Purification and analysis of a 5kDa component of enamel matrix derivative

Mumulidu, Alexandra; Hildebrand, B; Fabi, Beata; Hammarström, Lars; Cochran, David L.; Dard, Michel; Lemoult, Stephanie

doi:10.1016/j.jchromb.2007.07.017

Cited by 27 publications

(36 citation statements)

References 34 publications

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“…A variety of amelogenin peptides are produced in vivo during tooth morphogenesis by alternative splicing and proteolytic degradation [47,48], but the biological function of many of these peptides is not known. In contrast to the four peaks found in the original fractionation [29], here we found that Fraction C contained just two peaks, one of 43 amino acids and the other one corresponding to 45 amino acids in length, which correspond to the previously reported TRAP species [28]. The LRAP portion of amelogenin has been shown to induce osteogenesis in vivo [49] and osteoblast maturation in vitro [50,51], but the effect of Fraction C, containing the TRAP portion, was unclear.…”

Section: Discussioncontrasting

confidence: 70%

“…Recombinant human amelogenin (rhAmelogenin) and the 5 kDa peptides (Fraction C) were extracted and purified by Institut Straumann AG using a modification of previously described methods [29,39]. Briefly, Fraction C was extracted from EMD via size exclusion high-performance liquid chromatography (TSKgel SW3000, Tosoh Bioscience GmbH, Stuttgart, Germany) in 30% acetonitrile containing 0.9 mM NaCl resulting in one peak around 5 kDa.…”

Section: Proteinsmentioning

confidence: 99%

“…It is unclear whether the therapeutic effect of EMD is due to the full-length amelogenin protein, due to the splice variants, or to a combination of both. Three subfractions of EMD have been isolated by high-performance liquid chromatography: one containing mainly a 20 kDa peptide, one represented by two peptides of 12 kDa and 9 kDa, and one fraction identified as a single band at 5 kDa by SDS-PAGE analysis [29]. The 20 kDa peptide corresponds to the full-length amelogenin protein and the 5 kDa peptide corresponds to a portion of the N-terminus of the protein that includes peptides with the TRAP and LRAP sequences.…”

Section: Introductionmentioning

confidence: 99%

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Amelogenin Peptide Extract Increases Differentiation and Angiogenic and Local Factor Production and Inhibits Apoptosis in Human Osteoblasts

et al. 2013

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Enamel matrix derivative (EMD), a decellularized porcine extracellular matrix (ECM), is used clinically in periodontal tissue regeneration. Amelogenin, EMD's principal component, spontaneously assembles into nanospheres in vivo, forming an ECM complex that releases proteolytically cleaved peptides. However, the role of amelogenin or amelogenin peptides in mediating osteoblast response to EMD is not clear. Human MG63 osteoblast-like cells or normal human osteoblasts were treated with recombinant human amelogenin or a 5 kDa tyrosine-rich amelogenin peptide (TRAP) isolated from EMD and the effect on osteogenesis, local factor production, and apoptosis assessed. Treated MG63 cells increased alkaline phosphatase specific activity and levels of osteocalcin, osteoprotegerin, prostaglandin E 2 , and active/latent TGF-1, an effect sensitive to the effector and concentration. Primary osteoblasts exhibited similar, but less robust, effects. TRAP-rich 5 kDa peptides yielded more mineralization than rhAmelogenin in osteoblasts in vitro. Both amelogenin and 5 kDa peptides protected MG63s from chelerythrine-induced apoptosis. The data suggest that the 5 kDa TRAP-rich sequence is an active amelogenin peptide that regulates osteoblast differentiation and local factor production and prevents osteoblast apoptosis.

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Section: Discussioncontrasting

confidence: 70%

Section: Proteinsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Amelogenin Peptide Extract Increases Differentiation and Angiogenic and Local Factor Production and Inhibits Apoptosis in Human Osteoblasts

et al. 2013

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“…The exact mechanisms underlying the promoting effect of EMD on endothelial cells and angiogenesis remain unclear. EMD is composed mainly of amelogenin and amelogenin transcripts resulting of gene alternative splicing [31][32][33][34][35] . A recent study reported that different EMD components may possess different biological activity 31) , however their exact bioactive specificities in respect to different tissues and cells need to be investigated.…”

Section: Discussionmentioning

confidence: 99%

Effect of enamel matrix derivative on the angiogenic behaviors of human umbilical vein endothelial cells on different titanium surfaces

et al. 2017

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Angiogenesis play a crucial role in the regeneration of hard and soft tissue around dental titanium (Ti) implant. Enamel matrix derivative (EMD) promotes tissue regeneration and stimulates angiogenesis but its effect on the angiogenesis on Ti surfaces was never investigated. The effect of EMD on the angiogenic activity of endothelial cells cultured on pre-treated smooth Ti (PT), acidetched (A), coarse-grit blasted and acid-etched (SLA) surfaces and tissue culture plastic (TCP) in the presence or absence of EMD was investigated. EMD inhibited the proliferation/viability of human umbilical vein endothelial cells (HUVECs) growing on A and SLA Ti surfaces. EMD induced an increase in the expression of all these genes in HUVECs grown on SLA surface but not on other surfaces. Summarizing, our data show that EMD influences proliferation and expression of angiogenesis associated gene in HUVECs grown on moderately rough SLA surfaces, suggesting that EMD might promote angiogenesis following implantation.

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“…16 Both EMD and the two fractions LMW and HMW are naturally occurring, and all of them have been prepared from crude EMP extracts by a precisely controlled patented process that involves heat treatment of EMP. The LMW fraction comprises mainly an amelogenin-derived < 6-kDa TRAP and the HMW fraction contains > 6-kDa peptides, including the amelogenin peptide LRAP, sheathlin, and fulllength amelogenin.…”

Section: Introductionmentioning

confidence: 99%

Differential Effect of Amelogenin Peptides on Osteogenic DifferentiationIn Vitro: Identification of Possible New Drugs for Bone Repair and Regeneration

Amin

Olsen

Knowles

et al. 2012

Tissue Engineering Part A

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Enamel matrix proteins (EMP) have been shown to promote regeneration of periodontal ligament and root cementum, and sometimes to enhance the differentiation of bone-forming cells in vitro and new bone growth in vivo. However, the inconsistent and unpredictable effects of EMP that have been reported for bone regeneration may be due to the highly variable composition of this heterogeneous material, which is comprised mainly of amelogenin and amelogenin-derived peptides. The present study has therefore examined the effects of naturally occurring low-molecular-weight (LMW) and high-molecular-weight (HMW) fractions of Emdogain(®) (EMD; Institut Straumann, Basel, Switzerland), a commercially available form of EMP, on osteogenic differentiation of bone precursor cells in vitro. In addition, the effects of chemically synthesized specific components of LMW and HMW-namely, the tyrosine-rich amelogenin peptide (TRAP), a specific amelogenin isoform derived by proteolytic clipping, and a leucine-rich amelogenin peptide (LRAP), an isoform derived by alternative splicing-on bone-forming cell activity were also investigated. Our findings demonstrate that while TRAP suppressed the formation of bone-like mineralized nodules, LRAP upregulated osteogenic differentiation. Furthermore, synthetically produced TRAP and its unique C-terminal 12 amino acid sequence (TCT) also suppressed bone-forming cells, whereas LRAP and its unique C-terminal 23 amino acid sequence (LCT) markedly enhanced terminal differentiation of bone-forming cells. These findings suggest that the differential effects of amelogenin-derived peptide sequences present in EMP could be of potential clinical value, with the novel bioactive TCT peptide as a useful tool for limiting pathological bone cell growth and the unique LCT sequence having therapeutic benefits in the treatment of periodontal and orthopedic diseases.

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Purification and analysis of a 5kDa component of enamel matrix derivative

Cited by 27 publications

References 34 publications

Amelogenin Peptide Extract Increases Differentiation and Angiogenic and Local Factor Production and Inhibits Apoptosis in Human Osteoblasts

Amelogenin Peptide Extract Increases Differentiation and Angiogenic and Local Factor Production and Inhibits Apoptosis in Human Osteoblasts

Effect of enamel matrix derivative on the angiogenic behaviors of human umbilical vein endothelial cells on different titanium surfaces

Differential Effect of Amelogenin Peptides on Osteogenic DifferentiationIn Vitro: Identification of Possible New Drugs for Bone Repair and Regeneration

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