MUBANY - Mechanistically Understand the Bactericidal Action of Nanopillar Topography
Lead Research Organisation:
University of Bristol
Department Name: Oral and Dental Science
Abstract
Nature-inspired nanopillared surfaces demonstrate a means by which bacterial colonisation on surfaces can be controlled, and to do so is what this research aims to discover. It is the underpinning mechanism that is not yet fully resolved, impeding the development of novel antimicrobial surfaces and their implementation in real-world applications. MUBANY aims to investigate the kinetics of interactions between nanopillar surfaces and bacterial cell wall leading to cell death, focusing on deciphering time-resolved interactions at nanometer scales. For the first time, the metabolic activity of the surface attached bacteria will be assessed by Raman spectroscopy coupled with stable isotope probing in correlation with live/dead screening of bacteria. The project will combine a complementary suite of 2D/3D imaging modalities in a correlative workflow with a cryogenic sample preparation method, advancing the current state-of-the-art practices. This approach will establish ground truth data to elucidate the bactericidal mechanism of nanopillar topography. The most effective surface parameters to achieve optimal bactericidal activity will be determined using fabricated nanopillar polymer surfaces as test substrates. This fellowship will enable the applicant to conduct an original and robust research program, placing him at the forefront of an emerging field of bio-nanomaterials. An extensive training program will support the applicant to gain research skills in advanced nanofabrication, correlative microscopy, quantitative data analysis and improve his transferrable skills in communication, grant writing, and project management. The applicant will extend his professional network to the UK and EU. The socio-economic benefits of this research extend beyond the obvious scientific and health outcomes with planned public engagement activities raising citizen awareness of multidisciplinary research approaches to combat critical global challenges like antimicrobial resistance.
Organisations
- University of Bristol (Lead Research Organisation)
- Karlsruhe Institute of Technology (Collaboration)
- University of Nottingham (Collaboration)
- Helmholtz Association of German Research Centres (Collaboration)
- Institute of Molecular Genetics (Collaboration)
- Paul Scherrer Institute (Collaboration)
- Tescan (Collaboration)
| Description | Global Showcase Projects on Multimodal Imaging |
| Amount | $10,000 (USD) |
| Organisation | Chan Zuckerberg Initiative |
| Sector | Private |
| Country | United States |
| Start | 09/2023 |
| End | 12/2024 |
| Description | Confocal Raman analysis and Isotope Labelling |
| Organisation | Helmholtz Association of German Research Centres |
| Department | Helmholtz Centre for Environmental Research - UFZ |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | The researcher has brought his expertise on the subject and did the Raman and Helium ion measurements. |
| Collaborator Contribution | Institute provided access to Confocal Raman Spectroscope, Helium Ion Microscope and their expertise on Isotope labelling. |
| Impact | This is a research collaboration involved chemistry, isotope labelling, materials science, microbiology, spectroscopy and microscopy. Collaboration has successfully developed a methodology to characterise the bacterial live/dead status by measuring the metabolic activity of bacteria. Manuscript is currently being written. |
| Start Year | 2023 |
| Description | Correlative 3D-2D Microscopy To Understand Bacteria-Nanopillar Interactions |
| Organisation | Karlsruhe Institute of Technology |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | I have provided conceptualize the project and secured funding from NFFA. I have do bacteria cultures and prepare them for electron microscopy. Prepared FIB/SEM lift-outs for nanoCT analysis of several samples. |
| Collaborator Contribution | PSI: Fabricated nanopillar topography using EBL technique KIT: Prepare FIB/SEM liftouts and acquisition of nanoCT data of the samples. |
| Impact | Established a workflow to characterise bacteria using nanoCT. Results are to be published as a peer-review article. |
| Start Year | 2023 |
| Description | Correlative 3D-2D Microscopy To Understand Bacteria-Nanopillar Interactions |
| Organisation | Paul Scherrer Institute |
| Country | Switzerland |
| Sector | Academic/University |
| PI Contribution | I have provided conceptualize the project and secured funding from NFFA. I have do bacteria cultures and prepare them for electron microscopy. Prepared FIB/SEM lift-outs for nanoCT analysis of several samples. |
| Collaborator Contribution | PSI: Fabricated nanopillar topography using EBL technique KIT: Prepare FIB/SEM liftouts and acquisition of nanoCT data of the samples. |
| Impact | Established a workflow to characterise bacteria using nanoCT. Results are to be published as a peer-review article. |
| Start Year | 2023 |
| Description | Cryo NanoCT |
| Organisation | University of Nottingham |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Bring conceptualization, and develop workflow. |
| Collaborator Contribution | Nottingham provided cryo FIB/SEM facility and cryo fluorescence facility. Sample liftout is performed at nMRC. SiriusXt performed nanoCT measurements. |
| Impact | In progress. |
| Start Year | 2024 |
| Description | Develop cryo TEM workflow to characterise bacteria-material interactions |
| Organisation | Institute of Molecular Genetics |
| Country | Czech Republic |
| Sector | Public |
| PI Contribution | In this work, I have provided expertise on preparing bactericidal thin Ti substrates. |
| Collaborator Contribution | In this short venture, IMG provided access to high pressure freezing, cryo fluorescence microscope, cryo FIB/SEM, and cryo TEM. Tescan provided access to cryo Plasma FIB/SEM lamella lift-out. |
| Impact | Successfully developed a workflow to identify a single bacterium and lift-out for TEM tomography analysis. |
| Start Year | 2023 |
| Description | Develop cryo TEM workflow to characterise bacteria-material interactions |
| Organisation | Tescan |
| Country | Czech Republic |
| Sector | Private |
| PI Contribution | In this work, I have provided expertise on preparing bactericidal thin Ti substrates. |
| Collaborator Contribution | In this short venture, IMG provided access to high pressure freezing, cryo fluorescence microscope, cryo FIB/SEM, and cryo TEM. Tescan provided access to cryo Plasma FIB/SEM lamella lift-out. |
| Impact | Successfully developed a workflow to identify a single bacterium and lift-out for TEM tomography analysis. |
| Start Year | 2023 |
| Description | NanoCT Characterisation of Bacteria-nanopillar interaction |
| Organisation | Karlsruhe Institute of Technology |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | IN this collaboration researcher prepares the samples suitable for nanoCT characterisation. This requires the fabrication of surface, bacterial culture, fixation, dehydration. Finally this involves the FIB/SEM manipulation and lift-out preparation, as a step to getting samples ready for nanoCT. These samples are then shipped to Germany for characterisation. |
| Collaborator Contribution | KIT agreed to do nanoCT characterisation of my samples at 50 nm resolution. This equipment is not available at Bristol, hence collaboration is essential to complete the work proposed in the fellowship. Currently the workflow is established with the successful acquisition of first dataset. |
| Impact | This is a multidisciplinary collaboration which involves, materials science, microbiology, and microscopy. |
| Start Year | 2023 |
| Description | EuroBioimaging Interview on Czech Bioimaging on Accessing cryomicroscopy facilities to extend MUBANY research activities |
| Form Of Engagement Activity | Engagement focused website, blog or social media channel |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Media (as a channel to the public) |
| Results and Impact | The activity is about accessing research facilities and how would my research is benifitted acesscing Czech node of Euro bioimaging. |
| Year(s) Of Engagement Activity | 2024 |
| URL | https://youtu.be/36HNqmh1Sb8?si=-0K-vxjJ2aPA4CU8 |
| Description | FUTURES up late @ SS Great Britain |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | FUTURES at SS Great Britain is an event organized by the University of Bristol targeting general public and school children at premium location, SS Great Britain ship. Location is important as a preserved ship, and in premium tourist destination in Bristol. Therefore school children and local families are the targeted audience, and more than 100 people were visited our exhibit on dental heath, and microscopic investigation on bactericidal surfaces. The two kids activities; 1) Blast a biofilm, and 2) Blast a bacterium, which were developed during the fellowship as public reach out activities were was presented. "Blast a biofilm" which was targeting to teach importance of brushing teeth and maintaining oral hygiene. A 3D model of biofilm was prepared and displayed and basic oral hygiene related questions were asked from randomly selected participants to make it more interactive. Then, kids were given opportunity to create biofilm with hair-gel, and to embed microbes made out of clay. These were placed on water bucket representing teeth. Another set of bacteria were placed without gel indicating fresh bacteria attaching on the teeth. Then kids were given water guns to blow these away. As they blow-off bacteria, kids realised cleaning biofilms are difficult compared to early attached bacteria. So it was self-learning for kids that it is hard to clean and is best to brush their teeth twice a day to remove bacteria causing cavities. Many parents appreciated our approach as their kids are reluctant to brush teeth, but soon after participating our activity they are enthusiastic on brushing teeth. We have given away small sand timers to keep timing for brushing teeth for participants. Blast a bacterium", where resembling the MUBANY research's concepts at macro-scale, and show how researchers study nature to come up with innovative solutions to combat biofilms on implants. Here we have displayed implants, and also had an activity for kids. There was a bed of nails were prepared resembling a nanopillar surface of cicada wings, and water filled balloons were given to children asking them to drop on these bed of nails and on to flat surface. With micro-graphs we explained what we study and importance of research to solve medical challenges using innovative approaches. We had more than 200 people participated for this activity and most adults were fascinated about the research we do and kids enjoyed blasting bacteria on nanopillars. This was a satisfying two days where I could involve 4 more young PhD researchers from the Dental School for the activity and two staff members. |
| Year(s) Of Engagement Activity | 2024 |
| Description | Festival of Nature - Bristol City Centre as part of University Bristol Research Showcase |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Media (as a channel to the public) |
| Results and Impact | Festival of Nature is the UK's largest free celebration of the natural world held annually in Bristol and Bath. I have presented my research for general audience, along with two activities attracting kids. One activity named "Blast a biofilm" which was targeting to teach importance of brushing teeth and maintaining oral hygiene. Here kids were given opportunity to create biofilm with hair-gel, and to embed microbes made out of clay. These were placed on water bucket representing teeth. Another set of bacteria were placed without gel indicating fresh bacteria attaching on the teeth. Then kids were given water guns to blow these away. Kids learnt, once biofilms are formed inside the mouth, it is hard to clean and it is best brush their teeth twice a day to remove bacteria. Many parents appreciated our approach as their kids are reluctant to brush teeth, but soon after participating our activity they are enthusiastic on brushing teeth. More than 400 children and parents participated this activity over two days in a weekend. The second activity was "Blast a bacterium", where resembling the MUBANY research, and how researchers study nature to come up with innovative solutions to combat formation of biofilms on implants. Here we have displayed implants, and also had an activity for kids. There was a bed of nails were prepared resembling a nanopillar surface of cicada wings, and water filled balloons were given to children asking them to drop on these bed of nails and on to flat surface. With micro-graphs we explained what we study and importance of research to solve medical challenges using innovative approaches. We had more than 500 people participated for this activity and most adults were fascinated about the research we do and kids enjoyed blasting bacteria on nanopillars. This was a satisfying two days where I could involve 6 more young PhD researchers from the Dental School for the activity. |
| Year(s) Of Engagement Activity | 2024 |