Characterization of 3D Printed Poly(3-Hydroxybutyric-Co-3-Hydroxyvalerate) by Fused Granular Fabrication through Thermal and Mechanical Analyses

Poly[R-3-hydroxybutyrate-co-(R-3-hydroxyvalerate)] (PHBVs) copolymers are promising biopolymers, which could substitute petroleum-based plastics for various applications. PHB and PHBV pellets were processed on a customized 3D printer via Fused Granular Manufacturing (FGM) approach modified with a Mahor screw extruder. To anticipate the behaviour of PHBVs when transformed using conventional thermo-mechanical shaping processes, thermal and mechanical analyses were carried out in order to better understand the effect of annealing temperature on their crystallization behaviour and mechanical properties of PHB polymer and PHBV copolymer. The objectives of the present work were to propose an experimental strategy to study the melting and crystallization events, crystalline structure changes, and mechanical performances of both PHB homopolymer and PHBV copolymer according to identical thermal annealing treatments. A monitoring of 3D printed PHB and PHBV structures was achieved by coupling Differential Scanning Calorimetry (DSC) and tensile tests. .

Miscibility, Crystallization, and Rheological Behavior of Solution Casting Poly（3-hydroxybutyrate）/poly（ethylene succinate） Blends Probed by Differential Scanning Calorimetry, Rheology, and Optical Microscope Techniques

The miscibility and crystallization of solution casting biodegradable poly（3-hydroxybuty- rate）/poly（ethylene succinate） （PHB/PES） blends was investigated by differential scanning calorimetry, rheology, and optical microscopy. The blends showed two glass transition temperatures and a depression of melting temperature of PHB with compositions in phase diagram, which indicated that the blend was partially miscible. The morphology observation supported this result. It was found that the PHB and PES can crystallize simultaneously or upon stepwise depending on the crystallization temperatures and compositions. The spherulite growth rate of PHB increased with increasing of PES content. The influence of compositions on the spherulitic growth rate for the partially miscible polymer blends was discussed.

Biosynthesis of poly-3-hydroxybutyrate with a high molecular weight by methanotroph from methane and methanol

Poly-3-hydroxybutyrate （PHB） can be produced by various species of bacteria. Among the possible carbon sources, both methane and methanol could be a suitable substrate for the production of PHB. Methane is cheap and plentiful not only as natural gas, but also as biogas. Methanol can also maintain methanotrophic activity in some conditions. The methanotrophic strain Methylosinus trichosporium IMV3011 can accumulate PHB with methane and methanol in a brief nonsterile process. Liquid methanol （0.1%） was added to improve the oxidization of methane. The studies were carried out using shake flasks. Cultivation was performed in two stages： a continuous growth phase and a PHB accumulation phase under the conditions short of essential nutrients （ammonium, nitrate, phosphorus, copper, iron （Ⅲ）, magnesium or ethylenediamine tetraacetate （EDTA）） in batch culture. It was found that the most suitable growth time for the cell is 144 h. Then an optimized culture condition for second stage was determined, in which the PHB concentration could be much increased to 0.6 g/L. In order to increase PHB content, citric acid was added as an inhibitor of tricarboxylic acid cycle （TCA）. It was found that citric acid is favorable for the PHB accumulation, and the PHB yield was increased to 40% （w/w） from the initial yield of 12% （w/w） after nutrient deficiency cultivation. The PHB produced is of very high quality with molecular weight up to 1.5 × 10^6Da.

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) supports adhesion and migration of mesenchymal stem cells and tenocytes

AIM: To establish the potential of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) as a material for tendon repair. METHODS: The biocompatibility of PHBHHx with both rat tenocytes (rT) and human mesenchymal stem cells (hMSC) was explored by monitoring adhesive characteristics on films of varying weight/volume ratios coupled to a culture atmosphere of either 21% O2 (air) or 2% O2 (physiological normoxia). The diameter and stiffness of PHBHHx films was established using optical coherence tomography and mechanical testing, respectively. RESULTS: Film thickness correlated directly with weight/volume PHBHHx (r2 = 0.9473) ranging from 0.1 mm (0.8% weight/volume) to 0.19 mm (2.4% weight/volume). Film stiffness on the other hand displayed a biphasic response which increased rapidly at values > 1.6% weight/volume. Optimal cell attachment of rT required films of ≥ 1.6% and ≥ 2.0% weight/volume PHBHHx in 2% O2 and 21% O2 respectively. A qualitative adhesion increase was noted for hMSC in films ≥ 1.2% weight/volume, becoming significant at 2% weight/volume in 2% O2. An increase in cell adhesion was also noted with ≥ 2% weight/volume PHBHHx in 21% O2. Cell migration into films was not observed. CONCLUSION: This evaluation demonstrates that PHBHHx is a suitable polymer for future cell/polymer replacement strategies in tendon repair.

Production of Poly(3-Hydroxybutyrate)by Fed-Batch Culture of Alcaligenes Eutrophus at Phosphorus Limitation in 50 L fermenter

Poly(3-hydroxybutyrate) (PHB) is an intracellular carbon and energy storage material accumulated by many kinds of microorganism under unfavorable growth conditions. For the production of PHB, Alcaligenes eutrophus has been widely used because it is easy to grow, and its physiological and biochemical changes during the PHB synthesis is understood in details. A very high concentration and productivity of PHB could be obtained by fed-batch culture of Alcaligenes eutrophus with phosphate limitation in 50 L fermenter.

BIOSYNTHESIS AND THERMAL PROPERTIES OF POLY(3-HYDROXYBUTYRATE-co-3-HYDROXYVALERATE)WITH LARGE VARIETY OF HYDROXYVALERATE CONTENTS BY BACILLUS CEREUS

Biosynthesis and thermal properties of poly（3-hydroxybutyrate-co-3-hydroxyvalerate） （PHBV） with different HV （hydrovalerate） content produced by a Bacillus cereus strain were investigated. A large variety of HV contents （up to about 90 mol%） of PHBV could be produced by this strain. Combined nitrogen sources containing both yeast extract and ammonium sulphate were better for cell growth and polyhydroxyalkanoates （PHA） production than either yeast extract or ammonium sulphate alone. Propionic acid is more favorable for the production of HV content than that of valeric acid. Finally, thermal properties of PHBV produced by this strain are found close to the results of other groups.

Silk fibroins modify the atmospheric low temperature plasma-treated poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) film for the application of cardiovascular tissue engineering

Tissue engineered scaffold is one of the hopeful therapies for the patients with organ or tissue damages. The key element for a tissue engineered scaffold material is high biocompatibility. Herein the poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) film was irradiated by the low temperature atmospheric plasma and then coated by the silk fibroins (SF). After plasma treatment, the surface of PHBHHx film became rougher and more hydrophilic than that of original film. The experiment of PHBHHx flushed by phosphate buffer solution (PBS) proves that the coated SF shows stronger immobilization on the plasma-treated film than that on the untreated film. The cell viability assay demonstrates that SF-coated PHBHHx films treated by the plasma significantly supports the proliferation and growth of the human smooth muscle cells (HSMCs). Furthermore, the scanning electron microscopy and hemotoylin and eosin (HE) staining show that HSMCs formed a cell sub-monolayer and secreted a large amount of extracellular matrix (ECM) on the films after one week's culture. The silk fibroins modify the plasma-treated PHBHHx film, providing a material potentially applicable in the cardiovascular tissue engi-neering.

Complete Valorization of Olive Mill Wastewater through an Integrated Process for Poly-3-hydroxybutyrate Production

An economical and environmental sustainability of bioplastic production is dependent on the use of low cost and waste C-sources as raw materials. OMW （Olive Mill Wastewater） with its high organic load represents a dangerous polluting waste. Herein the authors present an integrated process for the simultaneous recovery of polyphenols, high value natural antioxidants, production of PHAs （polyhydroxyalkanotes）, thermoplastic bio-polymers, in particular of PHB （poly-3-hydroxybutyrate） starting from OMW. The combination of membrane filtration and bacterial digestion of OMW resulted in very high yields of polyphenols （3 2.5 g/L） and PHB （31.4 mg/L.h） if compared with the state of the art. These results make the technical approach described here effective for reducing the polluting effect of OMW and maximizing the valuable product yield. Moreover the process is readily suitable for an industrial scale PHB production from OMW.

STUDY ON BIODEGRADABILITY OF POLY (3-HYDROXYBUTYRATE-co-3-HYDROXYVALERATE)/ORGANOPHILIC MONTMORILLONITE NANOCOMPOSITES

Poly (3-hydroxybutyrate-co-3-hydroxyvalerate)/Organophilic montmorillonite (PHBV/OMMT) nanocomposites were prepared and the biodegradability of the PHBV/OMMT nanocomposites was studied by a cultivation degrading method in soil suspension. The relationship between structure and biodegradability of PHBV/OMMT nanocomposites was investigated. The results showed that the biodegradability of PHBV/OMMT nanocomposites decreased with increasing amount of OMMT and it was related to the number of PHBV degrading microorganisms in degradation environment, the anti-microbial property of OMMT and the degree of crystallinity of the nanocomposites.

THERMAL PROPERTY, MISCIBILITY AND CRYSTALLIZATION BEHAVIOR OF POLY(3-HYDROXYBUTYRATE-co-3-YDROXYHEXANOATE) AND POLY(BUTYLENE SUCCINATE-ADIPATE) (PHBHHX/PBSA) BLENDS

Blends of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) and poly(butylene succinate-adipate) (PBSA), both biodegradable semicrystalline polyesters, were prepared with the ratio of PHBHHx/PBSA ranging from 80/20 to 20/80 by melt mixing method. Differential scanning calorimetry (DSC), thermogravimetry analysis (TGA), dynamic mechanical thermal analysis (DMA), polarizing optical microscopy (POM) and wide angle X-ray diffractometer (WAXD) were used to study the miscibility and crystallization behavior of PHBHHx/PBSA blends. Experimental results indicate that PHBHHx is immiscible with PBSA as shown by the almost unchanged glass transition temperature and the biphasic melt.

Preparation of Poly(3-hydroxybutyrate) Porous Films by Microemulsion Templates Method

Porous poly(3 hydroxybutyrate)(PHB) membranes were prepared by microemulsion templates. The influence of microemulsion templates on pore size and porosity of the PHB membranes were studied. Preliminary studies showed that the pore size made by microemulsion templates were smaller and were estimated in the range of 100~500 nanometer. In this research, the ultraviolet/visible light spectrometer was applied in the investigation of H 2O/Span85 Tween60/chloroform reserve microemulsion. Surfactant content(P) and water content(R) were optimized. Furthermore, the effects of variation of P and R on the pore size and porosity of PHB films were discussed.

生物可降解PPC/P34HB共混物制备与性能

采用熔融共混的方法制备了不同质量比的聚碳酸亚丙酯/聚(3-羟基丁酸酯-co-4-羟基丁酸酯)共混物(PPC/P34HB),分析了P34HB添加量对共混物性能的影响。结果表明,PPC共混物的韧性随着P34HB的加入逐渐增大,当其添加量为30%时,冲击强度提升了271.4%。由复合材料流变性能测试可知,P34HB由于含有侧甲基,将其引入体系中提高了共混物的储能模量和损耗模量,降低了其复数黏度,改善了材料的加工性能。热失重结果表明,P34HB的引入,提高了共混物的初始分解温度。由其微观形貌可知,断面表面从光滑表面最后转变为“海岛”结构,证明材料韧性得到了改善。

聚(3-羟基丁酸酯-4-羟基丁酸酯)/聚乳酸纤维的制备及其性能

聚(3-羟基丁酸酯-4-羟基丁酸酯)(P34HB)是极具前景的生物基可降解材料,但结晶慢且加工黏度大等因素使其难以直接熔纺制备纤维。左旋聚乳酸(PLLA)与P34HB化学结构相似,加工温度接近,与P34HB具有协同增强作用。采用熔融纺丝法制备P34HB含量为30%的P34HB/PLLA纤维,研究纤维的力学性能、热稳定性能以及结晶性能。结果发现,2000 m/min的卷绕速率下制备的P34HB/PLLA预牵伸纤维(POY)经1.40、1.75倍牵伸和定形后得到的全牵伸纤维(FDY-1和FDY-2)的断裂强度分别为1.66、2.29 cN/dtex,断裂伸长率分别为33.3%、28.8%;差示扫描量热仪(DSC)测试结果显示,POY、FDY-1、FDY-2结晶度分别为62.51%、62.05%、61.36%;XRD衍射仪测试结果也表明POY的结晶度与FDY结晶度相近,这表明在POY成形过程中,纤维中的结晶已经接近完成,P34HB提高了PLLA结晶效率。

基于Caspase-3/PARP通路探究龟鹿二仙胶及拆方对IL-1β诱导的退变软骨细胞凋亡及细胞外基质的影响

目的 基于半胱氨酸蛋白酶3/多聚腺苷二磷酸核糖聚合酶(Caspase-3/PARP)通路探究龟鹿二仙胶及拆方对白细胞介素-1β(IL-1β)诱导的退变软骨细胞凋亡及细胞外基质(ECM)的影响。方法 采用SD大鼠制备空白血清、龟鹿二仙胶含药血清、龟甲胶含药血清及鹿角胶含药血清,采用酶消法体外培养C57BL/6小鼠软骨细胞。将软骨细胞分为5组,空白组加入空白血清培养,模型组加入IL-1β和空白血清培养,龟鹿组加入IL-1β和龟鹿二仙胶含药血清培养,龟板组加入IL-1β和龟甲胶含药血清培养,鹿角组加入IL-1β和鹿角胶含药血清培养,干预24 h后,采用CCK-8法检测软骨细胞活性,采用TUNEL法检测软骨细胞凋亡情况,采用免疫荧光染色法检测Caspase-3、PARP-1表达情况,分别采用Western blot和qRT-PCR法检测细胞中Caspase-3、PARP-1、基质金属蛋白酶-13(MMP-13)、聚集蛋白聚糖(Aggrecan)蛋白及mRNA表达情况。结果 与空白组比较,模型组软骨细胞活性明显降低(P<0.05),软骨细胞凋亡率明显升高(P<0.05);细胞中Caspase-3、PARP-1平均荧光强度和Caspase-3、PARP-1、MMP-13蛋白及mRNA相对表达量均明显升高(P均<0.05),细胞中Aggrecan蛋白及mRNA相对表达量均明显降低(P均<0.05)。与模型组比较,龟鹿组、龟板组及鹿角组软骨细胞活性均明显升高(P均<0.05),软骨细胞凋亡率均明显降低(P均<0.05);细胞中Caspase-3、PARP-1平均荧光强度和Caspase-3、PARP-1、MMP-13蛋白及mRNA相对表达量均明显降低(P均<0.05),细胞中Aggrecan蛋白及mRNA相对表达量均明显升高(P均<0.05)。与龟板组、鹿角组比较,龟鹿组软骨细胞活性更高(P均<0.05),软骨细胞凋亡率更低(P均<0.05);细胞中Caspase-3、PARP-1平均荧光强度和Caspase-3、PARP-1、MMP-13蛋白及mRNA相对表达量均更低(P均<0.05),细胞中Aggrecan蛋白及mRNA相对表达量均更高(P均<0.05)。与龟板组比较,鹿角组软骨细胞活性,软骨细胞凋亡率,细胞中Caspase-3、PARP-1平均荧光强度,细胞中Caspase-3、PARP-1、MMP-13、Aggrecan蛋白及mRNA相对表达量高均更高(P均<0.05)。结论 龟板胶可抑制软骨细胞凋亡及ECM降解,鹿角胶可促进软骨细胞增殖和ECM合成代谢,龟鹿二仙胶作用效果优于龟板胶及鹿角胶,作用机制可能与激活Caspase-3/PARP信号通路有关。

SYNTHESIS OF NOVEL BLOCK COPOLYMERS OF POLY(3-HYDROXYBUTYRIC ACID)WITH POLY(ETHYLENE GLYCOL)THROUGH ANIONIC POLYMERISATION

A novel kind of copolymer with ABA-type block structure was synthesized by anionic ring-opening polymerization of beta-butyrolactone (beta-BL) in the presence of a PEG-based dicarboxylates as macroinitiators which were prepared by the esterification of aliphatic cyclic anhydride and poly(ethylene glycol) (PEG) oligomers (M-n = 2000, 4000 and 6000) and conversion of potassium dicarboxylates. The resultant copolymers as well as the intermediates were characterized by IR, H-1-NMR and GPC.

Agro-Industrial Waste Materials as Substrates for the Production of Poly(3-Hydroxybutyric Acid)

Accumulation of recalcitrant plastics in the environment has become a world-wide problem in today’s societies. Rapid depletion of natural resources for synthetic plastics along with environmental concerns has directed research towards finding alternatives to petroleum-based polymers. Poly(3-hydroxybutyric acid) P(3HB), as one of these alternatives, have attracted much attention in recent years due to their varied mechanical properties, biocompatibility and biodegradability. The aim of this study was to identify an agro-industrial waste resource economically suitable for large-scale production of P(3HB), to optimize the production using Response Surface Methodology in small-scale and subsequently, to test the production in a continuously stirred tank reactor. Among a range of agro-industrial waste, orange peel was selected as the most suitable for P(3HB) production. P(3HB) concentration of 1.24 g P(3HB)/L culture broth with 41% P(3HB)/dcw yield was obtained using orange peel as the sole carbon source in optimized medium with a modified strain of Bacillus subtilis (B. subtilis OK2).

Novel sandwich structured glass fiber Cloth/Poly(ethylene oxide)-MXene composite electrolyte

Recently,poly(ethylene oxide)(PEO)-based solid polymer electrolytes have been attracting great attention,and efforts are currently underway to develop PEO-based composite electrolytes for next generation high performance all-solid-state lithium metal batteries.In this article,a novel sandwich structured solid-state PEO composite electrolyte is developed for high performance all-solid-state lithium metal batteries.The PEO-based composite electrolyte is fabricated by hot-pressing PEO,LiTFSI and Ti_(3)C_(2)T_(x) MXene nanosheets into glass fiber cloth(GFC).The as-prepared GFC@PEO-MXene electrolyte shows high mechanical properties,good electrochemical stability,and high lithium-ion migration number,which indicates an obvious synergistic effect from the microscale GFC and the nanoscale MXene.Such as,the GFC@PEO-1 wt%MXene electrolyte shows a high tensile strength of 43.43 MPa and an impressive Young's modulus of 496 MPa,which are increased by 1205%and 6048%over those of PEO.Meanwhile,the ionic conductivity of GFC@PEO-1 wt%MXene at 60℃ reaches 5.01×10^(-2) S m^(-1),which is increased by around 200%compared with that of GFC@PEO electrolyte.In addition,the Li/Li symmetric battery based on GFC@PEO-1 wt%MXene electrolyte shows an excellent cycling stability over 800 h(0.3 mA cm^(-2),0.3 mAh cm^(-2)),which is obviously longer than that based on PEO and GFC@PEO electrolytes due to the better compatibility of GFC@PEO-1 wt%MXene electrolyte with Li anode.Furthermore,the solid-state Li/LiFePO_(4) battery with GFC@PEO-1 wt%MXene as electrolyte demonstrates a high capacity of 110.2–166.1 mAh g^(-1) in a wide temperature range of 25–60C,and an excellent capacity retention rate.The developed sandwich structured GFC@PEO-1 wt%MXene electrolyte with the excellent overall performance is promising for next generation high performance all-solid-state lithium metal batteries.

Use of a Simple Fabrication Process to Produce a Biosensor： The 3-Hydroxybutyrate Dehydrogenase Case

This study was aimed to construct a biodegradable but reliable 3-β-hydroxybutymte biosensor. In this context a versatile paper based biosensor, quickly, easily and cheaply fabricated is reported. The procedure of fabrication is based on the assumption that the introduction of the enzyme in the carbon ink will allow enzyme stabilization and facilitate the study of the catalysis of enzymes and the detection of substrates. To prove this concept we use the enzyme 3-hydroxybutyrate dehydrogenase, in aqueous solution. This enzyme was chosen because it catalyzes the 3-β-hydroxybutyrate, which results from ketoacidosis. The quantification this substance in the diabetics＇ blood is very important as it can increase the reliability of the diagnosis of glycaemia. To prove the multi-use of this biosensor we not only study the redox process in steady state and during the catalytic process, but also detected and quantify the 3-β-hydroxybutyrate. Our results showed that it was possible to study the redox process that occurred during the catalysis and to confirm the amino acid residues that participate in it. It was also observed that glucose and ascorbic acid can interfere in the detection and quantification of the 3-β-hydroxybutyrate, what should be in mind when the quantification of the 3-β-hydroxybutyrate is made in blood samples.

Effect of Nanostructures Addition and Enhancement of Poly (Vinylidene Difluoride) (PVDF) Energy Harvesting

With concerns in energy crisis and global warming, researchers are actively investigating alternative energy renewable solutions. Among the various methods, piezoelectric transduction stands out due to its impressive electromechanical coupling factor and coefficient. As a result, piezoelectric energy harvesting has garnered significant attention from the scientific community. In this study, we explored methods to enhance the piezoelectric properties of polyvinylidene fluoride (PVDF) through two distinct approaches. The first approach involved applying external high voltages at various stages during the mixture reaction. The goal was to determine whether this voltage application could alter or enhance PVDF’s piezoelectric conformation by improving the alignment of polarized dipoles. In the second part of our study, we investigated the effects of incorporating various nanostructures (including Iron Oxide, Magnesium Oxide, and Zinc Oxide) into PVDF. To analyze changes in PVDF’s crystalline structure, we utilized Fourier Transform Infrared Spectroscopy (FTIR) and X-ray Diffraction (XRD) techniques. Additionally, we measured the electric polarization of samples using a Precision LC Meter and examined the morphology of nanofibers through Scanning Electron Microscopy (SEM).

Al(OH)_3、ISA206及Poly(I:C)免疫佐剂对灭活口蹄疫140S抗原免疫效果的影响

目的选取Al(OH)3、ISA206及Poly(I:C)3种佐剂,分别与灭活口蹄疫病毒(FMDV)配制成疫苗,并通过小鼠实验比较和评价3种佐剂的免疫效力。方法以口蹄疫O型病毒为毒种,经细胞培养传代,获得较高滴度的病毒培养物,并经化学方法灭活制成140S口蹄疫完整病毒粒子抗原。在蔗糖密度梯度定量后分别与Al(OH)3、ISA206及Poly(I:C)3种佐剂配合制作口蹄疫灭活疫苗。疫苗经肌肉注射免疫Balb/c小鼠,通过检测抗体效价评价机体产生的体液免疫反应。通过检测体外刺激免疫小鼠脾淋巴细胞后产生的细胞因子以及淋巴细胞增殖实验评价机体产生的细胞免疫反应。结果 ISA206口蹄疫灭活苗和Poly(I:C)口蹄疫灭活苗诱导小鼠产生的抗体效价较高;通过体外刺激实验发现,ISA206口蹄疫灭活苗免疫组小鼠脾淋巴细胞在抗原刺激后,产生的IFN-γ和IL-4含量最高;通过淋巴细胞增殖实验发现,ISA206口蹄疫灭活苗免疫组小鼠脾淋巴细胞的增殖能力最强(P<0.05)。结论ISA206佐剂在配合灭活口蹄疫140S完整病毒粒子进行免疫时所表现的免疫效力最高。