Limits...
New biomedical devices with selective peptide recognition properties. Part 1: Characterization and cytotoxicity of molecularly imprinted polymers.

Rechichi A, Cristallini C, Vitale U, Ciardelli G, Barbani N, Vozzi G, Giusti P - J. Cell. Mol. Med. (2007 Nov-Dec)

Bottom Line: The highly cross-linked polymers retained about 70% of the total template amount, against (20% for the less cross-linked ones).The extracted template amount and the rebinding capacity decreased with the cross-linking degree, while the selectivity showed the opposite behaviour.The PETRA cross-linked polymers showed the best recognition (MIP 2-, alpha=1.71) and selectivity (MIP 2+, alpha'=5.58) capabilities.

View Article: PubMed Central - PubMed

Affiliation: Department of Mechanical Engineering, Politecnico di Torino, Torino, Italy.

ABSTRACT
Molecular imprinting is a technique for the synthesis of polymers capable to bind target molecules selectively. The imprinting of large proteins, such as cell adhesion proteins or cell receptors, opens the way to important and innovative biomedical applications. However, such molecules can incur into important conformational changes during the preparation of the imprinted polymer impairing the specificity of the recognition cavities. The "epitope approach" can overcome this limit by adopting, as template, a short peptide sequence representative of an accessible fragment of a larger protein. The resulting imprinted polymer can recognize both the template and the whole molecule thanks to the specific cavities for the epitope. In this work two molecularly imprinted polymer formulations (a macroporous monolith and nanospheres) were obtained using the protected peptide Z-Thr-Ala-Ala-OMe, as template, and Z-Thr-Ile-Leu-OMe, as analogue for the selectivity evaluation, methacrylic acid, as functional monomer, and trimethylolpropane trimethacrylate and pentaerythritol triacrylate (PETRA), as cross-linkers. Polymers were synthesized by precipitation polymerization and characterized by standard techniques. Polymerization and rebinding solutions were analyzed by high performance liquid chromatography. The highly cross-linked polymers retained about 70% of the total template amount, against (20% for the less cross-linked ones). The extracted template amount and the rebinding capacity decreased with the cross-linking degree, while the selectivity showed the opposite behaviour. The PETRA cross-linked polymers showed the best recognition (MIP 2-, alpha=1.71) and selectivity (MIP 2+, alpha'=5.58) capabilities. The cytotoxicity tests showed normal adhesion and proliferation of fibroblasts cultured in the medium that was put in contact with the imprinted polymers.

Show MeSH

Related in: MedlinePlus

Cytotoxicity index of CP 2+ using NIH-3T3 cultured in conditioned media culture as function of cell culture time after incubation (at 24, 48 and 72 hrs). Number of cells cultured in medium not incubated with nanoparticles was used as control.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4401303&req=5

fig06: Cytotoxicity index of CP 2+ using NIH-3T3 cultured in conditioned media culture as function of cell culture time after incubation (at 24, 48 and 72 hrs). Number of cells cultured in medium not incubated with nanoparticles was used as control.

Mentions: Figure 6 presents the effects of CP 2+ nanoparticles incubated with media culture at several times (4 hrs, 24 hrs, 7 days and 15 days) time on the cell adhesion at proliferation as a function of incubation time (4, 24, 48 hrs). The ratio between the number of cells cultured in conditioned media and those in pure media was considered as an index of cytotoxicity.


New biomedical devices with selective peptide recognition properties. Part 1: Characterization and cytotoxicity of molecularly imprinted polymers.

Rechichi A, Cristallini C, Vitale U, Ciardelli G, Barbani N, Vozzi G, Giusti P - J. Cell. Mol. Med. (2007 Nov-Dec)

Cytotoxicity index of CP 2+ using NIH-3T3 cultured in conditioned media culture as function of cell culture time after incubation (at 24, 48 and 72 hrs). Number of cells cultured in medium not incubated with nanoparticles was used as control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4401303&req=5

fig06: Cytotoxicity index of CP 2+ using NIH-3T3 cultured in conditioned media culture as function of cell culture time after incubation (at 24, 48 and 72 hrs). Number of cells cultured in medium not incubated with nanoparticles was used as control.
Mentions: Figure 6 presents the effects of CP 2+ nanoparticles incubated with media culture at several times (4 hrs, 24 hrs, 7 days and 15 days) time on the cell adhesion at proliferation as a function of incubation time (4, 24, 48 hrs). The ratio between the number of cells cultured in conditioned media and those in pure media was considered as an index of cytotoxicity.

Bottom Line: The highly cross-linked polymers retained about 70% of the total template amount, against (20% for the less cross-linked ones).The extracted template amount and the rebinding capacity decreased with the cross-linking degree, while the selectivity showed the opposite behaviour.The PETRA cross-linked polymers showed the best recognition (MIP 2-, alpha=1.71) and selectivity (MIP 2+, alpha'=5.58) capabilities.

View Article: PubMed Central - PubMed

Affiliation: Department of Mechanical Engineering, Politecnico di Torino, Torino, Italy.

ABSTRACT
Molecular imprinting is a technique for the synthesis of polymers capable to bind target molecules selectively. The imprinting of large proteins, such as cell adhesion proteins or cell receptors, opens the way to important and innovative biomedical applications. However, such molecules can incur into important conformational changes during the preparation of the imprinted polymer impairing the specificity of the recognition cavities. The "epitope approach" can overcome this limit by adopting, as template, a short peptide sequence representative of an accessible fragment of a larger protein. The resulting imprinted polymer can recognize both the template and the whole molecule thanks to the specific cavities for the epitope. In this work two molecularly imprinted polymer formulations (a macroporous monolith and nanospheres) were obtained using the protected peptide Z-Thr-Ala-Ala-OMe, as template, and Z-Thr-Ile-Leu-OMe, as analogue for the selectivity evaluation, methacrylic acid, as functional monomer, and trimethylolpropane trimethacrylate and pentaerythritol triacrylate (PETRA), as cross-linkers. Polymers were synthesized by precipitation polymerization and characterized by standard techniques. Polymerization and rebinding solutions were analyzed by high performance liquid chromatography. The highly cross-linked polymers retained about 70% of the total template amount, against (20% for the less cross-linked ones). The extracted template amount and the rebinding capacity decreased with the cross-linking degree, while the selectivity showed the opposite behaviour. The PETRA cross-linked polymers showed the best recognition (MIP 2-, alpha=1.71) and selectivity (MIP 2+, alpha'=5.58) capabilities. The cytotoxicity tests showed normal adhesion and proliferation of fibroblasts cultured in the medium that was put in contact with the imprinted polymers.

Show MeSH
Related in: MedlinePlus