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Salwa Ibrahim,1,* Adam P Strange,2,* Sebastian Aguayo,2,3 Albatool Shinawi,1 Nabilah Harith,1 Nurjehan Mohamed-Ibrahim,1 Samera Siddiqui,2 Susan Parekh,1 Laurent Bozec4 1Department of Paediatric Dentistry, UCL Eastman Dental Institute, University College London, London, UK; 2Department of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, London, UK; 3School of Dentistry, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile; 4Faculty of Dentistry, University of Toronto, Toronto, Canada*These authors contributed equally to this workCorrespondence: Laurent BozecFaculty of Dentistry, University of Toronto, 124 Edward Street, Toronto ON M5G 1G6, CanadaEmail L.bozec@utoronto.caIntroduction: Dentinogenesis imperfecta type 1 (OIDI) is considered a relatively rare genetic disorder (1:5000 to 1:45,000) associated with osteogenesis imperfecta. OIDI impacts the formation of collagen fibrils in dentin, leading to morphological and structural changes that affect the strength and appearance of teeth. However, there is still a lack of understanding regarding the nanoscale characterization of the disease, in terms of collagen ultrastructure and mechanical properties. Therefore, this research presents a qualitative and quantitative report into the phenotype and characterization of OIDI in dentin, by using a combination of imaging, nanomechanical approaches.Methods: For this study, 8 primary molars from OIDI patients and 8 primary control molars were collected, embedded in acrylic resin and cut into longitudinal sections. Sections were then demineralized in 37% phosphoric acid using a protocol developed in-house. Initial experiments demonstrated the effectiveness of the demineralization protocol, as the ATR-FTIR spectral fingerprints showed an increase in the amide bands together with a decrease in phosphate content. Structural and mechanical analyses were performed directly on both the mineralized and demineralized samples using a combination of scanning electron microscopy, atomic force microscopy, and Wallace indentation.Results: Mesoscale imaging showed alterations in dentinal tubule morphology in OIDI patients, with a reduced number of tubules and a decreased tubule diameter compared to healthy controls. Nanoscale collagen ultrastructure presented a similar D-banding periodicity between OIDI and controls. Reduced collagen fibrils diameter was also recorded for the OIDI group. The hardness of the (mineralized) control dentin was found to be significantly higher (p

Tarek Ahmed,1 Anthony Nash,2 Kristina EN Clark,3 Marion Ghibaudo,4 Nora H de Leeuw,2 Anne Potter,4 Richard Stratton,3 Helen L Birch,5 Ramona Enea Casse,4 Laurent Bozec1 1Division of Biomaterials and Tissue Engineering, Eastman Dental Institute, University College London, 2Department of Chemistry, University College London, 3Centre for Rheumatology and Connective Tissue Diseases, Division of Medicine, University College London, London, UK; 4L’Oréal Research and Innovation, Aulnay-sous-Bois, France; 5Division of Surgery and Interventional Science, UCL Institute of Orthopaedics and Musculoskeletal Science, University College London, London, UK Abstract: The extracellular matrix of the dermis is a complex, dynamic system with the various dermal components undergoing individual physiologic changes as we age. Age-related changes in the physical properties of collagen were investigated in particular by measuring the effect of aging, most likely due to the accumulation of advanced glycation end product (AGE) cross-links, on the nanomechanical properties of the collagen fibril using atomic force microscope nano-indentation. An age-related decrease in the Young’s modulus of the transverse fibril was observed (from 8.11 to 4.19 GPa in young to old volunteers, respectively, P

Adam P Strange,1 Sebastian Aguayo,1 Tarek Ahmed,1 Nicola Mordan,1 Richard Stratton,2 Stephen R Porter,3 Susan Parekh,4 Laurent Bozec1 1Department of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, 2Centre for Rheumatology and Connective Tissue Diseases, Royal Free Hospital, UCL Medical School, 3UCL Eastman Dental Institute, 4Department of Pediatrics, UCL Eastman Dental Institute, London, UK Abstract: Scleroderma (or systemic sclerosis, SSc) is a disease caused by excess crosslinking of collagen. The skin stiffens and becomes painful, while internally, organ function can be compromised by the less elastic collagen. Diagnosis of SSc is often only possible in advanced cases by which treatment time is limited. A more detailed analysis of SSc may provide better future treatment options and information of disease progression. Recently, the histological stain picrosirius red showing collagen register has been combined with atomic force microscopy (AFM) to study SSc. Skin from healthy individuals and SSc patients was biopsied, stained and studied using AFM. By investigating the crosslinking of collagen at a smaller hierarchical stage, the effects of SSc were more pronounced. Changes in morphology and Young’s elastic modulus were observed and quantified; giving rise to a novel technique, we have termed “quantitative nanohistology”. An increase in nanoscale stiffness in the collagen for SSc compared with healthy individuals was seen by a significant increase in the Young’s modulus profile for the collagen. These markers of stiffer collagen in SSc are similar to the symptoms experienced by patients, giving additional hope that in the future, nanohistology using AFM can be readily applied as a clinical tool, providing detailed information of the state of collagen. Keywords: rheumatology, adjunct diagnosis, picrosirius red, collagen, nanohistology

Ensanya Ali Abou Neel,1–3 Anas Aljabo,3 Adam Strange,3 Salwa Ibrahim,3 Melanie Coathup,4 Anne M Young,3 Laurent Bozec,3 Vivek Mudera4 1Division of Biomaterials, Operative Dentistry Department, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia; 2Biomaterials Department, Faculty of Dentistry, Tanta University, Tanta, Egypt; 3Department of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, London, UK; 4UCL Institute of Orthopaedics and Musculoskeletal Sciences, Royal National Orthopaedic Hospital, Stanmore, London, UK Abstract: Biomineralization is a dynamic, complex, lifelong process by which living organisms control precipitations of inorganic nanocrystals within organic matrices to form unique hybrid biological tissues, for example, enamel, dentin, cementum, and bone. Understanding the process of mineral deposition is important for the development of treatments for mineralization-related diseases and also for the innovation and development of scaffolds. This review provides a thorough overview of the up-to-date information on the theories describing the possible mechanisms and the factors implicated as agonists and antagonists of mineralization. Then, the role of calcium and phosphate ions in the maintenance of teeth and bone health is described. Throughout the life, teeth and bone are at risk of demineralization, with particular emphasis on teeth, due to their anatomical arrangement and location. Teeth are exposed to food, drink, and the microbiota of the mouth; therefore, they have developed a high resistance to localized demineralization that is unmatched by bone. The mechanisms by which demineralization–remineralization process occurs in both teeth and bone and the new therapies/technologies that reverse demineralization or boost remineralization are also scrupulously discussed. Technologies discussed include composites with nano- and micron-sized inorganic minerals that can mimic mechanical properties of the tooth and bone in addition to promoting more natural repair of surrounding tissues. Turning these new technologies to products and practices would improve health care worldwide. Keywords: demineralization, remineralization, teeth, bone and calcium phosphates

Sebastian Aguayo,1 Nikolaos Donos,2 Dave Spratt,3 Laurent Bozec11Department of Biomaterials and Tissue Engineering, 2Periodontology Unit, 3Division of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, UKAbstract: As titanium (Ti) continues to be utilized in great extent for the fabrication of artificial implants, it is important to understand the crucial bacterium–Ti interaction occurring during the initial phases of biofilm formation. By employing a single-cell force spectroscopy technique, the nanoadhesive interactions between the early-colonizing Streptococcus sanguinis and a clinically analogous smooth Ti substrate were explored. Mean adhesion forces between S. sanguinis and Ti were found to be 0.32±0.00, 1.07±0.06, and 4.85±0.56 nN for 0, 1, and 60 seconds contact times, respectively; while adhesion work values were reported at 19.28±2.38, 104.60±7.02, and 1,317.26±197.69 aJ for 0, 1, and 60 seconds, respectively. At 60 seconds surface delays, minor-rupture events were modeled with the worm-like chain model yielding an average contour length of 668±12 nm. The mean force for S. sanguinis minor-detachment events was 1.84±0.64 nN, and Poisson analysis decoupled this value into a short-range force component of -1.60±0.34 nN and a long-range force component of -0.55±0.47 nN. Furthermore, a solution of 2 mg/mL chlorhexidine was found to increase adhesion between the bacterial probe and substrate. Overall, single-cell force spectroscopy of living S. sanguinis cells proved to be a reliable way to characterize early-bacterial adhesion onto machined Ti implant surfaces at the nanoscale.Keywords: atomic force microscopy, biophysics, bacterial adhesion, dental implants, titanium

Ensanya Ali Abou Neel,1–3 Laurent Bozec,3 Roman A Perez,4,5 Hae-Won Kim,4–6 Jonathan C Knowles3,5 1Division of Biomaterials, Operative Dentistry Department, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia; 2Biomaterials Department, Faculty of Dentistry, Tanta University, Tanta, Egypt; 3UCL Eastman Dental Institute, Biomaterials and Tissue Engineering, London, UK; 4Institute of Tissue Regenerative Engineering (ITREN), 5Department of Nanobiomedical Science and BK21 Plus NBM Global Research Center for Regenerative Medicine, 6Department of Biomaterials Science, College of Dentistry, Dankook University, Cheonan, Republic of Korea Abstract: Nanotechnology has rapidly expanded into all areas of science; it offers significant alternative ways to solve scientific and medical questions and problems. In dentistry, nanotechnology has been exploited in the development of restorative materials with some significant success. This review discusses nanointerfaces that could compromise the longevity of dental restorations, and how nanotechnolgy has been employed to modify them for providing long-term successful restorations. It also focuses on some challenging areas in dentistry, eg, oral biofilm and cancers, and how nanotechnology overcomes these challenges. The recent advances in nanodentistry and innovations in oral health-related diagnostic, preventive, and therapeutic methods required to maintain and obtain perfect oral health, have been discussed. The recent advances in nanotechnology could hold promise in bringing a paradigm shift in dental field. Although there are numerous complex therapies being developed to treat many diseases, their clinical use requires careful consideration of the expense of synthesis and implementation. Keywords: nanotechnology, nanointerfaces, biofilm-related oral diseases, tissue engineering, drug delivery, toxicity