Original Research Article Evaluation of in vitro cell and blood compatibility and in vivo analgesic activity of plant-derived dietary supplements Stefania Lamponi a,⇑ , Anna Maria Aloisi b , Claudia Bonechi a , Marco Consumi a , Alessandro Donati a , Gemma Leone a , Claudio Rossi a , Gabriella Tamasi a , Luana Ghiandai b,c , Ersilia Ferrini b,c , Paolo Fiorenzani d , Ilaria Ceccarelli d , Agnese Magnani a a Department of Biotechnology, Chemistry and Pharmacy, University of Siena, 53100 Siena, Italy b Union B.I.O. S.r.l., 52100 Arezzo, Italy c Labor Chimica S.r.l., 52100 Arezzo, Italy d Department of Medicine, Surgery and Neurosciences, University of Siena, 53100 Siena, Italy article info Article history: Received 22 August 2018 Accepted 29 December 2018 Available online xxxx Keywords: Plant extracts Fibroblasts Endothelial cells Cytotoxicity Cytocompatibility Haemocompatibility Pain reduction In vivo animal model abstract Objective: In vitro cell and blood compatibility of three dietary supplements, comprised of multiple plant extracts, Pneumo Go (PG), Green active (GA) and Equistasi (Eq), and their main component, the phyto- complex Matrix U.B. Ò (Union Bio S.r.l.) (M), were evaluated. Moreover, preliminary in vivo tests were performed on GA in order to assess its ability to reduce pain in an animal model. Methods: Cell compatibility was determined using fibroblasts (NIH3T3) and primary adult human microvascular endothelial cells (HMVECad) and the neutral red uptake test. Blood compatibility was eval- uated by analyzing blood parameters after incubation of the products with sodium citrate anticoagulated whole blood. Thrombin time was determined by adding thrombin to aliquots of human plasma contain- ing the samples. Clotting time was revealed by an automatic coagulometer. The in vivo analgesic effect of GA was evaluated in Wistar rats using the formalin test. Results: M and PG reduced the percentage of viable NIH3T3 cells, indicating their interference in the cell cycle. GA and Eq stimulated fibroblast proliferation and neutralized the toxic effect of M. M and PG reduced HMVECad cell viability. GA and Eq did not affect cell viability as well as negative control. The hemocom- patibility tests indicated that all the samples did not interfere with fibrinogen. The in vivo test carried out in male rats showed a significant analgesic effect of GA in all formalin-induced pain behaviors. Conclusion: No hemotoxicity and good cell compatibility were found for all the tested samples. GA and Eq were the best candidates for further biocompatibility testing. Moreover, GA reduced pain in the animal model. Please cite this article as: Lamponi S, Aloisi AM, Bonechi C, Consumi M, Donati A, Leone G, Rossi C, Tamasi G, Ghiandai L, Ferrini E, Fiorenzani P, Ceccarelli I, Magnani A. Evaluation of in vitro cell and blood compatibility and in vivo analgesic activity of plant-derived dietary supplements. J Integr Med. 2019; xx(x): xxx–xxx Ó 2019 Shanghai Changhai Hospital. Published by ELSEVIER B.V. All rights reserved. 1. Introduction Currently, traditional medicine is one of the therapeutic approaches available for treatment and prevention of numerous diseases. Phytotherapy and the use of dietary supplement products are considered to be complementary methods. In recent years, interest in drugs of plant origin and in nutraceutical compounds has increased for various reasons, including overcoming the limita- tions of synthetic drugs, which often can be inefficient or have negative side effects [1,2]. For example, in the past amphotericin B deoxycholate was the standard therapy for Candida infections, but it is often poorly tolerated and associated with nephrotoxicity and infection-related reactions [3]. One obstacle to the use of both plant-derived drugs and dietary supplement products is the lack of scientific information on these substances, such as pharmacologi- cal, phytochemical and in vitro and in vivo biological data [4]. For example, it is well known that essential oils from many plants possess antifungal activity and could be used as alternatives to synthetic products, but there is insufficient information on their https://doi.org/10.1016/j.joim.2019.02.004 2095-4964/Ó 2019 Shanghai Changhai Hospital. Published by ELSEVIER B.V. All rights reserved. ⇑ Corresponding author. E-mail address: stefania.lamponi@unisi.it (S. Lamponi). Journal of Integrative Medicine xxx (xxxx) xxx Contents lists available at ScienceDirect Journal of Integrative Medicine journal homepage: www.jcimjournal.com/jim www.journals.elsevier.com/journal-of-integrative-medicine Please cite this article as: S. Lamponi, A. M. Aloisi, C. Bonechi et al., Evaluation of in vitro cell and blood compatibility and in vivo analgesic activity of plant- derived dietary supplements, Journal of Integrative Medicine, https://doi.org/10.1016/j.joim.2019.02.004