Two poplar lignin-carbohydrate complexes (LCCs), LCC-48 and LCC-72, were isolated at different milling times, 48 h and 72 h, respectively. A new hydrogel carrier was prepared from these LCCs in the presence of polyethylene glycol diglycidyl ether as the crosslinker for cell (human hepatocyte (L-02)) culture. The effects of the structure of LCC on the carrier were investigated. The FT-IR spectra indicate that the two LCC samples were composed of lignin and polysaccharide, and showed a typical LCC structure. The galactose contents of LCC-48 and LCC-72 were 3.02% and 5.67%, respectively. The results of cell culture show that a large number of hepatocytes adhered to the porous carriers. Hepatocytes grown on the LCC carriers outperformed the control group in every observed category, including cell proliferation rate and metabolic activity. These results indicate that poplar LCC might be a great potential precursor of biological carriers for human hepatocytes culture.

Spherical biocarriers were prepared with lignin-carbohydrate complexes isolated from ginkgo (Ginkgo biloba L.) xylem. The specific surface and average pore size of the biocarriers were 17.15 m 2 g −1 and 21.59 nm, respectively. The carriers were stable in solution at pH 4.0∼9.5. Fourier transform infrared (FT-IR) spectrum indicated that the spherical carrier was composed of lignin and polysaccharides and had a typical lignin-carbohydrate complex (LCC) structure. The contents of galactose, lignin, and total sugar were 3.30%, 23.9%, and 64.62%, respectively, making the spherical biocarriers have good physical strength and compatible with hepatocytes. It was observed using a scanning electron microscopy (SEM) that liver cells adhered to the spherical biocarriers during culture. Cell counting indicated that the proliferation of liver cells in the experimental group was significantly higher than that of the control group. The albumin secretion (ALB) value and glucose consumption of the human hepatocytes were increased by 51.7% and 38.6%, respectively, by the fourth day when cultivated on the biocarriers. The results indicate that ginkgo LCC is very biocompatible and shows promise for the use as a biomaterial in the culture of human hepatocytes.

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Lignin-carbohydrate complexes (LCC) were isolated from poplar wood after ball-milling for 48 h and 72 h. The water-insoluble LCC-48 and LCC-72 were fractionated. Porous spherical composite bio-carriers were prepared by liquid-nitrogen freezing using the water-insoluble LCCs and cellulose nanocrystals (CNC). Fourier transform infrared spectroscopy (FT-IR) showed that the bio-carriers were composed of lignin moieties and polysaccharide units. Scanning electron microscopy (SEM) revealed that the outer surface and cut surface of the spherical carrier had many pores. The specific surface areas of the LCC-72/CNC and the LCC-48/CNC composite carriers were 24.2 m2/g and 28.9 m2/g, respectively, while their pore sizes were 28.6 nm and 33.6 nm, respectively. Both carrier samples had good stability at pH 4.6 to 9.5 for about 50 days. Human hepatocytes were cultured in vitro with the bio-carriers and the cells grew well. A large number of cells adhered to the porous bio-carriers, and the cells of the experimental group to which the carriers were added exhibited higher metabolic activities. The CNC improved the biocompatibility for human hepatocytes. In summary, spherical bio-carriers prepared from LCC/CNC composite displayed high biocompatibility and have potential applications in liver tissue engineering.