Exudates of Picea abies, Pinus nigra, and Larix decidua: Chromatographic Comparison and Pro-Migratory Effects on Keratinocytes In Vitro

Abstract

Balms and resins of Picea abies, Larix decidua, and Pinus nigra are traditionally used to treat wounds. Three chromatographic techniques differing in separation capacity and technical demands were employed to distinguish among these plant exudates. A TLC method was established for fingerprint comparison, providing a quick overview of a large number of samples at low cost. HPLC-DAD (RP18) and UHPSFC-DAD (Torus 2-Picolylamin), hyphenated to ESI-MS, represented orthogonal chromatographic systems with high separation performance. The developed methods allow for the separation and detection of major and minor constituents belonging to different compound classes (phenyl carboxylic acids, lignans, diterpene resin acids). The qualitative compositions of the diterpene resin acids, the main compounds in the exudates, were comparable in all three genera. Differences were detected in the distribution of hydroxylated diterpene resin acids, pinoresinol, and hydroxycinnamic acids. The three tested chromatographic systems with varying demands on lab equipment offer appropriate tools for the quality assessment of Picea abies, Larix decidua, and Pinus nigra. The extracts were furthermore tested at three different concentrations (10 µg/mL, 3 µg/mL, and 1 µg/mL) for boosted re-epithelialization, a crucial step in the wound-healing process, in an in vitro HaCaT keratinocyte-based scratch assay. Lysophosphatidic acid (LPA, 10 µM) and extracts of several medicinal plants well known for their wound-healing properties (birch, marigold, St. John’s wort, manuka honey) were used as positive controls. Picea abies and Pinus nigra showed concentration dependency; significant activity was measured for Larix decidua at 3 µg/mL.

Ultra High-Performance Supercritical Fluid Chromatography for the Quantitation of Diterpene Resin Acids in Norway Spruce Samples

“…The method presented allows for the determination of DRAs in pharmaceutical matrices like Vaseline (NSB-CP1, NSB-CP2, and NSB-CP3) and lard (NSB-CP4 and NSB-CP5). Thus, it represents a basic tool for quality control providing the differentiation of balms from different species of the Pinaceae family ( Goels et al, 2022 ). Moreover, it may serve as quantitation method, e.g., in ex vivo permeation experiments revealing information about the ability of DRAs to permeate skin into an acceptor medium.…”

“…Dehydroabietic acid ( 7 ) was shown to reverse the tumour necrosis factor-α (TNF-α)/forkhead box O1 (FOXO1) and transforming growth factor-β1 (TGF-β1)/Smads mediated decrease in diabetic wound healing in human adult dermal fibroblasts ( Wang et al, 2014 ). DRAs are therefore highly suitable as marker compounds for quality assessment of raw tree exudates ( Goels et al, 2022 ). Efficient GC-MS based quantitation methods have already been described in the literature ( Latorre et al, 2003 ; Holmbom et al, 2008 ; Alicandri et al, 2021 ).…”

A Scoping Review on the Therapeutic Potential of Resin From the Species Larix decidua Mill. [Pinaceae] to Treat Ulcerating Wounds

“…Sipponen et al (2012) included 23 patients in their study and saw a healing rate of complicated chronic surgical wounds of 100%, within a period of 43 ± 24 days. In addition, Goels et al (2022) compared the wound healing potential of P. abies, Pinus nigra J.F.Arnold and L. decidua in vitro. The reduction of cell-free area in a keratinocyte wound healing assay was significant for the balm from L. decidua (26%) when compared to the P. abies balm and resin (16.7% and 9.6%, respectively) and to the P. nigra resin (16.2%).…”

Analysis and quantitation of bioactive compounds in Norway spruce balm

“…The bioactive compounds are lignans like pinoresinol and different diterpene resin acids (DRAs). In order to guarantee consistent pharmaceutical quality of Norway spruce balm and commercial products thereof, it is necessary to provide analytical methods for the quantitation of bioactive substances 1 , 2 .…”