@article{Machla2024,

title = {Tissue-Engineered Oral Epithelium for Dental Material Testing: Toward In Vitro Biomimetic Models},

author = {Foteini Machla and Paraskevi Kyriaki Monou and Chrysanthi Bekiari and Dimitrios Andreadis and Evangelia Kofidou and Emmanuel Panteris and Orestis L. Katsamenis and Maria Kokoti and Petros Koidis and Imad About and Dimitrios Fatouros and Athina Bakopoulou},

url = {https://www.liebertpub.com/doi/abs/10.1089/ten.TEC.2024.0154},

doi = {10.1089/ten.tec.2024.0154},

issn = {1937-3392},

year  = {2024},

date = {2024-10-01},

journal = {Tissue Engineering Part C: Methods},

publisher = {Mary Ann Liebert Inc},

abstract = {Tissue-engineered oral epithelium (ΤΕΟΕ) was developed after comparing various culture conditions, including submerged (SUB) and air–liquid interface (ALI) human cell expansion options. Barrier formation was evaluated via transepithelial electrical resistance (TEER) and calcein permeation via spectrofluorometry. TEOE was further assessed for long-term viability via live/dead staining and development of intercellular connections via transmission electron microscopy. Tissue architecture was evaluated via histochemistry and the expression of pancytokeratin (pCK) via immunohistochemistry. The effect of two commonly used dental resinous monomers on TEOE was evaluated for alterations in cell viability and barrier permeability. ALI/keratinocyte growth factor-supplemented (ALI-KGS) culture conditions led to the formation of an 8–20-layer thick, intercellularly connected epithelial barrier. TEER values of ALI-KGS-developed TEOE decreased compared with all other tested conditions, and the established epithelium intensively expressed pCK. Exposure to dental monomers affected the integrity and architecture of TEOE and induced cellular vacuolation, implicating hydropic degeneration. Despite structural modifications, the permeability of TEOE was not substantially affected after exposure to the monomers. In conclusion, the biological properties of the TEOE mimicking the physiological functional conditions and its value as biocompatibility assessment tool for dental materials were characterized.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Cartlidge2024,

title = {In silico simulations of diffusion tensors and tortuosity in cells grown on 3D-printed scaffolds for tissue engineering.},

author = {Topaz A. A. Cartlidge and Yan Wu and Thomas B. R. Robertson and Orestis L. Katsamenis and Giuseppe Pileio},

url = {https://pubs.rsc.org/en/Content/ArticleLanding/2024/RA/D4RA05362A},

doi = {10.1039/d4ra05362a},

issn = {2046-2069},

year  = {2024},

date = {2024-10-01},

journal = {RSC advances},

volume = {14},

issue = {44},

pages = {32398–32410},

abstract = {Tissue engineering is set to revolutionise regenerative medicine, drug discovery, and cancer biology. For this to succeed, improved 3D imaging methods that penetrate non-invasively into the developing tissue is fundamental to guide the design of new and improved 3D supports. In particular, it is very important to characterise the time- and space-heterogeneous pore network that continuously changes as the tissue grows, since delivery of nutrients and removal of waste is key to avoid the development of necrotic tissues. In this paper, we combine high-resolution microfocus Computed Tomography (μCT) imaging and in silico simulations to calculate the diffusion tensor of molecules diffusing in the actual pore structure of a tissue grown on 3D-printed plastic scaffolds. We use such tensors to derive information about the changing pore network and derive tortuosity, a key parameter to understand how pore interconnection changes with cell proliferation. Such information can be used to improve the design of 3D-printed supports as well as to validate and improve cell culture protocols.},

keywords = {},

pubstate = {epublish},

tppubtype = {article}

}

@article{Laundon2024a,

title = {Quantitative microCT imaging of a whole equine placenta and its blood vessel network},

author = {Davis Laundon and Ella Proudley and Philip J. Basford and Orestis L. Katsamenis and David S. Chatelet and Jane K. Cleal and Neil J. Gostling and Pascale Chavatte-Palmer and Rohan M. Lewis},

url = {https://www.sciencedirect.com/science/article/pii/S0143400424006088},

doi = {10.1016/j.placenta.2024.07.313},

issn = {0143-4004},

year  = {2024},

date = {2024-09-01},

journal = {Placenta},

volume = {154},

pages = {216–219},

publisher = {Elsevier BV},

abstract = {Placental structure is linked to function across morphological scales. In the placenta, changes to gross anatomy, such as surface area, volume, or blood vessel arrangement, are associated with suboptimal physiological outcomes. However, quantifying each of these metrics requires different laborious semi-quantitative methods. Here, we demonstrate how, with minimal sample preparation, whole-organ computed microtomography (microCT) can be used to calculate gross morphometry of the equine placenta and a range of additional metrics, including branching morphometry of placental vasculature, non-destructively from a single dataset. Our approach can be applied to quantify the gross structure of any large mammalian placenta.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Chachlioutaki2024,

title = {Tailored Sticky Solutions: 3D-Printed Miconazole Buccal Films for Pediatric Oral Candidiasis},

author = {Konstantina Chachlioutaki and Anastasia Iordanopoulou and Orestis L. Katsamenis and Anestis Tsitsos and Savvas Koltsakidis and Pinelopi Anastasiadou and Dimitrios Andreadis and Vangelis Economou and Christos Ritzoulis and Dimitrios Tzetzis and Nikolaos Bouropoulos and Iakovos Xenikakis and Dimitrios Fatouros},

url = {https://link.springer.com/article/10.1208/s12249-024-02908-5},

doi = {10.1208/s12249-024-02908-5},

issn = {1530-9932},

year  = {2024},

date = {2024-08-01},

journal = {AAPS PharmSciTech},

volume = {25},

number = {7},

publisher = {Springer Science and Business Media LLC},

abstract = {In this research, 3D-printed antifungal buccal films (BFs) were manufactured as a potential alternative to commercially available antifungal oral gels addressing key considerations such as ease of manufacturing, convenience of administration, enhanced drug efficacy and suitability of paediatric patients. The fabrication process involved the use of a semi-solid extrusion method to create BFs from zein-Poly-Vinyl-Pyrrolidone (zein-PVP) polymer blend, which served as a carrier for drug (miconazole) and taste enhancers. After manufacturing, it was determined that the disintegration time for all films was less than 10 min. However, these films are designed to adhere to buccal tissue, ensuring sustained drug release. Approximately 80% of the miconazole was released gradually over 2 h from the zein/PVP matrix of the 3D printed films. Moreover, a detailed physicochemical characterization including spectroscopic and thermal methods was conducted to assess solid state and thermal stability of film constituents. Mucoadhesive properties and mechanical evaluation were also studied, while permeability studies revealed the extent to which film-loaded miconazole permeates through buccal tissue compared to commercially available oral gel formulation. Histological evaluation of the treated tissues was followed. Furthermore, in vitro antifungal activity was assessed for the developed films and the commercial oral gel. Finally, films underwent a two-month drug stability test to ascertain the suitability of the BFs for clinical application. The results demonstrate that 3D-printed films are a promising alternative for local administration of miconazole in the oral cavity.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Laundon2024b,

title = {Correlative three-dimensional X-ray histology (3D-XRH) as a tool for quantifying mammalian placental structure},

author = {Davis Laundon and Thomas Lane and Orestis L. Katsamenis and Jeanette Norman and Lois Brewer and Shelley E. Harris and Philip J. Basford and Justine Shotton and Danielle Free and Georgina Constable-Dakeyne and Neil J. Gostling and Pascale Chavatte-Palmer and Rohan M. Lewis},

url = {https://www.sciencedirect.com/science/article/pii/S0143400424006076},

doi = {10.1016/j.placenta.2024.07.312},

issn = {0143-4004},

year  = {2024},

date = {2024-07-01},

journal = {Placenta},

publisher = {Elsevier BV},

abstract = {Mammalian placentas exhibit unparalleled structural diversity, despite sharing a common ancestor and principal functions. The bulk of structural studies in placental research has used two-dimensional (2D) histology sectioning, allowing significant advances in our understanding of mammalian placental structure. However, 2D histology sectioning may be limited if it does not provide accurate information of three-dimensional (3D) tissue architecture. Here, we propose correlative 3D X-ray histology (3D-XRH) as a tool with great potential for resolving mammalian placental structures. 3D-XRH involves scanning a formaldehyde-fixed, paraffin embedded (FFPE) tissue block with 3D X-ray microscopy (microCT) prior to histological sectioning to generate a 3D image volume of the embedded tissue piece. The subsequent 2D histology sections can then be correlated back into the microCT image volume to couple histology staining (or immunolabelling) with 3D tissue architecture. 3D-XRH is non-destructive and requires no additional sample preparation than standard FFPE histology sectioning, however the image volume provides 3D morphometric data and can be used to guide microtomy. As such, 3D-XRH introduces additional information to standard histological workflows with minimal effort or disruption. Using primary examples from porcine, bovine, equine, and canine placental samples, we demonstrate the application of 3D-XRH to quantifying placental structure as well as discussing the limitations and future directions of the methodology. The wealth of information derived from 2D histological sectioning in the biomedical, veterinary, and comparative reproductive sciences provides a rich foundation from which 3D-XRH can build on to advance the study of placental structure and function.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Bielecka2024,

title = {Three-Dimensional Culture Modelling Reveals Divergent Mycobacterium tuberculosis Virulence and Antimicrobial Treatment Response},

author = {Magdalena K. Bielecka and Liku B. Tezera and Elena Konstantinopoulou and Nicola Casali and Orestis L. Katsamenis and Ximena Gonzalo and Francis Drobniewski and Paul T. Elkington},

editor = {Sanket Kaushik},

doi = {10.1155/2024/6458900},

issn = {1462-5814},

year  = {2024},

date = {2024-05-01},

journal = {Cellular Microbiology},

volume = {2024},

pages = {1–17},

publisher = {Hindawi Limited},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Laundon2024,

title = {Convergently evolved placental villi show multiscale structural adaptations to differential placental invasiveness.},

author = {Davis Laundon and Bram G. Sengers and James Thompson and Shelley E. Harris and Olivia Beasley and Philip J. Basford and Orestis L. Katsamenis and Patricia Goggin and Emilie Derisoud and Diana Fanelli and Carlotta Bocci and Francesco Camillo and Justine Shotton and Georgina Constable-Dakeyne and Neil J. Gostling and Pascale Chavatte-Palmer and Rohan M. Lewis},

doi = {10.1098/rsbl.2024.0016},

issn = {1744-957X},

year  = {2024},

date = {2024-03-01},

journal = {Biology letters},

volume = {20},

issue = {3},

pages = {20240016},

abstract = {Despite having a single evolutionary origin and conserved function, the mammalian placenta exhibits radical structural diversity. The evolutionary drivers and functional consequences of placental structural diversity are poorly understood. Humans and equids both display treelike placental villi, however these villi evolved independently and exhibit starkly different levels of invasiveness into maternal tissue (i.e. the number of maternal tissue layers between placental tissue and maternal blood). The villi in these species therefore serve as a compelling evolutionary case study to explore whether placentas have developed structural adaptations to respond to the challenge of reduced nutrient availability in less invasive placentas. Here, we use three-dimensional X-ray microfocus computed tomography and electron microscopy to quantitatively evaluate key structures involved in exchange in human and equid placental villi. We find that equid villi have a higher surface area to volume ratio and deeper trophoblastic vessel indentation than human villi. Using illustrative computational models, we propose that these structural adaptations have evolved in equids to boost nutrient transfer to compensate for reduced invasiveness into maternal tissue. We discuss these findings in relation to the 'maternal-fetal conflict hypothesis' of placental evolution.},

keywords = {},

pubstate = {ppublish},

tppubtype = {article}

}

@article{Laundon2023,

title = {Placental evolution from a three-dimensional and multiscale structural perspective},

author = {Davis Laundon and Neil J Gostling and Bram G Sengers and Pascale Chavatte-Palmer and Rohan M Lewis},

editor = {Tristan Stayton and Tracey Chapman},

url = {https://academic.oup.com/evolut/article/78/1/13/7425018},

doi = {10.1093/evolut/qpad209},

issn = {1558-5646},

year  = {2023},

date = {2023-11-01},

journal = {Evolution},

volume = {78},

number = {1},

pages = {13–25},

publisher = {Oxford University Press (OUP)},

abstract = {The placenta mediates physiological exchange between the mother and the fetus. In placental mammals, all placentas are descended from a single common ancestor and functions are conserved across species; however, the placenta exhibits radical structural diversity. The selective pressures behind this structural diversity are poorly understood. Traditionally, placental structures have largely been investigated by grouping them into qualitative categories. Assessing the placenta on this basis could be problematic when inferring the relative “efficiency” of a placental configuration to transfer nutrients from mother to fetus. We argue that only by considering placentas as three-dimensional (3D) biological structures, integrated across scales, can the evolutionary questions behind their enormous structural diversity be quantitatively determined. We review the current state of placental evolution from a structural perspective, detail where 3D imaging and computational modeling have been used to gain insight into placental function, and outline an experimental roadmap to answer evolutionary questions from a multiscale 3D structural perspective. Our approach aims to shed light on placental evolution, and can be transferred to evolutionary investigations in any organ system.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Keeling2023,

title = {68Ga-bisphosphonates for the imaging of extraosseous calcification by positron emission tomography},

author = {George P. Keeling and Friedrich Baark and Orestis L. Katsamenis and Jing Xue and Philip J. Blower and Sergio Bertazzo and Rafael T. M. Rosales},

url = {https://www.nature.com/articles/s41598-023-41149-7},

doi = {10.1038/s41598-023-41149-7},

year  = {2023},

date = {2023-09-01},

journal = {Scientific Reports},

volume = {13},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {Radiolabelled bisphosphonates (BPs) and [18F]NaF (18F-fluoride) are the two types of radiotracers available to image calcium mineral (e.g. bone), yet only [18F]NaF has been widely explored for the non-invasive molecular imaging of extraosseous calcification (EC) using positron emission tomography (PET) imaging. These two radiotracers bind calcium mineral deposits via different mechanisms, with BPs chelating to calcium ions and thus being non-selective, and [18F]NaF being selective for hydroxyapatite (HAp) which is the main component of bone mineral. Considering that the composition of EC has been reported to include a diverse range of non-HAp calcium minerals, we hypothesised that BPs may be more sensitive for imaging EC due to their ability to bind to both HAp and non-HAp deposits. We report a comparison between the 68Ga-labelled BP tracer [68Ga]Ga-THP-Pam and [18F]NaF for PET imaging in a rat model of EC that develops macro- and microcalcifications in several organs. Macrocalcifications were identified using preclinical computed tomography (CT) and microcalcifications were identified using µCT-based 3D X-ray histology (XRH) on isolated organs ex vivo. The morphological and mineral analysis of individual calcified deposits was performed using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX). PET imaging and ex vivo analysis results demonstrated that while both radiotracers behave similarly for bone imaging, the BP-based radiotracer [68Ga]Ga-THP-Pam was able to detect EC more sensitively in several organs in which the mineral composition departs from that of HAp. Our results strongly suggest that BP-based PET radiotracers such as [68Ga]Ga-THP-Pam may have a particular advantage for the sensitive imaging and early detection of EC by being able to detect a wider array of relevant calcium minerals in vivo than [18F]NaF, and should be evaluated clinically for this purpose.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Monou2023,

title = {Fabrication of 3D Printed Hollow Microneedles by Digital Light Processing for the Buccal Delivery of Actives},

author = {Paraskevi Kyriaki Monou and Eleftherios G. Andriotis and Konstantinos Tsongas and Emmanouil K. Tzimtzimis and Orestis L. Katsamenis and Dimitrios Tzetzis and Pinelopi Anastasiadou and Christos Ritzoulis and Ioannis S. Vizirianakis and Dimitrios Andreadis and Dimitrios G. Fatouros},

url = {https://pubs.acs.org/doi/full/10.1021/acsbiomaterials.3c00116},

doi = {10.1021/acsbiomaterials.3c00116},

year  = {2023},

date = {2023-08-01},

journal = {ACS Biomaterials Science & Engineering},

volume = {9},

number = {8},

pages = {5072–5083},

publisher = {American Chemical Society (ACS)},

abstract = {In the present study, two different microneedle devices were produced using digital light processing (DLP). These devices hold promise as drug delivery systems to the buccal tissue as they increase the permeability of actives with molecular weights between 600 and 4000 Da. The attached reservoirs were designed and printed along with the arrays as a whole device. Light microscopy was used to quality control the printability of the designs, confirming that the actual dimensions are in agreement with the digital design. Non-destructive volume imaging by means of microfocus computed tomography was employed for dimensional and defect characterization of the DLP-printed devices, demonstrating the actual volumes of the reservoirs and the malformations that occurred during printing. The penetration test and finite element analysis showed that the maximum stress experienced by the needles during the insertion process (10 N) was below their ultimate compressive strength (240–310 N). Permeation studies showed the increased permeability of three model drugs when delivered with the MN devices. Size-exclusion chromatography validated the stability of all the actives throughout the permeability tests. The safety of these printed devices for buccal administration was confirmed by histological evaluation and cell viability studies using the TR146 cell line, which indicated no toxic effects.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Katsamenis2023b,

title = {A high-throughput 3D X-ray histology facility for biomedical research and preclinical applications},

author = {Orestis L. Katsamenis and Philip J. Basford and Stephanie K. Robinson and Richard P. Boardman and Elena Konstantinopoulou and Peter M. Lackie and Anton Page and J. Arjuna Ratnayaka and Patricia M. Goggin and Gareth J. Thomas and Simon J. Cox and Ian Sinclair and Philipp Schneider},

url = {https://doi.org/10.12688/wellcomeopenres.19666.1},

doi = {10.12688/wellcomeopenres.19666.1},

year  = {2023},

date = {2023-08-01},

journal = {Wellcome Open Research},

volume = {8},

pages = {366},

publisher = {F1000 Research Ltd},

abstract = {Background: The University of Southampton, in collaboration with the University Hospital Southampton (UHS) NHS Foundation Trust and industrial partners, has been at the forefront of developing three-dimensional (3D) imaging workflows using X-ray microfocus computed tomography (μCT) -based technology. This article presents the outcomes of these endeavours and highlights the distinctive characteristics of a μCT facility specifically tailored for 3D X-ray Histology, with primary focus on applications in biomedical research and preclinical and clinical studies. 

Methods: The UHS houses a unique 3D X-ray Histology (XRH) facility, offering a range of services to national and international clients. The facility employs specialised μCT equipment designed specifically for histology applications, allowing whole-block XRH imaging of formalin-fixed and paraffin-embedded tissue specimens. It also enables correlative imaging by combining μCT imaging with other microscopy techniques, such as immunohistochemistry (IHC) and serial block-face scanning electron microscopy, as well as data visualization, image quantification, and bespoke analysis. 

Results: Over the past seven years, the XRH facility has successfully completed over 120 projects in collaboration with researchers from 60 affiliations, resulting in numerous published manuscripts and conference proceedings. The facility has streamlined the μCT imaging process, improving productivity, and enabling efficient acquisition of 3D datasets. 

Conclusions: The 3D X-ray Histology (XRH) facility at UHS is a pioneering platform in the field of histology and biomedical imaging. To the best of our knowledge, it stands out as the world's first dedicated XRH facility, encompassing every aspect of the imaging process, from user support to data generation, analysis, training, archiving, and metadata generation. This article serves as a comprehensive guide for establishing similar XRH facilities, covering key aspects of facility setup and operation. Researchers and institutions interested in developing state-of-the-art histology and imaging facilities can utilize this resource to explore new frontiers in their research and discoveries.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}