Pectin self-assembly and its disruption by water: Insights into plant cell wall mechanics

Plant cell walls undergo multiple cycles of dehydration and rehydration during their life. Calcium crosslinked low methoxy pectin is a major constituent of plant cell walls. Understanding the dehydration-rehydration behavior of pectin gels may shed light on the water transport and mechanics of plant cells. In this work, we report the contributions of microstructure to the mechanics of pectin-Ca gels subjected to different extents of dehydration and subsequent rehydration. This is investigated using a pectin gel composition that forms ‘egg-box bundles’, a characteristic feature of the microstructure of low methoxy pectin-Ca gels. Large Amplitude Oscillatory Shear (LAOS) rheology along with Small Angle Neutron Scattering and Near Infrared (NIR) spectroscopy on pectin gels are used to elucidate the mechanical and microstructural changes during dehydration-rehydration cycles. As the extent of dehydration increase, the reswelling ability, strain-stiffening behavior and the yield strain decreases. These effects are more prominent at faster rates of dehydration and are not completely reversible upon rehydration to the initial undried state. Microstructural changes due to the aggregation of egg-box bundles and single chains and the associated changes in the water configurations lead to these irreversible changes.

Pectin as Rheology Modifier of a Gelatin-Based Biomaterial Ink

Gelatin is a natural biopolymer extensively used for tissue engineering applications due to its similarities to the native extracellular matrix. However, the rheological properties of gelatin formulations are not ideal for extrusion-based bioprinting. In this work, we present an approach to improve gelatin bioprinting performances by using pectin as a rheology modifier of gelatin and (3-glycidyloxypropyl)trimethoxysilane (GPTMS) as a gelatin–pectin crosslinking agent. The preparation of gelatin–pectin formulations is initially optimized to obtain homogenous gelatin–pectin gels. Since the use of GPTMS requires a drying step to induce the completion of the crosslinking reaction, microporous gelatin–pectin–GPTMS sponges are produced through freeze-drying, and the intrinsic properties of gelatin–pectin–GPTMS networks (e.g., porosity, pore size, degree of swelling, compressive modulus, and cell adhesion) are investigated. Subsequently, rheological investigations together with bioprinting assessments demonstrate the key role of pectin in increasing the viscosity and the yield stress of low viscous gelatin solutions. Water stable, three-dimensional, and self-supporting gelatin–pectin–GPTMS scaffolds with interconnected micro- and macroporosity are successfully obtained by combining extrusion-based bioprinting and freeze-drying. The proposed biofabrication approach does not require any additional temperature controller to further modulate the rheological properties of gelatin solutions and it could furthermore be extended to improve the bioprintability of other biopolymers.

Pectin methylesterase selectively softens the onion epidermal wall yet reduces acid-induced creep

De-esterification of homogalacturonan (HG) is thought to stiffen pectin gels and primary cell walls by increasing calcium cross-linking between HG chains. Contrary to this idea, recent studies found that HG de-esterification correlated with reduced stiffness of living tissues, measured by surface indentation. The physical basis of such apparent wall softening is unclear, but possibly involves complex biological responses to HG modification. To assess the direct physical consequences of HG de-esterification on wall mechanics without such complications, we treated isolated onion (Allium cepa) epidermal walls with pectin methylesterase (PME) and assessed wall biomechanics with indentation and tensile tests. In nanoindentation assays, PME action softened the wall (reduced the indentation modulus). In tensile force/extension assays, PME increased plasticity, but not elasticity. These softening effects are attributed, at least in part, to increased electrostatic repulsion and swelling of the wall after PME treatment. Despite softening and swelling upon HG de-esterification, PME treatment alone failed to induce cell wall creep. Instead, acid-induced creep, mediated by endogenous α-expansin, was reduced. We conclude that HG de-esterification physically softens the onion wall, yet reduces expansin-mediated wall extensibility.

Preparation and Properties of Hydrogel Matrices based on Pectins from Callus Cultures

Hydrogel matrices have been obtained from pectins of Silene vulgaris (SVC), Tanacetum vulgare (TVC) and Lemna minor (LMC) callus cultures, and from apple pectin (AU701). They differed in gel strength, calcium ion content and surface morphology. During the incubation under simulated gastric conditions (pH 1.25), the gel strength and calcium content decreased. The degradation degree of hydrogels grew in the range TVC < SVC < LMC < AU701. The degradation degree of hydrogels from the SVC, LMC, and TVC pectins gradually increased over 4 hours, while hydrogels from the AU701 pectin were destroyed within 2 hours. The gels prepared from pectins with low methyl esterification, low branching and high linearity underwent less degradation. The TVC pectin gels had the highest strength and were most stable at pH 1.25. The data obtained can be used to produce hydrogel matrices with desired physicochemical and functional properties. callus culture, pectin polysaccharides, hydrogels, gastric medium

Gel granules from pectin callus Lupinus angustifolius prevent the premature release of mesalazine

The formation of cross-related pectin matrices on the base of physiologically active polysaccharides opens up new prospects for their application in medicine. The purpose of the study was to identify pectin gels that could retain the drag in an artificial gastric environment and did not have a cytotoxic effect. Here, pectins of apple, hogweed, thistle, rowan and lupine as well their gels were investigated. Pectin-gel granules were produced from 4% solutions of the pectins by ionotropic gelling. A metabolic activity of human cell cultures was evaluated in MTT test. The pectins at concentrations of less than 2 mg/mL were shown to be no cytotoxic. Pectin gels slightly reduced the metabolic activity of cells. The content of mesalazine in the gels and its releasing were evaluated by liquid chromatography-mass spectrometry method. Among pectines investigated, gel granules of lupine pectin retained the mesalazine most effectively. On the base of the results, one can consider the pectin polysaccharides of lupine as potential gel matrices for oral drag delivery systems. gel, pectins, HEK293, Caco-2, 5-aminosalicylic acid, gastric solution, chromatography-mass spectrometry

Pectin methylesterase selectively softens the onion epidermal wall yet reduces acid-induced creep

De-esterification of homogalacturonan (HG) is thought to stiffen pectin gels and primary cell walls by increasing calcium crosslinking between HG chains. Contrary to this idea, recent studies found that HG de-esterification correlated with reduced stiffness of living tissues, measured by surface indentation. The physical basis of such apparent wall softening is unclear, but possibly involves complex biological responses to HG modification. To assess the direct physical consequences of HG de-esterification on wall mechanics without such complications, we treated isolated onion (Allium cepa) epidermal walls with pectin methylesterase (PME) and assessed wall biomechanics with indentation and tensile tests. In nanoindentation assays, PME action softened the wall (reduced the indentation modulus). In tensile force/extension assays, PME increased plasticity, but not elasticity. These softening effects are attributed, at least in part, to increased electrostatic repulsion and swelling of the wall after PME treatment. Despite softening and swelling upon HG de-esterification, PME treatment alone failed to induce cell wall creep. Instead, acid-induced creep, mediated by endogenous α expansin, was reduced. We conclude that HG de-esterification physically softens the onion wall, yet reduces expansin-mediated wall extensibility.HighlightAfter enzymatic de-esterification without added calcium, the onion epidermal wall swells and becomes softer, as assessed by nanoindentation and tensile plasticity tests, yet exhibits reduced expansin-mediated creep.