Comparative study of three types of polymer materials co-cultured with bone marrow mesenchymal stem cells for use as a myocardial patch in cardiomyocyte regeneration

The purpose of this study was to investigate the most suitable polymer material for supporting stem cell growth as a myocardial patch. After cell isolation and expansion of mouse bone marrow mesenchymal stem cells (BMSC), the cells were induced to differentiate into cardiomyocytes with 5-azacytidine to determine their differentiation potential. BMSCs were also seeded onto three types of polymer material film, including polyurethane (PU), 3-hydroxybutyrate-co-4-hydroxybutyrate [P(3HB-co-4HB)], and polypropylene carbonate (PPC). The results revealed that cell numbers were more abundant on both the PU and P(3HB-co-4HB) material surfaces. Conversely, the surface of PPC was smooth with only cell lysate debris observed. The average cell counts were as follows: 143.78 ± 38.38 (PU group), 159.50 ± 33.07 [P(3HB-co-4HB) group], and 1.40 ± 0.70 (PPC group). There was no statistically significant difference in cell numbers between the PU and P(3HB-co-4HB) groups. A statistically significant difference was identified between the PPC group and both the PU (P1) and P(3HB-co-4HB) groups (P2). Polymer biomaterial patches composed of PU and P(3HB-co-4HB) permit good stem cell growth. P(3HB-co-4HB) has the potential for development as a clinical alternative to current treatment methods for the regeneration of cardiomyocytes in patients with myocardial infarction.     

聚丁二酸丁二醇酯/聚羟基烷酸酯熔融共混物的结晶及流变力学行为

用熔融共混法制备聚丁二酸丁二醇酯(PBS)/聚(3-羟基丁酸酯-co-4-羟基丁酸酯)[Poly(3HB-co-4HB)]复合降解材料,利用差示扫描量热(DSC)、旋转流变仪及万能拉力机对其结晶、流变行为及力学性能进行研究。结果表明,在PBS中加入Poly(3HB-co-4HB)后,发现结晶起始温度(To,c)、结晶峰温度(Tp,c)以及结晶结束温度(Te,c)有所提高,结晶度随着Poly(3HB-co-4HB)的增加呈先增大后下降的趋势;PBS/Poly(3HB-co-4HB)复合降解材料随着Poly(3HB-co-4HB)添加量的增大,断裂伸长率和拉伸强度却呈下降趋势;同时剪切储能模量(G′)、剪切损耗模量(G″)呈现出单增趋势。因此,在复合降解材料中添加适量的Poly(3HB-co-4HB)能改善PBS的结晶、流变及力学行为。     

聚(3-羟基丁酸酯-co-4-羟基丁酸酯)@海藻酸钠复合纤维膜的制备及其对Pb2+和Cu2+的吸附性

采用三氯甲烷和N,N二甲基甲酰胺（DMF）为溶剂,制备聚（3-羟基丁酸酯-co-4-羟基丁酸酯）（P（3HB-co-4HB））纺丝溶液,并通过静电纺丝技术制备P（3HB-co-4HB）纳米纤维膜,用海藻酸钠（SA）对P（3HB-co-4HB）纳米纤维膜进行包覆,获得P（3HB-co-4HB）@SA复合纤维膜。利用SEM、比表面积仪、原子吸收光谱分别表征了P（3HB-co-4HB）@SA复合纤维膜的纤维形态、比表面积、溶液残留液离子浓度。结果表明:纺丝液浓度在12%时,P（3HB-co-4HB）纤维成纤性好;随着静电压增大,P（3HB-co-4HB）纤维直径先减小后增大。P（3HB-co-4HB）支架材料可以使SA的比表面积提高约3.9倍,P（3HB-co-4HB）@SA复合纤维膜对Cu2+离子、Pb2+离子最大吸附量分别为26.25 mg/g和36.25 mg/g,折算为SA的吸附量分别为364.58 mg/g和503.47 mg/g。      

Isothermal crystallization kinetics and morphology of biodegradable poly(3-hydroxybutyrate-co-4-hydroxybutyrate)

Isothermal crystallization kinetics and morphology of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] with different 4-hydroxybutyrate (4HB) molar fraction were investigated by differential scanning calorimetry (DSC), wide angle X-ray diffraction (WAXD), and polarized optical microscopy (POM). The results show that the crystallization mechanism and crystal structure of P(3HB-co-4HB) copolymers are the similar as those of poly(3-hydroxybutyrate) (PHB). While the equilibrium melting point and crystallization rate decrease with the increase of 4HB molar fraction. Banded spherulites are observed in neat PHB and P(3HB-co-4HB) copolymers, and morphology is influenced apparently by the crystallization temperature and 4HB unit.     

Microparticles prepared from biodegradable polyhydroxyalkanoates as matrix for encapsulation of cytostatic drug

Microparticles made from degradable polyhydroxyalkanoates of different chemical compositions a homopolymer of 3-hydroxybutyric acid, copolymers of 3-hydroxybutyric and 4-hydroxybutyric acids (P3HB/4HB), 3-hydroxybutyric and 3-hydroxyvaleric acids (P3HB/3HV), 3-hydroxybutyric and 3-hydroxyhexanoic acids (P3HB/3HHx) were prepared using the solvent evaporation technique, from double emulsions. The study addresses the influence of the chemical compositions on the size and ξ-potential of microparticles. P3HB microparticles loaded with doxorubicin have been prepared and investigated. Their average diameter and ξ-potential have been found to be dependent upon the level of loading (1, 5, and 10 % of the polymer mass). Investigation of the in vitro drug release behavior showed that the total drug released from the microparticle into the medium increased with mass concentration of the drug. In this study mouse fibroblast NIH 3T3 cells were cultivated on PHA microparticles, and results of using fluorescent DAPI DNA stain, and MTT assay showed that microparticles prepared from PHAs of different chemical compositions did not exhibit cytotoxicity to cells cultured on them and proved to be highly biocompatible. Cell attachment and proliferation on PHA microparticles were similar to those on polystyrene. The cytostatic drug encapsulated in P3HB/3HV microparticles has been proven to be effective against HeLa tumor cells.     

4-羟基丁酸含量对聚 3-羟基丁酸 与 4-羟基丁酸共聚物性能和结构的影响

目的对3-羟基丁酸4-羟基丁酸共聚酯P(3HB-co-4HB)进行酶解测试。方法通过力学性能测试、失重率分析、热失重分析、X-射线衍射分析、扫描电镜等测试和表征手段对样品的物理性能及生物降解情况进行评价。结果 P(3HB-co-4HB)分子中4HB单体(文中均用摩尔分数表示)的引入提高了材料的柔韧性,材料的脆性下降;失重率方面,4种材料的降解速率从高到低依次为P(3HB)P(3HB-co-5%4HB)(3HB-co-10%4HB)P(3HB-co-15%4HB);酶解前期,材料的热稳定性增强,而酶解后期材料的热稳定性逐渐下降;XRD结果表明材料降解过程中结晶度的变化不明显;P(3HB-co-4HB)分子中随着4HB单体含量的增加,材料表面粗糙度降低,酶解后材料表面被侵蚀,降解速率与失重率结果一致。结论 P(3HB-co-4HB)分子中4HB单体的引入显著影响了材料的机械性能,随着4HB含量的增加,材料的失重率越来越大,热稳定性呈现先上升后下降的趋势,材料表面粗糙度逐渐降低,由于酶解过程属于从表面侵蚀开始,因此酶解过程中样品的结晶度变化不大。     

Medium chain length polyhydroxyalkanoates, promising new biomedical materials for the future

Medium chain length polyhydroxyalkanoates, mcl-PHAs (C₆-C₁₄ carbon atoms), are polyesters of hydroxyalkanoates produced mainly by fluorescent Pseudomonads under unbalanced growth conditions. These mcl-PHAs which can be produced using renewable resources are biocompatible, biodegradable and thermoprocessable. They have low crystallinity, low glass transition temperature, low tensile strength and high elongation to break, making them elastomeric polymers. Mcl-PHAs and their copolymers are suitable for a range of biomedical applications where flexible biomaterials are required, such as heart valves and other cardiovascular applications as well as matrices for controlled drug delivery. Mcl-PHAs are more structurally diverse than short chain length PHAs and hence can be more readily tailored for specific applications. Composites have also been fabricated using mcl-PHAs and their copolymers, such as poly (3-hydroxyoctanoate) [P(3HO)] combined with single walled carbon nanotubes and poly(3-hydroxbutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] combined with hydroxyapatite. Because of these attractive properties of biodegradability, biocompatibility and tailorability, Mcl-PHAs and their composites are being increasingly used for biomedical applications. However, studies remain limited mainly to P(3HO) and the copolymer P(3HB-co-3HHx), which are the only mcl-PHAs available in large quantities. In this review we have consolidated current knowledge on the properties and biomedical applications of these elastomeric mcl-PHAs, their copolymers and their composites.     

Experimental wound dressings of degradable PHA for skin defect repair

The present study reports construction of wound dressing materials from degradable natural polymers such as hydroxy derivatives of carboxylic acids (PHAs) and 3-hydroxybutyrate/4-hydroxybutyrate [P(3HB/4HB)] as copolymer. The developed polymer films and electrospun membranes were evaluated for its wound healing properties with Grafts—elastic nonwoven membranes carrying fibroblast cells derived from adipose tissue multipotent mesenchymal stem cells. The efficacy of nonwoven membranes of P(3HB/4HB) carrying the culture of allogenic fibroblasts was assessed against model skin defects in Wistar rats. The morphological, histological and molecular studies revealed the presence of fibroblasts on dressing materials which facilitated wound healing, vascularization and regeneration. Further it was also observed that cells secreted extracellular matrix proteins which formed a layer on the surface of membranes and promoted the migration of epidermal cells from the neighboring tissues surrounding the wound. The wounds under the P(3HB/4HB) membrane carrying cells healed 1.4 times faster than the wounds under the cell-free membrane and 3.5 times faster than the wounds healing under the eschar (control).The complete wound healing process was achieved at Day 14. Thus the study highlights the importance of nonwoven membranes developed from degradable P(3HB/4HB) polymers in reducing inflammation, enhancing angiogenic properties of skin and facilitating better wound healing process.     

成核剂对聚(3-羟基丁酸酯-co-4羟基丁酸酯)性能的影响

分别以氮化硼(BN)、BRUGGOLENP250、CaCO3和Tm-3为成核剂,用熔融模压法制备了聚(3-羟基丁酸酯-co-4羟基丁酸酯)[P(3HB-co-4HB)]样品,借用偏光显微镜(POM)、差示扫描量热(DSC)、热重分析(TGA)和扫描电镜(SEM)等考察了成核剂种类及用量对P(3HB-co-4HB)结晶形态、熔点、热分解温度、力学性能及断面形态的影响。结果表明,各种成核剂均能有效细化P(3HB-co-4HB)的球晶尺寸,提高其熔点及热分解温度;当成核剂BN的质量分数为5‰~8‰时,P(3HB-co-4HB)的综合性能最好。     

Layer‐by‐Layer Hydrogen‐Bonded Polymer Films: From Fundamentals to Applications

Recent years have seen increasing interest in the construction of nanoscopically layered materials involving aqueous‐based sequential assembly of polymers on solid substrates. In the booming research area of layer‐by‐layer (LbL) assembly of oppositely charged polymers, self‐assembly driven by hydrogen bond formation emerges as a powerful technique. Hydrogen‐bonded (HB) LbL materials open new opportunities for LbL films, which are more difficult to produce than their electrostatically assembled counterparts. Specifically, the new properties associated with HB assembly include: 1) the ease of producing films responsive to environmental pH at mild pH values, 2) numerous possibilities for converting HB films into single‐ or two‐component ultrathin hydrogel materials, and 3) the inclusion of polymers with low glass transition temperatures (e.g., poly(ethylene oxide)) within ultrathin films. These properties can lead to new applications for HB LbL films, such as pH‐ and/or temperature‐responsive drug delivery systems, materials with tunable mechanical properties, release films dissolvable under physiological conditions, and proton‐exchange membranes for fuel cells. In this report, we discuss the recent developments in the synthesis of LbL materials based on HB assembly, the study of their structure–property relationships, and the prospective applications of HB LbL constructs in biotechnology and biomedicine.     

GF/P(3HB-co-4HB)-PLA生物基复合材料的制备与性能

为了提高聚（3-羟基丁酸酯-co-4 羟基丁酸酯）-聚乳酸（P（3HB-co-4HB）-PLA）生物基共混材料的力学性能和尺寸稳定性,扩大应用领域,以P（3HB-co-4HB）和PLA共混物为基体,盐酸或偶联剂表面处理的玻璃纤维（GF）为增强材料,采用熔融共混法制备GF/P（3HB-co-4HB）-PLA复合材料。通过傅里叶变换红外光谱仪（FTIR）、扫描电子显微镜（SEM）、热失重分析仪（TGA）和万能电子拉力机等研究了GF表面处理方法对复合材料力学性能、热性能、尺寸稳定性及断面形态的影响。研究结果表明：表面改性GF的加入可显著提高P（3HB-co-4HB）-PLA共混材料的综合性能。经偶联剂表面接枝的GF可均匀分散在P（3HB-co-4HB）-PLA基体中并形成较强的界面结合。添加质量分数20%的偶联剂改性GF使复合材料的拉伸强度、弯曲强度、缺口冲击强度和硬度分别提高了29.38%、20.32%、41.38%和15.31%;初始热分解温度（IDT）和维卡软化温度（VST）分别提高了6.64 ℃和10.7 ℃;室温和60 ℃放置60 d后复合材料试样长度方向的尺寸稳定性分别提高了32.47%和33.70%。     

Incorporation of vitamin E in poly(3hydroxybutyrate)/Bioglass composite films: effect on surface properties and cell attachment

This study investigated the possibility of incorporating α-tocopherol (vitamin E) into poly(3hydroxybutyrate) (P(3HB))/Bioglass composites, which are being developed for bone tissue engineering matrices. P(3HB) films with 20 wt% Bioglass and 10 wt% vitamin E were prepared using the solvent casting technique. Addition of vitamin E significantly improved the hydrophilicity of the composites along with increasing the total protein adsorption. The presence of protein adsorbed on the composite surface was further confirmed using X-ray photoelectron spectroscopy analysis. Preliminary cell culture studies using MG-63 human osteoblasts showed that the addition of vitamin E in the P(3HB)/20 wt% Bioglass films significantly increased cell proliferation. The results achieved in this study confirmed the possibility of incorporating vitamin E as a suitable additive in P(3HB)/Bioglass composites to engineer the surface of the composites by promoting higher protein adsorption and increasing the hydrophilicity.     

聚(3-羟基丁酸酯-co-4-羟基丁酸酯)/POSS共混体系的性能

采用熔融模压法分别制备了聚(3-羟基丁酸酯-co-4-羟基丁酸酯)[P(3HB-co-4HB)]和两种多面体笼型硅氧烷(POSS)[八异丁基倍半硅氧烷(OIBS)和八氨基苯基倍半硅氧烷(OAPS)]的共混物,考察了不同含量的OIBS,OAPS对共混体系性能的影响。结果表明,两种POSS都起到成核剂的作用。OIBS,OAPS的质量分数小于1%时,可以提高体系结晶温度,力学性能;随着OIBS,OAPS质量分数的提高,成核性有所增加,但因分散性变差,体系热稳定性和力学性能变差。由于OAPS的活泼氨基可与P(3HB-co-4HB)发生化学反应,改性效果较OIBS优。     

SiO2/聚（3-羟基丁酸酯-co-4-羟基丁酸酯）薄膜研究

目的研究纳米SiO2对可生物降解聚(3-羟基丁酸酯-co-4-羟基丁酸酯)(P34HB)包装膜结晶行为和力学性能的影响。方法采用溶液浇铸法制备SiO_2/P34HB纳米复合薄膜,利用红外光谱仪(FTIR)、扫描电镜(SEM)、正置热台显微镜(POM)、差示扫描量热仪(DSC)和万能力学试验机等研究纳米SiO_2对P34HB结构、结晶性和力学性能等的影响。结果纳米SiO_2在P34HB中起到异相成核的作用,SiO2/P34HB复合膜的结晶速率和结晶度得到明显改善。相比P34HB包装膜,当纳米SiO_2质量分数为2%时,SiO_2/P34HB复合膜的弹性模量和拉伸强度分别提高了72.7%和60.9%。结论获得了纳米SiO2改善可生物降解聚(3-羟基丁酸酯-co-4-羟基丁酸酯)包装膜结晶度和力学性能的最佳掺杂比例参数。     

Effect of nanoparticulate bioactive glass particles on bioactivity and cytocompatibility of poly(3-hydroxybutyrate) composites

This work investigated the effect of adding nanoparticulate (29 nm) bioactive glass particles on the bioactivity, degradation and in vitro cytocompatibility of poly(3-hydroxybutyrate) (P(3HB)) composites/nano-sized bioactive glass (n-BG). Two different concentrations (10 and 20 wt %) of nanoscale bioactive glass particles of 45S5 Bioglass composition were used to prepare composite films. Several techniques (Raman spectroscopy, scanning electron microscopy, atomic force microscopy, energy dispersive X-ray) were used to monitor their surface and bioreactivity over a 45-day period of immersion in simulated body fluid (SBF). All results suggested the P(3HB)/n-BG composites to be highly bioactive, confirmed by the formation of hydroxyapatite on material surfaces upon immersion in SBF. The weight loss and water uptake were found to increase on increasing bioactive glass content. Cytocompatibility study (cell proliferation, cell attachment, alkaline phosphatase activity and osteocalcin production) using human MG-63 osteoblast-like cells in osteogenic and non-osteogenic medium showed that the composite substrates are suitable for cell attachment, proliferation and differentiation.     

Microbial Polyhydroxyalkanoates and Nonnatural Polyesters

Microorganisms produce diverse polymers for various purposes such as storing genetic information, energy, and reducing power, and serving as structural materials and scaffolds. Among these polymers, polyhydroxyalkanoates (PHAs) are microbial polyesters synthesized and accumulated intracellularly as a storage material of carbon, energy, and reducing power under unfavorable growth conditions in the presence of excess carbon source. PHAs have attracted considerable attention for their wide range of applications in industrial and medical fields. Since the first discovery of PHA accumulating bacteria about 100 years ago, remarkable advances have been made in the understanding of PHA biosynthesis and metabolic engineering of microorganisms toward developing efficient PHA producers. Recently, nonnatural polyesters have also been synthesized by metabolically engineered microorganisms, which opened a new avenue toward sustainable production of more diverse plastics. Herein, the current state of PHAs and nonnatural polyesters is reviewed, covering mechanisms of microbial polyester biosynthesis, metabolic pathways, and enzymes involved in biosynthesis of short-chain-length PHAs, medium-chain-length PHAs, and nonnatural polyesters, especially 2-hydroxyacid-containing polyesters, metabolic engineering strategies to produce novel polymers and enhance production capabilities and fermentation, and downstream processing strategies for cost-effective production of these microbial polyesters. In addition, the applications of PHAs and prospects are discussed.     

Biodegradation of non-porous films after submucoperiosteal implantation on the palate of Beagle dogs

The aim of this study is to investigate the in vivo behaviour of five different biodegradable films after submucoperiosteal implantation on the palate of Beagle dogs. Non-porous films of poly-(L-lactic) acid (=PLLA), high molecular weight poly-(L-lactic) acid (=HMW-PLLA), poly-(DL-lactic) acid (=PDLLA), poly(-caprolactone) (=PCL) and a copolymer of poly hydroxybutyrate 80%-hydroxyvalerate 20% (=PHB-co-HV 80/20) were implanted submucoperiostally on the palate of Beagle dogs. After 2, 4, 8 and 12 weeks in situ, the structure of the films and the tissue reactions were studied histologically. In terms of mechanical properties and tissue response, the PHB-co-HV film is the most suitable for use on dogs.     

可降解包装材料聚羟基烷酸酯合成工艺及影响因素的研究进展

目的综述可降解包装材料聚羟基烷酸酯(Polyhydroxyalkanoates,简称PHAs或PHA)的主要合成工艺、影响PHA产率的因素及其在包装工业领域上的应用研究进展。方法通过对国内外的研究现状和研究成果进行分析总结,介绍PHA的主要合成工艺,微生物法合成PHA产量的影响因素及PHA在包装方面的应用。结果 PHA合成工艺、碳源种类和浓度、氮磷源浓度、pH值、溶解氧浓度和温度对合成PHA的产量有很大影响,其中碳源种类不同还会使形成PHA的单体及单体在PHA中的比例不同,形成具有不同性质的PHA。PHA因其性能的多样性而广泛应用于包装领域。结论优化PHA的合成工艺及影响因素,能有效提高PHA产量或降低PHA生产成本。目前,以真实废弃物为底物的活性污泥系统生产PHA的研究还不够深入,由于PHA作为可降解的包装材料在包装领域具有显著的发展前景,因此,优化PHA合成工艺和影响因素是今后值得关注的重点方向。     

Pyroelectric polymer films

The objective of this investigation was to synthesize polymeric materials of high pyroelectric efficiency and to prepare thin test films suitable for use in pyroelectric vidicons.In our investigation of vinyl-addition polymers with highly polar pendant groups, we concentrated on atactic and isotactic substituted styrenes. Five atactic 4-substituted styrenes, poly(4-bromostyrene), poly(4-nitrostyrene), poly-(4-chlorostyrene), poly(4-methoxystyrene) and poly(4-fluorostyrene), were synthesized and evaluated, as were four isotactic 4-substituted styrenes, poly(4-bromostyrene), poly(4-chlorostyrene), poly(4-methoxystyrene) and poly(4-fluorostyrene).Hot-pressing was found to be a successful method of fabricating thick films of those polymers which did not undergo thermal decomposition near the melting point.Solvent casting of polymer films directly onto the test substrate proved to be the only practical method of handling brittle thin polymer films.Film formation by stretching and blowing was not found to be suitable for most of the polymers investigated.Laser evaporation was found to be a very useful technique for the preparation of polymer test films. Laser-evaporated poly(vinyl fluoride) showed the highest known pyroelectric coefficient for this material (1.8×10^-9C/cm² °K).None of the poly(4-substituted styrenes) showed any substantial pyro-electric behavior. This result can be attributed to their characteristics helical structure and lack of net permanent dipole moment.     

Polymer films with surfaces unmodified and modified by non-thermal plasma as new substrates for cell adhesion

The surface properties of biomaterials, such as wettability, polar group distribution, and topography, play important roles in the behavior of cell adhesion and proliferation. Gaseous plasma discharges are among the most common means to modify the surface of a polymer without affecting its properties. Herein, we describe the surface modification of poly(styrene) (PS) and poly(methyl methacrylate) (PMMA) films using atmospheric pressure plasma processing through exposure to a dielectric barrier discharge (DBD). After treatment the film surface showed significant changes from hydrophobic to hydrophilic as the water contact angle decreasing from 95° to 37°. All plasma-treated films developed more hydrophilic surfaces compared to untreated films, although the reasons for the change in the surface properties of PS and PMMA differed, that is, the PS showed chemical changes and in the case of PMMA they were topographical. Excellent adhesion and cell proliferation were observed in all films. In vitro studies employing flow cytometry showed that the proliferation of L929 cells was higher in the film formed by a 1:1 mixture of PS/PMMA, which is consistent with the results of a previous study. These findings suggest better adhesion of L929 onto the 1:1 PS/PMMA modified film, indicating that this system is a new candidate biomaterial for tissue engineering.