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Interaction of human plasma proteins with thin gelatin-based hydrogel films: a QCM-D and ToF-SIMS study.

Schönwälder SM, Bally F, Heinke L, Azucena C, Bulut ÖD, Heißler S, Kirschhöfer F, Gebauer TP, Neffe AT, Lendlein A, Brenner-Weiß G, Lahann J, Welle A, Overhage J, Wöll C - Biomacromolecules (2014)

Bottom Line: This technique enables the determination of adsorbant mass and changes in the shear modulus of the hydrogel layer upon adsorption of human proteins.Furthermore, Secondary Ion Mass Spectrometry and principal component analysis was applied to monitor the changed composition of the topmost adsorbate layer.This approach opens interesting perspectives for a sensitive screening of viscoelastic biomaterials that could be used for regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Karlsruhe Institute of Technology (KIT), Institute of Functional Interfaces (IFG) , 76344 Eggenstein-Leopoldshafen, Germany.

ABSTRACT
In the fields of surgery and regenerative medicine, it is crucial to understand the interactions of proteins with the biomaterials used as implants. Protein adsorption directly influences cell-material interactions in vivo and, as a result, regulates, for example, cell adhesion on the surface of the implant. Therefore, the development of suitable analytical techniques together with well-defined model systems allowing for the detection, characterization, and quantification of protein adsorbates is essential. In this study, a protocol for the deposition of highly stable, thin gelatin-based films on various substrates has been developed. The hydrogel films were characterized morphologically and chemically. Due to the obtained low thickness of the hydrogel layer, this setup allowed for a quantitative study on the interaction of human proteins (albumin and fibrinogen) with the hydrogel by Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D). This technique enables the determination of adsorbant mass and changes in the shear modulus of the hydrogel layer upon adsorption of human proteins. Furthermore, Secondary Ion Mass Spectrometry and principal component analysis was applied to monitor the changed composition of the topmost adsorbate layer. This approach opens interesting perspectives for a sensitive screening of viscoelastic biomaterials that could be used for regenerative medicine.

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IRRAS spectra of theamino-functionalized CVD polymer (1), coatedwith non-cross-linked gelatin after washing (2), or coated with cross-linkedgelatin prior to washing (3) and after washing (4).
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fig3: IRRAS spectra of theamino-functionalized CVD polymer (1), coatedwith non-cross-linked gelatin after washing (2), or coated with cross-linkedgelatin prior to washing (3) and after washing (4).

Mentions: Infrared Reflection Absorption Spectroscopy (IRRAS)48 was used to check the chemical composition ofthe surface layers. Cross-linked and non-cross-linked gelatin-basedfilms on amino-functionalized substrates were analyzed. Figure 3 shows that, even after intensive washing, the cross-linkedgelatin hydrogel film remains on the surface, previously modifiedwith amino-functionalized CVD polymer. Indeed, the broad hydroxylband at ∼3500 cm–1 and amide bands I to IIIat ∼1600 cm–1 on spectrum 4 indicate thepresence of the protein on the surface. If, however, the cross-linkingstep is omitted (see spectrum 2), the majority of the deposited gelatinis removed from the surface by washing at 37 °C. The absenceof a band at ∼2250 cm–1 indicates that theisocyanate groups were completely converted.21 Assignments of bands ascribed to gelatin, in accordance with a previousstudy,49 are given in Supporting Information, Table SI-1.


Interaction of human plasma proteins with thin gelatin-based hydrogel films: a QCM-D and ToF-SIMS study.

Schönwälder SM, Bally F, Heinke L, Azucena C, Bulut ÖD, Heißler S, Kirschhöfer F, Gebauer TP, Neffe AT, Lendlein A, Brenner-Weiß G, Lahann J, Welle A, Overhage J, Wöll C - Biomacromolecules (2014)

IRRAS spectra of theamino-functionalized CVD polymer (1), coatedwith non-cross-linked gelatin after washing (2), or coated with cross-linkedgelatin prior to washing (3) and after washing (4).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215905&req=5

fig3: IRRAS spectra of theamino-functionalized CVD polymer (1), coatedwith non-cross-linked gelatin after washing (2), or coated with cross-linkedgelatin prior to washing (3) and after washing (4).
Mentions: Infrared Reflection Absorption Spectroscopy (IRRAS)48 was used to check the chemical composition ofthe surface layers. Cross-linked and non-cross-linked gelatin-basedfilms on amino-functionalized substrates were analyzed. Figure 3 shows that, even after intensive washing, the cross-linkedgelatin hydrogel film remains on the surface, previously modifiedwith amino-functionalized CVD polymer. Indeed, the broad hydroxylband at ∼3500 cm–1 and amide bands I to IIIat ∼1600 cm–1 on spectrum 4 indicate thepresence of the protein on the surface. If, however, the cross-linkingstep is omitted (see spectrum 2), the majority of the deposited gelatinis removed from the surface by washing at 37 °C. The absenceof a band at ∼2250 cm–1 indicates that theisocyanate groups were completely converted.21 Assignments of bands ascribed to gelatin, in accordance with a previousstudy,49 are given in Supporting Information, Table SI-1.

Bottom Line: This technique enables the determination of adsorbant mass and changes in the shear modulus of the hydrogel layer upon adsorption of human proteins.Furthermore, Secondary Ion Mass Spectrometry and principal component analysis was applied to monitor the changed composition of the topmost adsorbate layer.This approach opens interesting perspectives for a sensitive screening of viscoelastic biomaterials that could be used for regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Karlsruhe Institute of Technology (KIT), Institute of Functional Interfaces (IFG) , 76344 Eggenstein-Leopoldshafen, Germany.

ABSTRACT
In the fields of surgery and regenerative medicine, it is crucial to understand the interactions of proteins with the biomaterials used as implants. Protein adsorption directly influences cell-material interactions in vivo and, as a result, regulates, for example, cell adhesion on the surface of the implant. Therefore, the development of suitable analytical techniques together with well-defined model systems allowing for the detection, characterization, and quantification of protein adsorbates is essential. In this study, a protocol for the deposition of highly stable, thin gelatin-based films on various substrates has been developed. The hydrogel films were characterized morphologically and chemically. Due to the obtained low thickness of the hydrogel layer, this setup allowed for a quantitative study on the interaction of human proteins (albumin and fibrinogen) with the hydrogel by Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D). This technique enables the determination of adsorbant mass and changes in the shear modulus of the hydrogel layer upon adsorption of human proteins. Furthermore, Secondary Ion Mass Spectrometry and principal component analysis was applied to monitor the changed composition of the topmost adsorbate layer. This approach opens interesting perspectives for a sensitive screening of viscoelastic biomaterials that could be used for regenerative medicine.

Show MeSH
Related in: MedlinePlus