospun scaffolds are able to emulate two dimensional (2D) ECMs, they cannot mimic the 3D ECM stroma. In this work, ultralow concentration phase separation (ULCPS) has been developed to fabricate gelatin scaffolds with 3D randomly oriented ultrafine fibers and loose structures. Besides, a non-toxic citric acid crosslinking system has been established for the ULCPS method. This system could endow the scaffolds with sufficient water stability, while maintain the fibrous structures of scaffolds. Comparing with electrospun scaffolds, the ULCPS scaffolds showed improved cytocompatibility and more importantly, cell infiltration. This research has proved the possibility of using gelatin ULCPS scaffolds as the substitutes of 3D ECMs.

Natural polysaccharides and proteins-based films for potential food packaging and mulch applications: A review.

Conventional nondegradable packaging and mulch films, after reaching the end of their use, become a major source of waste and are primarily disposed of in landfills. Accumulation of non-degradable film residues in the soil leads to diminished soil fertility, reduced crop yield, and can potentially affect humans. Application of degradable films is still limited due to the high cost, poor mechanical, and gas barrier properties of current biobased synthetic polymers. In this respect, natural polysaccharides and proteins can offer potential solutions. Having versatile functional groups, three-dimensional network structures, biodegradability, ease of processing, and the potential for surface modifications make polysaccharides and proteins excellent candidates for quality films. Besides, their low-cost availability as industrial waste/byproducts makes them cost-effective alternatives. This review paper covers the performance properties, cost assessment, and in-depth analysis of macromolecular structures of some natural polysaccharides and proteins-based films that have great potential for packaging and mulch applications. Proper dissolution of biopolymers to improve molecular interactions and entanglement, and establishment of crosslinkages to form an ordered and cohesive polymeric structure can help to obtain films with good properties. Simple aqueous-based film formulation techniques and utilization of waste/byproducts can stimulate the adoption of affordable biobased films on a large-scale.

Simultaneous toughness and stiffness of 3D printed nano-reinforced polylactide matrix with complete stereo-complexation via hierarchical crystallinity and reactivity.

A novel strategy adaptive to 3D printing of stereo-complexed polylactide matrix for simultaneous toughness and stiffness was designed. Stereo-complexation is a potent way to enhance both aqueous stability and heat resistance of polylactide, but also aggravates brittleness problem of polylactide. Though poly(butyleneadipate-co-terephthalate) elastomer with epoxidized compatibilizer improved stiffness and toughness of common polylactide, their effectiveness on mechanical and crystallization properties of stereo-complexed polylactide remained unknown. More importantly, incorporation of above techniques into 3D printing kept a fundamental challenge. Both stereo-complexation of polylactide and covalent coupling of polylactide and poly(butyleneadipate-co-terephthalate) by epoxidized compatibilizer are easy to occur when preparing the filaments for printing, impeding the following 3D printing procedure. The hypothesis for this research is that controlled hierarchical crystallization and reaction in three thermal processes could ensure simultaneous toughness and stiffness, and complete stereo-complexation in polylactide matrices. Reinforcing effects of a selected epoxidized compatibilizer, POSS(epoxy)8, on crystallinities, thermal properties, mechanical properties and morphologies were systematically studied. Such a strategy not only removed the obstacles in incorporating stereo-complexation and coupling techniques of polylactide into 3D printing, but also revealed the mechanism to produce high-performance 3D printed polylactide matrix via hierarchical crystallization and reaction.

Hierarchical crystallization strategy adaptive to 3-dimentional printing of polylactide matrix for complete stereo-complexation.

A novel strategy adaptive to 3D printing of PLA matrix for complete stereo-complexation was designed. Stereo-complexation has been demonstrated for its effectiveness in simultaneously improving aqueous stability and heat resistance of PLA. However, current techniques could not be directly incorporated into 3D printing of stereo-complexed PLA since stereo-complexed crystallites are easily formed before printing. High printing temperatures are thus required but decompose PLA materials at the same time. The hypothesis for this research is that controllable hierarchical crystallization in three thermal processes, the filament preparation, 3D printing and post annealing, could ensure feasibility of the strategy and a 100% stereo-complexation level in PLA matrices. Effects of extrusion, ambient and annealing temperatures on material structures were analyzed via WAXD, DSC and DMA. Resistance to hydrolysis and heat of the 3D printed PLA matrix was evaluated under practical conditions. It was showed that homo-crystallites anchored molecular chains of PLA during the post-annealing process for a high retention of tensile properties, while stereo-complexed crystallites provided stronger intermolecular interactions for improved hydrolytic and thermal resistance. This novel strategy via incorporating controlled hierarchical crystallization into 3D printing would enrich the fabrication and exploration of high-performance 3D printed PLA materials.

Ubiquitination Flow Repressors: Enhancing Wound Healing of Infectious Diabetic Ulcers through Stabilization of Polyubiquitinated Hypoxia-Inducible Factor-1α by Theranostic Nitric Oxide Nanogenerators.

Current treatments for diabetic ulcers (DUs) remain unsatisfactory due to the risk of bacterial infection and impaired angiogenesis during the healing process. The increased degradation of polyubiquitinated hypoxia-inducible factor-1α (HIF-1α) compromises wound healing efficacy. Therefore, the maintenance of HIF-1α protein stability might help treat DU. Nitric oxide (NO) is an intrinsic biological messenger that functions as a ubiquitination flow repressor and antibacterial agent; however, its clinical application in DU treatment is hindered by the difficulty in controlling NO release. Here, an intelligent near-infrared (NIR)-triggered NO nanogenerator (SNP@MOF-UCNP@ssPDA-Cy7/IR786s, abbreviated as SNP@UCM) is presented. SNP@UCM represses ubiquitination-mediated proteasomal degradation of HIF-1α by inhibiting its interaction with E3 ubiquitin ligases under NIR irradiation. Increased HIF-1α expression in endothelial cells by SNP@UCM enhances angiogenesis in wound sites, promoting vascular endothelial growth factor (VEGF) secretion and cell proliferation and migration. SNP@UCM also enables early detection of wound infections and ROS-mediated killing of bacteria. The potential clinical utility of SNP@UCM is further demonstrated in infected full-thickness DU model under NIR irradiation. SNP@UCM is the first reported HIF-1α-stabilizing advanced nanomaterial, and further materials engineering might offer a facile, mechanism-based method for clinical DU management.

Bioprinting of an osteocyte network for biomimetic mineralization.

Osteocytes, essential regulators of bone homeostasis, are embedded in the mineralized bone matrix. Given the spatial arrangement of osteocytes, bioprinting represents an ideal method to biofabricate a 3D osteocyte network with a suitable surrounding matrix similar to native bone tissue. Here, we reported a 3D bioprinted osteocyte-laden hydrogel for biomimetic mineralization in vitro with exceptional shape fidelity, a high cell density (107 cells per ml) and high cell viability (85%-90%). The bioinks were composed of biomimetic modified biopolymers, namely, gelatine methacrylamide (GelMA) and hyaluronic acid methacrylate (HAMA), with or without type I collagen. The osteocyte-laden constructs were printed and cultured in mineralization induction media. After 28 d, increased dendritic cell connections and enhanced mineralized matrix production were observed after the addition of type I collagen. These results were further confirmed by the expression of osteocyte-related genes, markers of osteocyte morphology (Connexin43 and E11/Podoplanin), markers of mineralization (dentin matrix acidic phosphoprotein 1 (Dmp1)) and the cellular response to parathyroid hormone (PTH). Moreover, the 3D bioprinting constructs outperformed the 2D monolayer culture and they were at least comparable to 3D casted hydrogels in mimicking the natural osteocyte phenotype. All results indicated that the 3D bioprinting osteocyte network shows promise for mechanistic studies and pharmaceutical screening in vitro.

Preparation and properties of starch acetate fibers for potential tissue engineering applications.

This article reports the development of fibers from starch acetates that have mechanical properties and water stability better than most polysaccharide-based biomaterials and protein fibers used in tissue engineering. In this research, starch acetates with three different degrees of substitution (DS) have been used to develop fibers for potential use as tissue engineering scaffolds. Varying the DS of starch acetate will provide fibers with different mechanical properties, hydrophilicity, and degradation behavior. Fibers made from DS 2.3 and 2.8 starch acetates have mechanical properties and water stability required for tissue engineering applications. The starch acetate fibers support the adhesion of fibroblasts demonstrating that the fibers would be suitable for tissue engineering and other medical applications.

Drug sorption onto and release from soy protein fibers.

Drug release in phosphate buffered saline (PBS pH 7.4) and artificial gastric juice (AGJ pH 1.2) and its relationship with kinetic and thermodynamic parameters of drug sorption onto soy protein (SP) fibers have been studied using Diclofenac, 5 Fluorouracil and Metformin as model drugs. Since SP is biodegradable, biocompatible, abundant and annually renewable, it has been widely used in medical applications. To understand drug release from SP fibers using sorption, kinetic and thermodynamic parameters have been investigated. Quantitative relationship between drug release and drug loading concentration, affinity, and activation energy for diffusion was established to predict initial bursts and later drug release. The study showed that Diclofenac had high initial bursts in PBS but more constant release in AGJ. It also has been found that drugs with lower diffusion coefficient and higher affinity (especially van der Waals force) on SP fiber are more suitable for sorption loading to achieve higher loading capacity and more constant releasing rate.

Submicron amino acid particles reinforced 100% keratin biomedical films with enhanced wet properties via interfacial strengthening.

Keratin films with wet stability and strength suitable for biomedical applications were developed via reinforcement with submicron cysteine particles for improved interfaces. Keratin products regenerated from wool or human hair were widely investigated as wound dressing and tissue engineering scaffolds for their satisfactory biomedical properties. However, regenerated keratin scaffolds usually did not have good mechanical properties, and also could not stand humid or wet biological environment due to poor moisture stability. Reinforcements for keratin materials were usually polysaccharides or synthetic polymers, and thus usually had non-ideal interfacial properties due to limited compatibility. In this research, submicron cystine particles were employed to reinforce keratin films for their high compatibility with keratin and bio-safety. Transition of primary and secondary structures of keratin due to matrix-reinforcement interaction was analyzed. The keratin films showed unprecedented pliancy, good tensile properties under humid conditions and biocompatibility, and thus had good potential for biomedical engineering applications.

Chitosan/gallnut tannins composite fiber with improved tensile, antibacterial and fluorescence properties.

Natural extracts gallnut tannins (GTs) were used as functional components to prepare chitosan/gallnut tannins (CS/GTs) composite fiber by blended solution spinning. Chitosan fiber has great potential to be used as absorbent suture and dressing due to its good biocompatibility. However, the weak mechanical properties limited its application. Chitosan and GTs were blended in aqueous solution of acetic acid to spin the composite fiber. The results indicated that CS/GTs fiber can be easily prepared due to the appropriate rheology characteristics for blended solution. Compared with pure chitosan fiber, CS/GTs fiber with 10% GTs showed lower hydrophilicity and higher dry, wet breaking strength by more than 40% due to ionic cross-linking between chitosan and GTs. The bacterial reduction to Staphylococcus aureus increased from 49.0 to 99.7% and about double green and red fluorescent intensity were observed for CS/GTs fiber. GTs have great potentiality in improving the properties of chitosan fiber.

Valorization of keratin from food wastes via crosslinking using non-toxic oligosaccharide derivatives.

Oligosaccharide derivatives were developed to crosslink keratin materials from poultry feathers, swine bristles and ox hairs to valorize these major wastes from meat industry. Global butchery generates more than 8,600,000 tons of keratinous wastes annually. Keratin was considered a promising resource for developing bio-based products as alternatives to petroleum products. Regenerated keratin products, such as films, usually showed insufficient mechanical properties, and required external crosslinking. However, most crosslinkers for proteins are either toxic, expensive, or with low efficiencies under mild conditions. In this research, regenerated keratin films were crosslinked by oxidized sucrose, a safe and potent bio-polyaldehyde. The crosslinker with verified low toxicity improved both tensile strength and elongation of keratin films, surpassing many other safe crosslinkers. Mechanism of the crosslinking reaction was proposed as forming Schiff bases and aminals and verified via 1H NMR and 13C NMR. Relationship between tensile properties and crosslinking degree of keratin films was also quantified.

Quantitation of fast hydrolysis of cellulose catalyzed by its substituents for potential biomass conversion.

This paper investigates the accelerated acidic hydrolysis of cellulose by its substituents for potential biomass conversion. Insufficient pretreatments and slow cellulose hydrolysis are major obstacles that impede efficient hydrolysis of cellulose. Substituted cellulose, such as dyed cotton, has large availability. It is susceptible to acidic hydrolysis and can be used for biomass conversion without any pretreatments. To understand the mechanism of accelerated hydrolysis of cellulose by its substituents is a prerequisite for cellulosic biomass conversion with high efficiency. Substituents with different charge properties were synthesized and their interactions with oxocarbenium ions were studied based on Density Functional Theory. Results indicate that hydrolysis rate is affected by field effect from substituents. Such field effect is dominated by amounts of negative charges on substituents and distance between negatively charged groups and oxocarbenium. Hydrolysis rate of dye-substituted cotton is higher than or comparable to that applied with other catalytic approaches.

Poly(l-lactic acid) bio-composites reinforced by oligo(d-lactic acid) grafted chitosan for simultaneously improved ductility, strength and modulus.

PLA bio-composites reinforced by oligo(d-lactic acid) grafted chitosan has been developed for simultaneously improved ductility, strength and modulus. Brittleness problem greatly limits the applications of PLA, a polymer derived from corn. Various methods have been developed to solve the brittleness problem. Unfortunately, these methods have their limitations, such as sacrifice of strength and modulus of PLA, use of toxic chemicals and high costs. Bio-based elastomers such as chitosan also have poor compatibility with PLA, leading to poor mechanical properties. The hypothesis for this research is that CS-g-oligo(D-LA) particles with good ductility could form strong interfacial interactions with PLLA matrix. Reinforcing effect of CS-g-oligo(D-LA) particles on PLLA matrix was systematically studied. Compatibility and intermolecular interactions between CS-g-oligo(D-LA) particles and PLLA matrix were studied by SEM, DSC and 13C NMR analyses. The reinforcing mechanism was summarized. Due to effective transfer of stress from PLLA matrix to the strong but ductile skeletons of CS-g-oligo(D-LA), ductility, strength and modulus of PLLA bio-composites were substantially improved. This novel reinforcing strategy via formation of strong interactions between enantiomeric lactyl units would enrich the fabrication and exploration of high-performance PLA-based bio-composites.

Characterizing natural cellulose fibers from velvet leaf (Abutilon theophrasti) stems.

Velvet leaf (Abutilon theophrasti) that is currently considered a weed and an agricultural problem could be used as a source for high quality natural cellulose fibers. The fibers obtained from the velvet leaf stems are mainly composed of approximately 69% cellulose and 17% lignin. The single cells in the fiber have lengths of approximately 0.9 mm, shorter than those in common bast fibers, hemp and kenaf. However, the widths of single cells in velvet leaf fibers are similar to the single cells in hemp and kenaf. The fibers exhibited breaking tenacity from 2.4 to 3.9 g/denier (325-500 MPa), breaking elongation of 1.6-2.4% and Young's modulus of 140-294 g/denier (18-38 GPa). Overall, velvet leaf fibers have properties similar to that of common bast fibers such as hemp and kenaf. Velvet leaves fibers could be processed on the current kenaf processing machineries for textile, composite, automotive and other fibrous applications.

Structure and properties of natural cellulose fibers obtained from sorghum leaves and stems.

For the first time, sorghum leaves and stems have been used to produce natural cellulose fibers with properties suitable for composite, textile, and other high-value fibrous applications. The leaf and stems fibers produced are multicellular and have similar cellulose contents. The breaking tenacity and elongation of the fibers are similar to that of natural cellulose fibers such as kenaf and cornstalk fibers. However, the sorghum fibers have a modulus of about 113 g/denier (15 GPa) similar to the modulus of cornstalk fibers but higher than that of cotton and cornhusk fibers. At least 7 million tons of natural cellulose fibers can be produced by using the sorghum stems and leaves available as byproducts every year. Using the sorghum byproducts as a source for cellulose fibers will help to add value to the sorghum crops and also make the fiber industry more sustainable.