Near infrared fluorescent nanoparticles based on hyaluronic acid

Self-assembly, optical properties, and cell interaction

Sneha S. Kelkar, Tanner K. Hill, Frank C. Marini, Aaron M Mohs

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Fluorescent imaging agents that can specifically highlight tumor cells could have a significant impact on image-guided tumor removal. Here, fluorescent nanoparticles (NPs) derived from hyaluronic acid (HA) are investigated. HA is a ligand for the receptor CD44, which is a common biomarker present on many primary tumor cells, cancer-initiating cells, and tumor-associated fibroblasts. In addition, a family of enzymes that degrade HA, called hyaluronidases (HYALs), are also overexpressed with increased activity in many tumors. We report the design and development of a panel of targeted imaging agents using the near-infrared (NIR) dye, Cy7.5, that was directly conjugated to hydrophobically-modified HA. Two different molecular weights of HA, 10 kDa and 100 kDa, and three different degrees of hydrophobic moiety conjugation (0, 10, and 30 mol%) were utilized to develop a panel of NPs with variable size that ranged from 50 to 400 nm hydrodynamic diameter (HD) depending HA molecular weight, extent of fluorescence quenching (25-50%), kinetics of cellular uptake, and targeting to CD44+ cells. The kinetics and energy-dependence of cellular uptake in breast and prostate cancer cell lines, MDA-MB 231 and PC-3 cells, respectively, showed increased uptake with longer incubation times (at 4 and 8 h compared to 1 h), as well as uptake at 37 °C but not 4 °C, which indicated energy-dependent endocytosis. NP uptake studies in the presence of excess free HA showed that pre-treatment of cells with excess high molecular weight (MW) free HA decreased NP uptake by up to 50%, while no such trend was observed with low MW HA. These data lay the foundation for selection of optimized HA-derived NPs for image-guided surgery. Statement of Significance Here, hyaluronic acid (HA), a well-studied biomacromolecule, is modified with a near infrared fluorophore and a hydrophobic moiety. The significance of this work, especially for imaging applications, is that the impact of HA molecular weight and the hydrophobic moiety conjugation degree on fluorescence and cell interaction can be predicted. With respect to existing literature, the eventual use of these HA-based NPs is image-guided surgery; thus, we focus on the dye, Cy7.5, for conjugation, which is more NIR than most existing HA literature. Furthermore, HA is a ligand for CD44, which is associated with cancer and tumor microenvironment cells. Systematic studies in this work highlight that HA can be tuned to maximize or minimize CD44 binding.

Original languageEnglish (US)
Pages (from-to)112-121
Number of pages10
JournalActa Biomaterialia
Volume36
DOIs
StatePublished - May 1 2016

Fingerprint

Hyaluronic acid
Hyaluronic Acid
Cell Communication
Nanoparticles
Self assembly
Optical properties
Infrared radiation
Tumors
Molecular Weight
Molecular weight
Cells
Computer-Assisted Surgery
Tumor Microenvironment
Imaging techniques
Surgery
Neoplasms
Coloring Agents
Dyes
Fluorescence
Ligands

Keywords

  • Cy7.5
  • Hyaluronic acid
  • Image-guided surgery
  • Intraoperative imaging
  • Near-infrared fluorescence

ASJC Scopus subject areas

  • Biomaterials
  • Biomedical Engineering
  • Biotechnology
  • Biochemistry
  • Molecular Biology

Cite this

Near infrared fluorescent nanoparticles based on hyaluronic acid : Self-assembly, optical properties, and cell interaction. / Kelkar, Sneha S.; Hill, Tanner K.; Marini, Frank C.; Mohs, Aaron M.

In: Acta Biomaterialia, Vol. 36, 01.05.2016, p. 112-121.

Research output: Contribution to journalArticle

@article{c66c0515801740c9950d70b64f7ec0fa,
title = "Near infrared fluorescent nanoparticles based on hyaluronic acid: Self-assembly, optical properties, and cell interaction",
abstract = "Fluorescent imaging agents that can specifically highlight tumor cells could have a significant impact on image-guided tumor removal. Here, fluorescent nanoparticles (NPs) derived from hyaluronic acid (HA) are investigated. HA is a ligand for the receptor CD44, which is a common biomarker present on many primary tumor cells, cancer-initiating cells, and tumor-associated fibroblasts. In addition, a family of enzymes that degrade HA, called hyaluronidases (HYALs), are also overexpressed with increased activity in many tumors. We report the design and development of a panel of targeted imaging agents using the near-infrared (NIR) dye, Cy7.5, that was directly conjugated to hydrophobically-modified HA. Two different molecular weights of HA, 10 kDa and 100 kDa, and three different degrees of hydrophobic moiety conjugation (0, 10, and 30 mol{\%}) were utilized to develop a panel of NPs with variable size that ranged from 50 to 400 nm hydrodynamic diameter (HD) depending HA molecular weight, extent of fluorescence quenching (25-50{\%}), kinetics of cellular uptake, and targeting to CD44+ cells. The kinetics and energy-dependence of cellular uptake in breast and prostate cancer cell lines, MDA-MB 231 and PC-3 cells, respectively, showed increased uptake with longer incubation times (at 4 and 8 h compared to 1 h), as well as uptake at 37 °C but not 4 °C, which indicated energy-dependent endocytosis. NP uptake studies in the presence of excess free HA showed that pre-treatment of cells with excess high molecular weight (MW) free HA decreased NP uptake by up to 50{\%}, while no such trend was observed with low MW HA. These data lay the foundation for selection of optimized HA-derived NPs for image-guided surgery. Statement of Significance Here, hyaluronic acid (HA), a well-studied biomacromolecule, is modified with a near infrared fluorophore and a hydrophobic moiety. The significance of this work, especially for imaging applications, is that the impact of HA molecular weight and the hydrophobic moiety conjugation degree on fluorescence and cell interaction can be predicted. With respect to existing literature, the eventual use of these HA-based NPs is image-guided surgery; thus, we focus on the dye, Cy7.5, for conjugation, which is more NIR than most existing HA literature. Furthermore, HA is a ligand for CD44, which is associated with cancer and tumor microenvironment cells. Systematic studies in this work highlight that HA can be tuned to maximize or minimize CD44 binding.",
keywords = "Cy7.5, Hyaluronic acid, Image-guided surgery, Intraoperative imaging, Near-infrared fluorescence",
author = "Kelkar, {Sneha S.} and Hill, {Tanner K.} and Marini, {Frank C.} and Mohs, {Aaron M}",
year = "2016",
month = "5",
day = "1",
doi = "10.1016/j.actbio.2016.03.024",
language = "English (US)",
volume = "36",
pages = "112--121",
journal = "Acta Biomaterialia",
issn = "1742-7061",
publisher = "Elsevier BV",

}

TY - JOUR

T1 - Near infrared fluorescent nanoparticles based on hyaluronic acid

T2 - Self-assembly, optical properties, and cell interaction

AU - Kelkar, Sneha S.

AU - Hill, Tanner K.

AU - Marini, Frank C.

AU - Mohs, Aaron M

PY - 2016/5/1

Y1 - 2016/5/1

N2 - Fluorescent imaging agents that can specifically highlight tumor cells could have a significant impact on image-guided tumor removal. Here, fluorescent nanoparticles (NPs) derived from hyaluronic acid (HA) are investigated. HA is a ligand for the receptor CD44, which is a common biomarker present on many primary tumor cells, cancer-initiating cells, and tumor-associated fibroblasts. In addition, a family of enzymes that degrade HA, called hyaluronidases (HYALs), are also overexpressed with increased activity in many tumors. We report the design and development of a panel of targeted imaging agents using the near-infrared (NIR) dye, Cy7.5, that was directly conjugated to hydrophobically-modified HA. Two different molecular weights of HA, 10 kDa and 100 kDa, and three different degrees of hydrophobic moiety conjugation (0, 10, and 30 mol%) were utilized to develop a panel of NPs with variable size that ranged from 50 to 400 nm hydrodynamic diameter (HD) depending HA molecular weight, extent of fluorescence quenching (25-50%), kinetics of cellular uptake, and targeting to CD44+ cells. The kinetics and energy-dependence of cellular uptake in breast and prostate cancer cell lines, MDA-MB 231 and PC-3 cells, respectively, showed increased uptake with longer incubation times (at 4 and 8 h compared to 1 h), as well as uptake at 37 °C but not 4 °C, which indicated energy-dependent endocytosis. NP uptake studies in the presence of excess free HA showed that pre-treatment of cells with excess high molecular weight (MW) free HA decreased NP uptake by up to 50%, while no such trend was observed with low MW HA. These data lay the foundation for selection of optimized HA-derived NPs for image-guided surgery. Statement of Significance Here, hyaluronic acid (HA), a well-studied biomacromolecule, is modified with a near infrared fluorophore and a hydrophobic moiety. The significance of this work, especially for imaging applications, is that the impact of HA molecular weight and the hydrophobic moiety conjugation degree on fluorescence and cell interaction can be predicted. With respect to existing literature, the eventual use of these HA-based NPs is image-guided surgery; thus, we focus on the dye, Cy7.5, for conjugation, which is more NIR than most existing HA literature. Furthermore, HA is a ligand for CD44, which is associated with cancer and tumor microenvironment cells. Systematic studies in this work highlight that HA can be tuned to maximize or minimize CD44 binding.

AB - Fluorescent imaging agents that can specifically highlight tumor cells could have a significant impact on image-guided tumor removal. Here, fluorescent nanoparticles (NPs) derived from hyaluronic acid (HA) are investigated. HA is a ligand for the receptor CD44, which is a common biomarker present on many primary tumor cells, cancer-initiating cells, and tumor-associated fibroblasts. In addition, a family of enzymes that degrade HA, called hyaluronidases (HYALs), are also overexpressed with increased activity in many tumors. We report the design and development of a panel of targeted imaging agents using the near-infrared (NIR) dye, Cy7.5, that was directly conjugated to hydrophobically-modified HA. Two different molecular weights of HA, 10 kDa and 100 kDa, and three different degrees of hydrophobic moiety conjugation (0, 10, and 30 mol%) were utilized to develop a panel of NPs with variable size that ranged from 50 to 400 nm hydrodynamic diameter (HD) depending HA molecular weight, extent of fluorescence quenching (25-50%), kinetics of cellular uptake, and targeting to CD44+ cells. The kinetics and energy-dependence of cellular uptake in breast and prostate cancer cell lines, MDA-MB 231 and PC-3 cells, respectively, showed increased uptake with longer incubation times (at 4 and 8 h compared to 1 h), as well as uptake at 37 °C but not 4 °C, which indicated energy-dependent endocytosis. NP uptake studies in the presence of excess free HA showed that pre-treatment of cells with excess high molecular weight (MW) free HA decreased NP uptake by up to 50%, while no such trend was observed with low MW HA. These data lay the foundation for selection of optimized HA-derived NPs for image-guided surgery. Statement of Significance Here, hyaluronic acid (HA), a well-studied biomacromolecule, is modified with a near infrared fluorophore and a hydrophobic moiety. The significance of this work, especially for imaging applications, is that the impact of HA molecular weight and the hydrophobic moiety conjugation degree on fluorescence and cell interaction can be predicted. With respect to existing literature, the eventual use of these HA-based NPs is image-guided surgery; thus, we focus on the dye, Cy7.5, for conjugation, which is more NIR than most existing HA literature. Furthermore, HA is a ligand for CD44, which is associated with cancer and tumor microenvironment cells. Systematic studies in this work highlight that HA can be tuned to maximize or minimize CD44 binding.

KW - Cy7.5

KW - Hyaluronic acid

KW - Image-guided surgery

KW - Intraoperative imaging

KW - Near-infrared fluorescence

UR - http://www.scopus.com/inward/record.url?scp=84961262520&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84961262520&partnerID=8YFLogxK

U2 - 10.1016/j.actbio.2016.03.024

DO - 10.1016/j.actbio.2016.03.024

M3 - Article

VL - 36

SP - 112

EP - 121

JO - Acta Biomaterialia

JF - Acta Biomaterialia

SN - 1742-7061

ER -