ABSTRACT
Bacterial resistance has become a serious public health problem in recent years, thus encouraging the search for new antimicrobial agents. Here, we report an antimicrobial peptide (AMP), called PEPAD, which was designed based on an encrypted peptide from a Kunitz-type plant peptidase inhibitor. PEPAD was capable of rapidly inhibiting and eliminating numerous bacterial species at micromolar concentrations (from 4 µM to 10 µM), with direct membrane activity. It was also observed that the peptide can act synergistically with ciprofloxacin and showed no toxicity in the G. mellonella in vivo assay. Circular dichroism assays revealed that the peptide's secondary structure adopts different scaffolds depending on the environment in which it is inserted. In lipids mimicking bacterial cell membranes, PEPAD adopts a more stable α-helical structure, which is consistent with its membrane-associated mechanism of action. When in contact with lipids mimicking mammalian cells, PEPAD adopts a disordered structure, losing its function and suggesting cellular selectivity. Therefore, these findings make PEPAD a promising candidate for future antimicrobial therapies with low toxicity to the host.
ABSTRACT
OBJECTIVE: This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1 mg/mL CaneCPI-5 (sugarcane-derive cystatin), 1.88 × 10- 5M StN15 (statherin-derived peptide) and 1.0 mg/mL hemoglobin (Hb) to change the protein profile of the acquired enamel pellicle(AEP) and the microbiome of the enamel biofilm. The in vitro, was designed to reveal the effects of Comb on the viability and bacterial composition of the microcosm biofilm, as well as on enamel demineralization. MATERIALS AND METHODS: In vivo study, 10 participants rinsed (10mL,1 min) with either deionized water (H2O-control) or Comb. AEP and biofilm were collected after 2 and 3 h, respectively, after rinsing. AEP samples underwent proteomics analysis, while biofilm microbiome was assessed via 16 S-rRNA Next Generation Sequencing(NGS). In vitro study, a microcosm biofilm protocol was employed. Ninety-six enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS(negative-control), 2)0.12%Chlorhexidine, 3)500ppmNaF and 4)Comb. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed. RESULTS: The proteomic results revealed higher quantity of proteins in the Comb compared to control associated with immune system response and oral microbial adhesion. Microbiome showed a significant increase in bacteria linked to a healthy microbiota, in the Comb group. In the in vitro study, Comb group was only efficient in reducing mineral-loss and lesion-depth compared to the PBS. CONCLUSIONS: The AEP modification altered the subsequent layers, affecting the initial process of bacterial adhesion of pathogenic and commensal bacteria, as well as enamel demineralization. CLINICAL RELEVANCE: Comb group shows promise in shaping oral health by potentially introducing innovative approaches to prevent enamel demineralization and deter tooth decay.
Subject(s)
Dental Caries , Tooth Demineralization , Humans , Dental Pellicle/chemistry , Dental Pellicle/microbiology , Dental Caries/prevention & control , Proteomics , Biofilms , Hemoglobins/analysis , Tooth Demineralization/prevention & controlABSTRACT
INTRODUCTION: This study investigated the changes in the acquired enamel pellicle (AEP) proteome when this integument is formed in vivo after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), and a statherin-derived peptide (StN15), or their combination and then exposed to an intrinsic acid challenge. The effectiveness of these treatments in preventing intrinsic erosion was also evaluated. METHODS: Ten volunteers, after prophylaxis, in 5 crossover phases, rinsed with the following solutions (10 mL, 1 min): control (deionized water-H2O) - group 1, 0.1 mg/mL CaneCPI-5 - group 2, 1.0 mg/mL HB - group 3, 1.88 × 10-5
Subject(s)
Calcium , Tooth Erosion , Humans , Calcium/metabolism , Dental Pellicle , Peptides , Proteome , Tooth Erosion/prevention & control , Hemoglobins/metabolismABSTRACT
OBJECTIVES: This study evaluated the protective impact of acquired enamel pellicle (AEP) engineering with statherin-derived peptide (StatpSpS), considering different AEP formation times. DESIGN: A total of 120 native human enamel specimens were divided into 2 main groups: 1) No AEP engineering and 2) AEP engineering with StatpSpS (pretreatment for 1 min; 37 °C, under agitation). Each group was further divided into 4 subgroups: No pellicle, or 1, 60-and-120 min AEP formation times (human saliva; 37 °C). The specimens were then subjected to an erosive challenge (1% citric acid; pH 3.6; 1 min; 25 °C). This procedure was repeated for 5 cycles. Relative surface reflection intensity (%SRI) was measured and scanning electron microscopy (SEM) of the enamel surface was done. RESULTS: All AEP engineering groups protected against initial dental erosion in comparison with No pellicle (p < 0.001), likewise all groups with AEP, independent of engineering or formation times (p 0.001). Furthermore, engineering with StatpSpS even without the presence of AEP protected the enamel when compared to the No engineering/No pellicle group (p < 0.0001). No difference was observed regarding the protection from the different AEP formation times (p > 0.05). Regarding the SEM analysis, in the "No AEP engineering & No AEP" group, a more severe effect of citric acid was observed, with more enamel prism heads and scratches on the surface when compared with the other groups. CONCLUSIONS: AEP provides almost instant protection at formation times even as short as 1 min, protecting the native enamel against erosion. Treatment with StatpSpS by itself provides similar protection as the AEP.
Subject(s)
Tooth Erosion , Humans , Dental Pellicle , Tooth Erosion/prevention & control , Dental Enamel , Peptides/pharmacology , Citric Acid/pharmacologyABSTRACT
The effect of solutions containing a statherin-derived peptide (Stn15pSpS) on the protection against enamel erosion in vitro was evaluated. Bovine enamel specimens were divided into 4 groups (n = 15/group): (1) deionized water (negative control), (2) Elmex Erosion Protection™ (positive control), (3) 1.88 × 10-5 M Stn15pSpS, and (4) 3.76 × 10-5 M Stn15pSpS. The solutions were applied on the specimens for 1 min. Stimulated saliva was collected from 3 donors and used to form a 2-h acquired pellicle on the specimens. Then, the specimens were submitted to an erosive pH-cycling protocol 4 times/day, for 7 days (0.01 M HCl pH 2.0/45 s, artificial saliva/2 h, and artificial saliva overnight). The solutions were applied again during pH-cycling, 2 times/day for 1 min after the first and last erosive challenges. Enamel loss (µm) was assessed by contact profilometry. Data were analyzed by Kruskal-Wallis and Dunn's test (p < 0.05). The best protection against erosion was conferred by Elmex Erosion Protection that significantly differed from all the other treatments, followed by the solutions containing Stn15pSpS, regardless of the concentration. However, 3.76 × 10-5 M Stn15pSpS did not differ from the negative control. The solution containing the lower concentration of Stn15pSpS protected against erosion in vitro, which should be confirmed using protocols that more closely resemble the clinical condition.
Subject(s)
Tooth Erosion , Animals , Cattle , Humans , Dental Enamel , Fluorides/pharmacology , Saliva, Artificial/pharmacology , Tooth Erosion/prevention & control , Salivary Proteins and Peptides/pharmacologyABSTRACT
The incorrect use of conventional drugs for both prevention and control of intestinal infections has contributed to a significant spread of bacterial resistance. In this way, studies that promote their replacement are a priority. In the last decade, the use of antimicrobial peptides (AMP), especially Ctx(Ile21)-Ha AMP, has gained strength, demonstrating efficient antimicrobial activity (AA) against pathogens, including multidrug-resistant bacteria. However, gastrointestinal degradation does not allow its direct oral application. In this research, double-coating systems using alginate microparticles loaded with Ctx(Ile21)-Ha peptide were designed, and in vitro release assays simulating the gastrointestinal tract were evaluated. Also, the AA against Salmonella spp. and Escherichia coli was examined. The results showed the physicochemical stability of Ctx(Ile21)-Ha peptide in the system and its potent antimicrobial activity. In addition, the combination of HPMCAS and chitosan as a gastric protection system can be promising for peptide carriers or other low pH-sensitive molecules, adequately released in the intestine. In conclusion, the coated systems employed in this study can improve the formulation of new foods or biopharmaceutical products for specific application against intestinal pathogens in animal production or, possibly, in the near future, in human health.
Subject(s)
Anti-Infective Agents , Chitosan , Animals , Humans , Chitosan/chemistry , Alginates/chemistry , Antimicrobial Peptides , Anti-Infective Agents/pharmacologyABSTRACT
The effect of gels containing a statherin-derived peptide (Stn) on the protection against enamel and dentin erosive tooth wear (ETW) in vitro was evaluated. Bovine enamel and dentin specimens were divided into 2 groups (n = 15 and 18/group for enamel and dentin, respectively) that were treated with Chitosan or Carboxymethylcellulose (CMC) gels containing Stn15pSpS at 1.88 × 10-5 M or 3.76 × 10-5 M. Chitosan or CMC gels without active ingredients served as negative controls, while chitosan gel containing 1.23% F (as NaF) and acidulated phosphate fluoride gel (1.23% F) served as positive controls. The gels were applied on the specimens for 4 min. Stimulated saliva was collected from 3 donors and used to form a 2-h acquired pellicle on the specimens. Then, the specimens were submitted to an erosive pH cycling protocol 4 times/day for 7 days (0.01 M HCl pH 2.0/45 s, artificial saliva/2 h, and artificial saliva overnight). The gels were applied again during pH cycling, 2 times/day for 4 min after the first and last erosive challenges. Enamel and dentin loss (µm) were assessed by contact profilometry. Scanning electron microscopy (SEM) was analyzed using a cold field emission. Data were analyzed by two-way ANOVA (for chitosan and CMC gels, separately) and Tukey's multiple comparison test. SEM images showed changes to enamel topography after application oft the gels containing Stn or F. Regarding CMC-based gels, for enamel, none of the treatments significantly reduced ETW in comparison with placebo; for dentin, however, gels containing Stn, regardless the concentration, significantly reduced the ETW. Moreover, Chitosan-based gels, regardless the Stn concentration, were able to protect enamel and dentin against ETW. Gels containing Stn might be a new approach to protect against ETW.
Subject(s)
Chitosan , Tooth Erosion , Tooth Wear , Cattle , Animals , Tooth Erosion/prevention & control , Tooth Erosion/drug therapy , Saliva, Artificial , Chitosan/pharmacology , Gels , Dentin , Peptides/pharmacology , Dental Enamel , FluoridesABSTRACT
Antimicrobial peptides (AMPs) are promising tools for developing new antibiotics. We described the design of IKR18, an AMP designed with the aid of computational tools. IKR18 showed antimicrobial activity against Gram-negative and Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). CD studies revealed that IKR18 assumes an alpha-helical structure in the membrane-mimetic environment. The action mechanism IKR18 involves damage to the bacteria membrane, as demonstrated by Sytox green uptake. Furthermore, IKR18 displayed synergic and additive effects in combination with antibiotics ciprofloxacin and vancomycin. The peptide showed anti-biofilm activity in concentration and efficiency compared with commercial antibiotics, involving the direct death of bacteria, as confirmed by scanning electron microscopy. The anti-infective activity of IKR18 was demonstrated in the Galleria mellonella model infected with S. aureus, MRSA, and Acinetobacter baumannii. The novel bioinspired peptide, IKR18, proved to be effective in the control of bacterial infection, opening opportunities for the development of further assays, including preclinical models.
Subject(s)
Anti-Infective Agents , Methicillin-Resistant Staphylococcus aureus , Moths , Animals , Antimicrobial Peptides , Staphylococcus aureus , Microbial Sensitivity Tests , Antimicrobial Cationic Peptides/pharmacology , Antimicrobial Cationic Peptides/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , BacteriaABSTRACT
OBJECTIVE: To study the proteomic alterations in the initial AEP after rinsing with CaneCPI-5, StN15 or Hb or their combination. MATERIALS AND METHODS: In five crossover phases, after prophylaxis, 10 volunteers in 5 consecutive days, rinsed (10 mL, 1 min) with the following solutions: deionized water (H2O- negative control- 1), 0.1 mg/mL CaneCPI-5 (2), 1.88×10-5 M StN15 (3), 1.0 mg/mL Hb (4) or their combination (5). The AEP formed after 3 min was collected with electrode filter papers soaked in 3% citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. RESULTS: Rinsing with the proteins/peptide increased the amounts of proteins in the AEP. The total numbers of proteins identified after rinsing with CaneCPI-5, StN15, Hb or their combination versus water, were 131, 167, 148 and 142, respectively. The treatment with the proteins/peptide or their combination increased proteins that bind calcium, phosphate and interact with distinct proteins, as well as proteins with antimicrobial and acid-resistant properties, such as, Cornifin-B (7.7, 12.6, and 4.3-fold for CaneCPI-5, StN15 and Hb, respectively), isoforms of Cystatin (2.2-2.4-fold for CaneCPI-5 and StN15), Proline-rich-protein 4 (4.3-fold; StN15), Histatin-1 (2.8-fold; StN15) and Hemoglobin (7.7-25-fold for Hb and Combination). Immunoglobulin, Keratin and Histone were exclusively identified upon treatment with the proteins/peptide, alone or combined. CONCLUSION: Rinsing with proteins/peptide, alone or combined, increased protective proteins in the initial AEP. CLINICAL RELEVANCE: Our results suggest that rinsing with the proteins/peptide or their combination increases the proteins capable of enhancing the protective function of the basal layer of AEP.
Subject(s)
Proteins , Proteomics , Dental Pellicle/chemistry , Humans , Peptides , WaterABSTRACT
Changes in the proteomic profile of the acquired enamel pellicle (AEP) formed for 3 min or 2 h after rinsing with a peptide containing the 15 N-terminal residues of statherin, with serines 2 and 3 phosphorylated (StatpSpS), were evaluated. Nine volunteers participated in 2 consecutive days. Each day, after professional tooth cleaning, they rinsed for 1 min with 10 mL of phosphate buffer containing 1.88 × 10-5 M StatpSpS or phosphate buffer only (control). The acquired pellicle formed on enamel after 3 min or 2 h was collected with electrode filter papers soaked in 3% citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. In the 3-min AEP, 19 and 131 proteins were uniquely identified in the StatpSpS and control groups, respectively. Proteins typically found in the AEP were only found in the latter. Only 2 proteins (neutrophil defensins) were increased upon treatment with StatpSpS, while 65 proteins (among which are several typical AEP proteins) were decreased. In the 2-h AEP, 50 and 108 proteins were uniquely found in StatpSpS and control groups, respectively. Hemoglobin subunits and isoforms of keratin were only found in the StatpSpS group, while cystatin-C, cathepsin D, and cathepsin G, isoforms of heat shock 70 and protocadherin were exclusively found in the control group. In addition, 23 proteins were increased upon treatment with StatpSpS, among which are histatin-1, serum albumin, and isoforms of neutrophil defensin and keratin, while 77 were decreased, most of them were typical AEP proteins. In both evaluated periods, rinsing with StatpSpS profoundly changed the proteomic profile of the AEP, which might impact the protective role of this integument against carious or erosive demineralization. This study provides important insights on the dynamics of the protein composition of the AEP along time, after rinsing with a solution containing StatpSpS.
Subject(s)
Proteome , Proteomics , Dental Enamel , Dental Pellicle , Humans , PeptidesABSTRACT
BACKGROUND: Antimicrobial peptides (AMPs) are molecules with potential application for the treatment of microorganism infections. We, herein, describe the structure, activity, and mechanism of action of RQ18, an α-helical AMP that displays antimicrobial activity against Gram-positive and Gram-negative bacteria, and yeasts from the Candida genus. METHODS: A physicochemical-guided design assisted by computer tools was used to obtain our lead peptide candidate, named RQ18. This peptide was assayed against Gram-positive and Gram-negative bacteria, yeasts, and mammalian cells to determine its selectivity index. The secondary structure and the mechanism of action of RQ18 were investigated using circular dichroism, large unilamellar vesicles, and molecular dynamic simulations. RESULTS: RQ18 was not cytotoxic to human lung fibroblasts, peripheral blood mononuclear cells, red blood cells, or Vero cells at MIC values, exhibiting a high selectivity index. Circular dichroism analysis and molecular dynamic simulations revealed that RQ18 presents varying structural profiles in aqueous solution, TFE/water mixtures, SDS micelles, and lipid bilayers. The peptide was virtually unable to release carboxyfluorescein from large unilamellar vesicles composed of POPC/cholesterol, model that mimics the eukaryotic membrane, indicating that vesicles' net charges and the presence of cholesterol may be related with RQ18 selectivity for bacterial and fungal cell surfaces. CONCLUSIONS: RQ18 was characterized as a membrane-active peptide with dual antibacterial and antifungal activities, without compromising mammalian cells viability, thus reinforcing its therapeutic application. GENERAL SIGNIFICANCE: These results provide further insight into the complex process of AMPs interaction with biological membranes, in special with systems that mimic prokaryotic and eukaryotic cell surfaces.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Cholesterol/pharmacology , Phospholipids/pharmacology , Pore Forming Cytotoxic Proteins/pharmacology , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/chemistry , Antifungal Agents/chemical synthesis , Antifungal Agents/chemistry , Candida/drug effects , Cholesterol/chemistry , Escherichia coli/drug effects , Eukaryotic Cells/drug effects , Humans , Microbial Sensitivity Tests , Molecular Dynamics Simulation , Phospholipids/chemistry , Pore Forming Cytotoxic Proteins/chemical synthesis , Pore Forming Cytotoxic Proteins/chemistry , Staphylococcus/drug effectsABSTRACT
OBJECTIVE: To compare bone marrow aspirate concentrate (BMAC) with the standard treatment for gluteal tendinopathies. METHODS: 48 patients diagnosed with gluteal tendinopathy at a university hospital were selected by a randomized clinical trial and divided into two groups: (G1) bone marrow aspirate concentrate and (G2) corticosteroid injections. RESULTS: 40 of the 48 selected patients were monitored for six months and both groups showed better scores. Visual analog scale (VAS) scores and Lequesne index were statistically significant higher in patients submitted to BMAC treatment when compared to standard treatment. Both groups improved their quality of life, without statistically significant difference. CONCLUSION: BMAC constitutes an alternative to gluteal tendinopathy standard treatment, proving to be a safe technique with promising results when combined with multidisciplinary team behavioral therapy. Level of Evidence II, Randomized Clinical Trial.
OBJETIVO: Estudo comparativo entre tratamento com corticóide e aspirado de medula óssea concentrado (BMAC) para o tratamento de tendinopatias glúteas. MÉTODOS: O ensaio clínico randomizado selecionou pacientes diagnosticados com tendinopatia glútea e os dividiu em dois grupos: (G1) aspirado de medula óssea concentrada e (G2) injeção de corticosteróide. RESULTADOS: Foram selecionados 48 pacientes, dos quais 40 foram monitorados por 6 meses, com melhora nos escores nos dois grupos. Os pacientes que foram submetidos ao tratamento com BMAC tiveram uma melhora estatisticamente significativa nos escores de EVA e nos escores de Lequesne em comparação ao tratamento padrão. Houve uma melhora na avaliação da qualidade de vida em ambos os grupos, sem diferença estatisticamente significativa. CONCLUSÃO: O aspirado de medula óssea concentrada surge como uma alternativa ao tratamento padrão da tendinopatia glútea, provando ser uma técnica segura e com resultados promissores quando combinada à terapia comportamental de equipe multidisciplinar. Nível de Evidência II, O ensaio clínico randomizado.
ABSTRACT
BACKGROUND: Antimicrobial resistance poses substantial risks to human health. Thus, there is an urgent need for novel antimicrobial agents, including alternative compounds, such as peptides derived from bacterial toxin-antitoxin (TA) systems. ParELC3 is a synthetic peptide derived from the ParE toxin reported to be a good inhibitor of bacterial topoisomerases and is therefore a potential antibacterial agent. However, ParELC3 is inactive against bacteria due to its inability to cross the bacterial membranes. To circumvent this limitation we prepared and used rhamnolipid-based liposomes to carry and facilitate the passage of ParELC3 through the bacterial membrane to reach its intracellular target - the topoisomerases. METHODS AND RESULTS: Small unilamellar liposome vesicles were prepared by sonication from three formulations that included 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and cholesterol. ParELC3 was loaded with high efficiency into the liposomes. Characterization by DLS and TEM revealed the appropriate size, zeta potential, polydispersity index, and morphology. In vitro microbiological experiments showed that ParELC3 loaded-liposomes are more efficient (29 to 11 µmol·L-1) compared to the free peptide (>100 µmol·L-1) at inhibiting the growth of standard E. coli and S. aureus strains. RL liposomes showed high hemolytic activity but when prepared with POPC and Chol this activity had a significant reduction. Independently of the formulation, the vesicles had no detectable cytotoxicity to HepG2 cells, even at the highest concentrations tested (1.3 mmol·L-1 and 50 µmol·L-1 for rhamnolipid and ParELC3, respectively). CONCLUSION: The present findings suggest the potential use of rhamnolipid-based liposomes as nanocarrier systems to enhance the bioactivity of peptides.
Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Carriers/chemistry , Glycolipids/chemistry , Nanoparticles/chemistry , Peptides/pharmacology , Toxin-Antitoxin Systems , Amino Acid Sequence , Cell Membrane Permeability/drug effects , Cell Survival/drug effects , Drug Liberation , Dynamic Light Scattering , Escherichia coli/drug effects , Hemolysis/drug effects , Hep G2 Cells , Humans , Hydrodynamics , Liposomes , Microbial Sensitivity Tests , Peptides/chemistry , Sonication , Staphylococcus aureus/drug effectsABSTRACT
OBJECTIVE: This study investigated the mechanism of action of different proteins/peptides (separately or in combination), focusing on how they act directly on the native enamel surface and on modifying the salivary pellicle. METHODS: A total of 170 native human enamel specimens were prepared and submitted to different treatments (2 h; 37 °C): with deionized water, CaneCPI-5, Hemoglobin, Statherin, or a combination of all three proteins/peptides. The groups were subdivided into treatment acting on the enamel surface (NoP - absence of salivary pellicle), and treatment modifying the salivary pellicle (P). Treatment was made (2 h; 37 °C) in all specimens, and later, for P, the specimens were incubated in human saliva (2 h; 37 °C). In both cases, the specimens were immersed in 1% citric acid (pH 3.6; 2 min; 25 °C). Calcium released from enamel (CaR) and its relative surface reflection intensity (%SRI) was measured after 5 cycles. Between-group differences were verified with two-way ANOVA, with "presence of pellicle" and "treatment" as factors (α = 0.05). RESULTS: The presence of pellicle provided better protection regarding %SRI (p < 0.01), but not regarding CaR (p = 0.201). In relation to treatment, when compared to the control group, all proteins/peptides provided significantly better protection (p < 0.01 for %SRI and Car). The combination of all three proteins/peptides demonstrated the best protective effect (p < 0.01 for %SRI). CONCLUSION: Depending on the protein or peptide, its erosion-inhibiting effect derives from their interaction with the enamel surface or from modifying the pellicle, so a combination of proteins and peptides provides the best protection. CLINICAL SIGNIFICANCE: The present study opens a new direction for a possible treatment with a combination of proteins for native human enamel, which can act directly on the enamel surface as well on the modification of the salivary pellicle, for the prevention of dental erosion.
Subject(s)
Tooth Erosion , Dental Enamel , Dental Pellicle , Humans , Peptides , Saliva , Tooth Erosion/prevention & controlABSTRACT
ABSTRACT Objective: To compare bone marrow aspirate concentrate (BMAC) with the standard treatment for gluteal tendinopathies. Methods: 48 patients diagnosed with gluteal tendinopathy at a university hospital were selected by a randomized clinical trial and divided into two groups: (G1) bone marrow aspirate concentrate and (G2) corticosteroid injections. Results: 40 of the 48 selected patients were monitored for six months and both groups showed better scores. Visual analog scale (VAS) scores and Lequesne index were statistically significant higher in patients submitted to BMAC treatment when compared to standard treatment. Both groups improved their quality of life, without statistically significant difference. Conclusion: BMAC constitutes an alternative to gluteal tendinopathy standard treatment, proving to be a safe technique with promising results when combined with multidisciplinary team behavioral therapy. Level of Evidence II, Randomized Clinical Trial.
RESUMO Objetivo: Estudo comparativo entre tratamento com corticóide e aspirado de medula óssea concentrado (BMAC) para o tratamento de tendinopatias glúteas. Métodos: O ensaio clínico randomizado selecionou pacientes diagnosticados com tendinopatia glútea e os dividiu em dois grupos: (G1) aspirado de medula óssea concentrada e (G2) injeção de corticosteróide. Resultados: Foram selecionados 48 pacientes, dos quais 40 foram monitorados por 6 meses, com melhora nos escores nos dois grupos. Os pacientes que foram submetidos ao tratamento com BMAC tiveram uma melhora estatisticamente significativa nos escores de EVA e nos escores de Lequesne em comparação ao tratamento padrão. Houve uma melhora na avaliação da qualidade de vida em ambos os grupos, sem diferença estatisticamente significativa. Conclusão: O aspirado de medula óssea concentrada surge como uma alternativa ao tratamento padrão da tendinopatia glútea, provando ser uma técnica segura e com resultados promissores quando combinada à terapia comportamental de equipe multidisciplinar. Nível de Evidência II, O ensaio clínico randomizado.
ABSTRACT
INTRODUCTION Adult gluteal tendinopathy is part of a group of pathologies of Greater Trochanteric Pain Syndrome (GTPS). This disorder is characterized by pain, functional limitation and loss of local strength. The diagnosis is made from clinical examination associated with complementary exams (Ultrasound or Magnetic Resonance). OBJECTIVE Comparative study between standard treatment and bone marrow aspirate concentrate (BMAC) for the treatment of gluteal tendinopathies. The randomized clinical trial selected patients diagnosed with gluteal tendinopathy at a university hospital and divided them into two groups: (G1) bone marrow aspirate concentrate and (G2) Corticosteroids injection. RESULTS 48 patients were selected, of which 40 were monitored for 06 months, with an improvement in scores in both groups. Patients who were submitted to the BMAC treatment had a statistically significant improvement in VAS scores and Lequesne scores compared to standard treatment. There was an improvement in the assessment of the quality of life in both groups with no statistically significant difference. CONCLUSION BMAC arises as an alternative to the standard treatment of gluteal tendinopathy, proving to be a safe technique and with promising results when combined with multidisciplinary team behavioral therapy.
INTRODUÇÃO A tendinopatia glútea adulta faz parte de um grupo de patologias da Síndrome Dolorosa Trocantérica (SDT). Esse distúrbio é caracterizado por dor, limitação funcional e perda de força local. O diagnóstico é feito a partir de exames clínicos associados a exames complementares (ultrassonografia ou ressonância magnética). OBJETIVO Estudo comparativo entre tratamento com corticóide e aspirado de medula óssea concentrado (BMAC) para o tratamento de tendinopatias glúteas. MATERIAL E MÉTODOS O ensaio clínico randomizado selecionou pacientes diagnosticados com tendinopatia glútea e os dividiu em dois grupos: (G1) aspirado de medula óssea concentrada e (G2) injeção de corticosteróide . RESULTADOS Foram selecionados 48 pacientes, dos quais 40 foram monitorados por 06 meses, com melhora nos escores nos dois grupos. Os pacientes que foram submetidos ao tratamento com BMAC tiveram uma melhora estatisticamente significativa nos escores de EVA e nos escores de Lequesne em comparação ao tratamento padrão. Houve uma melhora na avaliação da a qualidade de vida em ambos os grupos, sem diferença estatisticamente significativa. CONCLUSÃO O aspirado de medula óssea concentrada surge como uma alternativa ao tratamento padrão da tendinopatia glútea, provando ser uma técnica segura e com resultados promissores quando combinada à terapia comportamental de equipe multidisciplinar. Ensaio Clínico Randomizado.
ABSTRACT
OBJECTIVES: In the present study, we used an in vitro initial intrinsic erosion model to evaluate: (experiment 1) the influence of the degree of serine (Ser) phosphorylation of peptides containing the 15 N-terminal residues of statherin and (experiment 2) the effect of different concentrations of the peptide with the best performance in experiment 1 on initial enamel erosion. DESIGN: Bovine enamel specimens were divided into 6 groups (n = 15/group) for each experiment. In experiment 1, the peptides evaluated (at 1.88 × 10-5 M) were: not phosphorylated (StatSS), phosphorylated in Ser2 (StatpSS), phosphorylated in Ser3 (StatSpS) phosphorylated in Ser2 and Ser3 (StatpSpS). Phosphate buffer and human recombinant statherin were used as negative and positive controls, respectively. In experiment 2, StatpSpS was evaluated at different concentrations: 0.94, 1.88, 3.76 and 7.52 × 10-5 M. Phosphate buffer and 0.1 mg/mL CaneCPI-5 were employed as negative and positive controls, respectively. In each experiment, the specimens were incubated with the solutions for 2 h, then the AEP was allowed to form (under human pooled saliva) for 2 h. The specimens were then challenged with 0.01 M HCl for 10 s. Demineralization was evaluated by percentage of surface hardness change (%SHC). Data were analyzed by ANOVA and Tukey's test (p < 0.05). RESULTS: In experiment 1, only StatpSpS significantly reduced the % SHC in comparison with control. In experiment 2, 1.88 × 10-5 M StatpSpS significantly reduced the %SHC in comparison with control. CONCLUSIONS: This is the first study showing that statherin-derived peptide might protect against intrinsic erosion.
Subject(s)
Dental Enamel/chemistry , Salivary Proteins and Peptides/chemistry , Tooth Erosion , Animals , Cattle , Humans , In Vitro Techniques , Phosphorylation , Saliva , Serine/chemistry , Tooth Erosion/prevention & controlABSTRACT
OBJECTIVES: To evaluate, in vivo: 1) proteomic alterations in the acquired enamel pellicle (AEP) after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), statherin-derived peptide (StN15) or their combination before the formation of the AEP and subsequent erosive challenge; 2) the protection of these treatments against erosive demnineralization. MATERIALS AND METHODS: In 5 crossover phases, after prophylaxis, 10 volunteers rinsed (10â¯mL, 1â¯min) with: deionized water-1, 0.1â¯mg/mL CaneCPI-5-2, 1.0â¯mg/mL HB-3, 1.88â¯×â¯10-5 M StN15-4 or their combination-5. AEP was formed (2â¯h) and enamel biopsy (10⯵L, 1%citric acid, pH 2.5, 10â¯s) was performed on one incisor for calcium analysis. The same acid was applied on the vestibular surfaces of the remaining teeth. The acid-resistant proteins within the remaining AEP were collected. Samples were quantitatively analyzed by label-free proteomics. RESULTS: Treatment with the proteins/peptide, isolated or combined, increased several acid-resistant proteins in the AEP, compared with control. The highest increases were seen for PRPs (32-fold, StN15), profilin (15-fold, combination), alpha-amylase (9-fold; StN15), keratins (8-fold, CaneCPI-5 and HB), Histatin-1 (7-fold, StN15), immunoglobulins (6.5-fold, StN15), lactotransferrin (4-fold, CaneCPI-5), cystatins, lysozyme, protein S-100-A9 and actins (3.5-fold, StN15), serum albumin (3.5-fold, CaneCPI-5 and HB) and hemoglobin (3-fold, StN15). Annexin, calmodulin, keratin, tubulin and cystatins were identified exclusively upon treatment with the proteins/peptide, alone or combined. Groups 2, 3 and 4 had significantly lower Ca released from enamel compared to group 1 (Kruskal-Wallis/Dunn's, pâ¯<â¯0.05). CONCLUSIONS: Treatment with CaneCPI-5, HB or StN15 remarkably increases acid-resistant proteins in the AEP, protecting against erosion. CLINICAL SIGNIFICANCE: Our results show, for the first time, that treatment with proteins/peptide remarkably increases acid-resistant proteins in the AEP, protecting against erosive demineralization. These findings open an avenue for a new preventive approach for erosive demineralization, employing acquired pellicle engineering procedures that may in the future be incorporated into dental products.
Subject(s)
Tooth Demineralization , Tooth Erosion , Dental Enamel , Dental Pellicle , Humans , Peptides , Proteomics , Tooth Demineralization/prevention & controlABSTRACT
AIMS: This study aimed to evaluate the toxicity and humoral and cellular immune response of three heterologous vaccines against Leishmania infantum, yet containing synthetic peptides from Leishmania major in the experimental model in hamsters. METHODS AND RESULTS: Through bioinformatics analyses, two Leishmania major Gp63 peptides were predicted and selected for vaccine formulations. Hamsters were divided into four groups, with each group receiving doses of three vaccine formulations containing HLA-DR1 or HLA-A2 peptides plus MontanideTM or both associated with the adjuvant. The animals received three vaccine doses and were evaluated for toxicity after each dose, in addition to being analysed for the production of antibodies and lymphoproliferation on day 211 after the last vaccine dose. Peptides predicted in association with oily adjuvant induced a humoral response and strong lymphoproliferation to Leishmania infantum antigen-specific stimulation.
Subject(s)
Leishmania major/immunology , Leishmaniasis Vaccines/immunology , Leishmaniasis/immunology , Metalloendopeptidases/immunology , Peptides/immunology , Adjuvants, Immunologic/administration & dosage , Animals , Cross Protection , HLA-A2 Antigen/immunology , HLA-DR1 Antigen/immunology , Immunity, Cellular , Immunity, Humoral , Leishmania infantum/immunology , Leishmaniasis/prevention & control , Leishmaniasis Vaccines/administration & dosage , Leishmaniasis Vaccines/chemistry , Mesocricetus , Metalloendopeptidases/chemistry , Mineral Oil/administration & dosage , Peptides/administration & dosage , Peptides/chemistryABSTRACT
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