Investigating the residual effect of silver nanoparticles gel as an intra-canal medicament on dental pulp stromal cells

Background The aim of this study was to evaluate the indirect effects of residual silver nanoparticles (AgNPs) gel on human dental pulp stromal cells (DPSCs). Methods Ninety-five dentin discs (4x4x1 mm) were prepared from freshly extracted human single-rooted teeth following institutional ethical approval and informed consent. Samples were cleaned, autoclaved, and treated with: 1.5%NaOCl, Saline and 17% EDTA then randomly assigned to 5 groups that received 50 μl of one of the following treatments: 0.01%AgNPs, 0.015%AgNPs, 0.02%AgNPs, Calcium hydroxide (Ca (OH)2) or no treatment for 1 week. Discs were washed with Saline and 17%EDTA then seeded with DPSCs and incubated for 3 and 7 days. At 24 hours unattached cells were collected and counted. At each time point cytotoxicity (LDH assay), cell viability (live/dead staining and confocal microscopy) and cell proliferation (WST1 assay) were assessed. All experiments were repeated a minimum of 3 times using DPSCs isolated from 3 different donors for each time point assessed (n = 9/group). Statistical analysis was done using One-Way ANOVA followed by Tukey’s test and Kruskal Wallis followed by post-hoc comparisons with significance set at p ≤ 0.05. Results After 24 hours, the percentage of DPSCs attachment ranged between 92.66% ±4.54 and 95.08% ±1.44 with no significant difference between groups (P = 0.126). Cell viability was ≥92% at 24 hours for all groups. However this percentage dropped to less than 60% at 3 days then started to rise again at 7 days. There was no significant difference in cytotoxicity between different groups at all time points except for 0.01%AgNPs group which had the highest cytotoxicity. DPSCs proliferation increased significantly from 3 to 7 days in all groups except for Ca (OH)2 which showed lower proliferation rates at both 3 (45.89%) and 7 days (79.25%). Conclusion Dentin discs treated for 7 days with concentrations of AgNPs gel (0.01–0.02%) allowed more than 90% DPSCs cell attachment after 24 hours. The cytotoxicity and proliferation of DPSCs in response to AgNPs gel were comparable to those with calcium hydroxide. This suggests that AgNPs gel may represent a promising future candidate for clinical use in regenerative endodontics. However, its effects may be concentration-dependent warranting further investigation

A Modified Rabbit Ulna Defect Model for Evaluating Periosteal Substitutes in Bone Engineering: A Pilot Study

The present work defines a modified critical size rabbit ulna defect model for bone regeneration in which a non-resorbable barrier membrane was used to separate the radius from the ulna to create a valid model for evaluation of tissue-engineered periosteal substitutes. Eight rabbits divided into two groups were used. Critical defects (15 mm) were made in the ulna completely eliminating periosteum. For group I, defects were filled with a nanohydroxyapatite poly(ester urethane) scaffold soaked in PBS and left as such (group Ia) or wrapped with a tissue-engineered periosteal substitute (group Ib). For group II, an expanded-polytetrafluoroethylene (e-PTFE) (GORE-TEX\uae) membrane was inserted around the radius then the defects received either scaffold alone (group IIa) or scaffold wrapped with periosteal substitute (group IIb). Animals were euthanized after 12\u201316 weeks, and bone regeneration was evaluated by radiography, computed microtomography (\ub5CT), and histology. In the first group, we observed formation of radio-ulnar synostosis irrespective of the treatment. This was completely eliminated upon placement of the e-PTFE (GORETEX\uae) membrane in the second group of animals. In conclusion, modification of the model using a non-resorbable e-PTFE membrane to isolate the ulna from the radius was a valuable addition allowing for objective evaluation of the tissue-engineered periosteal substitut

Human Treated Dentin Matrix Hydrogel as a Drug Delivery Scaffold for Regenerative Endodontics

Introduction: The objective of the current study was to develop a human treated dentin matrix (hTDM) hydrogel for use as a scaffold to allow the controlled release of an antimicrobial agent for regenerative endodontics. Materials and Methods: Human extracted teeth were treated via chemical demineralization using ethylene diamine tetra-acetic acid solution to produce hTDM powder. Fourier transform infrared spectroscopy (FTIR) was conducted to determine the functional groups of hTDM, scanning electron microscopy (SEM) was used to define the morphology/particle size of hTDM, and energy dispersive X-ray analysis was performed to identify the superficial apatite groups. Prepared hTDM powder was added to the amoxicillin-clavulanate mixture with a mass ratio of 1:1. Then, the combination was dripped into a 5% (w/v) calcium chloride solution. Antibiotic release profiles were evaluated for 14 days via high performance liquid chromatography (HPLC). Hydrogel degradation properties were studied for 14 days using 10 mL of phosphate buffered saline (PBS). Encapsulation efficiency was determined by HPLC, while minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of amoxicillin-clavulanate were determined against Enterococcus faecalis (E. faecalis). The antibacterial activity of amoxicillin-clavulanate against E. faecalis was investigated for 14 days via agar diffusion test. Statistical analysis was performed with the Shapiro-Wilk test (P=0.05). Results: hTDM showed statistically a significant difference for percentage weight change (P=0.1). The encapsulation efficiencies for hTDM hydrogel with antibiotic and hydrogel with antibiotic was 96.08%±0.02 and 94.62%±0.11, respectively. MIC and MBC values of amoxicillin-clavulanate against E. faecalis were 2.4 µg/mL and 9.6 µg/mL, respectively. The antibacterial activity of antibiotic loaded hTDM hydrogels was significantly greater than loaded hydrogels alone by 31% after 4 and 100% at 14 days, respectively (P≤0.001). Conclusions: This in vitro study showed antibiotic-loaded injectable hTDM hydrogel could be an alternative system to transfer antibiotic-based intracanal medicaments for use in regenerative endodontics

Learning from Mother Nature: Innovative Tools to Boost Endogenous Repair of Critical or Difficult-to-Heal Large Tissue Defects

For repair of chronic or difficult-to-heal tissue lesions and defects, major constraints exist to a broad application of cell therapy and tissue engineering approaches, i.e., transplantation of "ex vivo" expanded autologous stem/progenitor cells, alone or associated with carrier biomaterials. To enable a large number of patients to benefit, new strategies should be considered. One of the main goals of contemporary regenerative medicine is to develop new regenerative therapies, inspired from Mother Nature. In all injured tissues, when platelets are activated by tissue contact, their released factors promote innate immune cell migration to the wound site. Platelet-derived factors and factors secreted by migrating immune cells create an inflammatory microenvironment, in turn, causing the activation of angiogenesis and vasculogenesis processes. Eventually, repair or regeneration of the injured tissue occurs via paracrine signals activating, mobilizing or recruiting to the wound site cells with healing potential, such as stem cells, progenitors, or undifferentiated cells derived from the reprogramming of tissue differentiated cells. This review, largely based on our studies, discusses the identification of new tools, inspired by cellular and molecular mechanisms overseeing physiological tissue healing, that could reactivate dormant endogenous regeneration mechanisms lost during evolution and ontogenesis

Dental Mesenchymal Stem Cell-Based Translational Regenerative Dentistry: From Artificial to Biological Replacement

Dentistry is a continuously changing field that has witnessed much advancement in the past century. Prosthodontics is that branch of dentistry that deals with replacing missing teeth using either fixed or removable appliances in an attempt to simulate natural tooth function. Although such “replacement therapies” appear to be easy and economic they fall short of ever coming close to their natural counterparts. Complications that arise often lead to failures and frequent repairs of such devices which seldom allow true physiological function of dental and oral-maxillofacial tissues. Such factors can critically affect the quality of life of an individual. The market for dental implants is continuously growing with huge economic revenues. Unfortunately, such treatments are again associated with frequent problems such as peri-implantitis resulting in an eventual loss or replacement of implants. This is particularly influential for patients having co-morbid diseases such as diabetes or osteoporosis and in association with smoking and other conditions that undoubtedly affect the final treatment outcome. The advent of tissue engineering and regenerative medicine therapies along with the enormous strides taken in their associated interdisciplinary fields such as stem cell therapy, biomaterial development, and others may open arenas to enhancing tissue regeneration via designing and construction of patient-specific biological and/or biomimetic substitutes. This review will overview current strategies in regenerative dentistry while overviewing key roles of dental mesenchymal stem cells particularly those of the dental pulp, until paving the way to precision/translational regenerative medicine therapies for future clinical use