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1.
J Orthop Surg Res ; 16(1): 580, 2021 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-34627321

RESUMO

BACKGROUND: Enterococcus faecalis (E. faecalis) is the most recovered species from the root canals after failed root canal treatment. Calcium phosphate bone cement (CPC) scaffold is promising for applications in endodontic treatment as a kind of root canal sealer. Graphene oxide (GO) has been extensively considered as a kind of promising nano-materials for antibacterial applications. In the present study, an injectable CPC-chitosan paste containing GO was developed for promising endodontic therapy. The antibacterial properties of this paste against E. faecalis biofilms as well as the support for human dental pulp stem cells (hDPSCs) were investigated. METHODS: CPC-chitosan composite with or without GO injectable scaffold was fabricated. The hDPSC growth and viability on scaffolds were investigated by live/dead assay. Antibacterial effects against E. faecalis biofilms were determined in clinical detin block samples. RESULTS: The antibacterial CPC-chitosan-GO disks had excellent hDPSC support with the percentages of live cells at around 90%. CPC-chitosan-GO also had greater antibacterial activity on E. faecalis than that of CPC-chitosan control using detin block models (p < 0.05). CONCLUSIONS: The injectable CPC-chitosan-GO paste had strong effects on inhibition E. faecalis and hDPSC support, which could fill the void of adjusting paste to the defect and shaping in situ for promising endodontic therapy.

2.
Mater Sci Eng C Mater Biol Appl ; 128: 112306, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34474857

RESUMO

Osteomyelitis is caused by Staphylococcus aureus (S. aureus), with associated progressive bone loss. This study developed for the first time a calcium phosphate cement (CPC) for delivery of doxycycline (DOX) and human platelet lysate (hPL) to fight against S. aureus infection and enhance the osteogenesis of human periodontal ligament stem cells (hPDLSCs). Chitosan-containing CPC scaffolds were fabricated in the absence (CPCC) or presence of DOX (CPCC+DOX). In addition, hPL was encapsulated in alginate microbeads and incorporated into CPCC+DOX (CPCC+DOX+ hPL). Flexural strength of CPCC+DOX + hPL was (5.56 ± 0.55) MPa, lower than (8.26 ± 1.6) MPa of CPCC+DOX (p < 0.05), but exceeding the reported strength of cancellous bone. CPCC+DOX and CPCC+DOX + hPL exhibited strong antibacterial activity against S. aureus, reducing biofilm CFU by 4 orders of magnitude. The hPDLSCs encapsulated in microbeads were co-cultured with the CPCs. The hPDLSCs were able to be released from the microbeads and showed a high proliferation rate, increasing by about 8 folds at 14 days for all groups. The hPL was released from the scaffold and promoted the osteogenic differentiation of hPDLSCs. ALP activity was 28.07 ± 5.15 mU/mg for CPCC+DOX + hPL, higher than 17.36 ± 2.37 mU/mg and 1.34 ± 0.37 mU/mg of CPCC+DOX and CPCC, respectively (p < 0.05). At 7 days, osteogenic genes (ALP, RUNX2, COL-1, and OPN) in CPCC+DOX + hPL were 3-10 folds those of control. The amount of hPDLSC-synthesized bone mineral with CPCC+DOX + hPL was 3.8 folds that of CPCC (p < 0.05). In summary, the novel CPC + DOX + hPL-hPDLSCs scaffold exhibited strong antibacterial activity, excellent cytocompatibility and hPDLSC osteogenic differentiation, showing a promising approach for treatment and prevention of bone infection and enhancement of bone regeneration.


Assuntos
Osteogênese , Ligamento Periodontal , Biofilmes , Fosfatos de Cálcio/farmacologia , Diferenciação Celular , Células Cultivadas , Humanos , Staphylococcus aureus , Células-Tronco
3.
Clin Oral Investig ; 2021 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-34392408

RESUMO

OBJECTIVES: A previous study showed that the combination of poly(amido amine) (PAMAM) and rechargeable composites with nanoparticles of amorphous calcium phosphate (NACP) induced dentin remineralization in an acidic solution with no initial calcium (Ca) and phosphate (P) ions, mimicking the oral condition of individuals with dry mouths. However, the frequent fluid challenge in the oral cavity may decrease the remineralization capacity. Therefore, the objective of the present study was to investigate the remineralization efficacy on dentin in an acid solution via PAMAM + NACP after fluid challenges for the first time. METHODS: The NACP nanocomposite was stored in a pH 4 solution for 77 days to exhaust its Ca and P ions and then recharged. Demineralized dentin samples were divided into four groups: (1) control dentin, (2) dentin coated with PAMAM, (3) dentin with recharged NACP composite, and (4) dentin with PAMAM + recharged NACP. PAMAM-coated dentin was shaken in phosphate-buffered saline for 77 days to desorb PAMAM from dentin. Samples were treated in pH 4 lactic acid with no initial Ca and P ions for 42 days. RESULTS: After 77 days of fluid challenge, PAMAM failed to prevent dentin demineralization in lactic acid. The recharged NACP nanocomposite raised the pH to above 6.5 and re-released more than 6.0 and 4.0 mmol/L Ca and P ions daily, respectively, which inhibited further demineralization. In contrast, the PAMAM + NACP combined method induced great dentin remineralization and restored the dentin microhardness to 0.54 ± 0.04 GPa, which approached that of sound dentin (P = 0.426, P > 0.05). CONCLUSIONS: The PAMAM + NACP combination achieved dentin remineralization in an acid solution with no initial Ca and P ions, even after severe fluid challenges. CLINICAL RELEVANCE: The novel PAMAM + NACP has a strong and sustained remineralization capability to inhibit secondary caries, even for individuals with dry mouths.

4.
J Dent ; 113: 103789, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34455017

RESUMO

OBJECTIVE: Composites with remineralizing and antibacterial properties are favorable for caries inhibition. The objectives of this study were to develop a new bioactive nanocomposite with remineralizing and antibiofilm properties by incorporating dimethylaminohexadecyl methacrylate (DMAHDM) and nano-calcium fluoride (nCaF2). METHODS: nCaF2 was produced via a spray-drying method and integrated at 15% mass fraction into composite. DMAHDM was added at 3% mass fraction. Mechanical properties and F and Ca ion releases were assessed. Colony-forming units (CFU), lactic acid and metabolic activity of biofilms on composites were performed. RESULTS: The new composites had flexural strengths of (95.28±6.32) MPa and (125.93±7.49) MPa, which were within the ISO recommendations. Biofilm CFU were reduced by 3-4 log (p<0.05). The composites achieved high F releases of (0.89±0.01) mmol/L and (0.44±0.01) mmol/L, and Ca releases of (1.46±0.05) mmol/L and (0.54±0.005) mmol/L. CONCLUSIONS: New nanocomposites were developed with good mechanical properties, potent antibacterial activity against salivary biofilms, and high F and Ca ion releases with potential for remineralization. CLINICAL SIGNIFICANCE: Novel nanocomposites using nCaF2 and DMAHDM were developed with potent antibacterial and remineralizing effects and high F and Ca ion releases. They are promising to inhibit recurrent caries, promote remineralization, and possess long-term sustainability.

5.
Acta Biomater ; 2021 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-34303014

RESUMO

The limited durability of dentin bonding harshly shortens the lifespan of resin composites restorations. The controlled, dynamic movement of materials through non-contacting forces provides exciting opportunities in adhesive dentistry. We, herein, describe comprehensive investigations of a new dental adhesive with superparamagnetic iron oxide nanoparticles (SPIONs) sensitive to magnetic fields for bonding optimization. This contribution outlines a roadmap of (1) designing and tuning of an adhesive formulation containing SPIONs to enhance penetrability into etched dentin guided by magnetic-field; (2) employing a clinically relevant model of simulated hydrostatic pulpal pressure on the microtensile bond to dentin; and (3) investigating a potential antibacterial effect of the formulated adhesives, and their biocompatibility. SPION-concentration-dependency chemical and mechanical behavior was shown via the degree of conversion, ultimate tensile strength, and micro shear bond strength to dentin. The effects of SPIONs carried on a dental adhesive on the bonding strength to dentin are studied in depth by combining experiments with in vitro simulated model. The results show that under the guided magnetic field, 0.07 wt.% of SPIONs-doped adhesive increased the bond strength that surpasses the reduction caused by hydrostatic pulpal pressure. Using a magnetic guide workflow during the bonding procedures, SPIONs-doped adhesives improved dentin's adhesion without changing adhesives' physicochemical properties. This outcome addresses the key challenge of poor resin infiltration of dentin's conventional total etching during the bonding procedure. The real-time magnetic motion of dental adhesives may open new paths to enhance resin-based restorations' longevity. STATEMENT OF SIGNIFICANCE: In this study, dental adhesives containing superparamagnetic iron oxide nanoparticles (SPIONs) were developed to enhance penetrability into dentin guided by a magnetic field. The adhesives were screened for physical, chemical, antibacterial properties, and cytotoxicity. For the first time, simulated pulpal pressure was used concurrently with the magnetic field to simulate a clinical setting. This approach showed that it is feasible to overcome pulpal pressure jeopardization on bond strength when SPIONs and a magnetic field are applied. The magnetic-responsive adhesives had great potential to improve bond strength, opening new paths to enhance resin-based restorations' longevity without affecting adhesives' biological properties. The use of magnetic-responsive particles and magnetically assisted motion is a promising strategy to improve the sealing ability of dental adhesives.

6.
Clin Oral Investig ; 25(9): 5375-5390, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33891172

RESUMO

OBJECTIVES: Dental caries is closely associated with acid-producing bacteria, and Streptococcus mutans is one of the primary etiological agents. Bacterial accumulation and dental demineralization lead to destruction of bonding interface, thus limiting the longevity of composite. The present study investigated remineralization effectiveness of adhesive containing nanoparticles of amorphous calcium phosphate (NACP) in a stimulated oral biofilm environment. METHODS: The enamel blocks were immersed in demineralization solution for 72 h to imitate artificial initial carious lesion and then subjected to a Streptococcus mutans biofilm for 24 h. All the samples then underwent 4-h demineralization in brain heart infusion broth with sucrose (BHIS) and 20-h remineralization in artificial saliva (AS) for 7 days. The daily pH of BHIS after 4-h incubation, lactic acid production, colony-forming unit (CFU) count, and content of calcium (Ca) and phosphate (P) in biofilm were evaluated. Meanwhile, the remineralization effectiveness of enamel was analyzed by X-ray diffraction (XRD), surface microhardness testing, transverse microradiography (TMR) and scanning electron microscopy (SEM). RESULTS: The NACP adhesive released abundant Ca and P, achieved acid neutralization, reduced lactic acid production, and lowered CFU count (P < 0.05). Enamel treated with NACP adhesive demonstrated the best remineralization effectiveness with remineralization value of 52.29 ± 4.79% according to TMR. Better microhardness recovery of cross sections and ample mineral deposits were also observed in NACP group. CONCLUSIONS: The NACP adhesive exhibited good performance in remineralizing initial enamel lesion with cariogenic biofilm. SIGNIFICANCE: The NACP adhesive is promising to be applied for the protection of bonding interface, prevention of secondary caries, and longevity prolonging of the restoration.


Assuntos
Cárie Dentária , Nanopartículas , Antibacterianos , Biofilmes , Fosfatos de Cálcio , Cárie Dentária/tratamento farmacológico , Suscetibilidade à Cárie Dentária , Cimentos Dentários , Humanos , Metacrilatos , Remineralização Dentária
7.
Dent Mater ; 37(6): 1009-1019, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33879343

RESUMO

OBJECTIVE: Fluoride (F)-releasing restoratives typically are either weak mechanically or release only low levels of F ions. The objectives of this study were to: (1) develop a novel photo-cured nanocomposite with strong mechanical properties and high levels of sustained F ion release via a two-step "co-precipitation + spray-drying" technique to synthesize CaF2 nanoparticles (nCaF2); and (2) investigate the effect of spray-drying treatment after co-precipitation of nCaF2 on mechanical properties and F ion release of composite. METHODS: Two types of CaF2 particles were synthesized: A co-precipitation method yielded CaF2cp; "co-precipitation + spray-drying" yielded nCaF2cpsd. Composites were fabricated with fillers of: (1) 0% CaF2 + 70% glass; (2) 10% CaF2cp + 60% glass; (3) 15% CaF2cp + 55% glass; (4) 20% CaF2cp + 50% glass; (5) 10% nCaF2cpsd + 60% glass; (6) 15% nCaF2cpsd + 55% glass; and (7) 20% nCaF2cpsd + 50% glass. A commercial F-releasing nanocomposite served as control. RESULTS: The nCaF2cpsd had much smaller particle size (median = 32 nm) and narrower distribution (22-57 nm) than CaF2cp (median = 5.25 µm, 162 nm-67 µm). The composite containing nCaF2cpsd had greater flowability, flexural strength, elastic modulus and hardness than CaF2cp composite and commercial control composite. At 84-day immersion in water, the nanocomposites containing 20% nCaF2cpsd had 65 times higher cumulative F release, and 77 times greater long-term F-release rate, than commercial control. CONCLUSIONS: A novel two-step "co-precipitation + spray-drying" technique of synthesizing nCaF2 was developed. The photo-cured nanocomposite containing 20% nCaF2cpsd possessed strong mechanical properties and excellent long-term F-release ability, and hence is promising for dental restoration applications to inhibit secondary caries.


Assuntos
Cárie Dentária , Nanocompostos , Resinas Compostas , Fluoretos , Cimentos de Ionômeros de Vidro , Humanos , Teste de Materiais
8.
Dent Mater J ; 40(3): 615-624, 2021 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-33814531

RESUMO

Human induced pluripotent stem cells (hiPSCs) are exciting for regenerative medicine due to their multi-potent differentiation. SB431542 bioactive molecule can activate bone morphogenetic protein-signalling in osteoblasts. The objectives were to: (1) develop a novel injectable calcium phosphate cement (CPC)-SB431542 scaffold for dental/craniofacial bone engineering; and (2) investigate cell proliferation and osteo-differentiation of hiPSC-derived mesenchymal stem cells (hiPSC-MSCs) on CPC-SB431542 scaffold. Three groups were tested: CPC control; CPC with SB431542 inside CPC (CPCSM); CPC with SB431542 in osteogenic medium (CPC+SMM). SB431542 in CPC promoted stem cell proliferation and viability. hiPSC-MSCs differentiated into osteogenic lineage and synthesized bone minerals. CPC with SB431542 showed much greater osteo-expressions and more bone minerals than those without SB431542. In conclusion, hiPSC-MSCs on CPC scaffold containing SB431542 showed excellent osteo-differentiation and bone mineral synthesis for the first time. CPC was a suitable scaffold for delivering stem cells and SB431542 to promote bone regeneration in dental/craniofacial applications.


Assuntos
Células-Tronco Pluripotentes Induzidas , Osteogênese , Cimentos Ósseos , Fosfatos de Cálcio/farmacologia , Diferenciação Celular , Células Cultivadas , Humanos , Engenharia Tecidual , Tecidos Suporte
9.
Materials (Basel) ; 14(5)2021 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-33668779

RESUMO

Candida albicans (C. albicans) biofilm is a common etiological factor in denture stomatitis. The purpose of this study was to investigate the effects of incorporating 2-methacryloyloxyethyl phosphorylcholine (MPC) as a protein repellent into a new high-impact denture acrylic (HIPA) resin on the surface roughness, solution pH, and C. albicans biofilm adhesion to the denture base. The new acrylic denture resin base was formulated by mixing MPC into HIPA resin at mass fractions of 1.5%, 3%, and 4.5%. Surface roughness was measured using a Mitutoyo surface roughness tester. C. albicans biofilm growth and viability were assessed via colony forming unit counts. The pH of the biofilm growth medium was measured using a digital pH meter. Adding MPC to the HIPA resin at percentages of 1.5% and 3% increased the roughness values significantly (p < 0.05), while adding 4.5% MPC resulted in no difference in roughness values to that of the control group (p > 0.05). All experimental groups demonstrated neutral pH values (pH ≅ 7) and were not significantly different from each other (p > 0.05). Incorporating 2-methacryloyloxyethyl phosphorylcholine at 4.5% resulted in a significant (≅1 log) colony-forming unit reduction compared with the control group with 0% MPC (p < 0.05). A fungal-retarding denture acrylic resin was developed through the incorporation of MPC for its protein-repelling properties. This newly developed denture acrylic material has the potential to prevent oral microbial infections, such as denture stomatitis.

10.
Mater Sci Eng C Mater Biol Appl ; 120: 111688, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33545850

RESUMO

Staphylococcus aureus (S. aureus) is the major pathogen for osteomyelitis, which can lead to bone necrosis and destruction. There has been no report on antibacterial calcium phosphate cement (CPC) against S. aureus. The aims of this study were to: (1) develop novel antibacterial CPC-chitosan-alginate microbead scaffold; (2) investigate mechanical and antibacterial properties of CPC-chitosan-penicillin-alginate scaffold; (3) evaluate the encapsulation and delivery of human umbilical cord mesenchymal stem cells (hUCMSCs). Flexural strength, elastic modulus and work-of-fracture of the CPC-chitosan-penicillin-alginate microbeads scaffold and CPC-chitosan scaffold were evaluated. Penicillin release profile and antibacterial effects on S. aureus were determined. The hUCMSC delivery and release from penicillin-alginate microbeads were investigated. Injectable CPC-chitosan-penicillin-alginate microbeads scaffold was developed for the first time. CPC-chitosan-penicillin-alginate microbeads scaffold had a flexural strength of 3.16 ± 0.55 MPa, matching that of cancellous bone. With sustained penicillin release, the new scaffold had strong antibacterial effects on S. aureus, with an inhibition zone diameter of 32.2 ± 2.5 mm, greater than that of penicillin disk control (15.1 ± 2.0 mm) (p < 0.05). Furthermore, this injectable and antibacterial scaffold had no toxic effects, yielding excellent hUCMSC viability, which was similar to that of CPC control without antibacterial activity (p > 0.05). CPC-chitosan-penicillin-microbeads scaffold had injectability, good strength, strong antibacterial effects, and good biocompatibility to support stem cell viability for osteogenesis. CPC-chitosan-penicillin-microbeads scaffold is promising for dental, craniofacial and orthopedic applications to combat infections and promote bone regeneration.


Assuntos
Células-Tronco Mesenquimais , Staphylococcus aureus , Antibacterianos/farmacologia , Cimentos Ósseos/farmacologia , Regeneração Óssea , Fosfatos de Cálcio/farmacologia , Diferenciação Celular , Humanos , Osteogênese , Células-Tronco , Engenharia Tecidual , Tecidos Suporte
11.
J Biomed Mater Res B Appl Biomater ; 109(8): 1124-1134, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33386668

RESUMO

The aims are: (a) To develop the first low-shrinkage-stress nanocomposite with antibacterial and remineralization capabilities through the incorporation of dimethylaminododecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP); (b) to investigate the effects of the new composite on biofilm inhibition, mechanical properties, shrinkage stress, and calcium (Ca) and phosphate (P) ion releases. The low-shrinkage-stress resin consisted of urethane dimethacrylate and triethylene glycol divinylbenzyl ether. Composite was formulated with 3% DMAHDM and 20% NACP. Mechanical properties, shrinkage stress, and degree of conversion were evaluated. Streptococcus mutans biofilm growth on composites was assessed. Ca and P ion releases were measured. The shrinkage stress of the low-shrinkage-stress composite containing 3% DMAHDM and 20% NACP was 36% lower than that of traditional composite control (p < 0.05), with similar degrees of conversion of 73.9%. The new composite decreased the biofilm colony-forming unit by 4 log orders and substantially reduced biofilm lactic acid production compared to control composite (p < 0.05). Incorporating DMAHDM to the low-shrinkage-stress composite did not adversely affect the Ca and P ion release. A novel bioactive nanocomposite was developed with low shrinkage stress, strong antibiofilm activity, and high levels of ion release for remineralization, without undermining the mechanical properties and degree of conversion.

12.
J Biomater Sci Polym Ed ; 32(7): 886-905, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33482702

RESUMO

A low-shrinkage-stress (LSS), antibacterial and remineralizing nanocomposite was recently developed; however, validation of its long-term antibacterial potency in modulating human salivary-derived biofilm is an unmet need. This study aimed to evaluate the antibacterial effect of the bioactive LSS composite before and after aging in acidic solution for 90 days using a multi-species biofilm model, and to evaluate its cytotoxicity. The LSS composite consisted of urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE), 3% dimethylaminohexadecyl methacrylate (DMAHDM) and 20% nanoparticles of amorphous calcium phosphate (NACP). Biofilm colony-forming units (CFU), lactic acid production, and confocal laser scanning microscopy (3D biofilm) were evaluated before and after three months of aging. Cytotoxicity was assessed against human gingival fibroblasts (HGF). The new LSS composite presented the lowest biofilm CFU, lactic acid and biofilm biomass, compared to controls (n = 6, p < 0.05). Importantly, the new composite exhibited no significant difference in antibacterial performance before and after 90-day-aging, demonstrating long-term antibacterial activity (p > 0.1). The LSS antibacterial and remineralizing composite presented a low cell viability at original extract that has increased with further dilutions. In conclusion, this study spotlighted that the new bioactive composite not only had a low shrinkage stress, but also down-regulated the growth of oral biofilms, reduced acid production, maintained antibacterial activity after the 90-day-aging, and did not compromise the cytocompatibility.

13.
J Tissue Eng Regen Med ; 15(3): 232-243, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33434402

RESUMO

Infectious bone defects remain a significant challenge in orthopedics and dentistry. Calcium phosphate cement (CPC) have attracted significant interest in use as local drug delivery system, which with great potential to control release of antibiotics for the treatment of infectious bone defects. Within the current study, a novel antibacterial scaffold of chitosan-reinforced calcium phosphate cement delivering doxycycline hyclate (CPCC + DOX) was developed. Furthermore, the capacity of CPCC + DOX scaffolds for bone regeneration was enhanced by the human periodontal ligament stem cells (hPDLSCs) encapsulated in alginate beads. CPCC + DOX scaffolds were fabricated to contain different concentrations of DOX. Flexural strength of CPCC + DOX ranged from 5.56 ± 0.70 to 6.2 ± 0.72 MPa, which exceeded the reported strength of cancellous bone. Scaffolds exhibited continual DOX release, reaching 80% at 21 days. Scaffold with 5 mg/ml DOX (CPCC + DOX5mg) had a strong antibacterial effect, with a 4-log colony forming unit reduction against S. aureus and P. gingivalis. The proliferation and osteogenic differentiation of hPDLSCs encapsulated in alginate hydrogel microbeads were investigated in culture with CPCC + DOX scaffolds. CPCC + DOX5mg had no negative effect on proliferation of hPDLSCs. Alkaline phosphatase activity, mineral synthesis, and osteogenic gene expressions for CPCC + DOX5mg group were much higher than control group. DOX did not compromise the osteogenic induction. In summary, the novel CPCC + DOX scaffold exhibited excellent mechanical properties and strong antibacterial activity, while supporting the proliferation and osteogenic differentiation of hPDLSCs. The CPCC + DOX + hPDLSCs construct is promising to enhance bone regeneration and combat bone infections in dental, craniofacial, and orthopedic applications.

14.
J Dent ; 104: 103529, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33189801

RESUMO

OBJECTIVES: This study aims to investigate the long-term demineralization-inhibition capability of a rechargeable adhesive with nanoparticles of amorphous calcium phosphate (NACP) on dentin in a biofilm-challenged environment. METHODS: The NACP adhesive was immersed in a pH 4 solution to exhaust calcium (Ca) and phosphate (P) ions and then recharged with Ca and P ions. Dentin samples were demineralized underStreptococcus mutans biofilms for 24 h and randomly divided into two groups: (1) dentin control, (2) dentin with recharged NACP adhesives. Each day, all the samples were immersed in brain heart infusion broth with 1% sucrose (BHIS) for 4 h, and then in artificial saliva (AS) for 20 h. This cycle was repeated for 10 days. The pH of BHIS, the Ca and P ions content of the BHIS and AS were measured daily. After 10 days, the lactic acid production and colony-forming units of the biofilms were tested. The changes of remineralization/demineralization were also analyzed. RESULTS: Dentin in the control group showed further demineralization. The recharged NACP adhesive neutralized acids, increasing the pH to above 5, and released large amounts of Ca and P ions each day. The recharged NACP adhesive decreased the production of lactic acid (P < 0.05), inhibited dentin demineralization and sustained the dentin hardness in the biofilm-challenged environment, showing an excellent long-term demineralization-inhibition capability. CONCLUSIONS: The NACP adhesive could continuously inhibit dentin demineralization in a biofilm-challenged environment by recharging with Ca and P ions. SIGNIFICANCE: The rechargeable NACP adhesive could provide long-term dentin bond protection.


Assuntos
Nanopartículas , Desmineralização do Dente , Antibacterianos , Biofilmes , Fosfatos de Cálcio/farmacologia , Cimentos Dentários/farmacologia , Dentina , Humanos , Metacrilatos , Desmineralização do Dente/prevenção & controle
15.
Tissue Eng Part A ; 2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33261521

RESUMO

Calcium phosphate cement (CPC) is promising for bone and dentin repair and regeneration. However, there has been no report of biphasic CPC for inducing dentin regeneration. The aim of this study was to develop a novel biphasic CPC containing ß-tricalcium phosphate (ß-TCP), and investigate its effects on odontogenic differentiation of human dental pulp stem cells (hDPSCs) and macrophage polarization. New biphasic CPC was formulated with different ratios of ß-TCP to an equimolar mixture of tetracalcium phosphate and dicalcium phosphate anhydrous. Mechanical properties, biocompatibility, and odontogenic differentiation induction ability of the cements and the inflammatory reaction to the cements were examined. A series of CPC containing ß-TCP were developed. CPC with 20% ß-TCP exhibited homogeneity and injectability, an acceptable setting time, and a twofold increase in compressive strength. Significant increases in hDPSCs' alkaline phosphatase activity, mineral deposit, DMP1 and DSPP gene, and protein expressions were obtained for 20% TCP-CPC, compared with traditional CPC (p < 0.01). The addition of ß-TCP did not promote macrophage polarization to the proinflammation phenotype. The addition of 10% and 20% ß-TCP promoted macrophage polarization to the anti-inflammatory phenotype. In conclusion, a biphasic ß-TCP-modified CPC was developed for the first time, demonstrating substantially increased dentin regeneration capability, while promoting macrophages to an anti-inflammation phenotype. The novel biphasic CPC is promising for tooth tissue engineering and dentin regeneration applications.

16.
Dent Mater ; 37(3): e182-e193, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33342588

RESUMO

OBJECTIVE: Various nanoparticles are currently under investigation to impart biointeractivity for dental materials. This study aimed to: (1) formulate an experimental dental adhesive containing ZnO nanoparticles; (2) evaluate its chemical and mechanical properties; and (3) assess the antibacterial response against oral microcosm biofilm. METHODS: Nanosized ZnO was chemically and morphologically evaluated. ZnO was incorporated at 0 (GCTRL), 2.5 (G2.5%), 5 (G5%) and 7.5 (G5%) wt.% in an experimental dental adhesive. The adhesives were evaluated for the degree of conversion (DC), flexural strength (FS), and elastic modulus (E). The antibacterial activity was evaluated using a 48h-microcosm biofilm model after the formation of acquired pellicle on samples' surfaces. Colony-forming units (CFU), metabolic activity, and live/dead staining were assessed. RESULTS: Nanosized ZnO presented characteristic peaks of Zn-O bonds, and the particles were arranged in agglomerates. The DC ranged from 62.21 (±1.05) % for GCtrl to 46.15 (±1.23) % for G7.5% (p<0.05). G7.5% showed lower FS compared to all groups (p<0.05). Despite achieving higher E (p<0.05), G2.5% did not show differences for GCtrl regarding the FS (p>0.05). G7.5% had lower CFU/mL compared to GCtrl for mutans streptococci (p<0.05) and total microorganisms (p<0.05), besides presenting lower metabolic activity (p<0.05) and higher dead bacteria via biofilm staining. SIGNIFICANCE: The dental adhesives' physicochemical properties were similar to commercial adhesives and in compliance with ISO recommendations. G7.5% restricted the growth of oral microcosm biofilm without impairing the physicochemical performance.


Assuntos
Óxido de Zinco , Antibacterianos/farmacologia , Biofilmes , Cimentos Dentários , Teste de Materiais , Metacrilatos , Cimentos de Resina , Streptococcus mutans , Óxido de Zinco/farmacologia
17.
Materials (Basel) ; 13(21)2020 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-33158111

RESUMO

Decays in the roots of teeth is prevalent in seniors as people live longer and retain more of their teeth to an old age, especially in patients with periodontal disease and gingival recession. The objectives of this study were to develop a biocompatible nanocomposite with nano-sized calcium fluoride particles (Nano-CaF2), and to investigate for the first time the effects on osteogenic and cementogenic induction of periodontal ligament stem cells (hPDLSCs) from human donors.Nano-CaF2 particles with a mean particle size of 53 nm were produced via a spray-drying machine.Nano-CaF2 was mingled into the composite at 0%, 10%, 15% and 20% by mass. Flexural strength (160 ± 10) MPa, elastic modulus (11.0 ± 0.5) GPa, and hardness (0.58 ± 0.03) GPa for Nano-CaF2 composite exceeded those of a commercial dental composite (p < 0.05). Calcium (Ca) and fluoride (F) ions were released steadily from the composite. Osteogenic genes were elevated for hPDLSCs growing on 20% Nano-CaF2. Alkaline phosphatase (ALP) peaked at 14 days. Collagen type 1 (COL1), runt-related transcription factor 2 (RUNX2) and osteopontin (OPN) peaked at 21 days. Cementogenic genes were also enhanced on 20% Nano-CaF2 composite, promoting cementum adherence protein (CAP), cementum protein 1 (CEMP1) and bone sialoprotein (BSP) expressions (p < 0.05). At 7, 14 and 21 days, the ALP activity of hPDLSCs on 20% Nano-CaF2 composite was 57-fold, 78-fold, and 55-fold greater than those of control, respectively (p < 0.05). Bone mineral secretion by hPDLSCs on 20% Nano-CaF2 composite was 2-fold that of control (p < 0.05). In conclusion, the novel Nano-CaF2 composite was biocompatible and supported hPDLSCs. Nano-CaF2 composite is promising to fill tooth root cavities and release Ca and F ions to enhance osteogenic and cementogenic induction of hPDLSCs and promote periodontium regeneration.

18.
J Tissue Eng Regen Med ; 14(12): 1779-1791, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33025745

RESUMO

Staphylococcus aureus (S. aureus) is a major pathogen for osteomyelitis. Calcium phosphate bone cement (CPC) paste is promising for orthopedic uses. Nanostructured graphene oxide (GO) showed antibacterial effect on Gram-positive bacteria. However, there has been no report of incorporating GO into CPC. The objectives of this study were to (a) develop an injectable and mechanically strong CPC-chitosan paste containing GO and (b) investigate the inhibition of S. aureus infection and the promotion of human umbilical cord mesenchymal stem cells (hUCMSCs) for bone regeneration. Injectable CPC-chitosan-GO paste was fabricated. Flexural strength, elastic modulus, and work-of-fracture of the CPC-chitosan and CPC-chitosan-GO bars were evaluated. Antibacterial effects against S. aureus biofilms were determined. hUCMSC growth and viability on disks were investigated. CPC-chitosan-GO bars had a flexural strength of 7.2 ± 1.6 MPa, matching that of CPC-chitosan control without GO. CPC-chitosan-GO had strong antibacterial effects on S. aureus, with an inhibition zone of 55.2 ± 2.5 mm, greater than that of CPC-chitosan control (30.1 ± 2.0 mm) (p < 0.05). CPC-chitosan-GO had potent antibacterial activity on S. aureus biofilms in vitro (p > 0.05). The injectable and antibacterial CPC-GO paste had no toxic effect, yielding excellent hUCMSC growth and viability on disks. The CPC-chitosan-GO had injectability, good strength, strong antibacterial effects, and excellent stem cell attachment and growth. CPC-chitosan-GO is promising for dental, craniofacial, and orthopedic applications to control infections and good biocompatibility to support stem cell viability to enhance bone regeneration.

19.
Int J Mol Sci ; 21(20)2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-33076241

RESUMO

Caries-related biofilms and associated complications are significant threats in dentistry, especially when biofilms grow over dental restorations. The inhibition of cariogenic biofilm associated with the onset of carious lesions is crucial for preventing disease recurrence after treatment. This in vitro study defined optimized parameters for using a photosensitizer, toluidine blue O (TBO), activated via a red light-emitting diode (LED)-based wireless device to control the growth of cariogenic biofilms. The effect of TBO concentrations (50, 100, 150, and 200 µg/mL) exposed to light or incubated in the dark was investigated in successive cytotoxicity assays. Then, a mature Streptococcus mutans biofilm model under sucrose challenge was treated with different TBO concentrations (50, 100, and 150 µg/mL), different light energy doses (36, 108, and 180 J/cm2), and different incubation times before irradiation (1, 3, and 5 min). The untreated biofilm, irradiation with no TBO, and TBO incubation with no activation represented the controls. After treatments, biofilms were analyzed via S. mutans colony-forming units (CFUs) and live/dead assay. The percentage of cell viability was within the normal range compared to the control when 50 and 100 µg/mL of TBO were used. Increasing the TBO concentration and energy dose was associated with biofilm inhibition (p < 0.001), while increasing incubation time did not contribute to bacterial elimination (p > 0.05). Irradiating the S. mutans biofilm via 100 µg/mL of TBO and ≈180 J/cm2 energy dose resulted in ≈3-log reduction and a higher amount of dead/compromised S. mutans colonies in live/dead assay compared to the control (p < 0.001). The light energy dose and TBO concentration optimized the bacterial elimination of S. mutans biofilms. These results provide a perspective on the determining parameters for highly effective photo-killing of caries-related biofilms and display the limitations imposed by the toxicity of the antibacterial photodynamic therapy's chemical components. Future studies should support investigations on new approaches to improve or overcome the constraints of opportunities offered by photodynamic inactivation of caries-related biofilms.


Assuntos
Biofilmes/efeitos da radiação , Luzes de Cura Dentária , Cárie Dentária/terapia , Streptococcus mutans/efeitos da radiação , Animais , Contagem de Colônia Microbiana , Cárie Dentária/microbiologia , Relação Dose-Resposta à Radiação , Camundongos , Fármacos Fotossensibilizantes/efeitos adversos , Células RAW 264.7 , Streptococcus mutans/patogenicidade , Streptococcus mutans/fisiologia , Cloreto de Tolônio/efeitos adversos
20.
Nanomaterials (Basel) ; 10(10)2020 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-33050559

RESUMO

Oral biofilm accumulation at the tooth-restoration interface often leads to recurrent dental caries and restoration failure. The objectives of this study were to: (1) develop a novel bioactive crown cement containing dimethylaminohexadecyl methacrylate (DMAHDM) and nano-sized amorphous calcium phosphate (NACP), and (2) investigate the mechanical properties, anti-biofilm activity, and calcium (Ca2+) and phosphate (PO43-) ion release of the crown cement for the first time. The cement matrix consisted of pyromellitic glycerol dimethacrylate and ethoxylated bisphenol-A dimethacrylate monomers and was denoted PEHB resin matrix. The following cements were tested: (1) RelyX luting cement (commercial control); (2) 55% PEHB + 45% glass fillers (experimental control); (3) 55% PEHB + 20% glass + 25% NACP + 0% DMAHDM; (4) 52% PEHB + 20% glass + 25% NACP + 3% DMAHDM; (5) 51% PEHB + 20% glass + 25% NACP + 4% DMAHDM; (6) 50% PEHB + 20% glass + 25% NACP + 5% DMAHDM. Mechanical properties and ion release were measured. Streptococcusmutans (S. mutans) biofilms were grown on cements, and colony-forming units (CFUs) and other biofilm properties were measured. The novel bioactive cement demonstrated strong antibacterial properties and high levels of Ca2+ and PO43- ion release to remineralize tooth lesions. Adding NACP and DMAHDM into the cement did not adversely affect the mechanical properties and dentin bonding strength. In conclusion, the novel NACP + DMAHDM crown cement has excellent potential for restoration cementation to inhibit caries by suppressing oral biofilm growth and increasing remineralization via Ca2+ and PO43- ions. The NACP + DMAHDM composition may have wide applicability to other biomaterials to promote hard-tissue formation and combat bacterial infection.

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