Analysis of the effects of Cu-MOFs on fungal cell inactivation

Three dimensional (3D) copper metal organic frameworks (Cu-MOFs) containing glutarates and bipyridyl ligands (bpa = 1,2-bis(4-pyridyl)ethane, bpe = 1,2-bis(4-pyridyl)ethylene, or bpp = 1,3-bis(4-pyridyl)propane) were synthesized by using previously reported hydrothermal reactions or a layering method. All three Cu-MOFs contained well-defined one dimensional (1D) channels with very similar pore shapes and different pore dimensions. The bulk purities of the Cu-MOFs were confirmed using powder X-ray diffraction (PXRD) and infrared spectroscopy (IR) spectra. When the three types of Cu-MOFs were applied to Candida albicans cells and Aspergillus niger spores, an average of about 50–80% inactivation was observed at the highest concentration of Cu-MOFs (2 mg mL−1). The efficiency of the fungal inactivation was not significantly different among the three different types (bpa, bpe, bpp). Treatment of the fungi using Cu-MOFs induced an apoptosis-like death and this was more severe in A. niger than C. albicans. This may be due to elevation of the intracellular level of reactive oxygen species (ROS) in A. niger. Generation of the reactive species in solution by Cu-MOFs was observed. However, there was a dramatic variation in the levels observed among the three types. Our results suggest that Cu-MOFs can produce antifungal effects and induce apoptosis-like death of the fungi, which was probably caused by the elevated level of intracellular reactive species.


Introduction
Metal-organic frameworks (MOFs) and coordination polymers (CPs) have attracted considerable interest in coordination and materials chemistry, 1 and well-known applications of MOFs include gas sorption and separation, 2 catalysis, 3 luminescence sensing, 4 and biomedical usage. 5 These MOFs can also be used as bioactive framework materials (BioMOFs) 5,6 and they have received considerable attention for the development of novel and efficient antimicrobial agents. Silver coordination networks have been used as effective antimicrobial agents, and have shown notable antimicrobial efficiency against a range of different bacteria (E. coli, P. aeruginosa and S. aureus) and yeast (C. albicans) by releasing Ag I ions from Ag-MOFs or Ag-CPs. 7 The water-stable Cu-BTC (1,3,5-benzenetricarboxylate) metal organic framework (MOF) can inhibit the rate of growth of C. albicans, 8 and another Cu-based HKUST-1 MOF can inactivate Saccharomyces cerevisiae, the antifungal property of which is a result of Cu II ions that are released from its structure. 9 A Cobased MOF (Co-TDM, TDM 8À ¼ [ (3,5-dicarboxyphenyl)oxamethyl]methane), was shown to be highly effective at inactivating E. coli. 10 Recently, Co II -coordination polymers (Co-CPs) containing glutarates and bipyridyl ligands (bpa ¼ 1,2-bis(4-pyridyl)ethane, bpe ¼ 1,2-bis(4-pyridyl)ethylene, or bpp ¼ 1, 3-bis(4-pyridyl) propane) have been prepared. 11 Co-CPs can inactivate C. albicans cells more efficiently than A. niger spores within the same treatment time, and the greater inactivation of C. albicans by Co-CPs may result from dramatic changes in the morphology of C. albicans. We concluded that the small amount of leached Co II ions and the robust Co-CPs themselves, as well as the reactive species generated by the Co-CPs, can actively participate in fungal inactivation.
As an extension of our previous work, we tested the antifungal properties of Cu-MOFs, with the same glutarate and bipyridyl ligand system (Scheme 1), against C. albicans and A. niger. In contrast to the Co-CPs, all three Cu-MOFs, with the formulation [Cu 2 (Glu) 2 (m-L)]$x(H 2 O) (L ¼ bpa, bpe, or bpp), have similar 3D frameworks with well-dened one dimensional (1D) channels and possess very similar pore shapes with different pore dimensions and void volumes. 12 These Cu-MOFs showed excellent antibacterial activities against the ve types of bacteria tested, including both Gram-positive bacteria (Staphylococcus aureus and MRSA) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa), with very low MBCs (minimal bactericidal concentrations). 12 The antifungal properties of the Cu-MOFs can be comparable to those of the Co-CPs containing the same ligand system.

Instrumentation
Powder X-ray diffraction (PXRD) patterns were recorded on a Bruker D8 Focus diffractometer (40 kV, 30 mA, step size ¼ 0.02 ). Infrared spectroscopy (IR) spectra were measured on a BIO RAD FTS 135 spectrometer using KBr pellets. Inductive coupled plasma mass spectrometer (ICP-MS) measurements were performed in the Seoul Center, Korea Basic Science Institute.

Fungal strains and culture conditions
The fungal strains used in the study were C. albicans KCTC7270 (yeast type) and A. niger which were kindly provided by the Korean Collection for Type Cultures in Korea Research Institute of Bioscience and Biotechnology (Jeongeup-si, Jeollabuk-do, Korea) and Dr. Seong-Hwan Kim's Laboratory in Dankook University (Cheonan-si, Chungnam, Korea), respectively. The fungi were grown and maintained in potato dextrose agar (PDA) at 30 C.

Treatment with Cu-MOFs and assay for fungal viability
C. albicans cells and A. niger spores were treated with Cu-MOFs containing glutarates and bpa (MOF 1), bpe (MOF 2) or bpp (MOF 3). C. albicans cells were prepared as follows, 1-2 colonies from the PDA culture plate were suspended in 1 mL of potato dextrose broth (PDB), and 10 mL of the suspension was inoculated into a further 15 mL of PDB. Aer incubation at 30 C with shaking for 20 h, the cells were pelleted down via centrifugation at 3134 Â g for 5 min. The liquid part was discarded, and the cell pellet was washed with 1Â PBS (phosphate buffered saline). Then, fresh PBS was added to the washed cell pellet and the concentration was adjusted to 10 8 per mL. To collect the spores of A. niger, about 10 mL PBS was added into a PDA culture plate grown for 2 weeks, and the fungal mycelia were scrapped using scrapper. The scrapped suspension was ltered through three layers of miracloth. The ltrate was centrifuged at 3134 Â g for 5 min and the liquid part was discarded. The spore pellet was washed with PBS and resuspended in new PBS to give a concentration of 10 8 per mL.
C. albicans cells and A. niger spores suspended in PBS at a concentration of 10 8 per mL were added to each well of a 24 well plate (1 mL per well). Stock suspensions of MOF 1, MOF 2 and MOF 3 were prepared by adding MOF powders in PBS (20 mg mL À1 ). Aer well mixed, an appropriate volume of MOF stock suspension was added into each well with nal concentrations 0, 0.125, 0.25, 0.5, 1, and 2 (mg mL À1 ). The plates were incubated at room temperature with shaking for 4 d.
Aer treatment with Cu-MOFs, 10 mL of the fungal suspension was removed and serially diluted, and then 100 mL of the diluted suspension was spread onto the PDA plate. The plates were incubated at 30 C for 2 d, and the number of colonies was counted. The number of colonies from nine replicate plates was averaged.

Scanning electron microscopy analysis
The surface morphology of the C. albicans cells and A. niger spores was analysed using scanning electron microscopy (SEM) aer treatment with Cu-MOFs (2 mg mL À1 ) at room temperature with shaking for 4 d. The untreated sample was used as a control. The treated and control fungal cells and spores were prepared for SEM analysis by following previously described procedures. 10 The prepared samples were examined under a scanning electron microscope (JEOL, Tokyo, Japan).

Measurement of the levels of reactive oxygen species and reactive nitrogen species
To measure the levels of the intracellular reactive species, C. albicans cells and A. niger spores in PBS (10 8 mL À1 ) were treated with Cu-MOFs (2 mg mL À1 ) for 4 d as described in an earlier section. An untreated sample was used as a control. The treated and control cells and spores were washed with PBS once and incubated in PBS containing 10 mM of H 2 DCFDA or DAF-FM DA at room temperature in the dark for 1 h. Aer incubation, the fungal cells and spores were washed with PBS twice and resuspended in 100 mL of PBS. The fungal suspensions were transferred to a black walled 96-well assay plate, and the uorescence was measured at 495 nm (excitation) and 515 nm (emission) using a Synergy HTX Multi-Mode Reader (BioTek Instruments, VT, USA).
To measure the levels of H 2 O 2 and NO x in the media, 1 mL of PBS was treated with the Cu-MOFs (2, 1, 0.5, 0.25, 0.125, and 0 mg mL À1 ) at room temperature with shaking. Aer treatment, the level of H 2 O 2 and NO x was measured immediately (0 d) and on the 4 th day. An Amplex™ Red Hydrogen Peroxide/Peroxidase Assay Kit (Molecular Probes, Eugene, OR, USA) and QuantiChrom™ Nitric Oxide Assay Kit (BioAssay Systems, Hayward, CA, USA) were used to determine the H 2 O 2 and NO levels, respectively, following the manufacturer's protocols.

TUNEL assay
Apoptosis-like fungal cell death was analysed using the TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labelling) assay. In the TUNEL assay, DNA fragmentation was detected by Scheme 1 Chemical structures of glutaric acid, bpa, bpe, and bpp ligands.
labelling the 3 0 -hydroxy terminals with uorescence. C. albicans cells and A. niger spores in PBS (10 8 mL À1 ) were treated with Cu-MOFs (2 mg mL À1 ) at room temperature with shaking for 4 d. An untreated sample was used as a control. The fungal cell wall was rst removed before the TUNEL assay. The treated and control fungal cells and spores were washed with PBS and then incubated in OM buffer (1 M MgSO 4 ,8.4 mM Na 2 HPO 4 , 1.6 mM NaH 2 PO 4 ) containing a lysing enzyme (10 mg mL À1 , Sigma, St. Louis, MO, USA) at 30 C for 2 h to digest the cell wall. Aer the cell wall had been digested the fungal protoplasts were washed with PBS once and xed with 1% (v/v) paraformaldehyde in PBS on ice for 15 min. Aer xation, the protoplasts were washed with PBS twice and resuspended in 0.1 mL PBS. 1 mL of ice-cold 70% (v/v) ethanol was added, this was incubated on ice for 30 min and then stored at À20 C until further use.
The fungal protoplasts were further processed using the TUNEL Assay Kit -FITC (Abcam, Cambridge, UK). Ethanol was removed and the protoplasts were washed twice with wash buffer. Aer washing, 51 mL of the DNA labelling solution containing 10 mL of the reaction buffer, 0.75 mL of the TdT enzyme, 8 mL of FITC-dUTP, and 32.25 mL of ddH 2 O were added to the samples and then incubated at 37 C for 3 h. Aer incubation, the protoplasts were washed twice with rinse buffer, resuspended in 0.1 mL of propidium iodide/RNase A and incubated at room temperature in the dark for 30 min. The uorescence was observed via a confocal laser scanning microscope (Olympus Corporation, Tokyo, Japan) at 488 nm (excitation) and 520 nm (emission).

Quantitative PCR for the metacaspase gene
The transcription levels of the metacaspase-1 (ref. 13) in C. albicans and metacaspase 1A and 1B in A. niger (homologs of CasA and CasB in A. fumigatus, respectively) 14 were assessed in the fungal samples treated with Cu-MOFs for 1 d using the quantitative polymerase chain reaction (QPCR). The total RNA extraction and QPCR were performed as described in a previous study. 15 The actin gene was used as a reference. The sequences of the primer used in the QPCR were obtained as follows: For A. niger: Metacaspase-1A: forward-ATCCTCAAGGAGCCCAACTT, reverse-CTGGTCTTGGTCTGGATGGT Metacaspase-1B: forward-CATCCAATTACCGCTTCCAG, reverse-CATCGGGTTCTTCTGATCGT Actin: forward-ACAGTCCAAGCGTGGTATCC, reverse-GCCTGGATGGAGACGTAGAA;and for C. albicans: Metacaspase-1: forward-CCACCAAGTCAACAAGGACA, reverse-GCCTTTTTCCGACCACTACA, Actin: forward-AGGTTTGGAAGCTGCTGGTA, reverse-GAAGATGGAGCCAAAGCAGT.

Structural properties of the Cu-MOFs containing glutarate and bipyridyl ligands
The 3D Cu-MOFs were synthesized by using previously reported hydrothermal reactions or a layering method. 12 The paddle-wheel Cu 2 dinuclear units in all three Cu-MOFs are connected by glutarates (Glu) to form 2D sheets, and these sheets are bridged by bipyridyl ligands (bpa, bpe, and bpp) to form the 3D frameworks (Fig. 1). The 3D frameworks had the formula [Cu 2 (Glu) 2 (m-L)]$x(H 2 O), in which L is bpa (MOF 1), bpe (MOF 2) or bpp (MOF 3). All three MOFs contained well-dened 1D channels, and they possessed very similar pore shapes with different pore dimensions and void volumes. 12 The bulk purities of the Cu-MOFs 1-3 were conrmed using PXRD (Fig. 2), and the IR spectra of Cu-MOFs 1-3 were found to be in good agreement with those reported previously (Fig. S1 †). 12 Very strong, slightly broader bands were observed at approximately 1620 and 1420 cm À1 , which represented the asymmetric and symmetric C]O stretching modes of the bound glutarate moieties. The absence of any bands in the area of approximately 1710 cm À1 indicated full deprotonation of all of the carboxylate groups in Cu-MOFs 1-3, 16 which is consistent with the results of the X-ray analyses. The structural stabilities of MOFs 1-3 aer 4 d in water or PBS solution were tested (Fig. 2). The PXRD patterns of MOFs 1-3 indicate that their structures are stable aer 4 d in water or PBS solution.
The degradation tests were carried out in the PBS solution used for antifungal testing at room temperature to conrm that the leaching of Cu II ions from Cu-MOFs was not signicant. The quantities of Cu II ions released from the Cu-MOFs were monitored using ICP-MS aer 1, 2, 3 and 4 d (Fig. 3). The concentration of the released Cu II ions from 1 mg of a compound in 1 mL of 1Â PBS aer 4 d was approximately 0.678 mg mL À1 for MOF 1. The amount of Cu II ions released from MOF 1 (0.170 mg mL À1 ¼ 0.00017 mg mL À1 ) is negligible compared to 2 mg mL À1 . The Cu-MOFs do not easily release Cu II ions and are stable in PBS.

Reduced fungal viability aer treatment with Cu-MOFs
Aer the C. albicans cells were treated with three types of Cu-MOF for 4 d, the CFU (colony forming unit) number was reduced in a MOF concentration dependent manner (Fig. 4). Approximately 35-50% of the cells were inactivated at 0.125 mg mL À1 , and a 50-70% reduction was observed at higher concentrations (Fig. 4). Although the efficiency of C. albicans inactivation was not signicantly different among the different MOFs, 1, 2 and 3, the CFU number was slightly lower aer treatment with MOF 3. In A. niger, about 60% of the spores were inactivated by 0.125 mg mL À1 MOF 1, whereas about 25-40% spores were inactivated by 0.125 mg mL À1 MOF 2 and MOF 3 (Fig. 5). A similar inactivation percentage was maintained in the treatment with 0.25-2.0 mg mL À1 MOF 1, and a further reduction in spore germination (50-80% reduction) was observed aer treatment with 0.25-2.0 mg mL À1 MOF 2 and MOF 3 (Fig. 5). Upon treatment with MOF 2, the inactivation rate was most signicantly decreased, by 80% at a concentration of 2.0 mg mL À1 (Fig. 5b). In both C. albicans and A. niger, no signicant difference in the cell inactivation was observed between the samples that were or were not shaken (Fig. S2 †).    The antifungal activity of Cu-based MOFs is occasionally reported. 8,9 Our results provide further evidence to prove that Cu-MOFs can inactivate yeast cells and fungal spores. The efficiency of the fungal inactivation seems to be reduced in our study, compared to that observed by Chiericatti et al. 9 and Bouson et al. 8 in their previously published studies, both found that a higher inactivation rate was achieved with a reduced amount of Cu-MOF in a shorter time. As the structure and properties of the MOFs and the fungal cell numbers used in the assay were different among studies, the fungal inactivation rate can be variable. The inactivation percentage was not signicantly different between C. albicans and A. niger, which showed an average of about 50-70% in this study. This result is different from that found in our previous study using Co-CPs. C. albicans showed a greater sensitivity to Co-CPs than A. niger under the same experimental conditions (fungal cell number, treatment time, MOF concentration). 11 A possible explanation for this difference is that overall level of reactive species generated by Cu-MOFs (about 0.05-9 mM for H 2 O 2 , 5-45 mM for NO x ) is lower than that by Co-CPs (0.2-12 mM for H 2 O 2 , 50-500 mM for NO x ). 11 Although reactive species are not the only factor for antifungal activity of Cu-MOFs, they may have contributed in part to generating the difference in antifungal activity between Co-CPs and Cu-MOFs. Another explanation is that 3D structures of Co-CPs and Cu-MOFs are different, and this may have caused the physical or chemical damage differently in fungal strains.

Apoptosis-like fungal death was observed aer treatment with Cu-MOFs
We observed apoptosis-like death in C. albicans cells and A. niger spores aer treatment with Cu-MOFs for 4 d (Fig. 6a, b, S3 and S4 †). The number of uorescent (DNA fragmentation) cells and spores were higher in the MOF treated samples compared to the control. The uorescence was more intense in A. niger than C. albicans indicating that apoptosis-like death might proceed more readily. In addition, the number of cells and the uorescence intensity were relatively higher aer treatment with MOF 3 compared to MOF 1 and MOF 2 for both C. albicans and A. niger (Fig. 6a, b, S3 and S4 †). We analysed the expression of the metacaspase gene that is known to be activated in fungal apoptosis-like death. 17 The level of mRNA for metacaspase 1 was much lower than that of the control in the samples treated with Cu-MOFs for 1 d (Fig. 6c). However, the amount of mRNA for metacaspase 1A and 1B was signicantly increased in the A. niger spores treated with the three types of Cu-MOFs. Treatment with MOF 2 showed the lowest increase in the amount of metacaspase mRNA (Fig. 6c).
Apoptosis-like cell death using MOFs is occasionally observed in human cells, and drug or photodynamic sensitizers encapsulated in MOFs can play a major role in inducing cell apoptosis. 18 In our results, the apoptosis-like fungal cell death seems to be caused directly by the Cu-MOFs, which is different from the results observed in previous studies. This is the rst time that MOFs have been observed to cause apoptosis-like cell death in microorganisms. Our results show that metacaspase transcription is promoted in MOF treated A. niger, but not C. albicans (Fig. 6c). This may be related to the reduced DNA fragmentation observed in C. albicans treated with MOFs in the TUNEL assay (Fig. 6).

Mechanism of the antifungal activity of the Cu-MOFs
As the antimicrobial activity of MOFs is known to be attributed to the free metals and organic ligands released from the framework, 19 we investigated whether free Cu 2+ and glutarate released from a MOF (Fig. 3) can produce an antifungal effect. Free Cu 2+ was obtained from the dissolved Cu(NO 3 ) 2 in PBS. When free Cu 2+ (as Cu(NO 3 ) 2 ) and glutarate was applied to C. albicans cells and A. niger spores with the released concentration for 4 d, no signicant difference in the fungal CFU number between the control and treatment groups was observed in both types of fungi, except for the treatment of C. albicans with glutarate in which the cell number almost doubled aer treatment (Fig. 7). This indicates that degeneration of metal and organic ligands is not a critical factor in the fungal inactivation observed aer treatment for 4 d. Although free Cu 2+ itself has antimicrobial effect, the amount released from Cu-MOFs in PBS is not enough to cause inactivation of C. albicans cells and A. niger spores. Cu-MOFs used in our study is water stable, and therefore, other factors may be more critical than free Cu ion and glutarate. Electric interaction of fungal cells with Cu and glutarate in MOF may possibly cause degeneration of biomolecules on fungal cell surface. Photocatalytic ROS generation from MOF can be another potential mechanism for fungal inactivation. 20 In the SEM analysis, many C. albicans cells treated with the three different types of Cu-MOFs were crushed, compared with the egg shaped control cells (Fig. 8a). No dramatic difference in the cell morphology was observed between the three types of Cu-MOFs. In A. niger, most of the spores became crumpled aer treatment with the three different types of Cu-MOFs (Fig. 8b). There were barely any crushed cells such as those observed in C. albicans (Fig. 8b). The spore surface layer (probably the cell wall) seemed to become rumpled (Fig. 8b).
MOFs are known to produce ROS, and this ROS can cause an antimicrobial effect. 20 To test the involvement of reactive species in fungal inactivation, production of ROS and RNS species (H 2 O 2 and NO x ) in PBS treated with the three different   types of Cu-MOFs was examined. Immediately aer treatment with the MOFs (0 d), the NO x level was found to increase with the increasing concentration of MOF 2 (from about 3 to 13 mM) but this increase was not detected aer treatment with MOFs 1 and 3 (Fig. 9a). A relatively low amount of H 2 O 2 (about 0.05-0.5 mM) was measured and this increased upon treatment with the MOFs in a concentration dependent manner (Fig. 9a). Aer 4 d, the NO x level in the MOF 2 treated PBS was elevated compared to 0 d and increased with upon treatment with the MOFs in a concentration dependent manner (Fig. 9a). The H 2 O 2 concentration was greater aer 4 d compared to 0 d and decreased upon treatment with the MOFs in a concentration dependent manner.
As apoptosis-like death was observed in both C. albicans and A. niger, the intracellular ROS and RNS level was assessed aer treatment with the MOFs. Intracellular NO was not generated aer treatment with the MOFs in either fungi (Fig. 9b). However, the level of intracellular ROS detected using H 2 DCFDA was signicantly increased in the A. niger spores aer treatment with all the MOFs, although a relatively lower increase was observed aer treatment with MOF 2 (Fig. 9b).
To determine the fungal responses to reactive species produced in PBS, we examined membrane lipid peroxidation. The amount of MDA (the product of membrane lipid peroxidation) was signicantly increased in C. albicans aer treatment with MOF 3 (Fig. 9c). However, it was decreased in A. niger aer treatment with all of the MOFs (Fig. 9c). This indicates that there was not a dramatic increase in the occurrence of membrane lipid oxidation aer treatment with the MOFs.
About the mechanism of the antimicrobial activity of the MOFs, metal ions and organic ligands released from the degraded MOFs are oen considered to be critical factors in microbial cell damage and death. 6,19 Chiericatti et al. 9 demonstrated that Cu ions released from a MOF crystal into the media were responsible for the antifungal effect. In our study, a small amount of Cu ions was released from the frameworks, but the amount was not sufficient to cause fungal inactivation. In addition, glutarate, an organic ligand chemical used in our study, was not found to be harmful to fungal cells and spores at the concentrations used in the construction of the MOF.
Several studies have demonstrated that the reactive species generated by MOF-mediated photooxidation are responsible for antimicrobial activity. 11,19,20 In our results, the H 2 O 2 level was increased immediately aer treatment with Cu-MOFs in a concentration dependent manner, however, the level was lower than that in the control aer 4 d. H 2 O 2 produced immediately aer treatment with MOFs may play a role in fungal inactivation. However, it may not be sufficient to cause fungal inactivation aer 4 d. The level of NO x in PBS was only found to be increased in MOF 2, and therefore, cannot be considered as a common factor generating an antifungal effect. In addition, membrane lipid peroxidation does not seem to signicantly occur in treated fungal cells. These results suggest that the reactive species generated in PBS by Cu-MOFs may not be critical to fungal inactivation. In addition, generation of H 2 O 2 and NO x by Cu-MOFs was much lower than that by  This may explain the high sensitivity of C. albicans to Co-MOFs than Cu-MOFs. 11 We observed a signicant increase in the level of intracellular ROS in A. niger spores. This may be related to the slightly higher level of apoptosis-like death observed in A. niger compared to C. albicans. In addition, transcription of the metacaspase gene was signicantly increased in A. niger treated with MOFs. Cu-MOF treatment may have caused the elevation of the intracellular ROS level in A. niger, and this could promote metacaspase gene expression, leading to apoptosis-like cell death. However, the higher rate of apoptosis-like death in the MOF 3 treated cells still cannot be explained. Further investigation on this is needed.

Conclusions
Three types of 3D Cu-MOFs containing glutarates and bipyridyl ligands (bpa, bpe, or bpp) were synthesized by using previously reported hydrothermal reactions or a layering method. All three Cu-MOFs contained well-dened 1D channels with very similar pore shapes and different pore dimensions. The bulk purities of the Cu-MOFs were conrmed using PXRD and IR spectra. The antifungal activity of the Cu-MOFs against C. albicans cells and A. niger spores showed a similar efficiency. However, our study also demonstrated that the molecular mechanisms for fungal inactivation of the Cu-MOFs were slightly distinct between the different fungal species. Although an average of about 50-80% fungal inactivation by Cu-MOFs was observed in our experimental conditions, our results suggest that Cu-MOFs have potential for application as antifungal agents that could be used in various contamination or infection situations.

Conflicts of interest
There are no conicts to declare.