Biomimetic Growth of Metal-Organic Frameworks for the Stabilization of the Dentin Matrix and Control of Collagenolysis.

The dentin matrix is a collagenous scaffold structurally involved in anchoring resin-based materials to the tooth. Time-dependent degradation of this scaffold at the resin-dentin interface remains a core problem in adhesive dentistry, limiting the service life of dental fillings. This study explored the use of emergent materials termed metal-organic frameworks (MOFs)─formed by the self-assembly of metal ions and organic building blocks─to safeguard the collagen integrity in the functional dentin matrix. We demonstrate that collagen fibrils (from demineralized human dentin) can induce the biomimetic growth of MOF crystals as protective coatings to strengthen and stabilize the fibrils. Zeolitic imidazolate framework-8 (ZIF-8), a zinc-based microporous MOF, was used to fabricate the MOF composites via a "one-pot" reaction in water. The ZIF-modified dentin matrix presented superior mechanical strength and resistance to proteolysis, which can positively affect the longevity of collagen as an anchoring substrate. This work identifies a potential biomedical application of biomimetically synthesized MOFs in repairing dental tissues critical to restorative therapies.

Pressure-Assisted Coating of Ceramics on 3D-Printed Polymeric Scaffolds.

Pressure-assisted coating (PAC) is introduced to coat 3D-printed polymeric scaffolds with ß-tricalcium phosphate (ß-TCP) for tissue engineering applications. The method consists of four steps: infiltration of ceramic particles into the porous structure of the polymeric scaffold, dehydration of the slurry, compaction of ceramic particles around the scaffold, and heat treatment. The optimal coating is obtained at an infiltration speed of 400 mm/min followed by complete dehydration, compaction under ca. 8 MPa pressure, and subsequent heat treatment at 65 °C. The outcome is a uniformly coated scaffold with no deformation or structural defects, as confirmed by micro-CT analysis and laser and scanning electron microscopy. Scaffolds coated using the PAC method present superior interface bonding strength compared to those coated with a biomimetic approach. The contact angle decreased from 75.2 ± 1.4° for the uncoated scaffold to 39.6 ± 9.6° for the PAC specimen. PAC also increased the surface roughness from 0.66 ± 0.08 to 6.89 ± 0.26 µm and doubled the number of attached cells on the 3rd day of culture. The described method is applicable to different structures, object sizes, pore sizes, and shapes. For instance, in-depth coating of a 10 mm × 10 mm (D × H) cone with a 58 ± 4 µm-thick layer of ß-TCP can be achieved using PAC. The method can be used to coat other polymers, such as poly(lactic-co-glycolic acid) (PLGA). Successful coating of ß-TCP on 3D-printed PLGA scaffolds is also presented as a proof of concept.

3D construct of hydroxyapatite/zinc oxide/palladium nanocomposite scaffold for bone tissue engineering.

The purpose of this study was to produce and characterize Hydroxyapatite/Zinc Oxide/Palladium (HA/0.05 wt% ZnO/0.1 wt% Pd) nanocomposite scaffolds and study their mechanical and antibacterial properties, biocompatibility and bioactivity. The initial materials were developed using sol-gel and precipitation methods. Scaffolds were characterized using atomic absorption analysis (AA), scanning electron microcopy (SEM), energy dispersive spectroscopy (EDS) and transmission electron microscopy (TEM), atomic force microscopy (AFM) and Brunauer-EmmeS-Teller (BET) method. Furthermore, the bioactivity of scaffolds in simulated body fluid (SBF) and the interaction of dental pulp stem cells (DPSCs) with the nanocomposite scaffolds were assessed. Our results showed that the HA/ZnO/Pd (H1), HA/ZnO/Pd coated by 0.125 g chitosan (H2) and HA/ZnO/Pd coated by 0.25 g chitosan (H3) scaffolds possess higher compressive strength and toughness and lower microhardness and density compared to the pure HA (H0) scaffolds. Immersion of samples in SBF showed the deposition of apatite on the surface of the scaffolds. The biocompatibility assay indicated lower cell proliferation on the H1, H2 and H3 in comparison to the H0. The antibacterial results obtained show a significant impact by loading Pd/ZnO on HA in the deactivation of microorganisms in vitro.

Effect of sodium chloride on gene expression of Streptococcus mutans and zeta potential of demineralized dentin.

PURPOSE: In this work, the effects of sodium chloride (NaCl) on gene expression of planktonic Streptococcus mutans cells are investigated. Also assessed are the effects of NaCl on zeta potential of sound and demineralized dentin. METHODS: The relative level of glucosyltransferase B (gtfB), gtfC and gtfD transcription of S. mutans in the presence of NaCl was evaluated by quantitative polymerase chain reaction (qPCR). The osmolality of varying salt (NaCl) concentrations and their influence on the zeta potential of sound and demineralized dentin was investigated as well. RESULTS: NaCl significantly reduced the expression of gtfB and C genes in planktonic S. mutans; whereas, gtf D gene expression significantly increased in the presence of NaCl (P < 0.05). NaCl at concentrations of 37.5 mg/ml reduced zeta potential of demineralized dentin, while no significant decrease of zeta potential was found when sound dentin was exposed to this concentration. CONCLUSION: NaCl reduces the expression of some gtfs in S. mutans and increases negative potential charge of demineralized dentin.

Dual Porosity Protein-based Scaffolds with Enhanced Cell Infiltration and Proliferation.

3D dual porosity protein-based scaffolds have been developed using the combination of foaming and freeze-drying. The suggested approach leads to the production of large, highly porous scaffolds with negligible shrinkage and deformation compared to the conventional freeze-drying method. Scanning electron microscopy, standard histological processing and mercury intrusion porosimetry confirmed the formation of a dual network in the form of big primary pores (243 ± 14 µm) embracing smaller secondary pores (42 ± 3 µm) opened onto their surface, resembling a vascular network. High interconnectivity of the pores, confirmed by micro-CT, is shown to improve diffusion kinetics and support a relatively uniform distribution of isolated human dental pulp stem cells within the scaffold compared to conventional scaffolds. Dual network scaffolds indicate more than three times as high cell proliferation capability as conventional scaffolds in 14 days.

Surface characteristics of three commercially available grafts and adhesion of stem cells to these grafts.

BACKGROUND: Reconstruction of bone defects is often performed using bone autografts. However, limitations associated with the use of autografts led to the use of bone substitute materials. OBJECTIVES: The purpose of this study was to compare the surface characteristics of three commercially available grafts namely allografts, xenografts and alloplasts. METHODS: This in vitro study was conducted on beta-tricalcium phosphate (ß-TCP) alloplast, a mixture of demineralized bone matrix and mineralized bone allograft (DBM&MBA) and natural bovine bone mineral (NBBM) xenograft. Presence of apatite groups on the surface of samples was assessed by X-ray diffraction (XRD) while the presence of functional groups was evaluated using Fourier transform infrared spectroscopy (FTIR). Also, dental pulp stem cells (DPSCs) were cultured on the surface of samples and their adhesion was evaluated under a scanning electron microscope (SEM). RESULTS: The FTIR showed a relatively similar pattern for NBBM and TCP samples and a different pattern in DBM&MBA. The results of XRD analysis also showed similarities between NBBM and TCP with sharper peaks than the DBM&MBA sample. The SEM micrographs showed that at 24 hours, no cell was detectable on the surface of NBBM sample; whereas, elongated cells were noted on the surface of TCP and DBM&MBA samples. CONCLUSIONS: The patterns of ß-TCP and NBBM samples in XRD and FTIR spectroscopy showed high resemblance but they had different behaviors with respect to cell adhesion.

Comparison of the Antimicrobial Efficacy of Calcium Hydroxide and Photodynamic Therapy Against Enterococcus faecalis and Candida albicans in Teeth With Periapical Lesions; An In Vivo Study.

Introduction: Elimination of pathological microflora of root canal systems is a major goal in endodontic treatment. This study aimed to compare the antimicrobial efficacy of calcium hydroxide as an intracanal medication and antibacterial photodynamic therapy (aPDT) against Enterococcus faecalis and Candida albicans in teeth with periapical (PA) lesions. Methods: This in vivo study was conducted on 20 patients with single-rooted mandibular premolar with previously failed endodontic treatment. This study was performed as a clinical trial (IRCTID: IRCT2016090429686N1). After conventional chemo-mechanical root canal preparation (hand and rotary instruments and 2.5% NaOCl), microbiological samples were obtained using sterile paper points, then stored in thioglycolate solution and transferred to a microbiology laboratory. Group 1 (n = 10) specimens underwent aPDT (diode laser 808 nm + 50 mg/mL methylene blue), while creamy calcium hydroxide paste was used in group 2 for a duration of 1 week. A control sample was taken with sterile paper points and F3 Protaper rotary file. The samples were dispersed in transport medium, serially diluted, and cultured on selective mediums to determine the number of colony forming units (CFUs). Data were analyzed by Mann-Whitney U test at 5% significance level. The significance level for all analyses was set at P < 0.05. Results: Number of CFU significantly decreased in both groups after the interventions (P < 0.001); however, there was no significant difference in the colony count between the 2 groups. Conclusion: aPDT and calcium hydroxide therapy showed the same antimicrobial efficacy on E. faecalis and C. albicans.

In vitro biological outcome of laser application for modification or processing of titanium dental implants.

There are numerous functions for laser in modern implant dentistry including surface treatment, surface coating, and implant manufacturing. As laser application may potentially improve osseointegration of dental implants, we systematically reviewed the literature for in vitro biological responses to laser-modified or processed titanium dental implants. The literature was searched in PubMed, ISI Web, and Scopus, using keywords "titanium dental implants," "laser," "biocompatibility," and their synonyms. After screening the 136 references obtained, 28 articles met the inclusion criteria. We found that Nd:YAG laser was the most commonly used lasers in the treatment or processing of titanium dental implants. Most of the experiments used cell attachment and cell proliferation to investigate bioresponses of the implants. The most commonly used cells in these assays were osteoblast-like cells. Only one study was conducted in stem cells. These in vitro studies reported higher biocompatibility in laser-modified titanium implants. It seems that laser radiation plays a vital role in cell response to dental implants; however, it is necessary to accomplish more studies using different laser types and parameters on various cells to offer a more conclusive result.

In vitro proliferation and osteogenic differentiation of endometrial stem cells and dental pulp stem cells.

Stem-cell-based therapies were introduced aiming to overcome the limitations of the existing procedures for regeneration of mineralized tissues. Stem cells isolated from the endometrial tissue and dental pulp have the capacity to differentiate into various functional cells including osteoblasts. However, studies comparing their ability to regenerate mineralized tissue are lacking. The purpose of this study was to compare the proliferation and osteogenic differentiation potential of endometrial stem cells (EnSCs) and dental pulp stem cells (DPSCs) using in vitro cell culture technique. The DPSCs and EnSCs were isolated from human dental pulp and endometrium, respectively. Their proliferation and osteogenic potential were compared in the same osteogenic medium (OM) after 3, 5, 7 and 10 days using the methyl thiazol tetrazolium assay, alizarin red staining, and real-time quantitative reverse transcription polymerase chain reaction (Real-Time qRT-PCR). The EnSCs showed higher proliferation rate compared to DPSCs. Regarding osteogenesis, alizarin red-positive colonies appeared earlier and in greater amounts in DPSCs group. The real-time qRT-PCR demonstrated significantly greater osteogenic potential of DPSCs compared to EnSCs. Our findings revealed significant differences in stem cell properties based on the tissue source. The EnSCs had greater proliferation rate than DPSCs, while DPSCs showed greater osteogenic potential compared to EnSCs in the same OM.

Surface characterization and biological properties of regular dentin, demineralized dentin, and deproteinized dentin.

Bone autografts are often used for reconstruction of bone defects; however, due to the limitations of autografts, researchers have been in search of bone substitutes. Dentin is of particular interest for this purpose due to high similarity to bone. This in vitro study sought to assess the surface characteristics and biological properties of dentin samples prepared with different treatments. This study was conducted on regular (RD), demineralized (DemD), and deproteinized (DepD) dentin samples. X-ray diffraction and Fourier transform infrared spectroscopy were used for surface characterization. Samples were immersed in simulated body fluid, and their bioactivity was evaluated under a scanning electron microscope. The methyl thiazol tetrazolium assay, scanning electron microscope analysis and quantitative real-time polymerase chain reaction were performed, respectively to assess viability/proliferation, adhesion/morphology and osteoblast differentiation of cultured human dental pulp stem cells on dentin powders. Of the three dentin samples, DepD showed the highest and RD showed the lowest rate of formation and deposition of hydroxyapatite crystals. Although, the difference in superficial apatite was not significant among samples, functional groups on the surface, however, were more distinct on DepD. At four weeks, hydroxyapatite deposits were noted as needle-shaped accumulations on DemD sample and numerous hexagonal HA deposit masses were seen, covering the surface of DepD. The methyl thiazol tetrazolium, scanning electron microscope, and quantitative real-time polymerase chain reaction analyses during the 10-day cell culture on dentin powders showed the highest cell adhesion and viability and rapid differentiation in DepD. Based on the parameters evaluated in this in vitro study, DepD showed high rate of formation/deposition of hydroxyapatite crystals and adhesion/viability/osteogenic differentiation of human dental pulp stem cells, which may support its osteoinductive/osteoconductive potential for bone regeneration.

Different methods of dentin processing for application in bone tissue engineering: A systematic review.

Dentin has become an interesting potential biomaterial for tissue engineering of oral hard tissues. It can be used as a scaffold or as a source of growth factors in bone tissue engineering. Different forms of dentin have been studied for their potential use as bone substitutes. Here, we systematically review different methods of dentin preparation and the efficacy of processed dentin in bone tissue engineering. An electronic search was carried out in PubMed and Scopus databases for articles published from 2000 to 2016. Studies on dentin preparation for application in bone tissue engineering were selected. The initial search yielded a total of 1045 articles, of which 37 were finally selected. Review of studies showed that demineralization was the most commonly used dentin preparation process for use in tissue engineering. Dentin extract, dentin particles (tooth ash), freeze-dried dentin, and denatured dentin are others method of dentin preparation. Based on our literature review, we can conclude that preparation procedure and the size and shape of dentin particles play an important role in its osteoinductive and osteoconductive properties. Standardization of these methods is important to draw a conclusion in this regard. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2616-2627, 2016.

Comparison of the Antibacterial Effect of 810 nm Diode Laser and Photodynamic Therapy in Reducing the Microbial Flora of Root Canal in Endodontic Retreatment in Patients With Periradicular Lesions.

INTRODUCTION: The aim of this study was to compare the antibacterial efficacy of diode laser 810nm and photodynamic therapy (PDT) in reducing bacterial microflora in endodontic retreatment of teeth with periradicular lesion. METHODS: In this in vivo clinical trial, 20 patients who needed endodontic retreatment were selected. After conventional chemo mechanical preparation of root canals, microbiological samples were taken with sterile paper point (PP), held in thioglycollate broth, and then were transferred to the microbiological lab. In the first group, PDT with methylene blue (MB) and diode laser (810 nm, 0.2 W, 40 seconds) was performed and in the second group diode laser (810 nm, 1.2 W, 30 seconds) was irradiated. Then second samples were taken from all canals. RESULTS: CFU/ml amounts showed statistically significant reduction in both groups (P < 0.001). CFU/ml amounts were compared between the two groups and there was no statistical difference. CONCLUSION: PDT and diode laser 810 nm irradiation are effective methods for root canal disinfection. PDT is a suitable alternative for diode laser 810 nm irradiation, because of lower thermal risk on root dentin.

Effects of Non-Collagenous Proteins, TGF-ß1, and PDGF-BB on Viability and Proliferation of Dental Pulp Stem Cells.

OBJECTIVES: The dentin matrix servers as a reservoir of growth factors, sequestered during dentinogenesis. The aim of this study was to assess the viability and proliferation of dental pulp stem cells in the presence of dentin matrix-derived non-collagenous proteins and two growth factors; platelet-derived growth factor BB and transforming growth factor beta 1. MATERIAL AND METHODS: The dental pulp cells were isolated and cultured. The dentin proteins were extracted and purified. The MTT assay was performed for assessment of cell viability and proliferation in the presence of different concentrations of dentin proteins and growth factors during 24 - 72 h post-treatment. RESULTS: The cells treated with 250 ng/mL dentin proteins had the best viability and proliferation ability in comparison with other concentrations (P < 0.05). The MTT assay demonstrated that cells cultured with 5 ng/mL platelet-derived growth factor BB had the highest viability at each time point as compared to other groups (P < 0.05). However, in presence of platelet-derived growth factor BB alone and in combination with transforming growth factor beta 1 and dentin proteins (10 ng/mL), significant higher viability was seen at all time points (P < 0.05). The least viability and proliferation at each growth factor concentration was seen in cells treated with combination of transforming growth factor beta 1 and dentin proteins at 72 h (P < 0.05). CONCLUSIONS: The results indicated that the triple combination of growth factors and matrix-derived non-collagenous proteins (especially at 10 ng/mL concentration) has mitogenic effect on dental pulp stem cells.

Effects of extracts of Salvadora persica on proliferation and viability of human dental pulp stem cells.

OBJECTIVES: Efficacy of an ideal antimicrobial agent depends on its ability to eliminate microorganisms while causing minimal toxicity to host cells. The purpose of this study was to assess the effect of ethanolic and water extracts of Salvadora persica (SP) on proliferation and viability of human dental pulp stem cells (hDPSCs). MATERIALS AND METHODS: In this in-vitro study, the effects of seven concentrations of ethanolic and water extracts of SP (ranging from 5.75 mg/ml to 0.08 mg/ml) on hDPSCs were evaluated using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. The results were analyzed using one-way ANOVA and Tukey's post-hoc test. P < 0.05 was considered statistically significant. RESULTS: Water extract of SP only had cytotoxic effect at 5.75 mg/ml concentration; and caused significant cell proliferation at 1.43-0.08 mg/ml concentrations at 24 h (P < 0.05). At 48 h, only 0.17 and 0.08 mg/ml concentrations caused significant cell proliferation (P < 0.05). Ethanolic extract of SP at 5.75-1.43 mg/ml concentrations showed severe cytotoxic effects at 24 and 48 h. Other concentrations had no significant effects on cells (P > 0.05). CONCLUSION: The highest concentrations of both water and ethanolic extracts of SP had cytotoxic effects on hDPSCs. Water extract of SP has favorable effects on cell proliferation at specific concentrations in a time-dependent manner.

Cytotoxic effects of various mineral trioxide aggregate formulations, calcium-enriched mixture and a new cement on human pulp stem cells.

INTRODUCTION: This in vitro study compared the cytotoxic effects of three commercially available MTA formulations naming ProRoot MTA (PMTA), Angelus MTA (AMTA), and Root MTA (RMTA), with calcium-enriched mixture (CEM) cement and a new nanohybrid MTA (NMTA) on human dental pulp stem cells (DPSC). METHODS AND MATERIALS: Four disc-shaped specimens of each material were prepared. After completion of setting, 2 different (neat and 1/2) elutes of the test materials were made. Then in each cavity of a 96-well plate, 3000 cells were seeded and incubated in a humidified incubator with 5% CO2 and 95% air at 37(°)C for 24 h. After this period, the culture medium of each well was replaced with 200 µL of test material elutes. Plain culture medium was used as the negative control and distilled water as the positive control group. Cell viability was assessed using 2, 5-diphenyl-SH-tetrazelium bromide colorimetric assay, aka Mosmann's tetrazolium toxicity (MTT) assay, at three time intervals (24, 48, and 72 h after mixing). Data were analyzed using the ANOVA and Tukey's post hoc test (P=0.05). RESULTS: After 24 h, the viability of cells in neat concentration had no significant differences (P>0.05) except for the NMTA. However, CEM and AMTA, at 1/2 concentration exerted significant proliferative effects on cells. At 48 and 72-h intervals, significant proliferation of DPSCs was seen in all samples, except for the NMTA which exerted toxic effects on cells. CONCLUSION: All of the three commercial MTAs and CEM cement showed comparative biocompatibility. However, NMTA had cytotoxic effects on DPSCs at all the time intervals.

Comparison of osteogenic medium and uniaxial strain on differentiation of endometrial stem cells.

BACKGROUND: Mechanotransduction plays a pivotal role in remodeling and repair of skeletal tissues. This mechanism has been widely used in bone tissue engineering especially under in vitro conditions. To date, various stem cells have been used for this purpose. The present study was the first to evaluate the effect of mechanical loading on differentiation of human endometrial stem cells (hESCs) to osteoblasts. MATERIALS AND METHODS: Adhesion of endometrial stem cells after isolation and culture on a silicone membrane covered with collagen was evaluated under scanning electron microscope (SEM). Twenty-four hours after cell culture on the membrane and ensuring appropriate cell adhesion, a group of cells in a conventional culture medium received 3% static uniaxial strain. In the positive control group, cells cultured on the membrane were placed in an osteogenic medium without receiving any mechanical strain. The negative control group was placed in a regular medium and received no strain either. Two weeks later, cultured cells were evaluated for expression of osteogenic markers using immunofluorescence staining and real-time polymerase chain reaction (PCR). Data of real-time PCR was analyzed by ANOVA. P < 0.05 was considered statistically significant. RESULTS: SEM analysis revealed adequate cell adhesion to the membrane after 24 h. Two weeks after loading, expression of markers in the positive control group was significantly higher compared to test group. CONCLUSION: We can conclude that static uniaxial strain exerted on hESCs results in their differentiation to osteoblasts. However, this magnitude of static strain in the tested time period cannot yield excellent differentiation when compared to the osteogenic medium.

In-vitro Comparison of the Antimicrobial Properties of Glass Ionomer Cements with Zinc Phosphate Cements.

White spot lesions are observed in nearly 50% of patients undergoing orthodontic treatment. Long-lasting antibacterial properties of orthodontic cements can reduce this phenomenon. The aim of this research was to compare antimicrobial activity of three commercial glass ionomer cements with three commercial zinc phosphate cements, over time, against streptococcus mutans and candida albicans. Direct contact test (DCT) was used to evaluate the antibacterial and antifungal activity of products after 48 h and 7 days of incubation. The results demonstrated that all the cements presented antibacterial activity but the antibacterial activity of glass ionomer cements was more than that of zinc phosphate cements. Counts of C. albicans after 48 h were lower and statistically different in the GIC group in relation to the control groups. But no differences were observed between GIC and control groups at 7 days. Based on the results of this study, the antimicrobial and mainly antifungal effects of all the cements were so short.

Evaluating the In-vitro Antibacterial Effect of Iranian Propolis on Oral Microorganisms.

Propolis has traditionally been used in curing infections and healing wounds and burns. Current researches have shown that propolis has antibacterial, antifungal and antiviral actions however, the pharmacological activity of propolis is highly variable depending on its geographic origin. There have been few studies on the effects of Iranian propolis on the oral microorganisms. In this in-vitro study, the antimicrobial activity of the ethanolic and water extracts of the Iranian propolis (10%, w/v) from north-east area of Tehran was evaluated. Susceptibility of the oral strains tested (Streptococcus mutans ATCC 35668; Streptococcus salivarius ATCC 9222; Staphylococcus aureus ATCC 25923; Enterococcus faecalis ATCC 9854 and Lactobacillus casei ATCC 39392) was evaluated using the agar diffusion method at a concentration of 20 mg/mL of propolis and the zones of growth inhibition were measured. Antibacterial activity was determined by using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) at different concentrations of propolis. The ethanolic extract showed bacteriostatic and bactericidal activity against all the strains, with MIC and MBC ranges of 250-500 µg/mL. The MIC concentration of the water extract was 500 µg/mL against S. mutans and E. faecalis. The water extract showed bactericidal activity only against S. mutans (20 mg/mL). These results indicate that the ethanolic extract is probably more useful in the control of oral biofilms and subsequent dental caries development. However, to determine the consequence of the ethanolic extract of Iranian propolis on the oral mucosa, in-vivo studies of its possible effects are needed.