Improved Protocol for Chondrogenic Differentiation of Bone Marrow Derived Mesenchymal Stem Cells -Effect of PTHrP and FGF-2 on TGFβ1/BMP2-Induced Chondrocytes Hypertrophy

Abstract

Growth factors have a pivotal role in chondrogenic differentiation of stem cells. The differential effects of known growth factors involved in the maintenance and homeostasis of cartilage tissue have been previously studied in vitro. However, there are few reported researches about the interactional effects of growth factors on chondrogenic differentiation of stem cells. The aim of this study is to examine the combined effects of four key growth factors on chondrogenic differentiation of mesenchymal stem cells (MSCs). Isolated and expanded rabbit bone marrow-derived MSCs underwent chondrogenic differentiation in a micromass cell culture system that used a combination of the following growth factors: transforming growth factor beta 1 (TGF-β1), bone morphogenetic protein 2 (BMP2), parathyroid hormone related protein (PTHrP), and fibroblast growth factor 2 (FGF2) according to a defined program. The chondrogenic differentiation program was analyzed by histochemistry methods, quantitative RT-PCR (qRT-PCR), and measurement of matrix deposition of sulfated glycosaminoglycan (sGAG) and collagen content at days 16, 23, and 30. The results showed that the short-term combination of TGF-β1 and BMP-2 increased sGAG and collagen content, Alkaline phosphates (ALP) activity, and type X collagen (COL X) expression. Application of either PTHrP or FGF2 simultaneously decreased TGF-β1/BMP-2 induced hypertrophy and chondrogenic markers (at least for FGF2). However, successive application of PTHrP and FGF2 dramatically maintained the synergistic effects of TGF-β1/BMP-2 on the chondrogenic differentiation potential of MSCs and decreased unwanted hypertrophic markers. This new method can be used effectively in chondrogenic differentiation programs.

Osteochondritis dissecans (OCD) in Horses – Molecular Background of its Pathogenesis and Perspectives for Progenitor Stem Cell Therapy

“…When mature tissue injury occurs during the development of OC(D), the damaged cartilage has only a very low intrinsic self-repair and regeneration potential, as long as chondrocytes are considered to not have the capacity for migration, proliferation and repair. In fact, although they have low metabolic activity these cells are mainly involved in the production of key elements essential to maintaining the integrity of cartilage, namely: type II collagen and aggrecan, mainly known to play a fundamental role in the mechanical resistance of articular cartilage tissue [ 36 – 38 ]. Moreover, chondrocytes tend to lose their reproductive potential of the extracellular matrix (ECM) by dedifferentiation rather quickly.…”

Effects of various antimicrobial agents on multi-directional differentiation potential of bone marrow-derived mesenchymal stem cells

“…As an important seed cell for local cartilage repair, the ability of bone marrow-derived MSCs to differentiate into chondrocytes in the direction of cartilage is essential[54]. After the cartilage tissue is damaged by factors such as trauma, inflammation, and infection, microfracture surgery is an important approach for clinical treatment of local cartilage defects[55].…”

Bone defect reconstruction via endochondral ossification: A developmental engineering strategy

“…For example, a combination of TGF-β 1 and BMP-2 has been shown to increase the GAG and collagen content and initiate robust endochondral lineage commitment. [158][159][160] The dual delivery of TGFβ 1 and BMP-2 within BMSC aggregates has been shown to result in enhanced chondrogenesis and an enhanced osteogenic phenotype, as well as a greater degree of mineralization and COL X expression. 161,162 Additionally, compared to TGF-β 3 alone, combinations of TGF-β 3 and BMP-6 can enhance the chondrogenic potential of BMSCs and ASCs.…”