Trzaskowska Marta, Vivcharenko Vladyslav, Kazimierczak Paulina, Wolczyk Agata, Przekora Agata
Independent Unit of Tissue Engineering and Regenerative Medicine, Medical University of Lublin, Chodzki 1, 20-093 Lublin, Poland.
Biomedicines. 2023 Apr 4;11(4):1095. doi: 10.3390/biomedicines11041095.
Over the years, essential oils (EOs) and their compounds have gained growing interest due to their anti-inflammatory, antimicrobial, antioxidant, and immunomodulatory properties. The aim of this study was to evaluate the effect of eight commercially available EO-derived compounds ((R)-(+)-limonene, (S)-(-)-limonene, sabinene, carvacrol, thymol, alpha-pinene (α-pinene), beta-pinene (β-pinene), and cinnamaldehyde) on the bone formation process in vitro to select the most promising natural agents that could potentially be used in the prevention or treatment of osteoporosis. Within this study, evaluation of cytotoxicity, cell proliferation, and osteogenic differentiation was performed with the use of mouse primary calvarial preosteoblasts (MC3T3-E1). Moreover, extracellular matrix (ECM) mineralization was determined using MC3T3-E1 cells and dog adipose tissue-derived mesenchymal stem cells (ADSCs). The two highest non-toxic concentrations of each of the compounds were selected and used for testing other activities. The conducted study showed that cinnamaldehyde, thymol, and (R)-(+)-limonene significantly stimulated cell proliferation. In the case of cinnamaldehyde, the doubling time (DT) for MC3T3-E1 cells was significantly shortened to approx. 27 h compared to the control cells (DT = 38 h). In turn, cinnamaldehyde, carvacrol, (R)-(+)-limonene, (S)-(-)-limonene, sabinene, and α-pinene exhibited positive effects on either the synthesis of bone ECM or/and mineral deposition in ECM of the cells. Based on the conducted research, it can be assumed that cinnamaldehyde and (R)-(+)-limonene are the most promising among all tested EO-derived compounds and can be selected for further detailed research in order to confirm their biomedical potential in the chemoprevention or treatment of osteoporosis since they not only accelerated the proliferation of preosteoblasts, but also significantly enhanced osteocalcin (OC) synthesis by preosteoblasts (the OC level was approx. 1100-1200 ng/mg compared to approx. 650 ng/mg in control cells) and ECM calcification of both preosteoblasts and mesenchymal stem cells. Importantly, cinnamaldehyde treatment led to a three-fold increase in the mineral deposition in ADSCs, whereas (R)-(+)-limonene caused a two-fold increase in the ECM mineralization of both MC3T3-E1 cells and ADSCs.
多年来，由于其抗炎、抗菌、抗氧化和免疫调节特性，精油（EOs）及其化合物越来越受到关注。本研究的目的是评估八种市售的源自EO的化合物（（R）-（+）-柠檬烯、（S）-（-）-柠檬烯、桧烯、香芹酚、百里香酚、α-蒎烯、β-蒎烯和肉桂醛）对体外骨形成过程的影响，以选择最有潜力用于预防或治疗骨质疏松症的天然药物。在本研究中，使用小鼠原代颅骨前成骨细胞（MC3T3-E1）对细胞毒性、细胞增殖和成骨分化进行了评估。此外，使用MC3T3-E1细胞和犬脂肪组织来源的间充质干细胞（ADSCs）测定细胞外基质（ECM）矿化。选择每种化合物的两种最高无毒浓度并用于测试其他活性。所进行的研究表明，肉桂醛、百里香酚和（R）-（+）-柠檬烯显著刺激细胞增殖。就肉桂醛而言，MC3T3-E1细胞的倍增时间（DT）与对照细胞（DT = 38小时）相比显著缩短至约27小时。反过来，肉桂醛、香芹酚、（R）-（+）-柠檬烯、（S）-（-）-柠檬烯、桧烯和α-蒎烯对细胞的骨ECM合成或/和ECM中的矿物质沉积均表现出积极作用。基于所进行的研究，可以假设肉桂醛和（R）-（+）-柠檬烯在所有测试的源自EO的化合物中最有前景，可以选择进行进一步的详细研究，以确认它们在骨质疏松症化学预防或治疗中的生物医学潜力，因为它们不仅加速了前成骨细胞的增殖，而且还显著增强了前成骨细胞的骨钙素（OC）合成（OC水平约为1100 - 1200 ng/mg，而对照细胞中约为650 ng/mg）以及前成骨细胞和间充质干细胞的ECM钙化。重要的是，肉桂醛处理使ADSCs中的矿物质沉积增加了三倍，而（R）-（+）-柠檬烯使MC3T3-E1细胞和ADSCs的ECM矿化增加了两倍。
