Evaluation of a cell-based osteogenic formulation compliant with good manufacturing practice for use in tissue engineering

Abstract

Proper bony tissue regeneration requires mechanical stabilization, an osteogenic biological activity and appropriate scaffolds. The latter two elements can be combined in a hydrogel format for effective delivery, so it can readily adapt to the architecture of the defect. We evaluated a Good Manufacturing Practice-compliant formulation composed of bone marrow-derived mesenchymal stromal cells in combination with bone particles (Ø = 0.25 to 1 µm) and fibrin, which can be readily translated into the clinical setting for the treatment of bone defects, as an alternative to bone tissue autografts. Remarkably, cells survived with unaltered phenotype (CD73+, CD90+, CD105+, CD31, CD45) and retained their osteogenic capacity up to 48 h after being combined with hydrogel and bone particles, thus demonstrating the stability of their identity and potency. Moreover, in a subchronic toxicity in vivo study, no toxicity was observed upon subcutaneous administration in athymic mice and signs of osteogenesis and vascularization were detected 2 months after administration. The preclinical data gathered in the present work, in compliance with current quality and regulatory requirements, demonstrated the feasibility of formulating an osteogenic cell-based tissue engineering product with a defined profile including identity, purity and potency (in vitro and in vivo), and the stability of these attributes, which complements the preclinical package required prior to move towards its use of prior to its clinical use.

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Data availability

All datasets generated for this study are included in the manuscript and the supplementary files.

Abbreviations

ALP:

Alkaline phosphatase

BM:

Bone Marrow

DMEM:

Dulbecco’s Modified Eagle’s Medium

EPC:

Endothelial progenitor cells

GMP:

Good Manufacturing Practice

HLA:

Human Leukocyte Antigen

HPC:

Hematopoietic stem progenitor cells

HSA:

Human serum albumin

MNC:

Mononuclear cell

MSC:

Multipotent Mesenchymal Stromal Cells

P:

Passage number

PBMC:

Peripheral Blood Mononuclear Cells

TEP:

Tissue Engineering Product

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Acknowledgements

The authors would like to acknowledge former members of Xcelia and current members of Servei de Teràpia Cel·lular (Banc de Sang i Teixits, Barcelona) for technical support and advice.

Funding

This work has been developed in the context of ADVANCE(CAT) with the support of ACCIÓ (Catalonia Trade & Investment; Generalitat de Catalunya) and the European Community under the Catalonian ERDF operational program (European Regional Development Fund) 2014-2020, Generalitat de Catalunya (Departament de Salut) PERIS Acció Instrumental de Programes de Recerca Orientats (SLT002/16/00234) and by the Spanish Cell Therapy Network (TerCel, expedient number: RD16/0011/0028). Project PI19/01788 is funded by Instituto de Salud Carlos III and co-funded by European Union (ERDF/ESF)—A way to build Europe. JV’s laboratory is awarded by the Generalitat de Catalunya as Consolidated Research Group (ref. 2017SGR719).

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Authors

Contributions

DV, MGV, IOV and JV performed experiments and analysed data; JGL and JV conceived the study; DV and JV revised data and wrote the manuscript. All authors revised and approved the final version of the manuscript.

Corresponding author

Correspondence to Joaquim Vives.

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All authors declares that they have no conflict of interest.

Ethical approval

Cells were sourced from Banc de Sang i Teixits’ Biobank (Barcelona, Spain) and had appropriate donor informed consent for use in research. Authorization issued by Hospital de la Vall d’Hebron’s Ethics Committee (Barcelona, Spain) to JV. All animal care and experimental procedures adhered to the recommendations of local, national, and European laws (Decret 214 de 1997, Real Decreto 53 de 2013, European directive 86/609/CEE of 1986, respectively) and were approved by the Universitat Autònoma de Barcelona’s Ethical Committee on Human and Animal Experimentation (Ref. No. CEAAH 1972), and registered by Generalitat de Catalunya (Reg. No. DMAiH 6967).

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Vivas, D., Grau-Vorster, M., Oliver-Vila, I. et al. Evaluation of a cell-based osteogenic formulation compliant with good manufacturing practice for use in tissue engineering. Mol Biol Rep (2020). https://doi.org/10.1007/s11033-020-05588-z

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Keywords

  • Multipotent mesenchymal stromal cell
  • Good manufacturing practice
  • Hydrogel
  • Bone remodelling
  • Cell therapy
  • Cell culture
  • Tissue engineering