In vitro comparison of the efficacy of TGF-β1 and PDGF-BB in combination with freeze-dried bone allografts for induction of osteogenic differentiation in MG-63 osteoblast-like cells

Abstract

Predictable regeneration of alveolar bone defects has always been a challenge in implant dentistry. Bone allografts are widely used bone substitutes with controversial osteoinductive activity. This in vitro study aimed to assess the osteogenic potential of some commercially available freeze-dried bone allografts supplemented with human recombinant platelet-derived growth factor-BB and transforming growth factor beta-1. Cell viability, mineralization, and osteogenic gene expression of MG-63 osteoblast-like cells were compared among the allograft alone, allograft/platelet-derived growth factor-BB, allograft/transforming growth factor beta-1, and allograft/platelet-derived growth factor-BB/transforming growth factor beta-1 groups. The methyl thiazol tetrazolium assay, real-time quantitative reverse transcription polymerase chain reaction and alizarin red staining were performed, respectively, for assessment of cell viability, differentiation, and mineralization at 24-72 h post treatment. The allograft with greater cytotoxic effect on MG-63 cells caused the lowest differentiation among the groups. In comparison with allograft alone, allograft/transforming growth factor beta-1, and allograft/transforming growth factor beta-1/platelet-derived growth factor-BB caused significant upregulation of bone sialoprotein and osteocalcin osteogenic mid-late marker genes, and resulted in significantly higher amounts of calcified nodules especially in mineralized non-cytotoxic allograft group. Supplementation of platelet-derived growth factor-BB alone in 5 ng/mL concentration had no significant effect on differentiation or mineralization markers. According to the results, transforming growth factor beta-1 acts synergistically with bone allografts to enhance the osteogenic differentiation potential. Therefore, this combination may be useful for rapid transformation of undifferentiated cells into bone-forming cells for bone regeneration. However, platelet-derived growth factor-BB supplementation did not support this synergistic ability to enhance osteogenic differentiation and thus, further investigations are required.

In vitro and in vivo effects of PDGF-BB delivery strategies on tendon healing: a review

“…For a comprehensive overview on the signalling pathways through which PDGF-BB elicits downstream cascades, refer to previous reviews on the topic (Andrae et al, 2008;Heldin and Westermark, 1999;Tallquist and Kazlauskas, 2004). PDGFR can also interact with integrins, through the Na + /H + exchanger regulatory factors (NHERFs) that link it to focal adhesion kinase and cytoskeleton (James et al, 2004;Veevers-Lowe et al, 2011). In turn, PDGFRs can be also affected by the ECM components (DeMali et al, 1999;Veevers-Lowe et al, 2011).…”

“…PDGFR can also interact with integrins, through the Na + /H + exchanger regulatory factors (NHERFs) that link it to focal adhesion kinase and cytoskeleton (James et al, 2004;Veevers-Lowe et al, 2011). In turn, PDGFRs can be also affected by the ECM components (DeMali et al, 1999;Veevers-Lowe et al, 2011).…”

“…In chronic foot ulcers, topical application of rhPDGF led to a significant increase (by 43 %) in the incidence of complete wound closure and decrease in healing time (by 32 %) over placebo-controlled wound care, resulting in the FDA approval of Regranex Gel ® (Wieman et al, 1998). The effect of PDGF-BB in supporting osteogenic differentiation has been shown in an in vitro study with MG63 cells (Vahabi et al, 2016). In addition, positive clinical results for GEM 21S ® were reported in regenerative www.ecmjournal.org O Evrova et al PDGF-BB delivery for tendon healing periodontal surgery (Singh and Suresh, 2012).…”

Freeze-Dried Platelet-Rich Plasma Induces Osteoblast Proliferation via Platelet-Derived Growth Factor Receptor-Mediated Signal Transduction

“…In previous studies, we showed that PRP accelerated bone union and induced a reduction of the bone union period in both rats and humans with spinal posterolateral fusion (PLF) [8][9][10]. It appears that platelet-derived growth factor (PDGF) plays an essential role in such effects as it has the ability to promote bone fusion [11]. PDGF is a member of a large family of GFs secreted by human vascular endothelial cells and fibroblasts to induce phosphorylation and activation of the PDGF receptor (PDGFR), which thereby activates downstream pathways that regulate cell growth and division.…”

Combination of Heel-strike like Mechanical Loading with Deproteinized Cancellous Bone Scaffold Implantation to Repair Segmental Bone Defects in Rabbits

“…The treatment of segmental bone defects caused by trauma, infection, or bone tumor resection remains a great challenge for orthopedic surgeons, and the growing clinical demands for bone substitutes highlights the need for researchers to both develop new bone scaffolds and improve the process of bone regeneration 1 . The latter has been the focus of many studies and several tissue-engineering techniques have been proposed to enhance osteogenesis inside bone scaffolds 2 , such as pre-seeding scaffolds with mesenchymal stem cells 3 , delivering growth factors like bone morphogenetic protein 2 (BMP-2) or transforming growth factor β1 (TGFβ-1) within scaffolds 4 - 5 , or pre-vascularization of scaffolds in vitro before implantation 6 . While each of these methods have been successful to some extent they all have disadvantages, including extended preparation time and a high cost for the final product.…”