Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT-Raman spectroscopy

Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37oC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

Vibrational Study (Raman, SERS, and IR) of Plant Gallnut Polyphenols Related to the Fabrication of Iron Gall Inks

FT-Raman, FTIR, and SERS spectra of the structurally related gallnut polyphenols tannic acid, gallic acid, pyrogallol, and syringic acid are reported in this work aiming at performing a comparative assignation of the bands and finding specific marker features that can identify these compounds in complex polyphenol mixtures. Tannic and gallic acids are the principal components in oak gallnuts, and they can be found in iron gall inks. The different functional groups existing in these molecules and their spatial distribution lead to slight changes of the vibrations. The Raman spectra are dominated by bands corresponding to the ring vibrations, but the substituents in the ring strongly affect these vibrations. In contrast, the FTIR spectra of these molecules are dominated by the peripheral oxygen-containing substituents of the aromatic ring and afford complementary information. SERS spectroscopy can be used to analyze trace amounts of these compounds, but the spectra of these polyphenols show strong changes in comparison with the Raman spectra, indicating a strong interaction with the metal. The most significant modification observed in the SERS spectra of these compounds is the weakening of the benzene 8a ring vibration and the subsequent intensification of the 19a mode of the benzene ring. This mode is also more intense in the FTIR spectra, and its intensification in the SERS spectra could be related to a drastic change in the molecular polarizability associated with the interaction of the polyphenol with the metal in Ag NPs.

Phytotoxic Effects of Selected Herbal Extracts on the Germination, Growth and Metabolism of Mustard and Oilseed Rape

The use of plants that have high allelopathic potential as natural herbicides in the form of aqueous extracts is gaining popularity in environmentally friendly agriculture. Usually, their effect on the germination and growth of weeds is investigated. However, less attention is paid to the effect of the allelopathic compounds from extracts on cultivated plants. Therefore, the aim of this study was to evaluate the impact of herbal extracts that have allelopathic properties on selected physiological and biochemical processes of two plants of great economic importance—white mustard (Sinapis alba L.) and oilseed rape (Brassica napus L. var. oleifera). The extracts were prepared from mountain arnica (Arnica montana L.), ribwort (Plantago lanceolata L.), hypericum (Hypericum perforatum L.), common milfoil (Achillea millefolium L.), sunflower (Helianthus annuus L.) and sage (Salvia officinalis L.). The germination of white mustard and oilseed rape was most inhibited by the extracts that were prepared from sage and sunflower. Additionally, in the germinating plants, the sunflower extracts increased the membrane permeability, which indicates membrane injuries. The metabolic changes in the plants were monitored using isothermal calorimetry and FT-Raman spectroscopy. The total heat production, which provided information about the metabolic activity of the white mustard and oilseed rape, was decreased the most by the sage extract but generally all of the tested extracts disturbed the shape of the heat emission curves compared to the water control. The impact of the allelopathic compounds that are present in the herbal extracts on the metabolism of the seedlings was clearly visible on the FT-Raman spectra—in the fatty acids and flavonoids range, confirmed using a cluster analysis. In conclusion, the herbal extracts from medicinal plants that have herbicidal activity could be used as a natural herbicide for weed control, but since they may also have negative impacts on cultivated plants, preliminary tests are advisable to find the extract from the species that has the least negative effect on a protected crop.