Norway Spruce Balm: Phytochemical Composition and Ability to Enhance Re-epithelialization In Vitro ^{[ # ]}

 

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Abstract

The balm of the Norway spruce (Picea abies) is a well-known traditional herbal medicine used to cure wounds. Even though clinical trials have confirmed its empirical use, the active constituents, their mode of action, and the exact composition of this natural product are still unknown. In this study, the balm was subjected to fractionated extraction and further purified employing flash chromatography, HPLC-PDA-ELSD, preparative and analytical TLC. Hydroxycinnamic acids (

1–

3), the lignan pinoresinol (

4), four hydroxylated derivatives of dehydroabietic acid (DHAA) (

5 – 

8), and dehydroabietic acid (

9) were isolated. Their structures were elucidated by LC-MS, 1D- and 2D-NMR. Four extracts, two commercially available resin acids–pimaric acid (

10) and isopimaric acid (

11)–and the isolated compounds were tested for increased re-epithelialization of cell-free areas in a human adult low calcium high temperature keratinocytes monolayer. Lysophosphatidic acid (10 µM) served as positive control and ranged between 100% and 150% rise in cell-covered area related to the vehicle control. Two extracts containing carboxylic acids and non-acidic apolar constituents, respectively, boosted wound closure by 47% and 36% at 10 and 3 µg/mL, respectively. Pinoresinol, DHAA, three of its hydroxylated derivatives, and pimaric and isopimaric acid as well as defined combinations of the hydroxylated DHAA derivatives led to a significantly enhanced wound closure by up to 90% at concentrations between 1 and 10 µM. Overall, lignans and diterpene resin acids, main constituents of Norway spruce balm, are able to increase migration or proliferation of keratinocytes in vitro. The presented data link the phytochemistry of this natural wound healing agent with boosted re-epithelialization.

^# Dedicated to emer. Univ.-Prof. Dr. Wolfgang Kubelka on behalf of his 85th birthday.

Publication History

Received: 23 January 2020

Accepted: 18 March 2020

Article published online:
21 April 2020

© 2020. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

• References

• 1 Han G, Ceilley R. Chronic wound healing: a review of current management and treatments. Adv Ther 2017; 34: 599-610
  CrossrefPubMedGoogle Scholar
• 2 Velnar T, Bailey T, Smrkolj V. The wound healing process: an overview of the cellular and molecular mechanisms. J Int Med Res 2009; 37: 1528-1542
  CrossrefPubMedGoogle Scholar
• 3 The Plant List. Version 1.1. (2013). Available at (Accessed January 7, 2020): http://www.theplantlist.org/tpl1.1/search?q=Picea+abies
  PubMedGoogle Scholar
• 4 Jokinen J, Sipponen A. Refined spruce resin to treat chronic wounds: rebirth of an old folkloristic therapy. Adv Wound Care (New Rochelle) 2016; 5: 198-207
  CrossrefPubMedGoogle Scholar
• 5 Sipponen A, Jokinen J, Sipponen P, Papp A, Sarna S, Lohi J. Beneficial effect of resin salve in treatment of severe pressure ulcers: a prospective, randomized and controlled multicentre trial. Br J Dermatol 2008; 158: 1055-1062
  CrossrefPubMedGoogle Scholar
• 6 Sipponen A, Kuokkanen O, Tiihonen R, Kauppinen H, Jokinen J. Natural coniferous resin salve used to treat complicated surgical wounds: pilot clinical trial on healing and costs. Int J Dermatol 2012; 51: 726-732
  CrossrefPubMedGoogle Scholar
• 7 Rautio M, Sipponen A, Peltola R, Lohi J, Jokinen J, Papp A, Carlson P, Sipponen P. Antibacterial effects of home-made resin salve from Norway spruce (Picea abies). APMIS 2007; 115: 335-340
  CrossrefPubMedGoogle Scholar
• 8 Sipponen A, Laitinen K. Antimicrobial properties of natural coniferous rosin in the European Pharmacopoeia challenge test. APMIS 2011; 119: 720-724
  CrossrefPubMedGoogle Scholar
• 9 Rautio M, Sipponen A, Lohi J, Lounatmaa K, Koukila-Kähkölä P, Laitinen K. In vitro fungistatic effects of natural coniferous resin from Norway spruce (Picea abies). Eur J Clin Microbiol Infect Dis 2012; 31: 1783-1789
  CrossrefPubMedGoogle Scholar
• 10 Holmbom T, Markku R, Pedro F. Composition of callus resin of Norway spruce, Scots pine, European larch and Douglas fir. Holzforschung 2008; 62: 417
  CrossrefPubMedGoogle Scholar
• 11 Thorlakson HH, Schreurs O, Schenck K, Blix IJS. Lysophosphatidic acid regulates adhesion molecules and enhances migration of human oral keratinocytes. Eur J Oral Sci 2016; 124: 164-171
  CrossrefPubMedGoogle Scholar
• 12 Wang XW, Yu Y, Gu L. Dehydroabietic acid reverses TNF-α-induced the activation of FOXO1 and suppression of TGF-β1/Smad signaling in human adult dermal fibroblasts. Int J Clin Exp Pathol 2014; 7: 8616-8626
  PubMedGoogle Scholar
• 13 Scheffler A. The wound healing properties of betulin from birch bark from bench to bedside. Planta Med 2019; 85: 524-527
  Article in Thieme ConnectPubMedGoogle Scholar
• 14 Haider N, Robien W. CSEARCH-Robot-Referee. Available at (Accessed March 3, 2020): http://nmrpredict.orc.univie.ac.at/c13robot/robot.php
  PubMedGoogle Scholar
• 15 Liang C, Park A, Guan J. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro . Nat Protoc 2007; 2: 329-333
  CrossrefPubMedGoogle Scholar
• 16 Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez J, White D, Hartenstein V, Eliceiri K, Tomancak P, Cardona A. Fiji: an open-source platform for biological-image analysis. Nat Methods 2012; 9: 676-682
  CrossrefPubMedGoogle Scholar
• 17 MRI Wound Healing Tool. Available at (Accessed January 21, 2020): http://dev.mri.cnrs.fr/projects/imagej-macros/wiki/Wound_Healing_Tool
  PubMedGoogle Scholar

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