# Antioxidant and Antibacterial Properties of Extracts and Bioactive Compounds in Bryophytes
## 3 Antioxidant Activity
Cell models are a valuable tool in the selection of compound bioactivities prior to clinical trials in animals and humans [43]. Furthermore, in vivo testing of pure isolated compounds is a mandatory step in the drug discovery process, supporting in vitro and cell studies [44]. Currently, no research has focused on in vivo experimentations of hyprophyritic antioxidant compounds. At the present stage, only a few studies have investigated the antioxidant activity of bryophytes extracts and pure compounds in cell models (phagocytes; murine fibroblasts) [45; 46; 47; 48; 49; 50]. [PERSON] et al. [45] studied the antioxidant activity of acetonic extracts of _Lumularia cruciata_ on phagocytes through the chemiluminescence inhibition test. In another study, _Leptodactylum riparian_ acetate extracts were used to treat whole blood phagocytes gathered from three healthy donors. Blood samples were treated with the ROS inducing agents opsonized zymosan (OZ) and phorbol myristate acetate (PMA). The authors found that the acetonic extracts of _L. riparian_ inhibited luminol-dependent chemiluminescence in samples treated with OZ and PMA in a dose-dependent manner [46]. In a study by [47], the authors treated murine fibroblast NIH-3T3 cells with water extract from the moss _Cryphae heteromalla_. NIH-3T3 cells exposed to 500 \(\mu\)M tert-butyl hydroperoxide (TBH) oxidative stress-inducing agent and treated with the water extract (0.5 \(\mu\)g/mL) showed a 50% inhibition of ROS generation for those treated only with TBH (500 \(\mu\)M) for 1 h.  
Other researchers have found dose-dependent Nrf-ARE system induction by diterponoids isolated from liverworks (Table 2) [48; 49; 50]. Nrf (nuclear factor erythroid 2- related factor) is a transcription factor responsive to cell redox status via Keep (Kelch-like ECH-associated protein 1), that bound the enhancer sequence ARE (antioxidant response elements). The ARE sequence is associated with several antioxidant cytoprotective proteins such as the NAD(P)H: quinone oxidoreductase 1 (NQO1) [13]. An investigation by [48] demonstrated that the diterponoid frullanian D isolated from the liverwort _Frullania__hamatiloba_ stimulated the nuclear translocation of Nrf2, inducing Nrf2-related enzymes (NAD(P)H:quinone oxidoreductase-1, \(\gamma\)-glutamyl cysteine synthetase) in MOVAS cells. Furthermore, 18 h pretreatment with frullanian D ameliorated the H2O2 oxidative damage in the model cells. Another study found a NAD(P)H:quinone oxidoreductase-1 (NQO1) inducing activity in _Diplophyllum taxifolium_ ethanolic extract. Researchers also isolated and characterized sixteen terpenoids and found that three of them, diplotaxifol A, diplotaxifol B, and atractynelonide III, induced NQO1 in a dose-dependent manner [49]. Similar results were obtained by [50] with the sacculatane diterpenoids eppyphyllin A-D and pellianolactone B. Therapeutic applications are also hampered by the need to collect large amounts of natural plant material, obtain pure material for high-value secondary metabolites, genetically stable populations and controlled growth conditions. As a result, in vitro cultures are the best way to develop large-scale production of bryophyte nutraceuticals [51]. At the current state of the research, methods to in vitro grow several bryophytes species have been well established [35]. Other studies have focused on the difference in antioxidant metabolites between in vitro grown and naturally grown bryophytes [30; 33; 37; 52; 53]. Interestingly, these studies have found no significant differences in antioxidant activity and metabolite composition in in vitro grown bryophytes. \begin{table}
\begin{tabular}{c c c c c c} \hline
**Extract** & & **Species** & **Method** & **Activity** & **References** \\ \hline Ethanol & Collected & _Marchuria_ & DPPH; ABTS; O2- & \ \begin{tabular}{c} TPC: 4.62 mg/g \\ DPPH \\ \end{tabular}  & [28] \\ \hline Water:methanol (3:7) & Collected &
\begin{tabular}{c} _Hypmam  
with no bacteria growth) is measured.