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Characterization of pore structure in biologically functional poly(2-hydroxyethyl methacrylate)-poly(ethylene glycol) diacrylate (PHEMA-PEGDA).

Zellander A, Zhao C, Kotecha M, Gemeinhart R, Wardlow M, Abiade J, Cho M - PLoS ONE (2014)

Bottom Line: The water and sucrose porogens were effective in creating porous and cytocompatible PHEMA-PEGDA scaffolds.The PHEMA-PEGDA scaffolds are easy to produce, non-toxic, and mechanically stable enough to resist fracture during routine handling.The PHEMA-PEGDA structures presented in this study may expedite the current research effort to engineer tissue scaffolds that provide both structural stability and biological activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of Illinois, Chicago, Illinois, United States of America.

ABSTRACT
A copolymer composed of poly(2-hydroxyethyl methacrylate) (PHEMA) and poly(ethylene glycol) diacrylate (PEGDA) (PHEMA-PEGDA) is structurally versatile. Its structure can be adjusted using the following porogens: water, sucrose, and benzyl alcohol. Using phase separation technique, a variety of surface architectures and pore morphologies were developed by adjusting porogen volume and type. The water and sucrose porogens were effective in creating porous and cytocompatible PHEMA-PEGDA scaffolds. When coated with collagen, the PHEMA-PEGDA scaffolds accommodated cell migration. The PHEMA-PEGDA scaffolds are easy to produce, non-toxic, and mechanically stable enough to resist fracture during routine handling. The PHEMA-PEGDA structures presented in this study may expedite the current research effort to engineer tissue scaffolds that provide both structural stability and biological activity.

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Collagen type I coating the PHEMA-PEGDA scaffolds was immunostained using collagen antibody.Scaffolds were removed from the tissue culture wells and rinsed with PBS. (A) Collagen gelled within the micron sized surface pores of PHEMA-PEGDA using the water porogen. Collagen fibers were evident in the sucrose and benzyl alcohol PHEMA-PEGDA scaffolds (B and C, respectively). At week 2, HCFs were detected in the three different PHEMA-PEDGA scaffold types, and average DNA contents were quantitatively measured (D; n = 6). “*” indicates p<0.05, and error bars represent standard error. Cell viability staining at day 7 shows both live (green) and a significant number of dead (red) HCFs in the benzyl alcohol PHEMA-PEGDA scaffold (E).
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pone-0096709-g004: Collagen type I coating the PHEMA-PEGDA scaffolds was immunostained using collagen antibody.Scaffolds were removed from the tissue culture wells and rinsed with PBS. (A) Collagen gelled within the micron sized surface pores of PHEMA-PEGDA using the water porogen. Collagen fibers were evident in the sucrose and benzyl alcohol PHEMA-PEGDA scaffolds (B and C, respectively). At week 2, HCFs were detected in the three different PHEMA-PEDGA scaffold types, and average DNA contents were quantitatively measured (D; n = 6). “*” indicates p<0.05, and error bars represent standard error. Cell viability staining at day 7 shows both live (green) and a significant number of dead (red) HCFs in the benzyl alcohol PHEMA-PEGDA scaffold (E).

Mentions: For tissue engineering applications, one of the important criteria for scaffolds is their capability to accommodate living cells. Because combinations of PEG and PHEMA may potentially be used in artificial corneas [17], we used human corneal fibroblasts (HCFs) to evaluate cytocompatibility. The PHEMA-PEGDA hydrogel was coated with collagen type I to promote cell adhesion to the surface features. Collagen antibody staining confirms the presence of collagen fibers in these scaffolds (Fig. 4A–C). It appears that the collagen fibers were incorporated within the surface pores of PHEMA-PEGDA. Since cell-free scaffolds were initially placed on the top of HCFs, the DNA quantification data showed that cells migrated into the collagen coated polymers (Fig. 4D). A comparison of the two porogen types, water and sucrose, demonstrated that the two structures supported essentially equivalent cell attachment. In contrast, a significantly fewer number of cells was attached to the structure made with the benzyl alcohol porogen. Figure 4E shows live and dead HCFs on the surface of scaffolds made with benzyl alcohol porogen, confirming the DNA measurement.


Characterization of pore structure in biologically functional poly(2-hydroxyethyl methacrylate)-poly(ethylene glycol) diacrylate (PHEMA-PEGDA).

Zellander A, Zhao C, Kotecha M, Gemeinhart R, Wardlow M, Abiade J, Cho M - PLoS ONE (2014)

Collagen type I coating the PHEMA-PEGDA scaffolds was immunostained using collagen antibody.Scaffolds were removed from the tissue culture wells and rinsed with PBS. (A) Collagen gelled within the micron sized surface pores of PHEMA-PEGDA using the water porogen. Collagen fibers were evident in the sucrose and benzyl alcohol PHEMA-PEGDA scaffolds (B and C, respectively). At week 2, HCFs were detected in the three different PHEMA-PEDGA scaffold types, and average DNA contents were quantitatively measured (D; n = 6). “*” indicates p<0.05, and error bars represent standard error. Cell viability staining at day 7 shows both live (green) and a significant number of dead (red) HCFs in the benzyl alcohol PHEMA-PEGDA scaffold (E).
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Related In: Results  -  Collection

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pone-0096709-g004: Collagen type I coating the PHEMA-PEGDA scaffolds was immunostained using collagen antibody.Scaffolds were removed from the tissue culture wells and rinsed with PBS. (A) Collagen gelled within the micron sized surface pores of PHEMA-PEGDA using the water porogen. Collagen fibers were evident in the sucrose and benzyl alcohol PHEMA-PEGDA scaffolds (B and C, respectively). At week 2, HCFs were detected in the three different PHEMA-PEDGA scaffold types, and average DNA contents were quantitatively measured (D; n = 6). “*” indicates p<0.05, and error bars represent standard error. Cell viability staining at day 7 shows both live (green) and a significant number of dead (red) HCFs in the benzyl alcohol PHEMA-PEGDA scaffold (E).
Mentions: For tissue engineering applications, one of the important criteria for scaffolds is their capability to accommodate living cells. Because combinations of PEG and PHEMA may potentially be used in artificial corneas [17], we used human corneal fibroblasts (HCFs) to evaluate cytocompatibility. The PHEMA-PEGDA hydrogel was coated with collagen type I to promote cell adhesion to the surface features. Collagen antibody staining confirms the presence of collagen fibers in these scaffolds (Fig. 4A–C). It appears that the collagen fibers were incorporated within the surface pores of PHEMA-PEGDA. Since cell-free scaffolds were initially placed on the top of HCFs, the DNA quantification data showed that cells migrated into the collagen coated polymers (Fig. 4D). A comparison of the two porogen types, water and sucrose, demonstrated that the two structures supported essentially equivalent cell attachment. In contrast, a significantly fewer number of cells was attached to the structure made with the benzyl alcohol porogen. Figure 4E shows live and dead HCFs on the surface of scaffolds made with benzyl alcohol porogen, confirming the DNA measurement.

Bottom Line: The water and sucrose porogens were effective in creating porous and cytocompatible PHEMA-PEGDA scaffolds.The PHEMA-PEGDA scaffolds are easy to produce, non-toxic, and mechanically stable enough to resist fracture during routine handling.The PHEMA-PEGDA structures presented in this study may expedite the current research effort to engineer tissue scaffolds that provide both structural stability and biological activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of Illinois, Chicago, Illinois, United States of America.

ABSTRACT
A copolymer composed of poly(2-hydroxyethyl methacrylate) (PHEMA) and poly(ethylene glycol) diacrylate (PEGDA) (PHEMA-PEGDA) is structurally versatile. Its structure can be adjusted using the following porogens: water, sucrose, and benzyl alcohol. Using phase separation technique, a variety of surface architectures and pore morphologies were developed by adjusting porogen volume and type. The water and sucrose porogens were effective in creating porous and cytocompatible PHEMA-PEGDA scaffolds. When coated with collagen, the PHEMA-PEGDA scaffolds accommodated cell migration. The PHEMA-PEGDA scaffolds are easy to produce, non-toxic, and mechanically stable enough to resist fracture during routine handling. The PHEMA-PEGDA structures presented in this study may expedite the current research effort to engineer tissue scaffolds that provide both structural stability and biological activity.

Show MeSH
Related in: MedlinePlus