A galloylated dimeric proanthocyanidin from grape seed exhibits dentin biomodification potential.

Grape seeds are a rich source of polyphenols, especially proanthocyanidins (PACs), and are also known for the presence of galloylated oligomeric PACs (OPACs). The present study focuses on the phytochemical methodology for grape seed (O)PACs and their potential role as dentin biomodifiers to be used in restorative and reparative dentistry. A new method using centrifugal partition chromatography (CPC) was developed for the preparative separation of the grape seed (O)PACs. Orthogonal phytochemical profiling of the resulting CPC fractions was performed using C18 and diol HPLC, normal phase HPTLC, and IT-TOF MS analysis. A galloylated procyanidin dimer (1) was isolated from a CPC fraction in order to evaluate its potential to enhance dentin bio-mechanical properties. Moreover, it helped to evaluate the impact of the galloyl moiety on the observed bioactivity. Structure elucidation was performed using ESI-MS, 1D and 2D NMR analyses. For the first time, (1)H iterative full spin analysis (HiFSA) was performed on this type of molecule, enabling a detailed proton chemical shift and coupling constant assignment. The CPC fractions as well as 1 showed promising results in the dentin stiffness bioassay and indicate that they may be used as dental intervention biomaterial.

[1]  G. D'andrea.  Pycnogenol: a blend of procyanidins with multifaceted therapeutic applications? , 2010, Fitoterapia.

[2]  H. Fujii,et al.  Syntheses of procyanidin B2 and B3 gallate derivatives using equimolar condensation mediated by Yb(OTf)3 and their antitumor activities. , 2013, Bioorganic & medicinal chemistry letters.

[3]  G. Pauli,et al.  Countercurrent separation of natural products. , 2008, Journal of natural products.

[4]  C. Kwik-Uribe,et al.  Method performance and multi-laboratory assessment of a normal phase high pressure liquid chromatography-fluorescence detection method for the quantitation of flavanols and procyanidins in cocoa and chocolate containing samples. , 2009, Journal of chromatography. A.

[5]  G. Pauli,et al.  G.U.E.S.S.—A Generally Useful Estimate of Solvent Systems for CCC , 2005 .

[6]  Smith Wr,et al.  Survival analysis of posterior restorations using an insurance claims database. , 2002 .

[7]  M. Cascante,et al.  The importance of polymerization and galloylation for the antiproliferative properties of procyanidin‐rich natural extracts , 2007, The FEBS journal.

[8]  Guido F Pauli,et al.  Quantitative 1H NMR: development and potential of a method for natural products analysis. , 2005, Journal of natural products.

[9]  N. Oberlies,et al.  HiFSA fingerprinting applied to isomers with near-identical NMR spectra: the silybin/isosilybin case. , 2013, The Journal of organic chemistry.

[10]  G. Pauli,et al.  qNMR--a versatile concept for the validation of natural product reference compounds. , 2001, Phytochemical analysis : PCA.

[11]  C. Duan,et al.  Biosynthesis and Genetic Regulation of Proanthocyanidins in Plants , 2008, Molecules.

[12]  E. Haslam,et al.  Polyphenol interactions. Part 2. Covalent binding of procyanidins to proteins during acid-catalysed decomposition; observations on some polymeric proanthocyanidins , 1985 .

[13]  Y. Ozeki,et al.  Structural characterization of a procyanidin tetramer and pentamer from the apple by low-temperature NMR analysis , 2008 .

[14]  S. Watson,et al.  Synthesis and preliminary anticancer activity studies of C4 and C8-modified derivatives of catechin gallate (CG) and epicatechin gallate (ECG). , 2006, The Journal of organic chemistry.

[15]  V. Wray,et al.  Structure elucidation of procyanidin oligomers by low-temperature 1H NMR spectroscopy. , 2011, Journal of agricultural and food chemistry.

[16]  S. Karol,et al.  Long-term stability of dentin matrix following treatment with various natural collagen cross-linkers. , 2011, Journal of the mechanical behavior of biomedical materials.

[17]  I. Medina,et al.  Contribution of galloylation and polymerization to the antioxidant activity of polyphenols in fish lipid systems. , 2010, Journal of agricultural and food chemistry.

[18]  Arvind Shenoy.  Is it the end of the road for dental amalgam? A critical review , 2008, Journal of conservative dentistry : JCD.

[19]  Judith Pfeiffer,et al.  Biosynthesis of flavan 3-ols by leucoanthocyanidin 4-reductases and anthocyanidin reductases in leaves of grape (Vitis vinifera L.), apple (Malus x domestica Borkh.) and other crops. , 2006, Plant physiology and biochemistry : PPB.

[20]  R. Latif.  Health benefits of cocoa , 2013, Current opinion in clinical nutrition and metabolic care.

[21]  J. McAlpine,et al.  Dentin biomodification: strategies, renewable resources and clinical applications. , 2014, Dental materials : official publication of the Academy of Dental Materials.

[22]  M. Yamauchi,et al.  Application of crosslinkers to dentin collagen enhances the ultimate tensile strength. , 2007, Journal of biomedical materials research. Part B, Applied biomaterials.

[23]  T. Shoji,et al.  Separation of proanthocyanidins by degree of polymerization by means of size-exclusion chromatography and related techniques. , 2003, Journal of biochemical and biophysical methods.

[24]  D. Ferreira,et al.  Oligomeric proanthocyanidins: naturally occurring O-heterocycles. , 2002, Natural product reports.

[25]  J. Napolitano,et al.  Dentin Biomodification Potential Depends on Polyphenol Source , 2014, Journal of dental research.

[26]  J. Lupton,et al.  Recommending flavanols and procyanidins for cardiovascular health: current knowledge and future needs. , 2010, Molecular aspects of medicine.

[27]  B. Bartolomé,et al.  Non-galloylated and galloylated proanthocyanidin oligomers in grape seeds from Vitus vinifera L. cv. Graciano, Tempranillo and Cabernet Sauvignon , 2006 .

[28]  R. Hemingway,et al.  Linkage isomerism in trimeric and polymeric 2,3-cis-procyanidins , 1982 .

[29]  J. Napolitano,et al.  Orthogonal analytical methods for botanical standardization: determination of green tea catechins by qNMR and LC-MS/MS. , 2014, Journal of pharmaceutical and biomedical analysis.

[30]  E. Çapanoğlu,et al.  A review on protein–phenolic interactions and associated changes , 2013 .

[31]  K. Faure,et al.  Solvent selection in countercurrent chromatography using small-volume hydrostatic columns , 2013 .

[32]  Shao-Nong Chen,et al.  Solubility study of phytochemical cross-linking agents on dentin stiffness. , 2010, Journal of dentistry.

[33]  N. Nakajima,et al.  Systematic synthesis of galloyl-substituted procyanidin B1 and B2, and their ability of DPPH radical scavenging activity and inhibitory activity of DNA polymerases. , 2005, Bioorganic & medicinal chemistry.

[34]  H. Akiyama,et al.  Isolation and structural elucidation of some procyanidins from apple by low-temperature nuclear magnetic resonance. , 2003, Journal of agricultural and food chemistry.

[35]  M. Herderich,et al.  Characterization of proanthocyanidins in grape seeds using electrospray mass spectrometry. , 2003, Rapid communications in mass spectrometry : RCM.

[36]  K. Miescke,et al.  Changes in stiffness of demineralized dentin following application of collagen crosslinkers. , 2008, Journal of biomedical materials research. Part B, Applied biomaterials.

[37]  M. Kelm,et al.  High-performance liquid chromatography separation and purification of cacao (Theobroma cacao L.) procyanidins according to degree of polymerization using a diol stationary phase. , 2006, Journal of agricultural and food chemistry.

[38]  J. McAlpine,et al.  Mimicking the hierarchical functions of dentin collagen cross-links with plant derived phenols and phenolic acids. , 2014, Langmuir : the ACS journal of surfaces and colloids.

[39]  Osamu Kawahara,et al.  Tannins and related compounds. XV. A new class of dimeric flavan-3-ol gallates, theasinensins A and B, and proanthocyanidin gallates from green tea leaf (1) , 1983 .

[40]  R. Grande,et al.  Mechanical characterization of proanthocyanidin-dentin matrix interaction. , 2010, Dental materials : official publication of the Academy of Dental Materials.