Transforming growth factor ß1-enriched secretome up-regulate osteogenic differentiation of dental pulp stem cells, and a potential therapeutic for gingival wound healing: A comparative proteomics study.

OBJECTIVES: Current study aimed at comparing the human dental pulp-derived stem cell (hDPSC) secretome (Control secretome) and transforming growth factor beta1 (TGF-ß1)-transfected hDPSC secretome (TGF-ß1 Secretome), which have the potential to be therapeutic in terms of regenerative dentistry, in terms of osteogenesis, adipogenesis and gingival wound healing with proteomic analyses. MATERIALS AND METHODS: pCMV-TGF-ß1 plasmid was transfected into hDPSCs by electroporation. hDPSC and TGF-ß1 transfected hDPSC secretomes were collected for LC-MS/MS. Protein contents in control secretome and TGF-ß1 secretome were analyzed by tandem mass spectrometry-based shotgun proteomic method. Bioinformatic evaluations for canonical pathways, upstream regulators and networks were completed via Ingenuity Pathway Analysis (IPA, QIAGEN) software. Surface marker expressions between groups, treated secretome were measured by flow cytometry. To support the proteomic data morphologically, we performed osteogenic-adipogenic differentiation in hDPSCs treated with control secretome and TGF-ß1 secretome, and scratch wound healing assay in gingival fibroblasts. Statistical analyses were performed by GraphPad Prism 8.02. RESULTS: Venn diagram classification showed us 174 common proteins were identified from each group. In the control secretome 140 unique proteins were identified and 66 entries were exclusive for TGF-ß1 secretome. TGF-ß1 secretome was found to have therapeutic effect on MSC-specific immunophenotypes. TGF-ß1 secretome was determined to up-regulate osteogenesis-related molecules and pathways while down-regulating adipogenesis-related pathways. Analysis of canonical pathways showed that TGF-ß1 secretome is associated with the wound healing pathway. CONCLUSION: Our study provided the first evidence that proteins identified in TGF-ß1-transfected hDPSC secretomes are potential regulators of osteogenic/adipogenic differentiation and fibroblast wound healing. CLINICAL SIGNIFICANCE: Based on these results, TGF-ß1 secretome may have a therapeutic effect in repairing osteoporosis-related bone injuries, wound healing of oral mucosa and gingival tissue. TGF-ß1 secretome may be a potential cell-free therapeutic in orthopedics and regenerative dentistry.

Long-term assessment of visual and refractive outcomes of laser in situ keratomileusis for hyperopia using the AMARIS® 750S Excimer laser.

PURPOSE: To evaluate the long-term efficacy and safety of hyperopic laser in situ keratomileusis (LASIK) using the AMARIS® 750S (Schwind, Eye-tech-solutions, GmbH) excimer laser. METHODS: The medical records of one hundred eleven eyes of 62 patients who underwent LASIK for hyperopia using the AMARIS® 750S excimer laser were reviewed retrospectively. Patients were divided into three groups based on preoperative spherical equivalent (SE) refraction: low hyperopia (less than +2.50 diopters [D]), moderate hyperopia (+2.75D to +4.00D), and high hyperopia (over +4.00D). Uncorrected and best corrected visual acuity (BCVA), long-term stability of refraction, and complications were evaluated. RESULTS: Of the entire sample, the mean preoperative SE was +3.64D±1.22D. The mean age was 37.4±11.2 years (20-59). The mean follow-up for all eyes was 51 months. At the last visit, the mean SE was +0.85D±0.34D (SD) in the low hyperopia group, +1.09D±0.43D in the moderate hyperopia group, and +1.63D±0.47D in the high hyperopia group. (+1.15D±0.49D overall). Preoperative uncorrected visual acuity (UCVA) was 0.52±0.34 logMAR and increased to 0.18±0.15 logMAR at 4 years follow-up (P<0.01). There was no statistically significant difference between preoperative and postoperative BCVA. The UCVA was 0.30 logMAR or better in 100% of eyes in the low hyperopia group, 93.7% in the moderate hyperopia group, and 69.9% in the high hyperopia group (%89.2 overall). CONCLUSIONS: LASIK is safe and effective for correcting hyperopia in the short term; however, the efficacy of the procedure is limited in the patients with high hyperopia and longer follow-up.

The effects of treatment in patients with childhood asthma on the elastic properties of the aorta

Introduction: The study aimed to investigate the effects of treatment in patients with childhood asthma on the elastic properties of the aorta and cardiovascular risk.Methods: The study was performed in 66 paediatric patients diagnosed with bronchial asthma (BA). All patients were administered the ß2 agonist, salbutamol, for seven days, followed by one month of montelukast and six months of inhaled steroid treatment. All patients underwent conventional transthoracic echocardiographic imaging before and after treatment. Aortic elasticity parameters were considered to be the markers of aortic function.Results: Aortic elasticity parameters, including aortic strain (15.2 ± 4.8 and 18.8 ± 9.5%, p = 0.043), aortic distensibility (7.26 ± 4.71 and 9.53±3.50 cm2/dyn, p = 0.010) and aortic stiffness index (3.2 ± 0.6 and 2.8 ± 0.5, p = 0.045 showed significant post-treatment improvement when compared to pre-treatment values. Tricuspid annular plane systolic excursion (TAPSE) was also observed to improve after treatment (1.81 ± 0.38 and 1.98 ± 0.43, p = 0.049). Conclusion: The study demonstrated that when provided at appropriate doses, medications used in BA may result in an improvement in aortic stiffness

In Vivo and In Vitro Effects of Chlorhexidine Pretreatment on Immediate and Aged Dentin Bond Strengths.

This study evaluated the effect of 2% chlorhexidine (CHX) pretreatment of dentin on the immediate and aged microtensile bond strength (µTBS) of different adhesives to dentin in vivo and in vitro. Class I cavities were prepared in 80 caries-free human third molars of 40 patients in a split-mouth fashion. In each tooth pair, one tooth received 2% CHX pretreatment after which both teeth were randomly assigned to one of the following groups with respect to the type of adhesive system applied: Adper Single Bond 2 (etch-and-rinse), Clearfil SE Bond (two-step self-etch), Clearfil S(3) Bond (one-step self-etch), and Adper Prompt-L-Pop (all-in-one self-etch). The teeth were restored with resin composite and extracted for µTBS testing either immediately or after six months in function. In vitro specimen pairs were prepared as with the clinical protocol in intact, freshly extracted human molars, and thereafter, subjected to testing immediately or after 5000× thermocycling. Data were analyzed with four-way analysis of variance (ANOVA). Bonferroni test was utilized for pair-wise comparisons. The immediate bond strength values were significantly higher than "aged" ones for all tested adhesives (p=0.00). The in vitro immediate bond strength values were statistically higher than in vivo bond strength values (p<0.05). While the bond strength of in vitro aged, CHX-treated samples were higher than their in vivo counterparts (p<0.05), no difference was observed in non-CHX treated groups (p>0.05). In the absence of CHX pretreatment, all adhesives showed significantly higher immediate bond strength values than CHX-treated groups, while all "aged", non-pretreated adhesives exhibited significantly lower bond strength values (both p<0.05). By contrast, chlorhexidine pretreatment resulted in significantly higher aged bond strengths, regardless of the adhesive system and testing condition. Aging-associated decline in dentin bond strength of etch-and rinse and self-etch adhesives can be counteracted by chlorhexidine application.

The role of neutrophil to lymphocyte ratio as a predictor of diastolic dysfunction in hypertensive patients.

OBJECTIVE: Neutrophil to lymphocyte ratio (NLR) is a novel parameter for cardiovascular research area. The higher values of NLR have been found to be associated with worse clinical outcomes in atherosclerotic heart disease, heart failure, heart valve disease and other various cardiovascular disorders. Although the relationship between NLR and almost all cardiovascular disorders have been investigated, the association between NLR and diastolic dysfunction remains unclear. We herein evaluated the association between NLR and diastolic dysfunction. PATIENTS AND METHODS: The study population consisted of 41 hypertensive patients with any grade of diastolic dysfunction and 41 hypertensive patients without diastolic dysfunction determined by echocardiographic evaluation constituted the control group. RESULTS: Mean NLR value was found to be 2.07 ± 0.82 in the diastolic dysfunction group while the control group had a mean value of 1.69 ± 0.60 (p = 0.020). The patients with diastolic dysfunction had significantly higher values of NLR. When grades of diastolic dysfunction were evaluated, NLR was 1.80 ± 0.82, 2.32 ± 0.73 and 2.75 ± 0.45 in patients with grade 1, grade 2 and grade 3 diastolic dysfunction, respectively. The patients with higher grade of diastolic dysfunction had higher values of NLR (p = 0.001). None of the other hematologic parameters differed significantly in patients with diastolic dysfunction when compared to controls. CONCLUSIONS: Patients with diastolic dysfunction had higher values of NLR compared to subjects without diastolic dysfunction. Furthermore higher grades of diastolic dysfunction were associated with higher levels of NLR. Further studies are needed to search the possible use of NLR as a marker for prognostic stratification in diastolic dysfunction which is associated with worse cardiovascular outcomes.