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Interfacial Interaction between Transmembrane Ocular Mucins and Adhesive Polymers and Dendrimers Analyzed by Surface Plasmon Resonance

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ABSTRACT

Purpose

Development of the first in vitro method based on biosensor chip technology designed for probing the interfacial interaction phenomena between transmembrane ocular mucins and adhesive polymers and dendrimers intended for ophthalmic administration.

Methods

The surface plasmon resonance (SPR) technique was used. A transmembrane ocular mucin surface was prepared on the chip surface and characterized by QCM-D (Quartz Crystal Microbalance with Dissipation) and XPS (X-ray photoelectron spectroscopy). The mucoadhesive molecules tested were: hyaluronic acid (HA), carboxymethyl cellulose (CMC), hydroxypropylmethyl cellulose (HPMC), chitosan (Ch) and polyamidoamine dendrimers (PAMAM).

Results

While Ch originated interfacial interaction with ocular transmembrane mucins, for HA, CMC and HPMC, chain interdiffusion seemed to be mandatory for bioadherence at the concentrations used in ophthalmic clinical practise. Interestingly, PAMAM dendrimers developed permanent interfacial interactions with transmembrane ocular mucins whatever their surface chemical groups, showing a relevant importance of co-operative effect of these multivalent systems. Polymers developed interfacial interactions with ocular membrane-associated mucins in the following order: Ch(1 %) > G4PAMAM-NH2(2 %) = G4PAMAM-OH(2 %) > G3.5PAMAM-COOH(2 %)>> CMC(0.5 %) = HA(0.2 %) = HPMC(0.3 %).

Conclusions

The method proposed is useful to discern between the mucin-polymer chemical interactions at molecular scale. Results reinforce the usefulness of chitosan and dendrimers as polymers able to increase the retention time of drugs on the ocular surface and hence their bioavailability.

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Scheme 1

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Abbreviations

Ch:

chitosan

CMC:

carboxymethyl cellulose

HA:

hyaluronic acid

HCLE:

telomerase-immortalized human corneal-limbal epithelial

HPMC:

hydroxypropylmethyl cellulose

IEP:

isoelectric point

PAMAM:

polyamidoamine dendrimers

QCM-D:

quartz crystal microbalance with dissipation

RU:

resonance units

SPR:

surface plasmon resonance

XPS:

x-ray photoelectron spectroscopy

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ACKNOWLEDGMENTS & DISCLOSURES

Dr. Bravo-Osuna would like to thank the Institute de Chimie du Centre National de Recherche Scientifique du France (CNRS) for financial support. She is also very thankful to Mrs. S. Mazzaferro (CNRS UMR 8612, Université Paris Sud) for the preparation of low molecular weight chitosan, to Dr. V. Andrés and Dr. M. Vicario (University Complutense of Madrid) for their useful comments, and to Dr. J.A. García (Surface Physics and Engineering Department–CSIC) for kindly help in XPS discussion. Dr. Bravo-Osuna, Dr. Herrero-Vanrell and Dr. Molina-Martínez would like to thank to Research Group UCM 920415 (GR35/10-A) and MAT2010-18242 for financial support. Dr. Argüeso would like to thank NIH/NEI Grant No. R01EY014847 (PA) for financial support. Authors would like to thank the IOTDYS (Université Paris VII) for XPS analysis.

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Authors and Affiliations

  1. Dept. of Pharmacy & Pharmaceutical Technology, School of Pharmacy, University Complutense of Madrid (UCM), Madrid, Spain

    I. Bravo-Osuna, I. T. Molina Martínez & R. Herrero-Vanrell

  2. CNRS UMR 8612, Université de Paris Sud Laboratoire de Physicochimie, Pharmacotechnie & Biopharmacie, Chatenay-Malabry, France

    M. Noiray, E. Briand & G. Ponchel

  3. Department of Ophthalmology, Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts, 02114, USA

    A. M. Woodward & P. Argüeso

Authors
  1. I. Bravo-Osuna
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  2. M. Noiray
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  3. E. Briand
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  4. A. M. Woodward
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  5. P. Argüeso
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  6. I. T. Molina Martínez
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  7. R. Herrero-Vanrell
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  8. G. Ponchel
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Corresponding author

Correspondence to I. Bravo-Osuna.

APPENDIX

APPENDIX

Chip Regeneration

Some authors have already observed the strong interaction established between mucins and polymers during SPR experiments, which makes difficult, even impossible to separate both ligand and analyte without the risk of altering mucin structure (10,11). Several detergents as well as acidic and basic solutions have been proposed to regenerate ligand surfaces in SPR studies (41), however, all recommended solutions were useless in the regeneration of a ligand layer in the present experimental work (data non shown). In order to reuse chips, it was necessary to develop a total regeneration method, which removed not only analyte but also ligand from the gold surface. The method selected, known as “basic piranha”, was able to eliminate all bound protein. This method, in combination with surface exposure to a UV-ozone chamber, was used to recondition the Au surface. XPS studies were performed on regenerated Au surfaces. According to Fig. 2b, c, the intensity of Au4f reached levels similar to the non-treated gold surfaces. Additionally in the core levels, neither N1s peak nor second maximum at 288 eV were recorded in the regenerated Au chip scan, indicative of the absence of proteins. Further, the cleaning procedure does not alter the chemical state since there is not energy displacement of peak positions.

Additionally, the baselines of “recycled” chips were monitored on Biacore® showing statistically similar (p > 0.05) values before and after regeneration.

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Bravo-Osuna, I., Noiray, M., Briand, E. et al. Interfacial Interaction between Transmembrane Ocular Mucins and Adhesive Polymers and Dendrimers Analyzed by Surface Plasmon Resonance. Pharm Res 29, 2329–2340 (2012). https://doi.org/10.1007/s11095-012-0761-1

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  • DOI: https://doi.org/10.1007/s11095-012-0761-1

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