The objective of the present studywas to investigate the properties of chitosan–gelatin membranes or scaffolds, which were modified byincorporation of hyaluronic acid in the surface or bulk phase through co-crosslinking with N;N-(3-dimethylaminopropyl)-N0-ethyl carbodiimide (EDC) and N-hydroxysuccinimide (NHS) in 2-morpholinoethane sulfonic acid (MES) buffer. The comparative studyon properties of surface modification (HA(S)) and polyblend membranes (HA(C)) revealed that gelatin was enriched on the surface of HA(C), while hyaluronic acid was enriched on the surface of the HA(S). The HA(S) membranes made by surface modification method had a characteristic surface morphology. The corresponding scaffolds were prepared through freezedrying. The incorporation of hyaluronic acid improved flexibility and fibroblasts adhesion, while slowing down the rate of biodegradation of chitosan-gelatin scaffold. Human fibroblasts adhered and proliferated well on the membranes or scaffolds in vitro.

Chitosan-gelatin hybrid polymer network scaffolds were prepared via the freeze-drying technique by using the ice microparticle as a porogen. Monolayer and bilayer scaffolds were obtained by using different pre-freezing methods. The novel bilayer scaffolds were prepared via contact with 561C lyophilizing plate directly, then lyophilized. The properties of chitosan-gelatin scaffolds, such as microstructure, physical and mechanical and degradable properties, were studied. These results suggested that the porosity and pore size of the scaffolds could be modulated with thermodynamic and kinetic parameters of ice formation. The scaffolds prepared from chitosan and gelatin can be utilized as a promising matrix for tissue engineering.

Glucose was oxidized to generate a glucose dialdehyde and chitosan (CS) was hydrophobically modified with butyl bromide, octyl bromide and dexyl bromide. The analysis of IR and X-ray diffraction results of CS derivatives confirms that the hydrogen bonds and crystallinity were weakened by the incorporation of pendant alkyl. The permeability coefficient P and diffusion coefficient D for model drug vitamin B2 through oxidized glucose-crosslinked alkylated CS membrane were determined under different pH media. The results show that for the same alkylated CS in different pH media, P and D decrease withan increase in pH; for different alkylated CSs in acidic media, P and D diminish with the increase in the length of alkyl side, which is supposedly originated from the enhancement of hydrophobicity. In alkali medium, P and D show a rising trend with the increase in the length of alkyl chain, which might be related to the loose stacking of network as it occurs to shrink in alkali medium. The preliminary cytotoxicity assay indicates that oxidized glucose-crosslinked alkylate CS membrane is non-toxic in vitro.

A novel biodegradable hydroxyapatite/chitosan-gelatin network (HA/CS-Gel) composite of similar composition to that of normal human bone was prepared as a three-dimensional biomimetic scaffold by phase separation method for bone tissue engineering. Changing the solid content and the compositional variables of the original mixtures allowed control of the porosities and densities of the scaffolds. The HA granules were dispersed uniformly in the organic network with intimate interface contact via pulverizing and ultrasonically treating commercial available HA particles. Scaffolds of 90.6% porosity were used to examine the proliferation and functions of the cells in this three-dimensional microenvironment by culturing neonatal rat caldaria osteoblasts. Histological and immunohistochemical staining and scanning electron microscopy observation ndicated that the osteoblasts attached to and proliferated on the scaffolds. Extracellular matrices including collagen I and proteoglycan-like substrate were synthesized, while osteoid and bone-like tissue formed during the culture period. Furthermore, the cell/scaffold constructs had good biomineralization effect after 3 weeks in culture.

The objective of this study was to modify the surface of poly(d,l-lactic acid) (PDLLA) with different molecular weight of silk fibroins, and assess the effects of the modified surfaces on the functions of rat osteoblasts cultured in vitro. The properties of the modified PDLLA surface and the control one were investigated by contact angle and electron spectroscopy for chemical analysis (ESCA). The former indicated the variation of hydrophilicity and the latter suggested that the modified PDLLA film using silk fibroin is enriched with nitrogen atoms. The biocompatibility of the PDLLA film may be altered and in turn affects the seeded cell functions Therefore, attachment and proliferation of osteoblasts seeded on the modified PDLLA films and the control one were examined. Cell morphologies on these films were studied by scanning electron microscopy (SEM) and cell viability was evaluated by MTT assay. In addition, differentiated cell function was assessed by measuring the alkaline phosphatase (ALP) activity. These results suggest that the silk fibroin-modified PDLLA surface can improve the interaction between osteoblasts and the PDLLA films.

The object of this study was to investigate the relationship between the concentrations of HA solutions and the physicochemical properties and the biocompatibility of Cs-Gel-HA membranes. The addition of different concentrations of HA not only improved the wettability significantly and extended the degradation time of Cs-Gel-HA membranes, but also changed their mechanical properties. The concentration of HA had a significant influence on the biocompatibility of Cs-Gel-HA membranes. Results demonstrated that it was only the concentrations of HA in a certain range (0.01-0.1%), that could promote the cell adhesion, migration and proliferation, while the concentration of HA was above 0.1% it would either reduce or even inhibit these behaviors.

The objective of this study was to investigate the efficiency of two treatments for poly(d,l-lactic acid) (PDLLA) surface modification using silk fibroin, one chemical treatment and one physical treatment: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (WSC) and entrapment. The properties of control films, WSC-modified and entrapment-treated PDLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The water-contact angle measurement indicated the change of hydrophilicity and the ESCA analysis suggested that the modified PDLLA film using silk fibroin became enriched with nitrogen atoms. The biocompatibility of PDLLA film might be altered, which in turn would affect the functions of cells that were seeded on it. Therefore, attachment and proliferation of osteoblasts that were seeded on modified PDLLA films and control films were examined. Cell viability was evaluated by the MTT assay and differentiated cell function was assessed by measuring alkaline phosphatase activity. These results suggested that silk fibroin was used to modify PDLLA surface via WSC and that entrapment could improve the interactions between osteoblasts and PDLLA films. The entrapment treatment was more effective than WSC treatment to accomplish the goal of surface modification.

The objective of this study was to investigate the efficiency of two treatments for poly(l-lactic acid) (PLLA) surface modification with chitosan, via entrapment and coupling by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide. The properties of original PLLA films, chitosan-entrapped and coupled PLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The contact angle indicated the change in hydrophilicity and the ESCA data suggested that the modified PLLA films became enriched with nitrogen atoms. The cytocompatibility of modified PLLA films might be improved. Therefore, the attachment and proliferation of bovine articular chondrocyte seeded on modified PLLA films and control one were examined. A whole cell enzyme-linked immunosorbent assay (Cell ELISA) that detects the BrdU incorporation during DNA synthesis and collagen type II secretion was applied to evaluate the chondrocytes on different PLLA films and tissue culture plates. Cell viability was estimated by the MTT assay and cell function were assessed by measuring sulfated glycosaminoglycan secreted by chondrocytes. These results implied that chitosan used to modify PLLA surface through entrapment and coupling could enhance the chondrocyte adhesion, proliferation and function.

Our objective in this study was to investigate the efficiency of two treatments for poly (l-lactic acid) (PLLA) surface modification with gelatin, via entrapment and coupling, using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The properties of original PLLA, gelatin-entrapped, and coupled PLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The water contact angle indicated that the incorporation of gelatin resulted in a change in hydrophilicity, and the ESCA data suggested that the modified PLLA films became enriched with nitrogen atoms. The cytocompatibility of modified PLLA films might be improved. Therefore, we examined the attachment and proliferation of bovine articular chondrocyte seeded on modified PLLA films and virgin films. A whole-cell enzyme-linked immunosorbent assay (cell ELISA) that detects 5-bromo-2-deoxyuridine (BrdU) incorporation during DNA synthesis and collagen type II secretion was applied to evaluate the chondrocytes on different PLLA films and tissue culture plates (TCPS). Cell viability was estimated by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, and cell function was assessed by measuring lycosaminoglycan (GAG) secreted by chondrocytes. These results implied that gelatin used to modify the PLLA surface through entrapment and coupling could enhance chondrocyte adhesion, proliferation, and function.

In this study, L929 cell adhesion, proliferation and apoptosis were investigated on chitosan (CS) and chitosan-gelatin (CS-Gel) membranes having different degrees of deacetylation (DD). The mitochondrial activity [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay results demonstrated that the DD played a key role in the L929 cell adhesion. Indeed, the higher the DD of a CS membrane, the stronger the cell adhesion. The cell cycle analysis showed that the CS surface inhibits most of the cells entering the cell cycle and the DD of CS had little effect on the cell cycle progression. On the other hand, this study also confirmed that CS, blended with Gel, induced L929 cells to enter the cell cycle, encouraged proliferation, decreased the degree of apoptosis on the CS membranes, and are comparable to the results from L929 cells on tissue culture plates. In brief, CS modified with Gel improves cytocompatibility by shielding the positively charged CS with a high charge density.