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Evaluation of Gelatin Microparticles as Adherent-Substrates for Mesenchymal Stem Cells in a Hydrogel Composite

  • Emerging Trends in Biomaterials Research
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Abstract

Due to the lack of cell-adhesive moieties in traditional synthetic hydrogels, the present work investigated the use of degradable gelatin microparticles (GMPs) as temporary adherent substrates for anchorage-dependent mesenchymal stem cells (MSCs). MSCs were seeded onto GMPs of varying crosslinking densities and sizes to investigate their role on influencing MSC differentiation and aggregation. The MSC-seeded GMPs were then encapsulated in poly(ethylene glycol)-based hydrogels and cultured in serum-free, growth factor-free osteochondral medium. Non-seeded MSCs co-encapsulated with GMPs in the hydrogels were used as a control for comparison. Over the course of 35 days, MSCs seeded on GMPs exhibited more cell–cell contacts, greater chondrogenic potential, and a down-regulation of osteogenic markers compared to the controls. Although the factors of GMP crosslinking and size had nominal influence on MSC differentiation and aggregation, GMPs demonstrate potential as an adherent-substrate for improving cell delivery from hydrogel scaffolds by facilitating cell–cell contacts and improving MSC differentiation.

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Abbreviations

ACAN :

Aggrecan

ALP:

Alkaline phosphatase

APS:

Ammonium persulfate

CC-PBS:

Collagenase-containing PBS

CDH2 :

N-cadherin

COL1 :

Type I collagen

COL2 :

Type II collagen

ddH2O:

Distilled, deionized water

FBS:

Fetal bovine serum

GAG:

Glycosaminoglycan

GMP:

Gelatin microparticle

HYP:

Hydroxyproline

LG-DMEM:

Low glucose-Dulbecco’s modified Eagle’s medium

MMP:

Matrix metalloproteinase

MSC:

Mesenchymal stem cell

OM:

Osteochondral medium

OPF:

Oligo(poly(ethylene glycol) fumarate

PBS:

Phosphate-buffered saline

PEG:

Poly(ethylene glycol)

RT-PCR:

Reverse transcription polymerase chain reaction

RUNX2 :

Runt-related transcription factor 2

SOX9 :

(Sex determining region Y)-box 9

TEMED:

N,N,N′,N′-tetramethylethylenediamine

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Acknowledgments

This work was supported by the National Institutes of Health (R01-AR048756 and R01-AR068073). The authors acknowledge the assistance of Brandon T. Smith with the rabbit bone marrow harvest, Marco Santoro and Tiffany N. Vo with MSC culture, and Kelsea M. Hubka with immunocytochemistry.

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Authors and Affiliations

  1. Department of Bioengineering, Rice University, P.O. Box 1892, MS-142, Houston, TX, 77005-1892, USA

    Steven Lu, Esther J. Lee, Johnny Lam & Antonios G. Mikos

  2. Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan

    Yasuhiko Tabata

Authors
  1. Steven Lu
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  2. Esther J. Lee
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  3. Johnny Lam
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  4. Yasuhiko Tabata
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Correspondence to Antonios G. Mikos.

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Associate Editor Michael S. Detamore oversaw the review of this article.

Invited submission to the special issue of “Biomaterials” in Annals of Biomedical Engineering, guest edited by Michael Detamore.

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Lu, S., Lee, E.J., Lam, J. et al. Evaluation of Gelatin Microparticles as Adherent-Substrates for Mesenchymal Stem Cells in a Hydrogel Composite. Ann Biomed Eng 44, 1894–1907 (2016). https://doi.org/10.1007/s10439-016-1582-x

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  • DOI: https://doi.org/10.1007/s10439-016-1582-x

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