Graphene-based materials: From single- to mixture ecotoxicity and challenges for nanoinformatics
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| Author | 0000-0002-0086-3055 (ORCID) | |
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Transcript: English(auto-generated)
00:00
Good morning, everyone. It's a big pleasure to stay here. And first of all, I'd like to thank you, Giorgia, Israel, Andreas, Michael, for our invitation. Today, I'm going to talk about graphene-based materials from single to mixture echotoxicity and some challenges
00:24
for non-informatics. This is the picture of our center in Brazil. The name is Brazilian Center for Research in Energy and Materials, CNP in Portuguese. We have here four national laboratories today. The Synchrotron Lab here is just three. It's the category in the world,
00:43
one of the most important today. Here National Laboratory, Nanotechnology Laboratory. Here Bioscience Lab and Bio-Enable Lab. And today, we have the ILUM, the School of Science, is the first other grad, just this course, to be the excellent teacher in Brazil. And it's important
01:03
to say, we are under construction, National Biosafety Lab, NIO4, because after COVID, the government said that it's very important for Latin America and to be the unique proposal in Latin America.
01:21
And where you are, right? This is the Brazilian map. We are in the southwest, in São Paulo State. We are in Campina City. Campina is over one million people living there. It's the second city in São Paulo State. We are just one hour from São Paulo, the capital. And the Brazilian Nanotechnology Laboratory is the youngest of the National Laboratory. We have
01:44
10 years of existence. And our focus are nanoscience, nanotechnology, innovation for sustainable development. Today, we are going in this direction, right? Everyone needs to go to sustainability. So we're looking for biodiversity, natural research, resources,
02:04
ecosystems, how to produce new materials using this, especially nanomaterials today. Looking for innovation and putting the balance with safety, of course. And this will be a contribution to sustainability. This is the current staff of the APIs at LNANO. We have three divisions, nanomaterials,
02:28
device and nanobiotechnology. I'm responsible for nanomaterials today. We have only existing researchers. He's not a big group yet. But very, very motivated people.
02:43
It's part to say that LNANO is the main lab for SISNANO. SISNANO is the national system of laboratories on nanotechnology in Brazil. We have different institutions associated with the system. It's a national program. And it's really direct from the Ministry of Science to do nanoscience.
03:00
And LNANO coordinating is the hub of this system. Today, together, the National Metrology Institute, who are participating in the CertificaNANO, is a program to certification of nanomaterials. And we started this four years
03:21
before. But the idea is to use the electronic notebook to manage the information of this important guideline, because labs need to know quality, need to do research with quality. And this is in progress. Professor Jose Mauro Granjero from IMETRO is responsible for this
03:43
project. And I'm collaborating with this. Basically, this ABENITE is the Brazilian National Standards Organization. And there is a project in the Senate to regulate nanotechnology and advanced materials in Brazil. And since NANO labs together international guidelines
04:03
and standards, we will develop these in Brazil. And we're very proud to be associated with the U.S. to construct a global management of nano and advanced materials. And looking for my talk today, excited to speak about the graphene. Everyone knows that in 2010,
04:21
it was an hour prize to Guy and those of the lab to discover of the graphene based from sticking from graphite, like it was in scot-tape, and was an impressive material in the very important type of nanomaterials in the nano today. After the discovery around 2002, 2004, different kinds of
04:45
two-dimensional materials have been developed. So it is very, very, very growing field to explore 2D materials for different applications and graphene decorated with gold nanoparticles,
05:01
with clays and different polymers, etc. And a very special of this is graphene oxide. It's a very versatile nanomaterial for innovation. You can use the graphite to exfoliate graphite, producing the graphene oxide. It's a single layer of carbon with oxygenate groups, different groups
05:23
on the surface. And there are many companies producing, commercializing these nanomaterials today for small to large scale, no regulations, no rules to commercialize these around the world. And you look for the literature and patents that are application in different sectors,
05:45
advanced materials, energy, electronics, health, and environment. So it's pretty much here to pay attention. And you look for a large scale, so more impact on the environment, right, production. It is a very interesting approach to produce graphene, exploring the mines from
06:06
graphite, natural research. It has a specific reaction, Raman's, then the name of this, you exfoliate the graphene, producing graphene oxide, and it could do a reaction to obtain the graphite. This is a typical industrial material production of the graphene oxide. And very
06:27
important to sustainability is to know where is the mines of graphite in the world, because it will be a starting material. And you look for the reserves in the world for graphite, two years before, the first one in Turkey, the second in China, and the third one in Brazil.
06:44
So due to this, the graphite is a strategy to do innovation, because we can produce different graphene-based materials, exploring graphene oxide and add value to this project.
07:01
Today, last year, we have a program, a specific program for the government to graphite in Brazil. It is the first meeting with Brazilian President Bolsonaro, the first prime minister for science, so promoting graphene, specifically research on graphene, because it's a strategy to Brazil today. And there are now big companies in Brazil producing, trying to put it in the market.
07:26
So that's the not so happy question, but companies don't like this in general, but we need to talk about. And we have three big, I say three big worlds, right? The world of the production of materials, the world of the human, and the world of the environment. So put all this
07:44
together is a big challenge to graphene-based material. We're doing many things on this direction also, and around the world. So the information are completely disorganized, and non-informatics need to help us. And starting a more complex, and adding more complexities,
08:04
is the change from analyzing the single toxic of each material, but in the environment everything is interacting. So how to assess the mixture of toxicity. This is an important topic in science recently, that we have a chemical with no toxins, no toxins, no toxins, but when you combine all
08:23
these mixtures, you can have a mixture risk. So how to manage it? That is the question. The OECD published a guideline, not standard yet, just a communication, how to assess the combined
08:41
risk of chemicals, right? And most very recently, this group publishing, Environmental Science Technology, a very interesting review article, then do a data mining research, and separating the effects of nanomaterial mixtures in different organ-use models, and different environments,
09:01
and we'll have synergies, we can have additivity, and you can have antagonism. So many factors influence all this response, and is critical to develop model to prediction of the mixture effects. And to gather this, environmental corona is coming, as a very, very important concept in
09:24
nanomaterial toxicity. So Corinne talked about this, and I liked too much of this review paper, my students love it, thank you. And we need to increase in the complexity of the direction to understand very complex surfaces, and how the biomolecules on the surface change the fate,
09:46
the toxicity, the biological response in the stable environment. In this direction, we try to explore biocoronas in environmental nano, reconnecting circular economy, metal remediation, just an example, okay, and echotaxis mitigation. In this publication,
10:04
for example, we're using the bovine plasma, in Brazil today has more coal than humans, more than 200 million of coals in Brazil, so you can measure the waste from the blood on this guy. So using this blood and special human plasma, bovine plasma, to produce a corona on carbon
10:27
nanotubes, and when you have this corona will increase for 400% of the copper remediation from water. So we're trying to use a resilient, abundant result to functionalize carbon nanotubes,
10:40
exploring the corona concept. And it was very interesting that this new material was very good to mitigate in vivo echotoxicity using Daphnia as model in aquatic echotoxicity. So we're looking to link the circular economy, metal remediation, and echotaxis mitigation. Here is a very interesting image using cryo-EM. Cryo-EM is very interesting because it's possible to see
11:06
the protein more close to the environment because it's in the hydrotrade system. And you look for this figure, the nanotube increases on the diameter and has an increase on the corona side.
11:23
So we now discuss with you to talk and how to develop a model to predict corona using cryo-EM as a correlative technique. Both. So considering the mixture echotoxicity and to do this translation from single to mixture,
11:43
this is a big picture that we'll do now. We have every time, we will have the materials, the water system, and the echotoxicity. We need to do a complete characterization of the material that we are studying. We have the technique that we are using. So the water is very complex system, depends on the approach. You can increase or decrease
12:01
the complexity of this, depends on the type of water. We need to understand colloidal stability and interaction, sometimes impossible to, indeed we don't have very good experimental techniques to assess dynamic systems like this. So computational simulation is very important.
12:21
And link it to echotoxicity. In my talk, I will talk about daphnia, elegance, and zebrafish, and how these interactions change the toxicity and naphthaneism of response. First, we start to study the toxins of graphene oxide and base wash graphene oxide, because
12:41
when you produce graphene oxide, it's not a poor material, right? You have a lot of debris on the surface of this material, and you compare the toxicity of graphene oxide with without oxidative debris. And we found using zebrafish model that the toxicity of graphene oxide is not high to zebrafish embryo, but when you remove the oxidative debris, these materials make more
13:03
hydrophobic, interact more with the eggs of the embryo, increasing the toxicity of the material. So we demonstrate that the surface byproduct is very important to interactions of the graphene oxide. Another example, now moving for more advanced materials, we complex graphene oxide
13:23
with silver to develop hybrid materials. Now you have the best of both worlds, from graphene and the silver. But the question is, how toxic is this? So the first, we perform a very good characterization of the hybrid system. Then we move to zebrafish embryo, but now consider a new
13:42
guideline that recommends to remove the membrane that protects the embryos, because this membrane is a poor membrane, and nanomaterials will be attached here, so not make a direct access to the embryo, but kill the embryo by appreciation, because it's a protection that poor and
14:02
nanomaterials interact there. So the guideline recommends to remove this membrane before to do the test, and you perform this, comparing the presence and the absence of the omic acid and chiral membrane, and you can see that both omic acid and chiral are influenced on the
14:22
response. So did a very basic but robust experiment, moving from the complete characterization of material, dispersions in studs, and ecotoxins assessment. And moving to now, to start understanding the interactions of the graphene oxide with
14:43
pollutants, this is a PD in progress. We compare, but the toxic on zebrafish, and now we mix with atrazine. Atrazine is an herbicide, very common to plantations and in agriculture. So we decide to
15:01
select the tanninic acid as our organic model, because more simple than omic acid, we start with this mixing. And first, we do the characterization of this complex, with Joel plus graphene oxide with tannic acid, using atomic photomeric acid, XPS characterization. However, to understand these
15:22
interactions is difficult, so we are losing now molecular simulations, like DFT, MV, and Vladimir will help us to increase the analysis of the system towards prediction, and how complex organic pollutants interact with the surface of the graphene oxide. And so we check the toxicity
15:48
is the preliminary result, but we can see that the atrazine is not so high, the toxicity to the embryos. However, when you mix with tanninic acid and graphene oxide, has a decrease of this
16:02
value demonstrate that are increasing, potentiating the effect of the atrazine, the graphene oxide and tanninic acid. However, when we mix the, looking for sub-lethal effect, when you have mix of tanninic acid and graphene oxide, plus atrazine, has a mitigation annihilation of the
16:25
effect. So, demonstrate that one plus one is not two, right? We need to understand the mixtures. And other interesting model is C. elegans. C. elegans is very, very important for biology
16:40
and ecotoxicology also. When you do research on toxicology, look at this, it's very simple, but when you're looking for the protocol that you use is a lot of variables to be considering, especially for modeling. So, what is our question here? The C. elegans to leave need to eat E. coli, right? So, the hypothesis was, E. coli in the gut will be degraded, releasing proteins. So,
17:10
when graphene oxide are exposed to C. elegans, it will not be bare, it will be corona coated inside. So, moving to understand these interactions. First, we perform a good characterization
17:24
of the corona associated with graphene oxide, okay? We are named E. coli corona on graphene oxide, hard corona. This is the proteins of the hard corona, the mixture of the E. coli on the surface of the graphene oxide. Then, we decide to mix this with silver, considering that
17:43
all materials in nanotechnology, the best are silver, and silver is that toxic for the environment, okay? And we demonstrate, when you have the GEO, GEO has a good capacity to interact with silver, but when you have a corona, increase a lot the adsorption of silver on the surface of
18:01
this material. So, consider mixture, graphene oxide, corona, and silver is a concern, right? Because now, we have more silver on the surface of corona. And the results confirm this. When you go to a good toxicity elegance, the silver has 12 microgram per liter, GEO with silver 5,
18:22
and after corona GEO 2.5. This is increasing in DLC50 value, demonstrate that the toxicity is increasing, depend of the graphene oxide and the corona, okay? And we did the silver quantification using ICP-MS recently, and it's very interesting. The corona increases the toxicity, but not improving
18:45
the silver inside the tissues. The mechanism is the corona dependence. Because when you compare the silver, there is much more ions inside the body, but the toxic is not high.
19:07
And to change everything, we are looking for a life cycle, okay? Degradation of the graphene oxide is very important, because it's a very metallostable system. So, looking for a real application, we decide to degradate graphene oxide using bleach, because bleach has been used everywhere,
19:26
right? Especially after COVID, right? And you're looking here is the graphene oxide in suspension of nine days is very stable in water, but when you incubate it, we have incubated in bleach, it's possible to see the degradation of the graphene over time. And looking for atomic
19:44
force microscopy, it's easy to see the decreasing of the size of the sheets, right? We have monitored the toxic C. elegans, and they are decreasing the toxicity due to the degradation of graphene dioxide. So, all major effects will be dependent of the degradation also, of course. We need to
20:04
study it. And now, it's a big challenge, and happy to stay here and know all people involved here of these papers, both papers, very important, right? And it's a big challenge, I know, right? But very important. And we start this looking for data life cycle, right?
20:27
If you don't look for this, we'll do mistakes, right? Because data is starting with data. Where data is coming? From labs, right? So, planning is very important. The planning of research is very
20:41
important. The next step is collecting the data. So, in my group, and focus on this, because I'm experimentalist, and I'll try to move together you to processing analysis of data, right? We have three different projects in progress, and two comes from lab to data, right? How can
21:02
you use non-informatics to do it, to help us to coordinate and activities in the lab? Well, the first project was in labware, an electronic notebook. We're using this to harmonize the methods for particle size, transmission, and scanning microscopy, other ISO 17025.
21:23
17025, right, is important on line for quality management system in the lab, right? We harmonized it in this project for this transmission and the scanning microscopy. Then, together in metro now,
21:41
and this is in progress, in my lab, you are harmonizing DLS and zebrafish model, okay? And together, Birmingham, we're using side note associated with nano commons project, trying to harmonize DAFNA method, right? So, I will talk about now just for DAFNA results, because more
22:03
close to mixture of toxicity using DAFNA, and a very simple experiment compare graphene mixed with cardamom and detoxes on DAFNA was 0.29. When you have a album in corona as a model, mixing the same concentration with cardamom, the toxicity enhanced for 0.61. So, demonstrating a
22:28
mitigation effect of the cardamom after corona formation was opposed to elegance and completely different on zebrafish. Demonstrating that there is no corona will always increase or
22:46
decrease in the toxicity. We don't have model, we don't have confirmations yet, but a complex system to continue study. And the next step after you have that good plan, right, is to elaborate
23:04
experimental workflow to make this data more organized and available to the community. And it's very good to now, we did, we separate this instance, right? I have time,
23:20
Corinne? Yeah. So, the first one is separate instance maps, separating different steps of the old experimental research that was done. Okay. After this, each information on this workflow
23:40
was translated to side note, electronic notebook. This is just an example. For instance, a graphene oxide synthase and characterization will have all information about the synthase and the characterization, but not here that we have used it. So, now we are translating to
24:00
monocomos platform. And more recently, doing research in Birmingham, we look for the literature and we decide to organize this information that Dafina, okay, and how nanomaterials are increasing, decreasing the toxicity after interaction with Polter. We did a systematic tutorial review
24:24
and a perspective on data-driven risk prediction. The first one is to find the problem after to compare the experimental research with theoretical methods available. Then it's possible to think about the matrix to machine learn predictions on this direction. So, training, validation and
24:43
to the prediction of new data. This is a big challenge and we are starting this. The first one, we try to prediction the pollutant interactions with EcoCorona because EcoCorona is a key in this process. And we are looking for different biomolecules on the surface of the
25:04
graphene oxide, how pollutants will be interacting, quantifying these pollutants and producing a data set for modeling to get Vladimir, Isilut, Antreya, Georgia, other comp-safe nano. So, we have students in this project going. So, a very new, also not published yet. The first one was Synchrotron.
25:27
Here, we're looking for Dafina, okay. It's a fluorescence micro, fluorescence synchrotron. So, it's very, very sensitive. The spatial resolution is 800 nanometers for analysis and impossible to check by fluorescence of the elements like calcium and copper. This is a control in head
25:45
is calcium, in green is copper, okay. After exposition to copper, it's possible to see now Dafina completely impregnated with copper. And when you mix with graphene oxide, look at them up for microscopy here. Graphene oxide inside the gut of the Dafina. And you can see the copper more
26:08
attracted on the surface of graphene oxide in the body. So, we have a big challenge because a lot of data to process, to have this imaging. It's not a direct analysis. So, the processing data is very complex and non-informatics will help us. And so, for conclusion now, we have graphene oxide. So,
26:27
from single to mature isotoxicity, this equation is very complex. So, we need to focus on this now. For conclusion, right, we have the surface of chemists demonstrating oxidative debris, silver decoration oxidation has a critical contribution to the colloidal
26:43
stability and toxicity of graphene oxide materials. This colloidal interactions in corona are essential elements to be considered in nature non-hypotoxicity. Here, graphene oxide increases the toxicity of agrochemicals and metals in vivo to in zebrafish and C. elegans. However, graphene oxide mitigates the occult toxicity of metals
27:04
to Dafina models. So, what was the effect? And furthermore, it's important to assess, considering long-term students and low-dose now, right, chronic and sub-lethal endpoints, and more biological, more models endpoints, including metal coils on this, right,
27:21
from cells to ecosystem that is the model of eco-toxicology. And it's very important to study other 2D materials than graphene oxide, because it's an increasing family of two-dimensional materials. And, in my view, it is a perspective from experimentalists, of the DAFTA lifecycle is start and plan in search and collect data.
27:44
So, both are in general neglected during experimental research, towards fair data. Implementation of quality systems and electronic laboratory books in experimental labs is a great challenge. Future, two minutes.
28:02
Time is different in university, industry, and the regulatory industry. Okay, protocols, OCL flows, databases are essential for the future of nano safety search, of course, including to build the link between single and mature ecosystem. And develop of non-informatics tools is very important,
28:22
but especially looking for beyond of the state of the art, like PRIM and synchro-conradiation. And the diversity of nano-biome interaction for calculations at multiple levels is impressive, and there are many gaps yet, but data size and non-informatics is a light at the end.
28:41
Linking, physics-based and data-deriving approach is a good strategy. Development of descriptors for materials and external conditions to traditional endpoints with machine learning algorithm for complex chemical mixtures. And it's crucial to enhance the collaboration between computational experimental research,
29:01
put multidisciplinary in practice, and non-informatics is a very good example of this. I'd like to invite the students of both of you to stay in Brazil next year in July. We are organizing a school focused on nano-agriculture and the environment.
29:22
This FAPESPI are sponsored this, and we have 50 places for international students. All expenses will be covered for this student by FAPESPI. So I will share the website with both of you, and please recommend your students to stay in Brazil with us next year.
29:41
Antreas and Iseult, we stay together. And I'd like to thank my group, and thank you all for the very nice days in Rubenstein. Thank you.