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Peptide Hydrogels - Versatile Matrices for 3D Cell Culture in Cancer Medicine.

Worthington P, Pochan DJ, Langhans SA - Front Oncol (2015)

Bottom Line: Traditional two-dimensional (2D) cell culture systems have contributed tremendously to our understanding of cancer biology but have significant limitations in mimicking in vivo conditions such as the tumor microenvironment.In addition, 3D cultures allow for the development of concentration gradients, including oxygen, metabolites, and growth factors, with chemical gradients playing an integral role in many cellular functions ranging from development to signaling in normal epithelia and cancer environments in vivo.One important area of synthetic materials currently available for 3D cell culture is short sequence, self-assembling peptide hydrogels.

View Article: PubMed Central - PubMed

Affiliation: Nemours Center for Childhood Cancer Research, Alfred I. duPont Hospital for Children , Wilmington, DE , USA ; Department of Biomedical Engineering, Delaware Biotechnology Institute, University of Delaware , Newark, DE , USA.

ABSTRACT
Traditional two-dimensional (2D) cell culture systems have contributed tremendously to our understanding of cancer biology but have significant limitations in mimicking in vivo conditions such as the tumor microenvironment. In vitro, three-dimensional (3D) cell culture models represent a more accurate, intermediate platform between simplified 2D culture models and complex and expensive in vivo models. 3D in vitro models can overcome 2D in vitro limitations caused by the oversupply of nutrients, and unphysiological cell-cell and cell-material interactions, and allow for dynamic interactions between cells, stroma, and extracellular matrix. In addition, 3D cultures allow for the development of concentration gradients, including oxygen, metabolites, and growth factors, with chemical gradients playing an integral role in many cellular functions ranging from development to signaling in normal epithelia and cancer environments in vivo. Currently, the most common matrices used for 3D culture are biologically derived materials such as matrigel and collagen. However, in recent years, more defined, synthetic materials have become available as scaffolds for 3D culture with the advantage of forming well-defined, designed, tunable materials to control matrix charge, stiffness, porosity, nanostructure, degradability, and adhesion properties, in addition to other material and biological properties. One important area of synthetic materials currently available for 3D cell culture is short sequence, self-assembling peptide hydrogels. In addition to the review of recent work toward the control of material, structure, and mechanical properties, we will also discuss the biochemical functionalization of peptide hydrogels and how this functionalization, coupled with desired hydrogel material characteristics, affects tumor cell behavior in 3D culture.

No MeSH data available.


Related in: MedlinePlus

Interaction between FEFK and enzyme. Reprinted with permission from Ref. (102). Copyright 2013 American Chemical Society.
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Figure 4: Interaction between FEFK and enzyme. Reprinted with permission from Ref. (102). Copyright 2013 American Chemical Society.

Mentions: FEFK and FEFKEFK form a hydrogel upon a unique enzymatic interaction with a metalloproteinase (99) (Figure 4). FEFK is a short chain peptide that does not form a gel on its own but in the presence of the metalloproteinase thermolysine, the peptide is broken down and rebuilt into longer chains that do gel (100). This is a different gel strategy compared to most peptide gels which depend on pH, ionic salts, or light (101). The final makeup of the gel is determined by the initial concentration of FEFK, and the mechanical properties are determined by the initial enzyme concentration (102). Gelation can also be controlled by the temperature used to initiate the reaction (103, 104). To prepare the cell–gel constructs, FEFK is dissolved in PBS, loaded into a syringe, and the enzyme is added. After incubation for 5 min, cells can be added and the solution is injected in a well, requiring frequent media change in the initial 1 h of incubation to remove the enzyme. The gel takes about 10 min for gelation and finishes around 2000 Pa (105, 106). The hydrogel construct has been used for successful culture of fibroblasts and osteoblasts and no negative effects of the enzyme used for gelation were observed although about 40% of the enzyme remained within the gel at day 7 (105). Multiple cell viability studies report biocompatibility of the gel but the most in depth biological investigation was on the ability of osteoblasts to mineralize the gel showing increased calcium phosphate deposits and an increase in gel stiffness as the cells deposit calcium and extracellular proteins, indicating bone formation (106). The cell–hydrogel constructs can be used for live dead staining and isolation of cells from the gel. Additional studies have shown that the peptide can be functionalized with polymers without affecting the gel (107).


Peptide Hydrogels - Versatile Matrices for 3D Cell Culture in Cancer Medicine.

Worthington P, Pochan DJ, Langhans SA - Front Oncol (2015)

Interaction between FEFK and enzyme. Reprinted with permission from Ref. (102). Copyright 2013 American Chemical Society.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4403249&req=5

Figure 4: Interaction between FEFK and enzyme. Reprinted with permission from Ref. (102). Copyright 2013 American Chemical Society.
Mentions: FEFK and FEFKEFK form a hydrogel upon a unique enzymatic interaction with a metalloproteinase (99) (Figure 4). FEFK is a short chain peptide that does not form a gel on its own but in the presence of the metalloproteinase thermolysine, the peptide is broken down and rebuilt into longer chains that do gel (100). This is a different gel strategy compared to most peptide gels which depend on pH, ionic salts, or light (101). The final makeup of the gel is determined by the initial concentration of FEFK, and the mechanical properties are determined by the initial enzyme concentration (102). Gelation can also be controlled by the temperature used to initiate the reaction (103, 104). To prepare the cell–gel constructs, FEFK is dissolved in PBS, loaded into a syringe, and the enzyme is added. After incubation for 5 min, cells can be added and the solution is injected in a well, requiring frequent media change in the initial 1 h of incubation to remove the enzyme. The gel takes about 10 min for gelation and finishes around 2000 Pa (105, 106). The hydrogel construct has been used for successful culture of fibroblasts and osteoblasts and no negative effects of the enzyme used for gelation were observed although about 40% of the enzyme remained within the gel at day 7 (105). Multiple cell viability studies report biocompatibility of the gel but the most in depth biological investigation was on the ability of osteoblasts to mineralize the gel showing increased calcium phosphate deposits and an increase in gel stiffness as the cells deposit calcium and extracellular proteins, indicating bone formation (106). The cell–hydrogel constructs can be used for live dead staining and isolation of cells from the gel. Additional studies have shown that the peptide can be functionalized with polymers without affecting the gel (107).

Bottom Line: Traditional two-dimensional (2D) cell culture systems have contributed tremendously to our understanding of cancer biology but have significant limitations in mimicking in vivo conditions such as the tumor microenvironment.In addition, 3D cultures allow for the development of concentration gradients, including oxygen, metabolites, and growth factors, with chemical gradients playing an integral role in many cellular functions ranging from development to signaling in normal epithelia and cancer environments in vivo.One important area of synthetic materials currently available for 3D cell culture is short sequence, self-assembling peptide hydrogels.

View Article: PubMed Central - PubMed

Affiliation: Nemours Center for Childhood Cancer Research, Alfred I. duPont Hospital for Children , Wilmington, DE , USA ; Department of Biomedical Engineering, Delaware Biotechnology Institute, University of Delaware , Newark, DE , USA.

ABSTRACT
Traditional two-dimensional (2D) cell culture systems have contributed tremendously to our understanding of cancer biology but have significant limitations in mimicking in vivo conditions such as the tumor microenvironment. In vitro, three-dimensional (3D) cell culture models represent a more accurate, intermediate platform between simplified 2D culture models and complex and expensive in vivo models. 3D in vitro models can overcome 2D in vitro limitations caused by the oversupply of nutrients, and unphysiological cell-cell and cell-material interactions, and allow for dynamic interactions between cells, stroma, and extracellular matrix. In addition, 3D cultures allow for the development of concentration gradients, including oxygen, metabolites, and growth factors, with chemical gradients playing an integral role in many cellular functions ranging from development to signaling in normal epithelia and cancer environments in vivo. Currently, the most common matrices used for 3D culture are biologically derived materials such as matrigel and collagen. However, in recent years, more defined, synthetic materials have become available as scaffolds for 3D culture with the advantage of forming well-defined, designed, tunable materials to control matrix charge, stiffness, porosity, nanostructure, degradability, and adhesion properties, in addition to other material and biological properties. One important area of synthetic materials currently available for 3D cell culture is short sequence, self-assembling peptide hydrogels. In addition to the review of recent work toward the control of material, structure, and mechanical properties, we will also discuss the biochemical functionalization of peptide hydrogels and how this functionalization, coupled with desired hydrogel material characteristics, affects tumor cell behavior in 3D culture.

No MeSH data available.


Related in: MedlinePlus