Tissue Engineering and Regenerative Medicine

, Volume 16, Issue 1, pp 69–80 | Cite as

Induction of Chondrogenic Differentiation in Human Mesenchymal Stem Cells Cultured on Human Demineralized Bone Matrix Scaffold under Hydrostatic Pressure

  • Saeid Reza Shahmoradi
  • Maryam Kabir SalmaniEmail author
  • Hamid Reza Soleimanpour
  • Amir Hossein Tavakoli
  • Kazem Hosaini
  • Nooshin Haghighipour
  • Shahin BonakdarEmail author
Original Article



Articular cartilage damage is still a troublesome problem. Hence, several researches have been performed for cartilage repair. The aim of this study was to evaluate the chondrogenicity of demineralized bone matrix (DBM) scaffolds under cyclic hydrostatic pressure (CHP) in vitro.


In this study, CHP was applied to human bone marrow mesenchymal stem cells (hBMSCs) seeded on DBM scaffolds at a pressure of 5 MPa with a frequency of 0.5 Hz and 4 h per day for 1 week. Changes in chondrogenic and osteogenic gene expressions were analyzed by quantifying mRNA signal level of Sox9, collagen type I, collagen type II, aggrecan (ACAN), Osteocalcin, and Runx2. Histological analysis was carried out by hematoxylin and eosin, and Alcian blue staining. Moreover, DMMB and immunofluorescence staining were used for glycosaminoglycan (GAG) and collagen type II detection, respectively.


Real-time PCR demonstrated that applying CHP to hBMSCs in DBM scaffolds increased mRNA levels by 1.3-fold, 1.2-fold, and 1.7-fold (p < 0.005) for Sox9, Col2, and ACAN, respectively by day 21, whereas it decreased mRNA levels by 0.7-fold and 0.8-fold (p < 0.05) for Runx2 and osteocalcin, respectively. Additionally, in the presence of TGF-β1 growth factor (10 ng/ml), CHP further increased mRNA levels for the mentioned genes (Sox9, Col2, and ACAN) by 1.4-fold, 1.3-fold and 2.5-fold (p < 0.005), respectively. Furthermore, in histological assessment, it was observed that the extracellular matrix contained GAG and type II collagen in scaffolds under CHP and CHP with TGF-β1, respectively.


The osteo-inductive DBM scaffolds showed chondrogenic characteristics under hydrostatic pressure. Our study can be a fundamental study for the use of DBM in articular cartilage defects in vivo and lead to production of novel scaffolds with two different characteristics to regenerate both bone and cartilage simultaneously.

Graphical abstract


Bone marrow mesenchymal stem cells Chondrogenic differentiation Hydrostatic pressure Demineralized bone matrix scaffold 



The authors would like to express their appreciation to Alborz Balk Pharmaceutical Company, Nano Zist Arrayeh, and Dr. Kayvan pathology lab for their generous support. This study was supported by Iran Pasteur Institute research (Grant No. 861).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

There are no animal experiments carried out in this article. However, tissue removal from donors was carried out according to Iran Brain death law and Iran Tissue Bank Standards. Ethical and technical proses had been approved by Iran Food and Drug Administration (664/188547).


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Copyright information

© The Korean Tissue Engineering and Regenerative Medicine Society and Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biomaterials and Tissue Engineering Department, Stem Cell DivisionNational Institute of Genetic Engineering and BiotechnologyTehranIran
  2. 2.Iranian Tissue Bank, Imam khomani HospitalUniversity of Medical SciencesTehranIran
  3. 3.National Cell Bank DepartmentPasteur Institute of Iran (IPI)TehranIran

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