Concentrated Growth Factors (CGF) Induce Osteogenic Differentiation in Human Bone Marrow Stem Cells

Abstract

Bone regeneration is a complex process regulated by several factors that control overlapping biological processes, coordinating interactions among distinct cell populations. There is a great interest in identifying new strategies for inducing osteogenesis in a safe and efficient manner. Concentrated Growth Factor (CGF) is an autologous blood derived product obtained by centrifugation of venous blood following the procedure set on the Silfradent device. In this study the effects of CGF on osteogenic differentiation of human Bone Marrow Stem Cells (hBMSC) in vitro have been investigated; hBMSC were cultured with CGF or osteogenic medium, for 21 days. The osteogenic differentiation was evaluated measuring alkaline phosphatase (ALP) enzyme activity, matrix mineralization by alizarin red staining and through mRNA and protein quantification of osteogenic differentiation markers by Real-time PCR and Western blotting, respectively. The treatment with CGF stimulated ALP activity and promoted matrix mineralization compared to control and seems to be more effective than osteogenic medium. Also, hBMSC lost mesenchymal markers and showed other osteogenic features. Our study showed for the first time that CGF alone is able to induce osteogenic differentiation in hBMSC. The application of CGF on hBMSC osteoinduction might offer new clinical and biotechnological strategies in the tissue regeneration field.

Angiogenic Properties of Concentrated Growth Factors (CGFs): The Role of Soluble Factors and Cellular Components

“…Venous blood (8 mL) from 5 healthy, non-smoking adult donors, two females and three males, aged between 27 and 50 years old, was collected and immediately centrifuged by a Medifuge device (Medifuge MF200; Silfradent srl, Forlì, Italy), at 25 °C, using a program with the following characteristics: 30 s acceleration, 2 min 2700 rpm, 4 min 2400 rpm, 4 min 2700 rpm, 3 min 3000 rpm, and 36 s deceleration and stop, to obtain CGF clot, as previously described [ 36 , 38 ]. Informed consent was obtained from the donors included in this study in accordance with Declaration of Helsinki.…”

“…Several studies have investigated the effects of CGFs on tissue regeneration of alveolar and sinus bone, fracture repair, and implant stability, displaying a good tissue regenerative property [ 29 , 33 , 34 , 35 ]. Some research also showed the effects of CGFs on the osteogenic differentiation of rabbit periosteum-derived cells and human bone marrow stem cells in vitro [ 36 , 37 ], and reported CD34 positive stem cells in CGFs [ 24 , 37 ]. However, to the best of our knowledge, the CGF effects on the neo-vascularization process have not yet been fully characterized.…”

The effects and potential applications of concentrated growth factor in dentin–pulp complex regeneration

“… Yu and Wang (2014) [ 40 ] Beagle-PDLSCs Proliferation, osteogenic differentiation Cell counting and an MTT assay; Mineralization nodule counts, ALP activity, western blotting, qPCR (BSP, OCN, COL1a1), and immunohistochemistry CGF promoted PDLSCs proliferation and osteogenic differentiation in a time- and dose-dependent manner. Rochira et al (2020) [ 41 ] h-BMSCs Osteogenic differentiation ALP activity, ARS, western blotting, qPCR (RUNX2, OSX, OPN, COL1a1) CGF alone can induce osteogenic differentiation in h-BMSCs. High RUNX2 expression and RUNX2 nuclear translocation are molecular mechanisms of h-BMSCs osteogenic differentiation induced by CGF.…”

“…However, some studies have reported a lack of dose dependence, which may be attributable to the different methods used to prepare CGF [ 34 , 38 ]. Three methods for preparing CGF have been described to date—namely, spontaneous release into a medium [ 41 ], freeze-drying [ 47 ], and freeze-thawing [ 16 ]. The first two methods are often used in in vitro studies of CGF and yield highly variable extract variable concentrations.…”

“…CGF has been shown to promote osteogenic/odontoblastic differentiation of DPSCs [ 37 ] and SCAPs [ 34 ] in vitro by inducing mineralised nodule formation and the expression of COL1a1 , ALP , OCN , DMP-1 , and DSPP genes, and osteogenic differentiation of PDLSCs [ 40 ] and BMSCs [ 41 ] by inducing the expression COL1a1 , ALP , OCN , and Osterix ( OSX ) genes. In general, MSCs treated with CGF undergo osteogenic differentiation, but this is inhibited at high concentrations by proinflammatory factors such as tumour necrosis factor (TNF)-α and interleukin (IL)-1 in CGF extract, which may exert more potent effects than GFs [ 34 , 42 ].…”