Nonuniformity in ligaments is a structural strategy for optimizing functionality.

Ligaments serve as compliant connectors between hard tissues. In that role, they function under various load regimes and directions. The 3D structure of ligaments is considered to form as a uniform entity that changes due to function. The periodontal ligament (PDL) connects the tooth to the bone and sustains different types of loads in various directions. Using the PDL as a model, employing a fabricated motorized setup in a microCT, we demonstrate that the fibrous network structure within the PDL is not uniform, even before the tooth becomes functional. Utilizing morphological automated segmentation methods, directionality analysis, as well as second harmonic generation imaging, we find high correlation between blood vessel distribution and fiber density. We also show a structural feature in a form of a dense collar around the neck of the tooth as well as a preferred direction of the fibrous network. Finally, we show that the PDL develops as a nonuniform structure, with an architecture designed to sustain specific types of load in designated areas. Based on these findings, we propose that ligaments in general should be regarded as nonuniform entities, structured already at developmental stages for optimal functioning under variable load regimes.

Tooth periodontal ligament: Direct 3D microCT visualization of the collagen network and how the network changes when the tooth is loaded.

The periodontal ligament (PDL), a soft tissue connecting the tooth and the bone, is essential for tooth movement, bone remodeling and force dissipation. A collagenous network that connects the tooth root surface to the alveolar jaw bone is one of the major components of the PDL. The organization of the collagenous component and how it changes under load is still poorly understood. Here using a state-of-the-art custom-made loading apparatus and a humidified environment inside a microCT, we visualize the PDL collagenous network of a fresh rat molar in 3D at 1 µm voxel size without any fixation or contrasting agents. We demonstrate that the PDL collagen network is organized in sheets. The spaces between sheets vary thus creating dense and sparse networks. Upon vertical loading, the sheets in both networks are stretched into well aligned arrays. The sparse network is located mainly in areas which undergo compressive loading as the tooth moves towards the bone, whereas the dense network functions mostly in tension as the tooth moves further from the bone. This new visualization method can be used to study other non-mineralized or partially mineralized tissues, and in particular those that are subjected to mechanical loads. The method will also be valuable for characterizing diseased tissues, as well as better understanding the phenotypic expressions of genetic mutants.

Tooth movements are guided by specific contact areas between the tooth root and the jaw bone: A dynamic 3D microCT study of the rat molar.

Teeth sustain high loads over a lifetime and yet intact tooth failure is rare. The different structures of the tooth, jaw bone and the intervening soft periodontal ligament enable the tooth to endure repeated loading during mastication. Although mechanical and functional properties of the different components are thoroughly investigated, the manner in which the whole tooth functions under load is still enigmatic. A custom-made loading system inside a microCT scanner was used to directly visualize the root movements in relation to the jaw bone as the rat molar tooth was loaded. At low loads no contact was observed between the root surface and the bone, whereas at higher loads three specific contact areas between the root surface and the jaw bone were observed. These contact areas restrict tooth movement in the buccal-lingual direction, but enable the tooth to rock in a "seesaw" like manner in the distal-mesial direction. The contact areas appear to play a role in determining tooth motion and in turn define the manner in which the whole tooth moves when loaded. These observations are important for understanding basic structure-function relations of the tooth-PDL-bone system, and have direct implications for better understanding pathological and therapeutic processes in orthodontics, periodontics and jaw bone regeneration.

3D Imaging of PDL Collagen Fibers during Orthodontic Tooth Movement in Mandibular Murine Model.

Orthodontic tooth movement is a complex biological process of altered soft and hard tissue remodeling as a result of external forces. In order to understand these complex remodeling processes, it is critical to study the tooth and periodontal tissues within their 3D context and therefore minimize any sectioning and tissue artefacts. Mouse models are often utilized in developmental and structural biology, as well as in biomechanics due to their small size, high metabolic rate, genetics and ease of handling. In principle this also makes them excellent models for dental related studies. However, a major impediment is their small tooth size, the molars in particular. This paper is aimed at providing a step by step protocol for generating orthodontic tooth movement and two methods for 3D imaging of the periodontal ligament fibrous component of a mouse mandibular molar. The first method presented is based on a micro-CT setup enabling phase enhancement imaging of fresh collagen tissues. The second method is a bone clearing method using ethyl cinnamate that enables imaging through the bone without sectioning and preserves endogenous fluorescence. Combining this clearing method with reporter mice like Flk1-Cre;TdTomato provided a first of its kind opportunity to image the 3D vasculature in the PDL and alveolar bone.

Functionalized Mouth-Conformable Interfaces for pH Evaluation of the Oral Cavity.

Oral health monitoring is highly desired, especially for in home use, however, current methods are not sensitive enough and technically convoluted for this purpose. This paper presents incorporation of bioactive materials and colorimetric chemical sensors into routinely used oral appliances transforming them into bioresponsive, conformable interfaces. Specifically, endodontic paper points and dental floss can be functionalized to locally sense and monitor pH variations within the oral cavity via color changes. Moreover, edible colorimetric indicators are developed and used to make sensing, edible devices in the form factor of candies that can dynamically and visually respond to acidity changes in saliva. These interfaces would enable early detection of caries (e.g., using dental floss and paper points) providing low-cost point of care devices that respond in real-time by detecting pH variations in biological fluids thus bringing monitoring to home settings .

In situ AFM-based nanoscale rheology reveals regional non-uniformity in viscoporoelastic mechanical behavior of the murine periodontal ligament.

The periodontal ligament (PDL) is a critical player in the maintenance of tooth health, acting as the primary stabilizer of tooth position. Recent studies have identified two unique regions within the PDL, the 'dense collar' region and the 'furcation' region, which exhibit distinct structural and compositional differences. However, specific functional differences between these regions have yet to be investigated. We adapted an AFM-based nanoscale rheology method to regionally assess mechanical properties and poroelasticity in the mouse PDL while minimizing the disruption of the 3-dimensional native boundary conditions, and then explored tissue mechanical function in four different regions within the dense collar as well as in the furcation region. We found significant differences between the collar and furcation regions, with the collar acting as a stabilizing ligamentous structure and the furcation acting as both a compressive cushion for vertical forces and a conduit for nutrient transport. While this finding supports our hypothesis, based on previous studies investigating structural and compositional differences, we also found surprising inhomogeneity within the collar region itself. This inhomogeneity supports previous findings of a tilting movement in the buccal direction of mandibular molar teeth and the structural adaptation to prevent lingual movement. Future work will aim to understand how different regions of the PDL change functionally during biological or mechanical perturbations, such as orthodontic tooth movement, development, or aging, with the ultimate goal of better understanding the mechanobiology of the PDL function in health and disease.

Tooth-PDL-bone complex: response to compressive loads encountered during mastication - a review.

The components of the tooth-periodontal ligament (PDL)-alveolar bone complex act in a synergistic manner to dissipate the loads incurred during mastication. The complex incorporates a diverse array of structural features for this purpose. These include the non-mineralized and hence soft PDL that absorbs much of the initial loads. The internal structure of the tooth also includes soft interphases that essentially surround the dentine core. These interphases, although stiffer than the PDL, still are more compliant than the dentine core, and are thus key components that allow the tooth itself to deform and hence help dissipate the compressive loads. There is also direct evidence that even under moderate compressive loads, when the tooth moves in the alveolar bone socket, this movement is guided by specific locations where the tooth comes into contact with the bone surface. The combination of all these responses to load is that each tooth type appears to move and deform in a specific manner when loaded. Much, however, still remains to be learned about these three-dimensional responses to load and the factors that control them. Such an understanding will have major implications for dentistry, that include a better understanding of phenomena such as abfraction, the manner in which tooth implants function even in the absence of a PDL-like tissue and the implications to bone remodelling of the movements imposed during orthodontic interventions.