Hybrid poly-l-lactic acid/poly(ε-caprolactone) nanofibrous scaffold can improve biochemical and molecular markers of human induced pluripotent stem cell-derived hepatocyte-like cells

A suitable alternative strategy for liver transplantation is the use of nanofibrous scaffolds together with stem cells. In this study, a random hybrid of poly-l -lactic acid (PLLA) and poly(ε-caprolactone) (PCL) was used as a three-dimensional (3D) culture for differentiation of hepatocyte-like cells and compared with routine culture (two-dimensional [2D]). The expression of the endodermal marker, forkhead box A2 (FOXA2), was assessed on Day 3 and the hepatic markers; albumin (ALB), α-1 antitrypsin (AAT), and cytokeratin-18 (CK-18) were evaluated on Day 18 using quantitative polymerase chain reaction qPCR. As well as, ALB, α-fetoprotein (AFP), and low-density lipoprotein (LDL) uptake were evaluated using immunocytochemistry; moreover, periodic acid-Schiff and Oil Red were done by cell staining. In addition, AFP and urea production were evaluated by chemiluminescence and colorimetric assays. Light and scanning electron microscopy (SEM) showed changes in the cells in 2D and 3D models. The gene expression of hepatic markers was significantly higher in the 3D cultures. In addition, immunocytochemistry and cell staining showed that ALB, AFP, LDL-uptake, periodic acid-Schiff, and Oil Red were expressed in both cells derived on 2D and 3D. Furthermore, the evaluation of AFP and urea secretion was significantly different between 2D and 3D strategies. These findings suggest that functionally cells cultured on a PLLA/PCL scaffold may be suitable for cell therapy and regenerative medicine.     

Purification and characterization of cutinase from Bacillus sp. KY0701 isolated from plastic wastes

A total of approximately 400 bacterial strains were isolated from 73 plastic wastes collected from 14 different regions. Nineteen isolates that form clear zones both on tributyrin and poly ε-caprolactone (PCL) agar, were identified based on 16S rRNA gene sequences. Among these, Bacillus sp. KY0701 that caused the highest weight loss of PCL films in minimal salt medium, was selected for cutinase production. The highest enzyme activity (15 U/mL) was obtained after 4 days of incubation at 50°C, pH 7.0 and 200?rpm in a liquid medium containing 1.5% (w/v) apple cutin and 0.1% (w/v) yeast extract. The purified enzyme was stable at high temperatures (50–70°C) and over a wide pH range (5.5–9.0). The relative activity of cutinase was at least 75% in the percent of various organic solvents. The apparent K_m and V_max values of the cutinase for p-nitrophenyl butyrate were 0.72?mM and 336.8?µmol p-nitrophenol/h/g, respectively. In addition, it showed high stability and compatibility with commercial detergents. These features of cutinase obtained from Bacillus sp. KY0701 make it a promising candidate for application in the detergent and chemical industries. In our best knowledge, this is the first report for cutinase production and characterization produced by a Bacillus strain.     

Promoting osteogenic differentiation of human-induced pluripotent stem cells by releasing Wnt/β-catenin signaling activator from the nanofibers

Implants that can enhance the stem cells differentiation in the absence of the chemical osteogenic growth factors will attract the great interest of orthopedic scientists. Inorganic polyphosphate (poly-P), as a ubiquitous biological polymer, is one of the factors that can be an alternative for osteogenic growth factors via activating Wnt/β-catenin signaling. In this study, poly-P was incorporated at the blend of polycaprolactone (PCL)/poly (l -lactic acid) (PLLA) electrospun nanofibers and then osteogenic differentiation potential of human-induced pluripotent stem cells (iPSCs) was investigated by the important bone markers. 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide (MTT) and scanning electron microscopy results confirmed the biocompatibility of the fabricated nanofibers, while higher proliferation rate of iPSCs was detected in PCL-PLLA(poly-P) group compared with the PCL-PLLA and tissue culture plate groups. Alkaline phosphatase activity, calcium content, and gene expression results demonstrated that osteogenic differentiation of iPSCs was increased when cultured on PCL-PLLA(poly-P) in comparison with other groups. According to the results, PCL-PLLA(poly-P) could be considered as a promising candidate for use as bone implants.     

Bioaugmentation of carbofuran residues in soil by Burkholderia cepacia PCL3: A small-scale field study

A small-scale field study was conducted in 1 m × 1.25 m × 20 cm plots. In the soil with only indigenous microorganisms, carbofuran degradation was slow with a long half-life (t_1/2) of 127 d. Bioaugmenting the soil with the immobilized PCL3 could shorten the t_1/2 of carbofuran to 16 d. The t_1/2 rose to a significantly longer 28 d when the free cells of PCL3 were used instead of the immobilized cells. Viable cell counting of carbofuran degraders in soil indicated that the carbofuran degradation efficiency directly correlated to the number of introduced carbofuran degraders that survived in the system. PCL3 in free-cell form did not survive in the soil during the field operation; this suggested that the immobilization of PCL3 is needed in order to implement the bioaugmentation technique in a real environment.     

膜性高分子材料在兔眼部的生物相容性

目的评价聚羟基脂肪酸(polyhydroxyalkanoates,PHA)、聚乳酸(polylacetic acid,PLA)和聚己内酯(polycaprolactone,PCL)三种膜性高分子材料在兔眼部的生物相容性。方法将24只新西兰兔随机分为4组,每组6只。PHA、PLA、PCL为实验组,材料植入兔右眼结膜下。假手术组结膜下钝性分离,但不植入任何高分子材料。使用裂隙灯显微镜观察并记录植入后不同时间手术眼的反应并评分。裂隙灯下观察材料的吸收时间。术后4周和16周取眼球,行HE染色、Masson染色和天狼猩红染色分别定性观察组织结构和炎症细胞、胶原纤维和胶原纤维的亚型与排列方向。结果术眼刺激性评分等级各组均不高于"轻度刺激性"。结膜下吸收时间PHA、PLA和PCL组分别是16周,12周和大于16周。组织学观察术后4周PHA、PLA和PCL组均形成材料包裹囊腔,囊壁以纤维组织为主,伴有毛细血管形成和炎性细胞浸润,以中性粒细胞为主。胶原纤维染色与假手术组无明显差异,以Ⅰ型和Ⅲ型为主,大致呈平行排列。术后16周PHA和PLA组材料已不可查及,包裹囊腔结构不规则,而PCL材料整体可查及,包裹囊腔规则。各组未见毛细血管,偶见淋巴细胞浸润。胶原纤维与假手术组无明显差异,以Ⅰ型和Ⅲ型为主,仍大致呈平行排列。结论 PHA、PLA和PCL三种膜性高分子材料在兔眼具有较好的生物相容性,结膜下吸收时间分别是16周,12周和大于16周。     

Molecular cloning and characterization of a mannose-binding lectin gene from <Emphasis Type="Italic">Pinellia cordata</Emphasis>

Using RNA extracted from Pinellia cordata young leaves and primers designed according to the conserved regions of Araceae lectins, the full-length cDNA of Pinellia cordata agglutinin (PCL) was cloned by rapid amplification of cDNA ends (RACE). The full-length cDNA of pcl was 1,182 bp and contained a 768 bp open reading frame (ORF) encoding a lectin precursor of 256 amino acids. Through comparative analysis of pcl gene and its deduced amino acid sequence with those of other Araceae species, it was found that pcl encoded a precursor lectin with signal peptide. PCL is a mannose-binding lectin with three mannose-binding sites. Semi-quantitative RT-PCR analysis revealed that pcl is expressed in all tested tissues including leaf, stem and bulbil, but with the highest expression in bulbil. PCL protein was successfully expressed in Escherichia coli with the molecular weight expected.     

Poly ε -Caprolactone Microparticles Containing Biosurfactants: Optimization of Formulation Factors

The objective of this research was to optimize the formulation factors and evaluate the release profiles of ε -polycaprolactone microparticles containing rhamnolipid biosurfactant (RhBS). Microparticles were prepared by a water-in-oil-in-water emulsion solvent evaporation technique. Optimization was studied through the effects of the volumes and concentrations of the internal and external phases of the microparticles on percent yield, particle size, encapsulation efficiency, and biosurfactant loading. Manipulation of the formulation factors yielded microparticles that were statistically the same size and generally classified as small. An increase in the volume of the internal phase above 1 ml caused a general decrease in yield and encapsulation efficiency and an increase in biosurfactant loading. When the volume of the external phase increased above 50 ml, decreases in percent yield and encapsulation efficiency and increases in biosurfactant loading were observed. Formulations with the highest encapsulation efficiencies and percentage yield and the lowest biosurfactant loading efficiencies were selected for further evaluation in release studies. Release studies were conducted in 15 and 32 ppt artificial seawater and deionized water. After 30 days microparticle formulations gradually released 80% to 100% of the encapsulated RhBS in all release media, with no significant differences in release rates in the different release media.