Induction of Chondrogenic Differentiation in Human Mesenchymal Stem Cells Cultured on Human Demineralized Bone Matrix Scaffold under Hydrostatic Pressure
10.1007/s13770-018-0164-4
- Author:
Saeid Reza SHAHMORADI
1
;
Maryam KABIR SALMANI
;
Hamid Reza SOLEIMANPOUR
;
Amir Hossein TAVAKOLI
;
Kazem HOSAINI
;
Nooshin HAGHIGHIPOUR
;
Shahin BONAKDAR
Author Information
1. Biomaterials and Tissue Engineering Department, Stem Cell Division, National Institute of Genetic Engineering and Biotechnology, Shahrak-e Pajoohesh, km 15, Tehran
- Publication Type:Original Article
- Keywords:
Bone marrow mesenchymal stem cells;
Chondrogenic differentiation;
Hydrostatic pressure;
Demineralized bone matrix scaffold
- MeSH:
Aggrecans;
Alcian Blue;
Bone Marrow;
Bone Matrix;
Cartilage;
Cartilage, Articular;
Collagen Type I;
Collagen Type II;
Eosine Yellowish-(YS);
Extracellular Matrix;
Fluorescent Antibody Technique;
Gene Expression;
Hematoxylin;
Humans;
Hydrostatic Pressure;
In Vitro Techniques;
Mesenchymal Stromal Cells;
Osteocalcin;
Real-Time Polymerase Chain Reaction;
RNA, Messenger
- From:
Tissue Engineering and Regenerative Medicine
2019;16(1):69-80
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
