Influence of transglutaminase treatment on the thermoreversible gelation of gelatin

Babin, Hélène; Dickinson, Eric

doi:10.1016/s0268-005x(01)00025-x

Cited by 158 publications

(110 citation statements)

References 11 publications

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“…Tensile and rheological testing of gelatin and collagen crosslinked with mTG has been analyzed previously 13,16,[22][23][24][25] along with gelation properties using thermal cycling. 16,22 For our purposes, only the elimination of the melting point at 37 uC in physical gels (gelatin that has been gelled by cooling, without enzyme present) is necessary for the integrity of the microchannels during incubation at 37 uC for cell culture and analysis.…”

Section: Verification Of Crosslinking and Materials Propertiesmentioning

confidence: 99%

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Gelatin based microfluidic devices for cell culture

Paguirigan¹,

Beebe²

2006

Lab Chip

145

139

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We have developed a technique for fabricating microfluidic devices from gelatin using a natural crosslinking process. Gelatin, crosslinked with the naturally occuring enzyme transglutaminase is molded to produce microchannels suitable for adherent cell culture and analysis. The autofluorescence of the material was shown to be minimal and within the range of typical background, ensuring utility with analyses using fluorescent dyes and labels would not be affected. Also, normal murine mammary epithelial cells were successfully cultured in the microchannels. The morphology of these adherent epithelial cells was shown to be significantly different for cells grown on rigid tissue culture plastic in either macro-or microscale cultures (even in the presence of a surface coating of gelatin) than those grown on the flexible crosslinked gelatin microchannels. Using these devices, the effects of both the extracellular matrix and soluble factors on cellular behavior and differentiation can be studied in microenvironments that more closely mimic the in vivo environment. IntroductionWhile microfluidics has shown considerable promise as a tool for studying cell biology, the potential for microfluidics to create more in vivo-like in vitro environments (or, in visio environments) is still largely untapped. It is becoming clear that the scale of the microenvironment provided by microchannels is an important biological parameter. Microchannels have been used for several steps in the in vitro production of embryos typically either matching or improving the performance of previous methods. 1,2 Insect cell cultures as well have shown very different dimension dependent growth kinetics in microscale cultures as compared to macroscale flask cultures. 3 However, much of the previous culture work in microfluidics has focused on non-adherent (e.g., embryos, insect cells) or 2-D adherent cell culture in the absence of extracellular matrix. While the effects of the extracellular matrix are of critical importance to in vivo tissue function, 4,5 the integration of extracellular matrix (ECM) materials into microfluidic systems is just emerging. 6,7 We have fabricated microdevices which can allow for control over the soluble microenvironment, while also providing a more in vivo-like substrate (flexible protein gels) as a step towards an in visio environment for cellular analysis. We present a fabrication protocol to produce enzymatically crosslinked gelatin microdevices composed of natural components, that are sterile and suitable for cell culture. Gelatin is a derivative of collagen, one of the most common extracellular matrix proteins. These devices were tested for their fluorescence properties to ensure that any autofluorescence of the material itself will not limit its utility while using fluorescent dyes for cellular analysis. Also, we have explored the potential for mammary epithelial cell culture in these devices and compared the cell morphology at multiple time points between macro-and microscale cultures on traditional rigid surfaces an...

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Section: Verification Of Crosslinking and Materials Propertiesmentioning

confidence: 99%

“…22 Both during commercial purification of gelatin and during processing for molding, thermal denaturation of the gelatin can occur. In addition, the formation of covalent crosslinked gelatin by mTG partially prevents the collagen-like tertiary structure from forming in the cooled crosslinked gelatin gels.…”

Section: Thermal Protection and Tertiary Structurementioning

confidence: 99%

Gelatin based microfluidic devices for cell culture

Paguirigan¹,

Beebe²

2006

Lab Chip

145

139

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“…The reaction catalyzed of glutamyltransferases is shown in Figure 2 (Yee et al, 1994) Polymerization using transglutaminase has been investigated with various protein sources including casein, soy proteins and gelatin, where different responses in gel strength were dependent on the reaction conditions and on the different protein sources (Sakamoto et al, 1994). The increase in gel strength of proteins submitted to the action of transglutaminase depended on the order and intensity by which the enzyme produced cross links, and the extent to which these new covalent linkages could impede the 'physical' cross-linkages occurring during renaturation and formation of the triple helix during gel formation (Babin & Dickinson, 2001). Larre et al (2000) reported that transglutaminase was effective in introducing covalent bonds into films obtained from slightly deamidated gluten.…”

Section: Modification Of Protein-based Edible Films By Enzymatic Treamentioning

confidence: 99%

Protein-Based Edible Films: Characteristics and Improvement of Properties

Wittaya¹

2012

Structure and Function of Food Engineering

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“…Nas temperaturas de 40 e 50 °C, os filmes modificados enzimaticamente apresentaram uma diminuição significativa da permeabilidade ao vapor de água em relação ao filme nativo (FN) e ao termicamente tratado (FC). Os resultados sugerem que, nestas temperaturas, a introdução de ligações cruzadas é favorecida, e que a introdução destas provocou mudanças morfológicas da matriz polimérica permitindo uma configuração intermediária com algum grau de renaturação das triplas hélices, favorecendo as propriedades de barreira à água desses filmes, embora o aumento do grau de ligações cruzadas pareça dificultar a formação de zonas de junção características da formação da matriz polimérica para a gelatina [4]. Estudos envolvendo a aplicação da enzima transglutaminase em géis [8] e em filmes à base de concentrado protéico de soro de leite [12] observaram diminuição da permeabilidade ao vapor de água da mesma forma como observado no presente trabalho.…”

Section: -Permeabilidade Ao Vapor De áGuaunclassified

Efeito do tratamento térmico e enzimático nas propriedades de filmes de gelatina

Carvalho¹,

Grosso²

2006

Ciênc. Tecnol. Aliment.

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RESUMODentre os fatores que afetam a atividade da enzima transglutaminase, a temperatura de reação ou incubação é um fator determinante no grau de reticulação. Por outro lado, para a gelatina, tipicamente a rede estrutural polimérica é estabilizada por forças secundárias, sendo que a formação da matriz polimérica envolve um delicado balanço entre interações polímero-polímero e polímero-solvente, e este balanço é fortemente dependente do histórico térmico da solução. Desta forma, o objetivo deste trabalho foi avaliar o efeito da temperatura na reação de modificação enzimática em relação às propriedades funcionais dos filmes modificados à base de gelatina (propriedades mecânicas, de barreira ao vapor de água, solubilidade em água e parâmetros de cor dos filmes). Viscosidade aparente das soluções filmogênicas foram também avaliadas. Foram produzidos filmes denominados nativo (FN), modificado enzimaticamente (FME) e termicamente tratado (FC). De acordo com os resultados obtidos, observou-se que a temperatura de reação não afetou as propriedades mecânicas e a solubilidade dos diferentes filmes estudados. Por outro lado, filmes modificados enzimaticamente (FME) na temperatura de 50 °C apresentaram permeabilidade ao vapor de água significantemente inferior aos produzidos nas demais temperaturas e tratamentos (FN e FC). O tratamento térmico também provocou redução da permeabilidade ao vapor de água. Palavras-chave: filmes, temperatura de reação, transglutaminase. SUMMARY EFFECT OF THERMAL AND ENZYMATIC TREATMENT ON THE PROPERTIES OF GELATIN FILMS.The activity of transglutaminase (one crosslinking enzyme) can be affected by the temperature of reaction. Gelatin matrix gel formation depends on the balance between polymer-polymer interactions and a polymer-solvent solution. This balance also depends on the amount and type of thermal treatment to the gelatin solution. The aim of this work is to evaluate the effect of the temperature on the crosslinking reaction using transglutaminase. Mechanical properties, water vapor permeability, water solubility and color/opacity of the films were evaluated. Viscosity of the solutions was also evaluated. Native film (FN), enzymatically modified film (FME) and thermal treated film (FC) were produced. According to the results, the temperature which was used did not significantly change the mechanical properties and the solubility of the films produced using different treatments. Enzymatically modified films at 50 °C showed the lowest water vapor permeability compared to the other temperatures and treatments (FN e FC). Thermal treatment also caused a reduction in the water vapor permeability.

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Influence of transglutaminase treatment on the thermoreversible gelation of gelatin

Cited by 158 publications

References 11 publications

Gelatin based microfluidic devices for cell culture

Gelatin based microfluidic devices for cell culture

Protein-Based Edible Films: Characteristics and Improvement of Properties

Efeito do tratamento térmico e enzimático nas propriedades de filmes de gelatina

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