Osseointegration of titanium implants after surface treatment with ultraviolet light or cold atmospheric plasma in vivo.

PURPOSE: To determine the histological effects of ultraviolet light and cold atmospheric plasma treatment on the osseointegration of titanium implants in vivo. MATERIALS AND METHODS: Six juvenile pigs were divided into three groups of two animals each. A total of 54 titanium implants were placed randomly in the pigs' calvarial bone (nine implants per pig). Of these, 18 implants served as untreated controls. The remaining 36 implants served as the experimental group and were treated with either ultraviolet light or argon plasma for 12 minutes each prior to insertion. Two pigs in each group were kept until 2, 4 and 8 weeks and then sacrificed. Resonance frequency analysis was conducted after implant placement and at the time of sacrifice. Osseointegration was evaluated using microcomputed tomography scans and histomorphometrical analysis. RESULTS: After initial loss, all implants showed a constant increase in implant stability quotient values over time without significant differences between the groups. The bone-implant contact values increased steadily for all implants over 8 weeks of healing. Surface-treated implants showed significantly higher bone-implant contact values compared to untreated implants at each time point. Bone area fraction occupancy values were almost always higher following both treatment methods; however, differences were only significant after 4 and 8 weeks for the cold atmospheric plasma group and after 4 weeks for the ultraviolet light group. CONCLUSIONS: Ultraviolet light and cold atmospheric plasma may improve histomorphometrical osseointegration of titanium implants significantly.

Comparison of functional and esthetic outcomes in digital versus analog rehabilitation of one-piece screw-retained implant crowns placed at second stage surgery.

PURPOSE: Analog and digital impressions are established procedures for restoration of single-tooth implants. In this study, single-tooth implants were restored with definitive restorations during second-stage surgery. Analog and digital workflows were compared. MATERIAL AND METHODS: Eighty single-tooth implants were examined in total. In 40 implants, an index was taken immediately after implant placement using composite resin to fabricate the final crowns (analog workflow). For the other 40 single-tooth implants, intraoral intraoperative scans were performed (digital workflow) during primary surgery. The custom-fabricated screw-retained crowns were placed during second-stage surgery. Photographs and examinations for the scores were taken at the time of the follow-up visit, 1-4 years after placement of the crowns. The number of treatment appointments required was recorded and the modified pink esthetic score (PES) was determined. Additionally, the functional implant prosthetic score (FIPS) was measured. RESULTS: The mean PES was 12.15/14 for the digital workflow and 11.95/14 for the analog workflow. The most common deficit was incomplete papillae for both workflows. Three treatment appointments were required for both workflows: (1) Scan and/or impressions making and patient consent, (2) implant placement, and (3) second-stage surgery with crown insertion. The FIPS was 9.1/10 for the digital workflow group and 9.2/10 for the analog workflow group. Common deficits presented as missing papillae as well as open approximal contacts. The FIPS was not significantly different between workflows (p = 0.679). The PES also did not show a statistically significant difference for both workflows (p = 0.654), however, the analog workflow showed better values for the papillae (p < 0.05). A significant difference was also found in the other PES values, with the digital workflow showing better results here (p < 0.05). A chronological analysis of the results of the digital technique showed that the cases treated last had significantly better values than the cases treated first. CONCLUSIONS: According to the results of this study, both workflows allowed placement of the definitive crowns on single-tooth implants during second-stage surgery. Both workflows were found to be equivalent in terms of esthetic results in this study, although the digital workflow demonstrated a learning curve.

Does placement of one-piece zirconia implants influence crestal bone loss? Retrospective evaluation 1 year after prosthetic loading.

PURPOSE: To evaluate the impact of immediate and delayed zirconia implant placement on crestal bone loss and the clinical outcomes achieved with this approach 1 year after prosthetic loading. Further objectives were to evaluate the influence of age, sex, smoking, implant dimensions, platelet-rich fibrin application and location of the implants in the jawbone on the crestal bone level. MATERIALS AND METHODS: Clinical and radiographic analysis were performed to evaluate the success rates for both groups. The data were analysed statistically through a linear regression analysis. RESULTS: No significant difference was found between immediate and delayed implant placement concerning the amount of crestal bone loss. Sex, age, bone augmentation, diabetes and prosthetic complications did not have a statistically significant influence; only smoking had a negative and statistically significant influence on crestal bone loss (P < 0.05). CONCLUSIONS: Immediate or delayed placement of one-piece zirconia implants may be a good alternative to the use of titanium implants concerning success and survival.

Do Titanium Dioxide Particles Stimulate Macrophages to Release Proinflammatory Cytokines and Increase the Risk for Peri-implantitis?

PURPOSE: Titanium dioxide (TiO2) particles detached from titanium dental implants by tribocorrosion can be phagocytosed by macrophages, releasing various proinflammatory cytokines at the implant sites that may trigger peri-implantitis. The study objective was to measure the association between peri-implantitis and the presence of non-allergy-related proinflammatory cytokines associated with TiO2 particles. METHODS: The investigators implemented a retrospective cross-sectional study and enrolled a sample of 60 subjects from a dental practice. Subjects were excluded if the plaque index was grade 3 (Silness and Löe). The predictor variable was a positive or negative TiO2 stimulation test, an in vitro macrophage proinflammatory response test. The outcome variable was peri-implantitis status defined as present or absent. Three groups were considered: control group with 20 patients without dental implants (group 1), 2 groups of patients with titanium dental implants, one without peri-implantitis (group 2), and the other with peri-implantitis (group 3) (n = 20 each). For patients with implants, depth of the gingival pockets of the implants were measured, and existing bleeding and suppuration were determined to assess peri-implantitis. Radiographs were taken if one or more factors applied to confirm the diagnosis of peri-implantitis. Further covariates were age, sex, duration of implant wear, and number of implants which were analyzed descriptively. Inferential analyses were undertaken using χ2 test, Kruskal-Wallis-, Wilcoxon-two-sample tests, and logistic regressions. RESULTS: The sample was composed of 35 female and 25 male patients with a mean age of 54.2 years (standard deviation = 14.76). The overall TiO2 stimulation test positivity frequency was 28.3% and were 30.0%, 5.0%, and 50.0% in the control, implants without peri-implantitis, and implants with peri-implantitis groups. No statistically significant differences could be seen in the frequencies of the TiO2 stimulation test results between control group and combined groups 2 and 3 (P-value = .84). The risk for positive TiO2 patients with a titanium implant of developing peri-implantitis was statistically significant and higher compared to negative TiO2 patients (odds ratio, 19.0 with 95% confidence interval [2.12,170.38]; P-value< .01). CONCLUSIONS: The data in this study showed a statistically significant relationship between a positive TiO2 stimulation test and peri-implantitis. Further studies with larger numbers of subjects are recommended to confirm this result.

New and innovative biomaterials, techniques and therapy concepts: Biologization in maxillofacial surgery, oral surgery and dentistry is in full swing. PRF, PRGF, PRP, blood plasma-stabilized augmentations, supplementation of micronutrients and vitamins - what opportunities do such "biological" approaches actually offer? We introduce them here.

Biomaterials of natural origin have recently gained increasing attention in the field of dental implantology. The requirements for such materials, however, are very high. In addition to high clinical efficiency in tissue regeneration, wound healing should be demonstrably positively influenced. The translational division for regenerative orofacial medicine of the Clinic and Polyclinic for Oral and Maxillofacial Surgery of the University Medical Center Hamburg-Eppendorf (UKE) is examining this research topic by investigating which innovative treatment methods for the reconstruction of bone defects or for augmentative procedures can be applied in the future or are already being applied in the field of oral and maxillofacial surgery.

Tissue Adhesives in Reconstructive and Aesthetic Surgery-Application of Silk Fibroin-Based Biomaterials.

Tissue adhesives have been successfully used in various kind of surgeries such as oral and maxillofacial surgery for some time. They serve as a substitute for suturing of tissues and shorten treatment time. Besides synthetic-based adhesives, a number of biological-based formulations are finding their way into research and clinical application. In natural adhesives, proteins play a crucial role, mediating adhesion and cohesion at the same time. Silk fibroin, as a natural biomaterial, represents an interesting alternative to conventional medical adhesives. Here, the most commonly used bioadhesives as well as the potential of silk fibroin as natural adhesives will be discussed.

Horizontal augmentation techniques in the mandible: a systematic review.

PURPOSE: Placement of dental implants has evolved to be an advantageous treatment option for rehabilitation of the fully or partially edentulous mandible. In case of extensive horizontal bone resorption, the bone volume needs to be augmented prior to or during implant placement in order to obtain dental rehabilitation and maximize implant survival and success. METHODS: Our aim was to systematically review the available data on lateral augmentation techniques in the horizontally compromised mandible considering all grafting protocols using xenogeneic, synthetic, or allogeneic material. A computerized and manual literature search was performed for clinical studies (published January 1995 to March 2021). RESULTS: Eight studies ultimately met the inclusion criteria comprising a total of 276 procedures of xenogeneic, allogeneic, or autogenous bone graft applications in horizontal ridge defects. Particulate materials as well as bone blocks were used as grafts with a mean follow-up of 26.0 months across all included studies. Outcome measures, approaches and materials varied from study to study. A gain of horizontal bone width of the mandible with a mean of 4.8 mm was observed in seven of eight studies. All but one study, reported low bone graft failure rates of 4.4% in average. CONCLUSIONS: Only limited data are available on the impact of different horizontal augmentation strategies in the mandible. The results show outcomes for xenogeneic as well as autologous bone materials for horizontal ridge augmentation of the lower jaw. The use of allogeneic bone-block grafts in combination with resorbable barrier membranes must be re-evaluated. Randomized controlled clinical trials are largely missing.

Success and patient satisfaction of immediately loaded zirconia implants with fixed restorations one year after loading.

BACKGROUND: There is limited evidence for the use of zirconium dioxide implants in immediate implant placement as well as for related immediate loading protocols. The aim of this retrospective study was to investigate the survival rate, success and patient satisfaction of immediately placed zirconia implants compared to delayed placed implants. METHODS: The study included 58 partially edentulous patients who were treated between 2013 and 2015 with immediate and delayed transgingival healing zirconium dioxide implants (SDS, Kreuzlingen/ Switzerland). In addition to survival and success rate, marginal bone loss was assessed using radiographs and soft tissue was evaluated using Pink Esthetic Score. Oral health-related quality of life was investigated prospectively using a modified OHIP questionnaire. RESULTS: The cumulative survival rate of all implants included was 92% with 88% classified as full success. No significant difference was found between the bone levels of immediately and delayed placed and immediately and delayed loaded implants. The mean Pink Esthetic Score after final prosthetic rehabilitation was 12.2/14 points indicating excellent esthetic clinical results. Analysis of the OHIP questionnaire showed a mean value of 0.54/100 points reflecting a high patient satisfaction. CONCLUSIONS: Immediate and delayed placed as well as loaded zirconium dioxide implants showed excellent results regarding implant success and survival in this study. Zirconium dioxide implants may ensure excellent esthetic results and high patient satisfaction.

Attachment and Osteogenic Potential of Dental Pulp Stem Cells on Non-Thermal Plasma and UV Light Treated Titanium, Zirconia and Modified PEEK Surfaces.

Ultraviolet (UV) light and non-thermal plasma (NTP) treatment are chairside methods that can efficiently improve the biological aging of implant material surfaces caused by customary storage. However, the behaviors of stem cells on these treated surfaces of the implant are still unclear. This study aimed to investigate the effects of UV light and NTP treated surfaces of titanium, zirconia and modified polyetheretherketone (PEEK, BioHPP) on the attachment and osteogenic potential of human dental pulp stem cells (DPSCs) in vitro. Machined disks were treated using UV light and argon or oxygen NTP for 12 min each. Untreated disks were set as controls. DPSCs were cultured from the wisdom teeth of adults that gave informed consent. After 24 h of incubation, the attachment and viability of cells on surfaces were assessed. Cells were further osteogenically induced, alkaline phosphatase (ALP) activity was detected via a p-Nitrophenyl phosphate assay (day 14 and 21) and mineralization degree was measured using a Calcium Assay kit (day 21). UV light and NTP treated titanium, zirconia and BioHPP surfaces improved the early attachment and viability of DPSCs. ALP activity and mineralization degree of osteoinductive DPSCs were significantly increased on UV light and NTP treated surfaces of titanium, zirconia and also oxygen plasma treated Bio-HPP (p < 0.05). In conclusion, UV light and NTP treatments may improve the attachment of DPSCs on titanium, zirconia and BioHPP surfaces. Osteogenic differentiation of DPSCs can be enhanced on UV light and NTP treated surfaces of titanium and zirconia, as well as on oxygen plasma treated Bio-HPP.

Impact of the Veneering Technique and Framework Material on the Failure Loads of All-Ceramic Computer-Aided Design/Computer-Aided Manufacturing Fixed Partial Dentures.

OBJECTIVES: Zirconia (Y-TZP) ceramics are considered as posterior fixed partial denture (FPD) materials; however, their applications are limited due to chipping. The use of monolithic lithium disilicate (LiDi) glass ceramics in posterior FPDs can be advantageous. This in vitro study aims to compare the loads until failure of posterior Y-TZP-FPDs and LiDi-FPDs before and after aging.

Osseointegration of Zirconia Implants after UV-Light or Cold Atmospheric Plasma Surface Treatment In Vivo.

The influence of UV light and non-thermal plasma on the osseointegration of yttria-stabilized zirconia implants (Y-TZP) comparing the two methods is unclear. The aim of this study was to show the influence of these methods on the osseointegration of dental zirconia implants in an animal model. A total of 54 implants were either untreated, treated with UV light (UV), or non-thermal oxygen plasma for 12 min and inserted into the parietal bones of six domestic pigs. The animals were sacrificed after a healing interval of two, four, and nine weeks. The degree of osseointegration was determined using histomorphometric determination of bone-to-implant contact values (BIC) and the bone-to-implant contact values within the retentive parts of the implants (BAFO). BIC values decreased in all groups after four weeks of healing and re-increased after nine weeks in all groups. BAFO increased significantly over time in all groups. However, there were no statistically significant differences in BIC and BAFO values between the control group and the test groups and over time. Clinical studies may follow to confirm the influence of cold plasma and UV light on the healing and survival of zirconia implants.

Effects of a Novel Cold Atmospheric Plasma Treatment of Titanium on the Proliferation and Adhesion Behavior of Fibroblasts.

Cold plasma treatment increases the hydrophilicity of the surfaces of implants and may enhance their integration with the surrounding tissues. The implaPrep prototype device from Relyon Plasma generates cold atmospheric plasma via dielectric barrier discharge (DBD). In this study, titanium surfaces were treated with the implaPrep device for 20 s and assessed as a cell culture surface for fibroblasts. One day after seeding, significantly more cells were counted on the surfaces treated with cold plasma than on the untreated control titanium surface. Additionally, the viability assay revealed significantly higher viability on the treated surfaces. Morphological observation of the cells showed certain differences between the treated and untreated titanium surfaces. While conventional plasma devices require compressed gas, such as oxygen or argon, the implaPrep device uses atmospheric air as the gas source. It is, therefore, compact in size and simple to handle, and may provide a safe and convenient tool for treating the surfaces of dental implants, which may further improve the implantation outcome.

Time Dependency of Non-Thermal Oxygen Plasma and Ultraviolet Irradiation on Cellular Attachment and mRNA Expression of Growth Factors in Osteoblasts on Titanium and Zirconia Surfaces.

Ultraviolet (UV) light and non-thermal plasma (NTP) are promising chair-side surface treatment methods to overcome the time-dependent aging of dental implant surfaces. After showing the efficiency of UV light and NTP treatment in restoring the biological activity of titanium and zirconia surfaces in vitro, the objective of this study was to define appropriate processing times for clinical use. Titanium and zirconia disks were treated by UV light and non-thermal oxygen plasma with increasing duration. Non-treated disks were set as controls. Murine osteoblast-like cells (MC3T3-E1) were seeded onto the treated or non-treated disks. After 2 and 24 h of incubation, the viability of cells on surfaces was assessed using an MTS assay. mRNA expression of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were assessed using real-time reverse transcription polymerase chain reaction analysis. Cellular morphology and attachment were observed using confocal microscopy. The viability of MC3T3-E1 was significantly increased in 12 min UV-light treated and 1 min oxygen NTP treated groups. VEGF relative expression reached the highest levels on 12 min UV-light and 1 min NTP treated surfaces of both disks. The highest levels of HGF relative expression were reached on 12 min UV light treated zirconia surfaces. However, cells on 12 and 16 min UV-light and NTP treated surfaces of both materials had a more widely spread cytoskeleton compared to control groups. Twelve min UV-light and one min non-thermal oxygen plasma treatment on titanium and zirconia may be the favored times in terms of increasing the viability, mRNA expression of growth factors and cellular attachment in MC3T3-E1 cells.

Effect of process parameters on additive-free electrospinning of regenerated silk fibroin nonwovens.

Silk fibroin is a biomaterial with multiple beneficial properties for use in regenerative medicine and tissue engineering. When dissolving and processing the reconstituted silk fibroin solution by electrospinning, the arrangement and size of fibers can be manifold varied and according fiber diameters reduced to the nanometer range. Such nonwovens show high porosity as well as potential biocompatibility. Usually, electrospinning of most biomaterials demands for the application of additives, which enable stable electrospinning by adjusting viscosity, and are intended to evaporate during processing or to be washed out afterwards. However, the use of such additives increases costs and has to be taken into account in terms of biological risks when used for biomedical applications. In this study, we explored the possibilities of additive-free electrospinning of pure fibroin nonwovens and tried to optimize process parameters to enable stable processing. We used natural silk derived from the mulberry silkworm Bombyx mori. After degumming, the silk fibroin was dissolved and the viscosity of the spinning solution was controlled by partial evaporation of the initial solving agent. This way, we were able to completely avoid the use of additives and manufacture nonwovens, which potentially offer higher biocompatibility and reduced immunogenicity. Temperature and relative humidity during electrospinning were systematically varied (25-35 °C, 25-30% RH). In a second step, the nonwovens optionally underwent methanol treatment to initiate beta-sheet formation in order to increase structural integrity and strength. Comprehensive surface analysis on the different nonwovens was performed using scanning electron microscopy and supplemented by additional mechanical testing. Cytotoxicity was evaluated using BrdU-assay, XTT-assay, LDH-assay and live-dead staining. Our findings were, that an increase of temperature and relative humidity led to unequal fiber diameters and defective nonwovens. Resistance to penetration decreased accordingly. The most uniform fiber diameters of 998 ± 63 nm were obtained at 30 °C and 25% relative humidity, also showing the highest value for resistance to penetration (0.20 N). The according pure fibroin nonwoven also showed no signs of cytotoxicity. However, while the biological response showed statistical evidence, the material characteristics showed no statistically significant correlation to changes of the ambient conditions within the investigated ranges. We suggest that further experiments should explore additional ranges for temperature and humidity and further focus on the repeatability of material properties in dependency of suitable process windows.

Cytocompatibility of Titanium, Zirconia and Modified PEEK after Surface Treatment Using UV Light or Non-Thermal Plasma.

A number of modifications have been developed in order to enhance surface cytocompatibility for prosthetic support of dental implants. Among them, ultraviolet (UV) light and non-thermal plasma (NTP) treatment are promising methods. The objective of this study was to compare the effects of UV light and NTP on machined titanium, zirconia and modified polyetheretherketone (PEEK, BioHPP) surfaces in vitro. Machined samples of titanium, zirconia and BioHPP were treated by UV light and NTP of argon or oxygen for 12 min each. Non-treated disks were set as controls. A mouse fibroblast and a human gingival fibroblast cell line were used for in vitro experiments. After 2, 24 and 48 h of incubation, the attachment, viability and cytotoxicity of cells on surfaces were assessed. Results: Titanium, zirconia and BioHPP surfaces treated by UV light and oxygen plasma were more favorable to the early attachment of soft-tissue cells than non-treated surfaces, and the number of cells on those treated surfaces was significantly increased after 2, 24 and 48 h of incubation (p < 0.05). However, the effects of argon plasma treatment on the cytocompatibility of soft tissue cells varied with the type of cells and the treated material. UV light and oxygen plasma treatments may improve the attachment of fibroblast cells on machined titanium, zirconia and PEEK surfaces, that are materials for prosthetic support of dental implants.

Production and Characterization of Porous Fibroin Scaffolds for Regenerative Medical Application.

BACKGROUND/AIM: Silk is a natural biomaterial with several superior features for applications in regenerative medicine. In the present study an optimized process for manufacturing porous scaffolds out of the silk protein fibroin was developed. MATERIALS AND METHODS: The silk protein fibroin was dissolved in Ajisawa's reagent and the resulting fibroin solution was used to produce scaffolds by means of freeze-thawing cycling. Porosity, pressure and stab resistance as well as degradation behavior were assessed in order to characterize the physical properties of the resulting scaffolds. RESULTS: The resulting sponge-like fibroin scaffolds were highly porous while the porosity correlated inversely with the concentration of the starting fibroin solution. Increased initial fibroin concentrations of the scaffolds resulted in increased compressive and cannulation resistance. The majority of the fibroin scaffolds were digested by 1 mg/ml protease XIV in 3 weeks, indicating their biodegradability. CONCLUSION: The production of scaffolds made of varying fibroin concentrations by means of freeze-thawing, following dissolution using Ajisawa's reagent, provides a simple and straightforward strategy for adjusting the physical and chemical properties of fibroin scaffolds for various medical applications.

Extent of Inflammation and Foreign Body Reaction to Porous Polyethylene In Vitro and In Vivo.

BACKGROUND/AIM: High-density porous polyethylene (PP) offers possibilities for reconstruction in craniofacial surgery. The purpose of this study was to evaluate the extent of inflammation and foreign body reactions to PP in vitro and in vivo. MATERIALS AND METHODS: Cell attachment, proliferation and expression of inflammatory cytokines were assessed using murine macrophages (RAW 264.7) on two different PP materials in vitro. In vivo, Balb/c mice received PP implants at their dorsum. After sacrifice, samples were analyzed histologically and real-time PCR was used to assess expression of inflammatory cytokines. RESULTS: Cells showed a significantly decreased proliferation (p<0.001) after 48 h and a significantly increased expression of TNF-α (p<0.05) at 24, 48 and 72 h. All animals showed foreign body cell reactions and signs of chronic inflammation. Expression of all but one of the investigated cytokines dropped to non-significant levels after an initial increase. CONCLUSION: Application of porous polyethylene can cause local chronic inflammatory reactions. Although clinical application seems to be immunologically safe, indication and risks should be evaluated carefully when using PP implants.

Improved In Vitro Test Procedure for Full Assessment of the Cytocompatibility of Degradable Magnesium Based on ISO 10993-5/-12.

Magnesium (Mg)-based biomaterials are promising candidates for bone and tissue regeneration. Alloying and surface modifications provide effective strategies for optimizing and tailoring their degradation kinetics. Nevertheless, biocompatibility analyses of Mg-based materials are challenging due to its special degradation mechanism with continuous hydrogen release. In this context, the hydrogen release and the related (micro-) milieu conditions pretend to strictly follow in vitro standards based on ISO 10993-5/-12. Thus, special adaptions for the testing of Mg materials are necessary, which have been described in a previous study from our group. Based on these adaptions, further developments of a test procedure allowing rapid and effective in vitro cytocompatibility analyses of Mg-based materials based on ISO 10993-5/-12 are necessary. The following study introduces a new two-step test scheme for rapid and effective testing of Mg. Specimens with different surface characteristics were produced by means of plasma electrolytic oxidation (PEO) using silicate-based and phosphate-based electrolytes. The test samples were evaluated for corrosion behavior, cytocompatibility and their mechanical and osteogenic properties. Thereby, two PEO ceramics could be identified for further in vivo evaluations.

Influence of UV Irradiation and Cold Atmospheric Pressure Plasma on Zirconia Surfaces: An In Vitro Study

PURPOSE: To compare the influence of ultraviolet (UV) irradiation and cold atmospheric pressure plasma (CAP) treatment on surface structure, surface chemistry, cytocompatibility, and cell behavior on zirconia in vitro. MATERIALS AND METHODS: Zirconia samples (TZ-3YSBE) were treated by UV irradiation, oxygen plasma, or argon plasma for 12 minutes each and compared with the nontreated samples. Surface analysis was conducted using scanning electron microscopy, roughness analysis, and x-ray photoelectron spectroscopy. Cell proliferation, viability, and cell attachment as well as cytotoxicity were evaluated using MC3T3-E1 murine osteoblasts cultivated directly on the zirconia samples. RESULTS: Surface structure and roughness were not affected by the surface treatments. CAP and UV irradiation significantly reduced organic material and increased the surface oxidation on the zirconia samples. Furthermore, CAP and UV treatment significantly decreased the contact angle on the zirconia samples, indicating superhydrophilicity. Cell attachment was significantly increased on oxygen plasma-treated zirconia samples compared with the nontreated samples at all times (P < .001). After 24 and 48 hours, cell proliferation and viability (P < .001) were significantly increased on oxygen plasma-treated samples in comparison with the nontreated, UV-treated, and argon plasma-treated samples. Neither UV nor CAP treatment led to cytotoxicity. CONCLUSION: In vitro, surface treatment by UV irradiation or CAP causes a significant reduction of organic material, increases the hydrophilicity of zirconia, and improves the conditions for osteoblasts. Results stipulate that treatment of zirconia surfaces with oxygen plasma may favor cell proliferation.

Enriching Stem/Progenitor Cells from Dental Pulp Cells by Low-density Culturing.

BACKGROUND/AIM: Clonogenicity is a key feature of stem/progenitor cells. The present study aimed to enrich stem/progenitor cells from dental pulp cells by means of culturing the cells at a low clonal density with spatial separation and the evaluate differentiation potential of the surviving cells. MATERIALS AND METHODS: Pulp cells derived from wisdom teeth were seeded into wells of a 96-plate at a mean density of 1 cell/well and cultured for 2 weeks. Surviving cells were harvested, pooled together and subjected to differentiation into adipocytes, osteoblasts and neurons using respective inducing conditions for 3 weeks. The former two types of cells were examined by staining with Oil Red O and Alizarin Red, respectively. Neuron-like cells were inspected for their morphology and immunostained for microtubule-associated protein 2 and ß-tubulin III. RESULTS: Vital cells were obtained in eight wells of a 96-well plate, corresponding to a survival rate of 8%. Since fewer than two wells would be expected to contain more than four cells at seeding, the majority of surviving cells likely grew from 1-3 cells, which is a very low density. These cells differentiated into functional adipocytes and osteoblasts, and morphologically neuron-like cells. CONCLUSION: Low-density seeding with spatial separation enables statistical estimation of cell number in wells and provides an effective strategy for enriching stem/progenitor cells and for isolating clonal dental pulp cells.