Biomaterial and Biofilm Interactions with the Pulp-Dentin Complex-on-a-Chip.

Calcium silicate cements (CSCs) are the choice materials for vital pulp therapy because of their bioactive properties, promotion of pulp repair, and dentin bridge formation. Despite the significant progress made in understanding CSCs' mechanisms of action, the key events that characterize the early interplay between CSC-dentin-pulp are still poorly understood. To address this gap, a microfluidic device, the "tooth-on-a-chip," which was developed to emulate the biomaterial-dentin-pulp interface, was used to test 1) the effect of CSCs (ProRoot, Biodentine, and TheraCal) on the viability and proliferation of human dental pulp stem cells, 2) variations of pH, and 3) release within the pulp chamber of transforming growth factor-ß (TGFß) as a surrogate of the bioactive dentin matrix molecules. ProRoot significantly increased the extraction of TGFß (P < 0.05) within 24 to 72 h and, along with Biodentine, induced higher cell proliferation (P > 0.05), while TheraCal decreased cell viability and provoked atypical changes in cell morphology. No correlation between TGFß levels and pH was observed. Further, we established a biofilm of Streptococcus mutans on-chip to model the biomaterial-biofilm-dentin interface and conducted a live and dead assay to test the antimicrobial capability of ProRoot in real time. In conclusion, the device allows for direct characterization of the interaction of bioactive dental materials with the dentin-pulp complex on a model of restored tooth while enabling assessment of antibiofilm properties at the interface in real time that was previously unattainable.

Immunolocalization and distribution of proteoglycans in carious dentine.

BACKGROUND: Collagen type I, proteoglycans (PG) and non-collagenous proteins represent important building blocks of the dentine matrix. While different PGs have been identified in dentine, changes in the distribution of these macromolecules with the progression of caries have been poorly characterized. The aim of this study was to compare the immunolocalization of three small collagen-binding PGs (biglycan, fibromodulin and lumican) as well as collagen (types I and VI) in healthy versus carious dentine. METHODS: Longitudinal demineralized sections of extracted teeth were stained with antibodies recognizing specific PG core proteins and collagens, as well as glycosaminoglycans (GAGs) with toluidine blue. RESULTS: In healthy dentine, PGs appeared to be more abundant near the tubule walls and directly under the cusps. Conversely, in carious dentine, specific locations appeared to be more prone to PG degradation than others. These degradation patterns were well correlated with the progression of caries into the tissue, and also appeared to trigger interesting morphological changes in the tissue structure, such as the deformation of dentine tubules near highly infected areas and the lower concentration of PG in tertiary dentine. CONCLUSIONS: This study presents new insights into the involvement of PGs in the progression of caries.

Three-Dimensional Bioprinting for Regenerative Dentistry and Craniofacial Tissue Engineering.

Craniofacial tissues are organized with complex 3-dimensional (3D) architectures. Mimicking such 3D complexity and the multicellular interactions naturally occurring in craniofacial structures represents one of the greatest challenges in regenerative dentistry. Three-dimensional bioprinting of tissues and biological structures has been proposed as a promising alternative to address some of these key challenges. It enables precise manufacture of various biomaterials with complex 3D architectures, while being compatible with multiple cell sources and being customizable to patient-specific needs. This review describes different 3D bioprinting methods and summarizes how different classes of biomaterials (polymer hydrogels, ceramics, composites, and cell aggregates) may be used for 3D biomanufacturing of scaffolds, as well as craniofacial tissue analogs. While the fabrication of scaffolds upon which cells attach, migrate, and proliferate is already in use, printing of all the components that form a tissue (living cells and matrix materials together) to produce tissue constructs is still in its early stages. In summary, this review seeks to highlight some of the key advantages of 3D bioprinting technology for the regeneration of craniofacial structures. Additionally, it stimulates progress on the development of strategies that will promote the translation of craniofacial tissue engineering from the laboratory bench to the chair side.

Biomechanical perspective on the remineralization of dentin.

The objective of this article is to critically evaluate the methods that are used to assess outcomes of remineralization of dentin. Currently, the most used assessment methods fall either into quantitative analysis of the mineral content of the remineralized structures or dry measurements of their mechanical properties. Properties obtained from the dehydrated organic dentin matrix may not reflect the true mechanical behavior of the remineralized tissue under physiological and hydrated conditions. Here we seek to clarify the biomechanical aspects of remineralization of dentin, pointing out the effects of hydration and dehydration on the mechanical properties of treated tissues. We also emphasize that a more appropriate endpoint to evaluate the effectiveness of remineralization in dentin should be associated with the recovery of the mechanical properties of the hydrated tissue, which is presumed to correlate well with its overall functionality.

Effects of benzodiazepine and pilocarpine on rat parotid glands: histomorphometric and sialometric study.

Benzodiazepines are among the most frequently prescribed drugs and are often related with dry mouth. Pilocarpine is a cholinergic agonist that increases salivary flow rate and has been used to treat xerostomia. This study aimed to measure salivary flow rate of rats under chronic treatment with benzodiazepine (Diazepam), to analyze by histomorphometry the effects of the drug in the parotids glands and to verify the effect of the pilocarpine in glandular parenchyma and in the salivary flow rate. Seventy-two male Wistar rats were allocated to four groups. Control groups received saline during 60 days (C60) and pilocarpine (Pilo) during 60 days. Experimental groups were dealt with Diazepam associated with saline (DS), and Diazepam associated with pilocarpine (DP) during 60 days. The stimulated salivary flow rate was obtained by using the gravimetric method. After the animals were killed, parotid glands were removed and mass and size were determined. The specimens were processed and stereological analysis revealed cell volume. Mean values of size and salivary flow rate varied from 9.007 mm and 0.015 mg/min in DS to 7.854 mm and 0.029 mg/min in DP, respectively. ANOVA showed statistically significant differences between groups for size (p=0.0028) and salivary flow rate (p=0.0003). Psychotropic drugs caused hyposalivation in rats and acinar hypertrophy in their parotid glands. Pilocarpine, a cholinergic agonist with topical appliance, showed significant secretagogue action in the treatment of hyposalivation induced by Diazepam chronic use.

Papain-gel degrades intact nonmineralized type I collagen fibrils.

Papain-gel has been utilized as a chemo-mechanical material for caries removal due to its ability to preserve underlying sound dentin. However, little is known about the effect of the papain enzyme on intact type I collagen fibrils that compose the dentin matrix. Here we sought to define structural changes that occur in intact type I collagen fibrils after an enzymatic treatment with a papain-gel. Intact and nonmineralized type I collagen fibrils from rat tail were obtained and treated with a papain-gel (Papacarie) for 30 s, rinsed with water and imaged using an atomic force microscope (AFM). Additionally, polished healthy dentin specimens were also treated using the same protocol described above and had their elastic modulus (E) and hardness (H) measured by means of AFM-based nanoindentation. AFM images showed that the papain-gel induced partial degradation of the fibrils surface, yet no rupture of fibrils was noticed. The distinction between gap and overlap zones of fibrils vanished in most regions after treatment, and overlap zones appeared to be generally more affected. Mechanical data suggested a gradual decrease in E and H after treatments. A significant two-fold drop from the values of normal dentin (E=20+/-1.9, H=0.8+/-0.08 GPa) was found after four applications (E=9.7+/-3.2, H=0.24+/-0.1 GPa) (P<0.001), which may be attributed to the degradation of proteoglycans of the matrix. In summary, this study provided novel evidence that intact nonmineralized type I collagen fibrils are partially degraded by a papain-gel.