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Effects of Nanoparticle Properties on Kartogenin Delivery and Interactions with Mesenchymal Stem Cells

  • S.I. : Biomaterials - Engineering Cell Behavior
  • Published:
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Abstract

Clinical trials with mesenchymal stem cells (MSCs) have demonstrated potential to treat osteoarthritis, a debilitating disease that affects millions. However, these therapies are often less effective due to heterogeneous MSC differentiation. Kartogenin (KGN), a synthetic small molecule that induces chondrogenesis, has recently been explored to decrease this heterogeneity. KGN has been encapsulated in nanoparticles due to its hydrophobicity. To explore the effect of nanoparticle properties on KGN and MSC interactions, here we fabricated three nanoparticle formulations that vary in hydrophobicity, size, and surface charge using nanoprecipitation: KGN-loaded poly(lactic acid-co-glycolic acid) (PLGA) nanoparticles (hydrophobic surface, negative charge, ~ 167 nm), PLGA–poly(ethylene glycol) (PEG) nanoparticles (hydrophilic surface, positive charge, ~ 297 nm), and PLGA–PEG–hyaluronic acid (HA) nanoparticles (hydrophilic surface, negative charge, ~ 507 nm). We observed differences in KGN loading, release, and suspension stability, with the PLGA particles exhibiting ~ 50% drug loading and PLGA–PEG–HA particles releasing the most KGN. All nanoparticles were found to interact with MSCs with evidence of increased uptake in PLGA–PEG and PLGA–PEG–HA compared with surface association of PLGA particles. Over short times (~ 7 days), MSCs incubated with all KGN-loaded formulations exhibited a similar increase in sulfated glycosaminoglycans, characteristic of chondrogenic differentiation, compared with non-KGN loaded formulations.

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Abbreviations

OA:

Osteoarthritis

ECM:

Extracellular matrix

MSCs:

Mesenchymal stem cells

hMSCs:

Human mesenchymal stem cells

KGN:

Kartogenin

CBFβ:

Core-binding factor subunit β

RUNX:

Runt-related transcription factor

TGF-β1:

Transforming growth factor beta 1

EC50 :

Half maximal effective concentration

MW:

Molecular weight

PLGA:

Poly(lactic-co-glycolic acid)

PLGA–PEG:

PLGA–poly(ethylene glycol)

PLGA–PEG–HA:

PLGA–PEG–hyaluronic acid

GAG:

Glycosoaminoglycan

sGAG:

Sulfated glycosaminoglycan

PEG-bis-NH2 :

PEG-bis-amine

PVA:

Poly(vinyl alcohol)

Sulfo-NHS:

N-hydroxysulfosuccinimide

DCC:

N,N′-dicyclohexylcarbodiimide

EDC:

N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride

DCM:

Dichloromethane

ACN:

Acetonitrile

MES:

2-(N-morpholino)ethanesulfonic acid

FITC:

Fluorescein isothiocyanate

CTAB:

Hexadecyltrimethylammonium bromide

PBS:

Phosphate buffered saline

DMSO:

Dimethyl sulfoxide

EDTA:

Ethylenediaminetetraacetic acid disodium salt

HCl:

Hydrochloric acid

TFA:

Trifluoroacetic acid

CCK-8:

Cell Counting Kit-8

DMEM:

Dulbecco’s modified Eagle’s medium

ITS+:

Insulin/transferrin/selenium

HPLC:

High performance liquid chromatography

N2 :

Nitrogen

RT:

Room temperature

1H-NMR:

Proton nuclear magnetic resonance

DLS:

Dynamic light scattering

PDI:

Polydispersity index

TEM:

Transmission electron microscopy

EE%:

Encapsulation efficiency

DL:

Drug loading

ANOVA:

Analysis of variance

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Acknowledgments

The authors acknowledge financial support from Brown University. We thank Dr. Alessia Battigelli from Brown University for her helpful guidance on the chemical syntheses presented. We also thank Kevin Carlson from the Flow Cytometry Core at Brown University and Professor Robert Hurt at Brown for use of his DLS. Finally, we thank Anthony McCormick from Brown University’s Electron Microscopy Core Facility for assistance with TEM.

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Author notes

  1. Bethany Almeida and Yingying Wang are co-first authors and contributed equally to this work.

Authors and Affiliations

  1. School of Engineering, Center for Biomedical Engineering, Institute for Molecular and Nanoscale Innovation, Brown University, 184 Hope Street, Providence, RI, USA

    Bethany Almeida & Anita Shukla

  2. Department of Chemistry, Brown University, 324 Brook Street, Providence, RI, USA

    Yingying Wang

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  1. Bethany Almeida
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Correspondence to Anita Shukla.

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Almeida, B., Wang, Y. & Shukla, A. Effects of Nanoparticle Properties on Kartogenin Delivery and Interactions with Mesenchymal Stem Cells. Ann Biomed Eng 48, 2090–2102 (2020). https://doi.org/10.1007/s10439-019-02430-x

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