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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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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DOI: https://doi.org/10.1007/s10439-019-02430-x