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Chondrogenic Commitment of human Bone  Marrow Mesenchymal Stem Cells cultured under perfusion within a 3D collagen  environment releasing hTGF\(\beta\)1
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  • Erwin Lamparelli,
  • Joseph Lovecchio,
  • Luigi Marino,
  • Maria Ciardulli,
  • Carmine Selleri,
  • Nicholas Forsyth,
  • Emanuele Giordano,
  • Nicola Maffulli,
  • Giovanna Della Porta
Erwin Lamparelli
University of Salerno - Baronissi Campus

Corresponding Author:elamparelli@unisa.it

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Joseph Lovecchio
University of Bologna
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Luigi Marino
University of Salerno - Baronissi Campus
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Maria Ciardulli
University of Salerno - Baronissi Campus
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Carmine Selleri
University of Salerno - Baronissi Campus
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Nicholas Forsyth
Keele University
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Emanuele Giordano
Università di Bologna
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Nicola Maffulli
University of Salerno
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Giovanna Della Porta
University of Salerno - Baronissi Campus
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Abstract

The optimal growth, maturation and function of bioengineered tissues are mediated by both biochemical and physical cues. We here describe a 3D biomimetic environment directing stem cells towards a chondrogenic phenotype. This system comprises a collagen hydrogel and poly-lactic-co-glycolic acid microcarriers (PLGA-MCs) engineered to protect, carry and release a human Transforming Growth Factor b1 (hTFGb1) payload. PLGA-MCs were prepared using supercritical emulsion extraction technology and integrated into a collagen hydrogel co-seeded with human Bone Marrow Mesenchymal Stem Cells (hBM-MSCs). Testing different concentrations of hTFGb1 supplemented to cell monolayer cultures suggested 10 ng/mL as the most appropriate concentration to promote upregulation of SRY-Related HMG-BOXGene 9 (4-fold) and collagen type II (2-fold) specific markers, at Day 16. A similar growth factor concentration was delivered within the 3D bioengineered environment cultured in a dynamic via a custom perfusion bioreactor. A chondrogenic commitment was obtained as indicated by upregulation of collagen type II (5-fold) and downregulation of collagen types I and III (both 0.1-fold) at Day 16. Histological analysis confirmed the remodeling of the synthetic extracellular matrix in where an enhanced mass exchange was described by FEM analysis of fluid-dynamics and related nutrient mass transfer within the 3D construct. This study supports the use of 3D bioengineered scaffolds cultured in a dynamic environment as a suitable tissue engineered model to study chondrogenic differentiation in vitro and opens perspectives for an injectable collagen-based advanced therapy system.
24 Jun 2020Submitted to Biotechnology and Bioengineering
24 Jun 2020Submission Checks Completed
24 Jun 2020Assigned to Editor
29 Jun 2020Reviewer(s) Assigned
07 Aug 2020Review(s) Completed, Editorial Evaluation Pending