Agar-gelatin hybrid sponge-induced three-dimensional<i>in vitro</i>‘liver-like’ HepG2 spheroids for the evaluation of drug cytotoxicity

Verma, Poonam; Verma, Vipin; Ray, Pratima; Ray, Alok R.

doi:10.1002/term.172

Cited by 17 publications

(20 citation statements)

References 25 publications

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“…Another important parameter for improved cell function is appropriate 3D microenvironmental cues. Hence, it may be essential to develop methods to generate pores that allow simultaneous encapsulation of cells inside the hydrogels using mild cell-compatible conditions and form 3D structure in short time [9, 18, 23, 29, 30]. Here, we used gelatin beads as template for creating pores in cell-laden alginate hydrogel.…”

Section: Resultsmentioning

confidence: 99%

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Fabrication of three-dimensional porous cell-laden hydrogel for tissue engineering

et al. 2010

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For tissue engineering applications, scaffolds should be porous to enable rapid nutrient and oxygen transfer while providing a three-dimensional (3D) microenvironment for the encapsulated cells. This dual characteristic can be achieved by fabrication of porous hydrogels that contain encapsulated cells. In this work, we developed a simple method that allows cell encapsulation and pore generation inside alginate hydrogels simultaneously. Gelatin beads of 150–300 μm diameter were used as a sacrificial porogen for generating pores within cell-laden hydrogels. Gelation of gelatin at low temperature (4 °C) was used to form beads without chemical crosslinking and their subsequent dissolution after cell encapsulation led to generation of pores within cell-laden hydrogels. The pore size and porosity of the scaffolds were controlled by the gelatin bead size and their volume ratio, respectively. Fabricated hydrogels were characterized for their internal microarchitecture, mechanical properties and permeability. Hydrogels exhibited a high degree of porosity with increasing gelatin bead content in contrast to nonporous alginate hydrogel. Furthermore, permeability increased by two to three orders while compressive modulus decreased with increasing porosity of the scaffolds. Application of these scaffolds for tissue engineering was tested by encapsulation of hepatocarcinoma cell line (HepG2). All the scaffolds showed similar cell viability; however, cell proliferation was enhanced under porous conditions. Furthermore, porous alginate hydrogels resulted in formation of larger spheroids and higher albumin secretion compared to nonporous conditions. These data suggest that porous alginate hydrogels may have provided a better environment for cell proliferation and albumin production. This may be due to the enhanced mass transfer of nutrients, oxygen and waste removal, which is potentially beneficial for tissue engineering and regenerative medicine applications.

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Section: Resultsmentioning

confidence: 99%

“…It has already been reported that multicellular spheroid formation with 3D structures is an important step in maintaining hepatocellular functions [9, 18, 23, 29, 30]. This cell aggregation is mediated by stronger cell–cell interactions compared to cell–matrix interactions.…”

Section: Resultsmentioning

confidence: 99%

Fabrication of three-dimensional porous cell-laden hydrogel for tissue engineering

et al. 2010

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“…), evaluation of the toxicity of drugs and chemicals (Verma et al . ) and drug delivery (Saxena et al . ; Shome et al .…”

Section: Introductionmentioning

confidence: 99%

In vitro antistaphylococcal effects of a novel 45S5 bioglass/agar- gelatin biocomposite films

et al. 2013

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Aims To assess the antibacterial efficacy of new composite materials developed from microparticles of 45S5 bioactive glass (BG) and agar–gelatin films. Methods and Results In vitro antibacterial activity was evaluated against Staphylococcus spp. because of the importance of this pathogen in damaged tissues and in failures associated with biomaterial implants. To our knowledge, this is the first paper reporting on the suitable combination of BG and agar–gelatin for bioactive and antibacterial films. Bacterial suspensions up or below 105 CFU ml−1 reflecting situations of wound infection and of noninfection, respectively, were prepared and then put in contact with the biomaterials at 37°C. After 24 and 48 h of incubation, the pH value was measured and the staphylococci strains viability was determined by counting in Mueller–Hinton agar plates. Moreover, the biomaterials were prepared for observation under scanning electron microscopy (SEM). Biocomposites (BCs) showed a strong antibacterial effect against all staphylococci strains tested. Some differences were found depending on the strain, the inoculum size and the contact time. This effect was correlated with an alkalinization of the media. By SEM analyses, no bacterial presence was observed on the surface of BCs in any of the cell concentrations tested at any time. Conclusions Overall, the coating of 45S5 BG on agar–gelatin films promoted BCs with strong antistaphylococcal activity. The effect was efficient under bacterial concentration up or below 105 CFU ml−1. Additionally, none of the strains were found on BCs surfaces. Significance and Impact of Study 45S5 bioglass/agar–gelatin biocomposite films are reported for the first time. The results suggest a potential application as wound dressing.

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“…On the TCPS, the cells lost the activity with culture period. Some studies have reported that human hepatocellular cells lose their liver specific functions in two dimensional (2D) culture; due to monolayer fails to accurately the native microenvironment [8][9]. From the result, the PMBV/PVA hydrogel did not induce the reduction of cell-specific function, such as the albumin synthetic function of HepG2 cells.…”

Section: Analysis Of the Distribution Of Cell Cyclementioning

confidence: 59%

Controlling cell functions by encapsulation with cytocompatible phospholipid polymer hydrogel

Fukuei

Konno

Ishihara

2010

Trans. Mat. Res. Soc. Japan

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To control functions of cells during their culture period in vitro is important for providing cells in the cell engineering fields. We designed a novel soft biodevice for realizing cell engineering. That is, spontaneously forming and highly cytocompatible hydrogel system composed of poly (2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate-co-p-vinylphenylboronic acid) (PMBV) and poly (vinyl alcohol) (PVA), was prepared to encapsulate cells as "Cell Container". This hydrogel was reversibly dissociated by addition of sugar derivatives. When mouse fibroblast (L929) cell lines and human hepatocellular liver carcinoma (HepG2) cell lines were encapsulated in the PMBV/PVA hydrogel, the cells could proliferate slowly compared with those on conventional tissue culture plate and form cell clusters derived by expansion of the single cells, without forming aggregations. After the dissociation of the PMBV/PVA hydrogel, the distribution of cell cycle phase of L929 cells was analyzed. It was found that the PMBV/PVA hydrogel lead a synchronization of cell cycle phase of L929 cells to G0/G1 phase. As other cell-specific function of HepG2, albumin production and releasing from HepG2 was evaluated. The PMBV/PVA hydrogel did not induce a significant reduction of albumin production from HepG2. The PMBV/PVA hydrogel could preserve cells for long term inhibiting the excessive proliferation, provide normalized cells with uniformly cell cycle phase, and maintain the cell-specific function. Thus, we concluded that the PMBV/PVA hydrogel will be a novel and powerful device to control cell functions in cell engineering fields.

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Agar-gelatin hybrid sponge-induced three-dimensionalin vitro‘liver-like’ HepG2 spheroids for the evaluation of drug cytotoxicity

Cited by 17 publications

References 25 publications

Fabrication of three-dimensional porous cell-laden hydrogel for tissue engineering

Fabrication of three-dimensional porous cell-laden hydrogel for tissue engineering

In vitro antistaphylococcal effects of a novel 45S5 bioglass/agar- gelatin biocomposite films

Controlling cell functions by encapsulation with cytocompatible phospholipid polymer hydrogel

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