Effect of Concentrated Growth Factor (CGF) on the Promotion of Osteogenesis in Bone Marrow Stromal Cells (BMSC) in vivo

Chen, Xia; Wang, Jian; Li, Yu; Zhou, Jia; Zheng, Duo; Zhang, Bo

doi:10.1038/s41598-018-24364-5

Cited by 62 publications

(56 citation statements)

References 26 publications

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“…Due to the advanced extraction technology and specialized equipment (Medifuge, Silfradent Srl, Italy), the preparation process is simple and does not require any synthetic or catalytic substances. In the present study, the SEM results showed that CGF has a complex porous three-dimensional brin structure, which is consistent with others' results [14,32,33]. This dense brin network can provide a matrix for cell adherence and migration [33,34].…”

Section: Discussionsupporting

confidence: 93%

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Effects of Concentrated Growth Factor on the Proliferation, Migration, and Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells: An in vitro study

Yu¹,

Ren²,

Shang³

et al. 2020

Preprint

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Background Concentrated growth factor (CGF) has been reported to be effective in bone formation or soft/hard tissue healing in recent years. Despite a few studies regarding the effects of CGF on the proliferation, migration, and osteogenic differentiation of BMSCs, their underlying mechanisms are not fully understood. The purpose of this study is to investigate the effects and possible mechanisms of CGF on the proliferation, migration, and osteogenic differentiation of rat-derived bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods CGF was extracted from the Sprague Dawley (SD) rats by venipuncture of the abdominal aortic vein, and scanning electron microscopy (SEM) was used for the structural characterization. The release of bone morphogenetic protein 2 (BMP-2) from CGF was measured over the periods of 1 ~ 14 days, using the enzyme-linked immunosorbent (Elisa) assay. Cell Counting Kit-8 (CCK-8) assay was used to measure cell proliferation. Migration capacity was analyzed using the transwell assay. The osteogenic differentiation and mineralization ability were determined by Alkaline phosphatase activity (ALP) staining and Alizarin Red staining respectively. Quantitative real-time PCR (RT-qPCR), was used to evaluate the mRNA expression levels of Runx2, Ocn, Smad1, and Smad5 after culture for 14 days. Further, the protein expression of BMP-2, phosphorylated-Smad1/5 (p-Smad1/5), and Smad1/5/8 was determined by Western blot after a 14-day cell culture. Results The SEM analysis showed a porous and dense three-dimensional fibrin network in CGF. The Elisa assay showed that BMP-2 was released from CGF extract for more than 14d, and it reached a peak at the time point of 5d. The cell densities of the CGF group at the different concentrations (5%, 10%, and 20%) were significantly higher than that of the control group at the periods of day 1 to day 5 (p < 0.05). Moreover, the number of migratory cells of the CGF group was greater than that of the control group at 24 h. ALP activity analysis and Alizarin Red staining results demonstrated that CGF may successfully induce osteogenic differentiation of BMSCs. Moreover, the RT-qPCR results showed that CGF extracts dramatically enhanced the mRNA expression levels of Runx2, Ocn, Smad1, and Smad5 in BMSCs at days 14 (p < 0.05). Furthermore, Western blot results showed that CGF extracts markedly up-regulated the protein expression levels of BMP-2, p-Smad1/5, and Smad1/5/8. Conclusions CGF can promote the proliferation, migration, and promote the osteogenic differentiation potential of BMSCs in vitro. The BMP-2/Smad signaling pathway was involved in the osteogenic differentiation and mineralization of BMSCs induced by CGF. Therefore, CGF has good application potential in tissue engineering for bone regeneration and repair.

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Section: Discussionsupporting

confidence: 93%

“…It appears to have more abundant growth factors and a more rigid brin structure than PRP and PRF, due to the special centrifugation process [10,11]. A number of previous studies have demonstrated that CGF's potentials to promote bone formation, tissue repair and regeneration [6,[11][12][13][14][15][16][17][18].…”

Section: Introductionmentioning

confidence: 99%

Effects of Concentrated Growth Factor on the Proliferation, Migration, and Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells: An in vitro study

Yu¹,

Ren²,

Shang³

et al. 2020

Preprint

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“…In the study of Honda et al, concentrations of 1–10% of CGF extract had a positive effect on proliferation and differentiation in a dose‐dependent manner, while high concentration (20%) had an inhibitory effect (Honda et al, ). Chen et al, by studying the effect of 5, 10, 20, and 40% CGF, demonstrated more proliferation and differentiation in 10% CGF (Chen et al, ). No dose‐dependent effect was also stated by Hong et al in investigation of the effect of three different concentration of CGF–CCM (1, ½, ¼) (Hong et al, ).…”

Section: Resultsmentioning

confidence: 99%

“…As seen in the result of this review, CGF was used alone in in vitro studies, but in in vivo studies, the combination of CGF with stem cells or biomaterials has been reported (Chen et al, ; Durmuslar et al, ; Masuki et al, ; Qiao et al, ; Qiao & An, ; Rodella et al, ). Tissue engineering is a complex of stem cells, scaffold, and growth factors.…”

Section: Discussionmentioning

confidence: 99%

In vitro and in vivo effects of concentrated growth factor on cells and tissues

Tabatabaei

Aghamohammadi

Tayebi

2020

J Biomedical Materials Res

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This article reviews the biological outcome of the concentrated growth factor (CGF), a new platelet derivative used for tissue regeneration, in published articles related to the use of this product in basic and clinical studies. An electronic literature research using PubMed and SCOPUS was performed using combination of keywords: “concentrated growth factor” (OR “CGF”), AND “stem cells,” AND “cells” OR “cell proliferation” OR “cell migration” OR “cell differentiation,” AND “repair” OR “survival” OR “revitalization,” AND “tissue” OR “bone.” Forty‐five articles that were published between 2012 and 2020 met the inclusion criteria. These studies have used CGF as fresh solid form, freeze‐dried, membrane, extract, or exudate. Most studies demonstrate the positive effects of CGF in a dose‐dependent manner under certain concentrations. Studies comparing CGF with other platelet concentrates, report lower efficiency, no statistically significant differences, or better results for CGF. Combination of CGF with stem cells and biomaterials significantly improves bone regeneration and the effect of allograft or collagen membrane is better than CGF alone. For a better examination of the biological outcomes of CGF, the standardization of CGF preparation regarding the choice of the test tube material for blood collection, the required volume of blood, the necessary count of platelets in CGF, and the most appropriate type of CGF are recommended.

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“…A part of BMSCs were collected at passage 2 and cryopreservation. Another part was cultured until passage 3, and the BMSCs (passage 3) and its culture supernatant were collected used for the following experiments [22].…”

Section: Isolation and Culture Of Rat Bone Marrow-derived Mesenchymalmentioning

confidence: 99%

Comparison of the therapeutic effect of BMSCs and culture supernatant injection therapy on osteoarthritis and mechanism exploration

Peng

Zhao

Huang

et al. 2020

Preprint

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Background Osteoarthritis (OA) is a chronic disease that can cause disability. The prevalence of osteoarthritis has increased year by year, become a disease that mainly causes the elderly to suffer. With the increasing understanding of bone marrow mesenchymal stem cell (BMSC) in tissue repair, BMSCs shown good therapeutic effects in OA, it may be due to BMSCs exerted their capacities of differentiation, secretion and immunoregulatory in the joints. As previous describe, BMSCs culture supernatant is proved that rich in multiple cytokines. Therefore, how BMSCs injection therapy exerts its ability to exert therapeutic effects, and whether its therapeutic effect can be replaced by its culture supernatant has become the focus of this article. Methods In vitro, we designed a co-culture system to deepen understand this new stem cell therapy. Through two kinds of cells cultured alone, direct and indirect cell contact co-culture to observe changes in cell morphology, quantity and cytoplasmic glycoprotein of these two types of cells and changes of the level of growth factor in the culture supernatant to explore the interaction of these two types of cells; in vivo, we induced an OA model and Injected Saline solution, BMSCs and their culture supernatants respectively for treatment, then we compared the improvement results of OA by evaluating changes in the cartilage layer after different treatments. And determine the changes of some growth factors and inflammatory factors in synovial fluid to analyze the possible mechanisms of multiple treatments. Result In the co-culture system, it was found that the direct co-culture of BMSCs and OCs can enhance the proliferation ability of OCs, OCs can retain more cytoplasmic glycoprotein, and BMSCs did not occurred abnormal differentiation during the co-culture. In animal experiments, it was found that the ability of BMSCs injection treatment has obvious therapeutic effect on OA, and the effect is better than its supernatant injection treatment. BMSCs therapy reduced matrix loss in articular cartilage cells, and reduced type I collagen production and fibrosis on articular cartilage, effectively regulating EGF and TGF-β1change and inhibited intra-articular inflammation. Supernatant injection treatment will not significantly delay the progress of OA, and it cannot replace BMSCs for the treatment of OA. Conclusion BMSCs therapy is a potential new therapy for OA. The secretion and regulation ability of BMSCs plays an important role in the treatment process. BMSCs are activated by the intra-articular environment of OA, regulating growth factors such as EGF and TGFβ to promote articular cartilage regeneration, and reducing intra-articular inflammation by regulating inflammatory factors and delaying the progression of OA. These effects cannot be replaced by the culture supernatant which is rich in multiple factors, and its regulation function requires the presence of BMSCs. These results provide a relatively comprehensive understanding of BMSCs cell therapy in OA and provide a new explanation for the possible anti-inflammatory effects of BMSCs in the joint.

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Effect of Concentrated Growth Factor (CGF) on the Promotion of Osteogenesis in Bone Marrow Stromal Cells (BMSC) in vivo

Cited by 62 publications

References 26 publications

Effects of Concentrated Growth Factor on the Proliferation, Migration, and Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells: An in vitro study

Effects of Concentrated Growth Factor on the Proliferation, Migration, and Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells: An in vitro study

In vitro and in vivo effects of concentrated growth factor on cells and tissues

Comparison of the therapeutic effect of BMSCs and culture supernatant injection therapy on osteoarthritis and mechanism exploration

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