Effects of enamel matrix derivative and transforming growth factor‐β1 on human periodontal ligament fibroblasts

Abstract

Aim: The objective of this study was to evaluate the effects of enamel matrix derivative (EMD), transforming growth factor‐β1 (TGF‐β1), and a combination of both factors (EMD+TGF‐β1) on periodontal ligament (PDL) fibroblasts. Material and methods: Human PDL fibroblasts were obtained from three adult patients with a clinically healthy periodontium, using the explant technique. The effects of EMD, TGF‐β1, or a combination of both were analysed on PDL cell proliferation, adhesion, wound healing, and total protein synthesis, and on alkaline phosphatase (ALP) activity and bone‐like nodule formation. Results: Treatment with EMD for 4, 7, and 10 days increased cell proliferation significantly compared with the negative control (p<0.05). At day 10, EMD and EMD+TGF‐β1 showed a higher cell proliferation compared with TGF‐β1 (p<0.01). Cell adhesion was significantly up‐regulated by TGF‐β1 compared with EMD and EMD+TGF‐β1 (p<0.01). EMD enhanced in vitro wound healing of PDL cells compared with the other treatments. Total protein synthesis was significantly increased in PDL cells cultured with EMD compared with PDL cells treated with TGF‐β1 or EMD+TGF‐β1 (p<0.05). EMD induced ALP activity in PDL fibroblasts, which was associated with an increase of bone‐like nodules. Conclusion: These findings support the hypothesis that EMD and TGF‐β1 may play an important role in periodontal regeneration. EMD induced PDL fibroblast proliferation and migration, total protein synthesis, ALP activity, and mineralization, while TGF‐β1 increased cellular adhesion. However, the combination of both factors did not positively alter PDL fibroblast behaviour.

Morinda citrifolia leaves enhance osteogenic differentiation and mineralization of human periodontal ligament cells

“…There was also an absence of nodule formation and matrix mineralization: only aggregated cells with light alizarin red S staining was observed for hPDL cells treated with ascorbic acid and β-GP. These results well agreed with previous studies which suggested that nodule formation of hPDL cells mostly occurred in certain condition such as in the presence of dexamethasone, estradiol, or EMD with TGF-β1 13,29,30) . To confirm the potency of noni leaf extract in inducing osteogenic differentiation and matrix mineralization, a mouse preosteoblast cell line (MC3T3-1) was treated with the optimal concentration of 2.5% of noni leaf extract.…”

“…The ability to grow and manipulate stem cells within the periodontal ligament is of considerable clinical significance, especially for developing novel mechanisms to achieve bone and periodontal tissue regeneration 28) . Previous findings have indicated that hPDL cells possess several osteoblast-like properties, including the ability to form mineralized nodules in vitro 13,29) when cultured in the presence of growth factors such as enamel matrix derivative (EMD) and transforming growth factor-beta1 (TGF-β1) 30) . However, not only is EMD costly, clinical and cell culture studies have yielded controversial and conflicting results 32) .…”

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“…It has been shown that EMD has no effect on ALP activity (59); however, EMD both decreases (57) and increases (62,73,74) ALP activity in PDL fibroblasts. The production of osteopontin and osteoprotegerin has been shown to be increased by EMD (60,74).…”

Root Biomodification in Periodontics - The Changing Concepts

“…EMD induced PDL fibroblast proliferation and migration, total protein synthesis, ALP activity, and mineralization, while TGF-β1 increased cellular adhesion. However, the combination of both factors did not positively alter PDL fibroblast behaviour [59]. PDGF-BB in concentrations equal to or greater than 50 ng/ml demonstrates a significant stimulation of PDL cells adherence to periodontal diseased root surface.…”

“…It is extracted from developing embryonal enamel of porcine origin and contains several matrix proteins from the amelogenin family. Several studies have shown that EMD influences the migration, attachment, proliferative capacity and biosynthetic activity of periodontal ligament cells [59,[67][68][69].…”