3-D culture of human umbilical vein endothelial cells with reversible thermosensitive hydroxybutyl chitosan hydrogel.

The aim of this study was to present a non-trypsin 3D cell culture method with a reversible thermosensitive HBCS hydrogel. In this study, hydroxybutyl chitosan (HBCS) was synthesized by grafting hydroxybutyl groups on chitosan molecule chains. The prepared HBCS was water-soluble, and the reversible phase transformation temperature was 26 °C. Scanning electron microscope images illuminated the 3-D network of hydrogel formed irregular porous structure which ranged from 50-250 µm. Cell viability assay indicated that HBCS solution could promote the proliferation of human umbilical vein endothelial cells (HUVECs), and the boost of proliferation was enhanced with the increase of HBCS concentration. HBCS had no harm to the nitric oxide (NO) synthesis functionality of HUVECs. HUVECs could grow and reproduce inside the hydrogel, and showed good vitality after 14-days culture. Meanwhile, cells cultured inside the hydrogel could be passaged successively through the reversible phase transformation process of HBCS. The results revealed that HBCS have the potential to be used for 3-D cell culture without the use of trypsin.

Structural characterization of lignin-carbohydrate complexes from Chinese quince fruits extracted after enzymatic hydrolysis pretreatment.

Chinese quince fruit (CQF) contains abundant pectin; however, the pectin cannot be efficiently separated by conventional approaches because of strong lignin-carbohydrate complexes (LCC). In this study, to elucidate the structural characteristics of the original LCC formed by lignin and pectin in CQF, single and multiple enzymatic hydrolysis pretreatments were innovatively performed, and the resulting LCC preparations were comprehensively characterized using a series of techniques. The enzymatic hydrolysis pretreatments significantly increase the LCC yield, releasing LCC fractions with low molecular weights (Mw = 4660-8288 Da). LCC-4, isolated by pretreatment with cellulase plus xylanase, had the highest galacturonic acid content (15.5 %), followed by LCC-2 (isolated by xylanase pretreatment) of 14.0 %. In CQF, lignin develops lignin-carbohydrate (LC) bonds with pectin to form LCC, with phenyl-glycoside bond being the dominant linkage. Although the pectinase pretreatment reduced the pectin content, signals of the LC linkages in the 2D-HSQC spectra were enhanced. LCC-4 could be considered as the most representative of the original LCC in CQF due to its high pectin content and multiple LCC signals in the 2D-HSQC spectrum. The structural understanding of the original LCC in CQF will lay a foundation for designing appropriate methods for extracting pectin from CQF.

Structural changes of lignin-carbohydrate complexes (LCCs) from Chinese quince fruits during the sequential fractionation of pectic and hemicellulosic polysaccharides.

Chinese quince (Chaenomeles sinensis) fruits offer a potential source of pectin and hemicellulose. However, the existence of lignin-carbohydrate complexes (LCCs) can negatively impact the extraction of pectin and hemicellulose. In this work, LCCs were sequentially fractionated from Chinese quince during the removal of pectin and hemicellulose. The structures of LCCs were characterized by HPAEC, FT-IR, GPC, Py-GC/MS, TGA and 2D HSQC NMR. The results showed that the carbohydrate content and molecular weight of LCCs was found to be changed significantly after the removal of hemicellulose (KSH). The lignin in Björkman LCCs was found to be linked mainly to galactan and fructan, whereas the lignin LCC-AcOHs was found to be linked mainly to arabinan after the removal of KSH. The isolation of carbonate-soluble pectin (NSP) increased thermal stability of Björkman LCC fraction, however, the isolation of chelator-soluble pectin (CSP) increased the thermal stability of LCC-AcOHs. The S/G ratios of LCC-AcOHs increased and large amounts of S-type lignin released during sequential fractionation of pectin and hemicellulose. These results will be beneficial for understanding the mechanisms of pectin and hemicellulose isolation, thereby facilitating the potential application of Chinese quince as a valuable natural resource for food and other industries.

[Cone-beam computed tomography-synthesized cephalograms for evaluating the vertical dimension of occlusions].

OBJECTIVE: The accuracy of the occlusion vertical dimensions of edentulous Han patients from Yunnan province was compared and analyzed on the basis of cone-beam computed tomography (CBCT)-synthesized cephalograms, closest speaking space method, and interocclusal distance. METHODS: A database correlating the CBCT head lateral images of Han patients from Yunnan province with normal occlusal conditions was first constructed. Then, five edentulous Han patients aged 63-78 years old from Yunnan Province were selected. NNT.View software was used to measure and analyze hard tissue cephalometric radiographs that had been transformed by the CBCT marker. The radiographs were then combined with the normal population database for the assessment of occlusion vertical dimensions. The occlusion vertical dimensions determined on the basis of CBCT-synthesized cephalograms, the closest speaking space method, and the free-way space were analyzed. RESULTS: The closest speaking space method was used as the standard control group, the differences between seven methods and the closest speaking space method were analyzed. The seven methods include free-way space method and six CBCT-synthesized cephalograms methods (N-ANS/ANS-Me, S-Go/N-Me, ANS-Gn/N-ANS, ANS-FH/Me-FH, ANS-Xi-Pm, and CA/LA). The seven methods were highly consistent with the closest speaking space method (intraclass correlation coefficient>0.986). The absolute values of the differences between the methods of free-way space, N-ANS/ANS-Me, S-Go/N-Me and the closest speaking space method were lower than those of the other four groups (P<0.05), while the differences between ANS-FH/Me-FH and the closest speaking space method was higher than those other groups (P<0.05). CONCLUSIONS: CBCT-synthesized cephalograms, with the exception of ANS-FH/Me-FH, can provide references for the clinical evaluation of the occlusion vertical dimensions of patients with edentulous jaws.

Preparation of biocompatible chitosan grafted poly(lactic acid) nanoparticles.

Chitosan grafted poly(lactic acid) (CS-g-PLA) copolymer was synthesized and characterized by FT-IR and elemental analysis. The degree of poly(lactic acid) substitution on chitosan was 1.90 ± 0.04%. The critical aggregation concentration of CS-g-PLA in distilled water was 0.17 mg/ml. Three methods of preparing CS-g-PLA nanoparticles (diafiltration method, ultrasonication method and diafiltration combined with ultrasonication method) were investigated and their effect was compared. Of the three methods, diafiltration combined with ultrasonication method produced nanoparticles with optimal property in terms of size and morphology, with size ranging from 133 to 352 nm and zeta potential from 36 to 43 mV. Also, the hemolytic activity and cytotoxicity of the CS-g-PLA based nanoparticles was tested, and results showed low hemolysis rate (<5%) and no significant cytotoxicity effect of these nanoparticles.