3D-printing-assisted fabrication of chitosan scaffolds from different sources and cross-linkers for dental tissue engineering.

The aim of the present study was to fabricate and characterise chitosan scaffolds from animal and fungal sources, with or without gelatine as a co-polymer, and cross-linked to 3-glycidyloxyproply trimethoxysilane (GPTMS) or genipin for application in dental root tissue engineering. Chitosan-based scaffolds were prepared by the emulsion freeze-drying technique. Scanning electron microscopy (SEM) and nano-focus computed tomography (nano-CT) were used to characterise scaffold microstructure. Chemical composition and cross-linking were evaluated by Fourier transform infrared-attenuated total reflectance spectroscopy. Compression tests were performed to evaluate scaffold mechanical properties. Scaffold degradation was evaluated by gravimetric method and SEM. Scaffold bioactivity immersed in simulated body fluid was evaluated by SEM, with associated electron dispersive X-ray spectroscopy, and apatite formation was examined by X-ray diffraction. Finally, human dental pulp stem cells (hDPSCs) viability was evaluated. The fabrication method used was successful in producing scaffolds with organised porosity. Chitosan source (animal vs. fungal), co-polymerisation with gelatine and cross-linking using GPTMS or genipin had a significant effect on scaffold properties and hDPSCs response. Chitosan-genipin (CS-GEN) scaffolds had the largest pore diameter, while the chitosan-gelatine-GPTMS (CS-GEL-GPTMS) scaffolds had the smallest. Animal chitosan-gelatine co-polymerisation increased scaffold compressive strength, while fungal chitosan scaffolds (fCS-GEL-GPTMS) had the fastest degradation rate, losing 80 % of their weight by day 21. Gelatine co-polymerisation and GPTMS cross-linking enhanced chitosan scaffolds bioactivity through the formation of an apatite layer as well as improved hDPSCs attachment and viability. Tailored chitosan scaffolds with tuned properties and favourable hDPSCs response can be obtained for regenerative dentistry applications.

Use of CBCT Guidance for Tooth Autotransplantation in Children.

Tooth autotransplantation (TAT) offers a viable biological approach to tooth replacement in children and adolescents. The aim of this study was to evaluate the outcome of the cone-beam computed tomographic (CBCT)-guided TAT compared to the conventional TAT protocol and to assess the 3-dimensional (3D) patterns of healing after CBCT-guided TAT (secondary aim). This study included 100 autotransplanted teeth in 88 patients. Each experimental group consisted of 50 transplants in 44 patients (31 males and 19 females). The mean (SD) age at the time of surgery was 10.7 (1.1) y for the CBCT-guided group. This was 10.6 (1.3) y for the conventional group. The mean (SD) follow-up period was 4.5 (3.1) y (range, 1.1 to 10.4 y). Overall survival rate for the CBCT-guided TAT was 92% with a success rate of 86% compared to an 84% survival rate and a 78% success rate for the conventional group ( P > 0.005). The following measurements were extracted from the 3D analysis: root hard tissue volume (RV), root length (RL), apical foramen area (AFA), and mean and maximum dentin wall thickness (DWT). Overall, the mean (SD) percentage of tissue change was as follows: RV gain by 65.8% (34.6%), RL gain by 37.3% (31.5%), AFA reduction by 91.1% (14.9%), mean DWT increase by 107.9% (67.7%), and maximum DWT increase by 26.5% (40.1%). Principal component analysis (PCA) identified the mean DWT, RV, and maximum DWT as the parameters best describing the tissue change after TAT. Cluster analysis applied to the variables chosen by the PCA classified the CBCT group into 4 distinct clusters (C1 = 37.2%, C2 = 17.1%, C3 = 28.6%, C4 = 17.1%), revealing different patterns of tissue healing after TAT. The CBCT-guided approach increased the predictability of the treatment. The 3D analysis provided insights into the patterns of healing. CBCT-guided TAT could be adopted as an alternative for the conventional approach. (Clinical trial center and ethical board University Hospitals, KU Leuven: S55287; ClinicalTrials.gov Identifier: NCT02464202).

Implanted Dental Pulp Cells Fail to Induce Regeneration in Partial Pulpotomies.

Cell-based partial pulp regeneration is one of the promising approaches to obtain newly formed functional dentin-pulp complex. It relies on the preservation of the healthy tissue while regenerating the damaged pulp. The aim of this study was to investigate whether this regenerative process could be achieved by implanting porcine dental pulp cells (pDPCs) in pulp defects in the minipig. By split-mouth model, self-assembling injectable nanopeptide hydrogel, with and without pDPCs, was implanted after cameral pulpotomy in premolars and molars. At day 21 after surgery, 3-dimensional morphometric characterization, Masson's trichrome staining, and immunolabeling for DSP and BSP (dentin sialoprotein and bone sialoprotein) were performed on treated teeth. This study demonstrated no pulp regeneration but systematic reparative dentinogenesis. In fact, regardless of the presence of pDPCs in the scaffold, an osteodentin bridge-the microarchitecture of which significantly differed from the native dentin-was systematically obtained. Furthermore, the presence of pDPCs significantly affected the microstructure of the dentin bridges. In the radicular area of each treated tooth, hyperemia in the remaining pulp and external root resorptions were observed. Under the conditions tested in this work, pulp regeneration was not achieved, which highlights the need of further investigations to develop favorable regenerative microenvironment.

Artefact expression associated with several cone-beam computed tomographic machines when imaging root filled teeth.

AIM: To evaluate the characteristic artefact patterns associated with teeth root filled with Gutta-percha when scanned with four cone-beam CT devices. METHODOLOGY: Whilst using soft tissue simulation, ten root filled human premolars were placed in empty sockets in a dry human skull. Subsequently, the skull was scanned using 3D Accuitomo 170(®) , WhiteFox(®) , Cranex 3D(®) and Scanora 3D(®) following clinical protocols with the highest resolution and artefact reduction. After proper image registration in OnDemand3D(®) software (Cybermed, Seoul, Korea), each image slice was evaluated by three trained and calibrated dentomaxillofacial radiologists, which scored absence (0) and presence (1) of cupping artefact, hypodense halos and streak artefacts. Kappa test was performed for intra- and interobserver agreement. RESULTS: A moderate to perfect agreement for each observer (intra-observer κ = 0.5-1.0) was found. Agreement between the different observers was moderate to almost perfect for the different artefact patterns (interobserver κ = 0.55-0.9). Cupping artefact was the most prevalent (70%), followed by a hypodense halo (35%) and streak artefacts (16%). The Chi-squared test revealed significantly more streaks in axial slices (P < 0.0001), with some CBCT systems yielding significantly inferior results to others (P < 0.05). The dedicated EndoMode and artefact reduction did not improve the result significantly. CONCLUSIONS: The variation of artefact expression was significantly different amongst CBCT machines for root filled teeth. Continuous efforts are needed to improve CBCT reconstruction algorithms, with a specific focus on reducing artefacts induced by dense dental materials, whilst striving for enhanced image quality at low-radiation doses.

Long-term outcome of oral health in patients with early childhood caries treated under general anaesthesia.

AIM: Reports on the long-term outcome of oral health in children with early childhood caries (ECC) treated under general anaesthesia (GA) are lacking. The aim of this study was to assess oral health in adolescents with history of ECC treated under GA at young age. METHODS: A total of 98 children treated under GA because of ECC at the University Hospitals of Leuven (Belgium) (1995-1996) were included in a follow-up study. Information was collected at baseline (GA) and 1 and 12 years after treatment. The children were examined clinically [oral hygiene, caries experience (CE)] and oral health behaviour was recorded using a questionnaire. RESULTS: At second recall, 46 (48 %) adolescents could be contacted; 21 of these attended the dental clinic. Their mean age was 17.5 years (SD 1.4) with mean time span since GA of 13.1 years (SD 0.8); 91 % presented with CE. Mean D3MFT was 8.2 (SD 5.6) (median 7; range 0-18); 71 % showed untreated decay. A considerable gap between knowledge of content and actual use of fluoride in toothpaste was evident. CONCLUSIONS: Individuals with a history of ECC remained at high risk for caries in their permanent dentition. There is a need for well-designed long-term studies to explore associated factors.