Determination of the decay rate constant for hepatocytes immobilized in alginate microcapsules

Leal‐Egaña, Aldo; Díaz‐Cuenca, Aránzazu; Bader, Augustinus

doi:10.3109/02652040903050550

Cited by 7 publications

(5 citation statements)

References 20 publications

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“…Several methods can be used to this aim, each of them having advantages and disadvantages and the choice is taken considering how close the conditions of this determinations are with respect to the effective conditions in which the hydrogel is design to operate. Methods such as N 2 physisorption and mercury intrusion porosimetry are usually applied to xerogel, a colloidal state far from the typical operative conditions used in controlled tissue growth or in drug delivery. Other “chemical” methods are based on titration of the crosslinks.…”

Section: Resultsmentioning

confidence: 99%

Multiresponsive Hyaluronan‐p(NiPAAm) “Click”‐Linked Hydrogels

Pasale

Cerroni

Ghugare

et al. 2014

Macromolecular Bioscience

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Combined reversible addition-fragmentation chain transfer (RAFT) and chemoselective "click" chemistry are used for assembling two polymeric chains into a hybrid network capable to respond simultaneously or separately to different external stimuli. An azido-derivative of hyaluronate is clicked together with a new telechelic RAFT-generated p(NiPAAm), carrying a propargyl function at both ends, suitable as macromolecular "clickable" cross-linker with controlled molecular weight. This hybrid system displays a multiresponsive behavior versus temperature, pH, and ionic strength, maintaining cumulative as well as separate sensitivities to the external stimuli. Hyaluronidase catalyzed degradation of the hydrogels, mimicking the extracellular matrix degradation process, is an additional asset for the use of this class of hydrogels as scaffold. Tumor cells, HT-29, grow on the surface of these hybrid hydrogels more than the healthy ones, as NIH3T3. This finding opens a road to micro- and nano-devices based on hyaluronic acid as a promising biopolymer to pursue localized drug delivery.

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Section: Resultsmentioning

confidence: 99%

Multiresponsive Hyaluronan‐p(NiPAAm) “Click”‐Linked Hydrogels

Pasale

Cerroni

Ghugare

et al. 2014

Macromolecular Bioscience

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“…HepG 2 cells were immobilized in 0.8% and 1.4% w/v alginate-CaCl 2 microcapsules of 500µm diameter according to methods described previously [4,8]. A commercially available encapsulation system (Innotech, IE-50R) with a 250µm nozzle was used.…”

Section: Resultsmentioning

confidence: 99%

“…Some methods to characterize the porous structure of the 3D networks have been previously reported, such as mercury intrusion porosimetry [3], nitrogen physisorption [4], and the diffusion kinetics of relevant solutes [5]. Nevertheless, these techniques cannot be applied in the presence of cells, nor do they give information about modifications produced at the cell-biomaterial interface due to cell proliferation.…”

Section: Introductionmentioning

confidence: 99%

Determination of pore size distribution at the cell-hydrogel interface

Leal‐Egaña

Braumann

Díaz‐Cuenca

et al. 2011

Journal of Nanobiotechnology

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BackgroundAnalyses of the pore size distribution in 3D matrices such as the cell-hydrogel interface are very useful when studying changes and modifications produced as a result of cellular growth and proliferation within the matrix, as pore size distribution plays an important role in the signaling and microenvironment stimuli imparted to the cells. However, the majority of the methods for the assessment of the porosity in biomaterials are not suitable to give quantitative information about the textural properties of these nano-interfaces.FindingsHere, we report a methodology for determining pore size distribution at the cell-hydrogel interface, and the depth of the matrix modified by cell growth by entrapped HepG2 cells in microcapsules made of 0.8% and 1.4% w/v alginate. The method is based on the estimation of the shortest distance between two points of the fibril-like network hydrogel structures using image analysis of TEM pictures. Values of pore size distribution determined using the presented method and those obtained by nitrogen physisorption measurements were compared, showing good agreement. A combination of these methodologies and a study of the cell-hydrogel interface at various cell culture times showed that after three days of culture, HepG2 cells growing in hydrogels composed of 0.8% w/v alginate had more coarse of pores at depths up to 40 nm inwards (a phenomenon most notable in the first 20 nm from the interface). This coarsening phenomenon was weakly observed in the case of cells cultured in hydrogels composed of 1.4% w/v alginate.ConclusionsThe method purposed in this paper allows us to obtain information about the radial deformation of the hydrogel matrix due to cell growth, and the consequent modification of the pore size distribution pattern surrounding the cells, which are extremely important for a wide spectrum of biotechnological, pharmaceutical and biomedical applications.

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“…These assays were carried out in 3D, using methodologies already published. [28,29,33] With respect to the generation of polyploid cells, our results show that the enhanced number of multinucleated MCF7 found in the microcapsules seems to be strongly related to their doubling capabilities, as shown in Figure 5. [59,60] It is relevant to mention that the low proliferation rate described in MCF7 in suspension (Figure 5A, red curve) have been previously reported, showing that this cell line exhibits a weaker duplication in absence of matrix-anchorage, compared to very pathogenic breast cancer cells (i.e.…”

Section: Resultsmentioning

confidence: 62%

“…Studies of cell duplication were carried out with cells cultured in 3D using published methods. [33] Briefly, around microcapsules and spheroids were placed in a 48 well plate. These were cultured in presence of WST-8 (10% v/v, Thermofisher, Germany), for three hours, before we carried out the measurements, in accordance with the provider instructions.…”

Section: Cell Proliferation Assaysmentioning

confidence: 99%

When Mechanical Stress Matters: Generation of Polyploid Giant Cancer Cells in Tumor-like Microcapsules

Buehler

Krueger

Monavari

et al. 2022

Preprint

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In this work, we studied the generation and rising of polyploid cancer cells as a product of mechanical stress. To this purpose, MCF7 breast cancer cells were cultured on 2D (i.e. flasks, or flat hydrogels), and in 3D milieus (i.e. Spheroids, or immobilized within alginate-gelatin microbeads, named in this work as tumor-like microcapsules), and further analyzed by biophysical and genetic methods (i.e. single-cell Traction Force Microscopy and RNA-seq respectively). Our results show that MCF7 cells preconditioned onto 2D surfaces exhibit a low number of polynucleated cells, while their culture in 3D environments triggered their progressive generation with time. Genetic studies enabled us to determine that polyploid cells found in tumor-like microcapsules are likely originated by cell-cell fusion and disrupted cytokinesis, showing most of the genetic markers for Polyploid Giant Cancer Cell, while cells cultured as spheroids seem to be likely generated by other mechanisms, such as cell cannibalisms, entosis, or emperipolesis. Our outcomes strongly suggest that both mechanical stress and confinement are required to stimulate cell polyploidy, which can be easily addressed by the immobilization of breast cancer cells in tumor-like microcapsules.

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Determination of the decay rate constant for hepatocytes immobilized in alginate microcapsules

Cited by 7 publications

References 20 publications

Multiresponsive Hyaluronan‐p(NiPAAm) “Click”‐Linked Hydrogels

Multiresponsive Hyaluronan‐p(NiPAAm) “Click”‐Linked Hydrogels

Determination of pore size distribution at the cell-hydrogel interface

When Mechanical Stress Matters: Generation of Polyploid Giant Cancer Cells in Tumor-like Microcapsules

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