Mostafa EzEldeen

Aim: This study aims to comprehensively investigate the dynamic expression pattern of CXCR4 in human dental pulp stromal cells (hDPSCs) and stromal cells from the apical papilla (SCAPs) and assess the impact of different cell dissociation methods on its precise assessment. On this basis, a standardised CXCR4 detection and cell sorting strategy can be established to ensure the high purity and viability of the sorted population. Methodology: hDPSCs and SCAPs were collected by the outgrowth method from human third molars from three independent healthy (ASA I) patients (< 18 years old). Cells were characterised as mesenchymal stromal cells (MSCs) and expanded to passage 3–6 for experiments. Three different dissociation methods (Enzymatic digestion, Non‐enzymatic digestion, Mechanical collection) were used to collect single‐cell suspensions from culture flasks. Afterwards, the cells were resuspended in culture medium and allowed to recover at 37°C/5% CO ₂ for 0–4 h. Surface expression of CXCR4 on viable cells was assessed using flow cytometry. Cell metabolic activity and senescence after dissociation treatment were tested by XTT Assay and senescence staining. Cell migration capacity was evaluated by wound healing assay. The data were statistically analysed using p < 0.05 as a statistical significance reference. Results: hDPSCs and SCAPs exhibited typical MSC profiles and comparable cell viability under the same dissociation methods and time points. hDPSCs showed higher CXCR4 expression than SCAPs. CXCR4 expression displayed time‐dependent fluctuations in both cell types and cell viability was significantly affected by dissociation method and recovery period. Enzymatic digestion resulted in higher CXCR4 expression after 2‐h recovery and preserved over 90% cell viability. In contrast, non‐enzymatic dissociation and mechanical scraping impaired cell metabolic activity, migration capacity, and induced premature senescence. Conclusion: In in vitro expanded hDPSCs and SCAPs, the cell‐detachment method and recovery time can significantly affect the CXCR4 membrane detection and subsequent cell‐survival performance. To balance optimal cell viability and detection reliability, enzymatic cell dissociation followed by a 2‐h recovery time seems to be the best protocol for CXCR4 detection and subsequent downstream experiments within the 0–4 h time frame.

The history of stromal-derived factor-1 (SDF-1), alias CXCL12, started serendipitously and relatively late in the cytokine cDNA cloning era (1975–2000) and evolved at the biological level from progenitor cell-specific chemokine in the bone marrow to multifunctional cytokine with growth factor-like and tissue-regenerative activities. This evolution was parallelled by the integration of SDF-1/CXCL12 within the protein families of chemokines, cytokines and cell growth-promoting recombinant products having the potential for clinical applications. Here, we use this central position of CXCL12 as small signaling protein as an example for future developments in regenerative medicine. We provide context about SDF-1 biology within the field of skin wound healing research and how this compares with studies of other cytokines and growth factors. We also discuss whether SDF-1 formulations may be exemplary for other cytokines used for tissue regeneration. Normal skin wound healing is fraught with delays and complications in patients with specific underlying diseases, such as diabetes, hypertension and other elderly-related comorbidities, skin infections and accidental physical insults. Except for platelet-derived growth factor (PDGF), many cytokines, including vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), have failed so far in clinical studies of skin wound healing. This is in part due to the fact that (i) the biology of tissue regeneration is complex and insufficiently studied, (ii) in vitro approaches hardly mimic in vivo situations and (iii) commonly used animal models of acute and chronic wounding do not perfectly match human skin wound regeneration. A review of critical cells and molecules in normal skin and their actions in wounded tissue and a balanced comparison of the recent literature are preambles for progress in wound repair. We define advantages and limitations of recent approaches and appeal for more research. In particular, the possibilities of cellular immunomodulation mediated by endogenous and exogenous SDF-1/CXCL12 as a key molecule for skin regeneration are reviewed. Furthermore, biomaterials and scaffolds for the delivery and use of cytokines in precision medicine and aspects of their biofabrication are outlined with SDF-1 as an example. Finally, we indicate how applications of dermatological SDF-1 formulations for skin wound healing may be tailored for applications in other acute and chronic inflammatory conditions and regenerative medicine. Thereby, SDF-1/CXCL12 is placed at the crossroads between recombinant products, cytokines, chemokines and growth factors and occupies a central position between regenerative biology and medicine.

While available treatments have addressed a variety of complications in the dentoalveolar region, associated challenges have resulted in exploration of tissue engineering techniques. Often, scaffold biomaterials with specific properties are required for such strategies to be successful, development of which is an active area of research. This study focuses on the development of a copolymer of poly (N-isopropylacrylamide) (pNIPAM) and chitosan, used for 3D printing of scaffolds for dentoalveolar regeneration. The synthesized material was characterized by Fourier transform infrared spectroscopy, and the possibility of printing was evaluated through various printability tests. The rate of degradation and swelling was analyzed through gravimetry, and surface morphology was characterized by scanning electron microscopy. Viability of dental pulp stem cells seeded on the scaffolds was evaluated by live/dead analysis and DNA quantification. The results demonstrated successful copolymerization, and three formulations among various synthesized formulations were successfully 3D printed. Up to 35% degradability was confirmed within 7 days, and a maximum swelling of approximately 1200% was achieved. Furthermore, initial assessment of cell viability demonstrated biocompatibility of the developed scaffolds. While further studies are required to achieve the tissue engineering goals, the present results tend to indicate that the proposed hydrogel might be a valid candidate for scaffold fabrication serving dentoalveolar tissue engineering through 3D printing.

The aim of this study was to compare the quantification of hard-tissue debris by using micro-computed tomography (micro-CT) and nano-focus computed tomography (nano-CT) after root canal instrumentation. Ten mandibular molars containing an isthmus in the mesial root were scanned in a SkyScan 1172 micro-CT device with a voxel size of 12.8 µm and in a NanoTom nano-CT device with 5.5 µm. The mesial root canals were irrigated with 5 mL of saline solution at the orifice level, instrumented with Reciproc R25 files and a second scanning was performed by micro-CT and nano-CT devices for post-instrumentation images. DataViewer software was used for registering the pre- and post-operative micro-CT and nano-CT images. The root canal and the debris were segmented for quantitative analysis of the volume of the canal and volume of debris using CTAn software. Statistical analysis was performed using the T test for comparison between volume of the canal after instrumentation and volume of debris in both image modalities. The level of significance was set at p < 0.05. Nano-CT images showed higher values of debris when compared with micro-CT (p < 0.05) after root canal instrumentation. No difference was observed between the volume of the root canal after instrumentation in the two imaging methods used (p > 0.05). Nano-CT technology can be recommended as a more precise method for quantitative analysis of hard-tissue debris. Moreover, in Endodontic research it is a promising method, as it is capable of providing higher spatial and contrast resolution, faster scanning and higher image quality.

The recent advances in the field of cell-based therapeutics open promising perspectives for oral tissue regeneration. The development of large animal models, which overcome the limits of the rodent models and allow to emulate clinical situations, is crucial for the validation of regenerative strategies to move toward clinical application. Currently, porcine, canine, and ovine models are mainly developed for oral regeneration and their specific characteristics have an impact on the outcomes of the studies. Thus, this systematic review investigates the application of porcine, canine, and ovine models in present cell-based oral regeneration, according to the species characteristics and the targeted tissue to regenerate. A customized search of PubMed, EMBASE, Scopus, and Web of Science databases from January 2015 to March 2020 was conducted. Relevant articles about cell-based oral tissues engineering in porcine, canine, and ovine models were evaluated. Among the evaluated articles, 58 relevant studies about cell-based oral regeneration in porcine, canine, and ovine models matched the eligibility criteria and were selected for full analysis. Porcine models, the most similar species with humans, were mostly used for bone and periodontium regeneration; tooth regeneration was reported only in pig, except for one study in dog. Canine models were the most transversal models, successfully involved for all oral tissue regeneration and notably in implantology. However, differences with humans and ethical concerns affect the use of these models. Ovine models, alternative to porcine and canine ones, were mainly used for bone and, scarcely, periodontium regeneration. The anatomy and physiology of these animals restrain their involvement. If consistency was found in defect specificities and cell trends among different species animal models of bone, dentin-pulp complex, or tooth regeneration, variability appeared in periodontium. Regeneration assessment methods were more elaborate in porcines and canines than in ovines. Risk of bias was low for selection, attrition and reporting, but unclear for performance and detection. Overall, if none of the large animal models can be considered an ideal one, they are of deemed importance for oral cell-based tissue engineering and researchers should consider their relevance to establish favorable conditions for a given preclinical cell-based therapeutics. Impact statement: This systematic review investigates porcine, canine, and ovine models for current oral cell-based regeneration procedures, and researchers could refer to it for the choice of the most pertinent preclinical model for a given cell-based therapeutics.

Objectives: The present study aims to describe the relationship between upper first molar roots and maxillary sinus, for the first time with a truly three‐dimensional approach.Methods: From a retrospective cone‐beam computed tomography (CBCT) sample of the upper jaw, a total of 105 upper first molars in contact with maxillary sinus from 74 patients (male 24, female 50, mean age 42) were included in the present study. Segmentation of the upper first molar and maxillary sinus in CBCT was performed utilizing a semiautomatic livewire segmentation tool in MeVisLab v.3.1. The segmentations were analyzed in 3‐matic Medical 20.0 for root volume and the contact area between upper first molar roots and maxillary sinus. Analysis of variance test was applied to detect statistically significant differences between the roots.Results: The palatal root had the largest contact area with maxillary sinus 27.8 ± 21.4 mm² (20% of the root area) followed by the mesiobuccal 20.5 ± 17.9 mm² (17% of the root area) and distobuccal root 13.7 ± 12 mm² (14% of the root area). A significant difference in the contact area of the different roots of the upper first molar was seen.Conclusions: This study showed that 70% of the upper first molars were in contact with the maxillary sinus. The palatal root had on average a fifth of its root surface in contact with the sinus, while for mesiobuccal this was a sixth of its root surface and distobuccal roots this was somewhat less. The true 3D relationship could help to better understand maxillary anatomy in relation to occurring pathologies and treatment planning in this area.