Safety and effectiveness of sodium colistimethate-loaded nanostructured lipid carriers (SCM-NLC) against P. aeruginosa: in vitro and in vivo studies following pulmonary and intramuscular administration.

The usefulness of nanotechnology to increase the bioavailability of drugs and decrease their toxicity may be a tool to deal with multiresistant P. aeruginosa (Mr-Pa) respiratory infections. We describe the preparation and the in vivo efficacy and safety of sodium colistimethate-loaded nanostructured lipid carriers (SCM-NLC) by the pulmonary and intramuscular routes. Nanoparticles showed 1-2 mg/L minimum inhibitory concentration against eight extensively drug-resistant P. aeruginosa strains. In vivo, SCM-NLC displayed significantly lower CFU/g lung than the saline and similar to that of the free SCM, even the dose in SCM-NLC group was lower than free SCM. There was no tissue damage related to the treatments. Biodistribution assessments showed a mild systemic absorption after nebulization and a notorious absorption after IM route. Altogether, it could be concluded that SCM-NLC were effective against P. aeruginosa in vivo, not toxic and distribute efficiently to the lung and liver after pulmonary or intramuscular administrations.

Two cycles of plasma rich in growth factors (PRGF-Endoret) intra-articular injections improve stiffness and activities of daily living but not pain compared to one cycle on patients with symptomatic knee osteoarthritis.

PURPOSE: To assess the clinical efficacy and safety of a treatment based on one cycle versus two cycles of intra-articular injections of plasma rich in growth factors (PRGF-Endoret) on patients with knee osteoarthritis (OA). METHODS: Ninety patients with knee OA were included and evaluated. A total of 48 patients received one cycle (OC group) (3 injections on a weekly basis), while 42 patients received two cycles of PRGF-Endoret (TC group) spaced 6 months between them. Patients were evaluated with LEQUESNE and WOMAC scores before treatment and after 48 weeks. Safety assessment was also performed. RESULTS: A significant reduction of all assessed outcome measures was shown for both groups at 48 weeks compared with baseline values (P < 0.001). Patients of TCs group showed a significantly higher reduction (P < 0.05) in WOMAC stiffness subscales. Regarding LEQUESNE INDEX, a significantly higher reduction was observed in the TC group in all subscales except in pain score. In the maximum walking distance subscale (MCD), the improvement rate was 31.8% higher for the TCs group compared with the OC group (P < 0.01). In addition, the TC group showed a significant improvement in LEQUESNE activities of daily living (ADV) and global subscales of 14.7 and 11.8% (P < 0.05) higher, respectively, than the OC group. CONCLUSIONS: Treatment with two cycles of PRGF did not show a significantly higher pain reduction compared with one cycle treatment. However, two cycles of PRGF showed a significant improvement in WOMAC stiffness, LEQUESNE MCD, LEQUESNE ADV and LEQUESNE global subscales. Therefore, patients treated with two cycles present an improvement in quality of life. LEVEL OF EVIDENCE: II.

Efficacy and safety of plasma rich in growth factors intra-articular infiltrations in the treatment of knee osteoarthritis.

PURPOSE: The goal of this study was to systematically review the efficacy and safety of plasma rich in growth factors (PRGF) as a treatment for reducing symptoms in patients with knee osteoarthritis. METHODS: A comprehensive and systematic literature search was conducted for PRGF treatment of knee osteoarthritis following the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. All the studies had to include a PRGF group and a control group. Pre- and post-treatment measures of joint pain, reduced function, and stiffness were evaluated using the Western Ontario and McMaster Universities Osteoarthritis Index, Knee Injury and Osteoarthritis Outcome Score, International Knee Documentation Committee score, Lequesne index, or number of Outcome Measures for Rheumatology Committee and Osteoarthritis Research Society International Standing Committee for Clinical Trials Response Criteria Initiative (OMERACT-OARSI) responders, with a follow-up period of at least 4 weeks. An assessment of both the quality and risk of bias of the studies was conducted. RESULTS: The literature search yielded 91 citations, but only 5 were eligible publications that met the inclusion criteria (2 randomized controlled trials, 2 prospective studies, and 1 retrospective analysis). Two studies were rated as having a low risk of bias whereas 3 had a high risk. In both randomized controlled trials, it was observed that after 6 months of treatment, the number of patients with a pain reduction of more than 50% was significantly higher in the PRGF group. In 2 other studies, the patients treated with PRGF showed a significant pain reduction compared with the control group. The remaining variables (Western Ontario and McMaster Universities Osteoarthritis Index scale for pain, function, and stiffness; Lequesne index; Knee Injury and Osteoarthritis Outcome Score scale; and number of OMERACT-OARSI responders) showed a statistically significant superiority of the group treated with PRGF. CONCLUSIONS: The current clinical evidence suggests that PRGF intra-articular infiltrations in patients with knee osteoarthritis reduce pain and therefore are clinically efficacious in osteoarthritis treatment. LEVEL OF EVIDENCE: Level III, systematic review of Level I, II, and III studies.

Soy protein/ß-chitin sponge-like scaffolds laden with human mesenchymal stromal cells from hair follicle or adipose tissue promote diabetic chronic wound healing.

Chronic wounds are a worldwide problem that affect >40 million people every year. The constant inflammatory status accompanied by prolonged bacterial infections reduce patient's quality of life and life expectancy drastically. An important cell type involved in the wound healing process are mesenchymal stromal cells (MSCs) due to their long-term demonstrated immunomodulatory and pro-regenerative capacity. Thus, in this work, we leveraged and compared the therapeutic properties of MSCs derived from both adipose tissue and hair follicle, which we combined with sponge-like scaffolds (SLS) made of valorized soy protein and ß-chitin. In this regard, the combination of these cells with biomaterials permitted us to obtain a multifunctional therapy that allowed high cell retention and growing rates while maintaining adequate cell-viability for several days. Furthermore, this combined therapy demonstrated to increase fibroblasts and keratinocytes migration, promote human umbilical vein endothelial cells angiogenesis and protect fibroblasts from highly proteolytic environments. Finally, this combined therapy demonstrated to be highly effective in reducing wound healing time in vivo with only one treatment change during all the experimental procedure, also promoting a more functional and native-like healed skin.

A randomized clinical trial evaluating plasma rich in growth factors (PRGF-Endoret) versus hyaluronic acid in the short-term treatment of symptomatic knee osteoarthritis.

PURPOSE: This multicenter, double-blind clinical trial evaluated and compared the efficacy and safety of PRGF-Endoret (BTI Biotechnology Institute, Vitoria-Gasteiz, Spain), an autologous biological therapy for regenerative purposes, versus hyaluronic acid (HA) as a short-term treatment for knee pain from osteoarthritis. METHODS: We randomly assigned 176 patients with symptomatic knee osteoarthritis to receive infiltrations with PRGF-Endoret or with HA (3 injections on a weekly basis). The primary outcome measure was a 50% decrease in knee pain from baseline to week 24. As secondary outcomes, we also assessed pain, stiffness, and physical function using the Western Ontario and McMaster Universities Osteoarthritis Index; the rate of response using the criteria of the Outcome Measures for Rheumatology Committee and Osteoarthritis Research Society International Standing Committee for Clinical Trials Response Criteria Initiative (OMERACT-OARSI); and safety. RESULTS: The mean age of the patients was 59.8 years, and 52% were women. Compared with the rate of response to HA, the rate of response to PRGF-Endoret was 14.1 percentage points higher (95% confidence interval, 0.5 to 27.6; P = .044). Regarding the secondary outcome measures, the rate of response to PRGF-Endoret was higher in all cases, although no significant differences were reached. Adverse events were mild and evenly distributed between the groups. CONCLUSIONS: Plasma rich in growth factors showed superior short-term results when compared with HA in a randomized controlled trial, with a comparable safety profile, in alleviating symptoms of mild to moderate osteoarthritis of the knee. LEVEL OF EVIDENCE: Level I, randomized controlled multicenter trial.

Multiclass classification of breast cancer histopathology images using multilevel features of deep convolutional neural network.

Breast cancer is a common malignancy and a leading cause of cancer-related deaths in women worldwide. Its early diagnosis can significantly reduce the morbidity and mortality rates in women. To this end, histopathological diagnosis is usually followed as the gold standard approach. However, this process is tedious, labor-intensive, and may be subject to inter-reader variability. Accordingly, an automatic diagnostic system can assist to improve the quality of diagnosis. This paper presents a deep learning approach to automatically classify hematoxylin-eosin-stained breast cancer microscopy images into normal tissue, benign lesion, in situ carcinoma, and invasive carcinoma using our collected dataset. Our proposed model exploited six intermediate layers of the Xception (Extreme Inception) network to retrieve robust and abstract features from input images. First, we optimized the proposed model on the original (unnormalized) dataset using 5-fold cross-validation. Then, we investigated its performance on four normalized datasets resulting from Reinhard, Ruifrok, Macenko, and Vahadane stain normalization. For original images, our proposed framework yielded an accuracy of 98% along with a kappa score of 0.969. Also, it achieved an average AUC-ROC score of 0.998 as well as a mean AUC-PR value of 0.995. Specifically, for in situ carcinoma and invasive carcinoma, it offered sensitivity of 96% and 99%, respectively. For normalized images, the proposed architecture performed better for Makenko normalization compared to the other three techniques. In this case, the proposed model achieved an accuracy of 97.79% together with a kappa score of 0.965. Also, it attained an average AUC-ROC score of 0.997 and a mean AUC-PR value of 0.991. Especially, for in situ carcinoma and invasive carcinoma, it offered sensitivity of 96% and 99%, respectively. These results demonstrate that our proposed model outperformed the baseline AlexNet as well as state-of-the-art VGG16, VGG19, Inception-v3, and Xception models with their default settings. Furthermore, it can be inferred that although stain normalization techniques offered competitive performance, they could not surpass the results of the original dataset.

Bioactive and degradable hydrogel based on human platelet-rich plasma fibrin matrix combined with oxidized alginate in a diabetic mice wound healing model.

In the present study we developed an injectable, bioactive and degradable hydrogel composed of alginate at 2.5% oxidation degree and calcium-activated platelet rich plasma (PRP) for wound healing applications (PRP-HG-2.5%). The alginate gives mechanical support to the hydrogel while the activated PRP provides growth factors that enhance wound healing and fibrin which creates an adequate microenvironment for cell migration and proliferation. The rheological and mechanical properties of the hydrogel were characterized. Further characterization revealed that PRP-HG-2.5% showed a faster hydrolitic degradation rate than unmodified alginate and a similar platelet derived growth factor (PDGF-BB) release profile. In vitro efficacy studies, carried out in human fibroblasts and keratinocytes, showed that PRP-HG-2.5% was not cytotoxic and that it was able to promote cell adhesion and proliferation. Thereafter, in an in vivo full thickness wound healing study conducted in diabetic mice, no differences were found among PRP-HG-2.5% and its counterpart without PRP, likely due to the xenogeneic origin of the PRP. This hypothesis was validated in vitro, since a cytotoxic effect was observed after human PRP application to mouse fibroblasts. Therefore, PRP-HG-2.5% might be a promising strategy for chronic woundstreatment, although its effectiveness should be evaluated in a more reliable preclinical model.

Lymph Node Tumor Burden Correlates With Tumor Budding and Poorly Differentiated Clusters: A New Prognostic Factor in Colorectal Carcinoma?

INTRODUCTION: Molecular lymph node (LN) staging in early colorectal cancer (CRC) has demonstrated to be more precise than conventional histopathology pN staging. Tumor budding (TB) and poorly differentiated clusters (PDCs) are associated with LN metastases, recurrences, and lower survival in CRC. We evaluated the correlation between the total tumor load (TTL) in LNs from CRC surgical specimens with patient outcome, TB, and PDC. METHODS: In this retrospective multicentre study, 5,931 LNs from 342 stage I-III CRC were analyzed by both hematoxylin and eosin and molecular detection of tumor cytokeratin 19 mRNA by one-step nucleic acid amplification. TB and PDC were evaluated by hematoxylin and eosin and cytokeratin 19 immunohistochemistry. RESULTS: One-step nucleic acid was positive in 38.3% patients (n = 131). Tumor Budding was low in 45% cases, intermediate in 25%, and high in 30%. Poorly Differentiated Clusters were low-grade G1 in 53%, G2 in 32%, and G3 in 15%. TB and PDC correlated with TTL, high-grade, lymphovascular and perineural invasion, pT, pN and stage (P < 0.001). TB, PDC, and TTL ≥ 6,000 copies/µL were associated with worse overall survival (P = 0.002, P = 0.013, and P = 0.046) and disease-free survival (P < 0.001). DISCUSSION: The implementation of more sensitive molecular methods to assess LN status is a promising alternative approach to pN staging, which could be integrated to other factors to help risk stratification and management of patients with early-stage CRC. This study demonstrates the correlation of the amount of LN tumor burden with TB and PDCs. TTL is related to the outcome and could be used as a new prognostic factor in CRC (see Visual Abstract, Supplementary Digital Content 2, http://links.lww.com/CTG/A512).

Miki (Mitotic Kinetics Regulator) Immunoexpression in Normal Liver, Cirrhotic Areas and Hepatocellular Carcinomas: a Preliminary Study with Clinical Relevance.

Hepatocellular carcinoma (HCC) is the most common type of primary malignant tumor in the liver. One of the main features of cancer survival is the generalized loss of growth control exhibited by cancer cells, and Miki is a protein related to the immunoglobulin superfamily that plays an important role in mitosis. We aim to study protein expression levels of Miki in non-tumoral liver and 20 HCCs recruited from a Pathology Department. Clinical information was also obtained. A tissue microarray was performed, and immunohistochemical techniques applied to study protein expression levels of Miki. In normal liver, Miki was weakly expressed, showing nuclear staining in the hepatocytes. Cirrhotic areas and HCCs showed a variety of staining patterns. Most HCC samples showed positive expression, with three different staining patterns being discernible: nuclear, cytoplasmic and mixed. Statistical analysis showed a significant association between grade of differentiation, Ki-67 proliferative index, survival rates and staining patterns. This study has revealed the positive expression of Miki in normal liver, cirrhotic areas and HCCs. Three different staining patterns of Miki expression with clinical relevance were noted in HCCs.

Composite nanofibrous membranes of PLGA/Aloe vera containing lipid nanoparticles for wound dressing applications.

Electrospun nanofibrous dressings present suitable characteristics to be used in wound healing, such as high porosity and high surface area-to-volume ratio. In this study, a wound dressing based on PLGA and Aloe vera containing lipid nanoparticles (NLCs) was developed. NLCs were added in order to add a lipid component that could avoid the adhesion of the dressing to the wound and improve its handling. Membranes with and without NLCs were composed of uniform fibers of about 1 µm in diameter. Their porosity was above 80% and their thickness was about 160 µm. Both dressings showed similar water vapour transmission rate 1100 g/m2day. The formulation containing NLCs presented a higher ultimate tensile strength (2.61 ±â€¯0.46 MPa) and a higher water uptake. Both formulations were biocompatible in vitro. Furthermore, the cell adhesion assay demonstrated that both membranes had a low adherence profile, although it was lower with the dressing containing NLCs. Finally, their efficacy was evaluated in a full thickness wound healing assay conducted in db/db mice, where both enhanced healing similarly. Accordingly, the PLGA-AV-NLC membrane might be a promising strategy for the treatment of chronic wounds, since it improved handling in comparison to the formulation without NLCs.

5-year clinical experience with BTI dental implants: risk factors for implant failure.

OBJECTIVES: The aims of this study were to identify with appropriate statistical tests the risk factors associated with implant failure and to evaluate the long-term survival of dental implants using implant loss as an outcome variable and performing an implant-, surgery- and patient-based analysis of failures. MATERIAL AND METHODS: A retrospective cohort study design was used. One thousand sixty patients received 5787 BTI implants during the years of 2001-2005 in Vitoria, Spain. The potential influence of demographic items, clinical items, surgery-dependent items and prosthetic variables on implant survival was studied. Implant survival was analysed using a life-table analysis. Cox proportional hazards regression was used to identify risk factors related to implant failure. RESULTS: Smoking habits, implant position, implant staging (two-stage implants) and the implementation of special techniques were statistically correlated with lower implant survival rates. Two risk factors associated with implant failure were detected in this study: implant staging (two-stage implants) and the use of special techniques. Additionally, the overall survival rates of BTI implants were 99.2%, 96.4% and 96% for the implant-, surgery- and patient-based analysis, respectively. Totally, 28 out from 5787 implants (0.48%) were lost during the observation period. Most of the patients with implant failure (69.6%) presented chronic or aggressive periodontitis. CONCLUSIONS: Implant staging and the use of special techniques are risk factors for implant failure.

Five-year clinical evaluation of short dental implants placed in posterior areas: a retrospective study.

BACKGROUND: The aims of this study were to evaluate the long-term survival rates of short dental implants in posterior areas and to analyze the influence of different factors on implant survival. METHODS: A retrospective cohort study design was used. A total of 293 subjects received 532 short implants between 2001 and 2004. All implants were placed by two experienced surgeons, and rehabilitations were done by three prosthodontists. Each implant failure was analyzed carefully. The potential influence of demographic factors, clinical factors, surgery-dependent factors, and prosthetic variables on implant survival was studied. Implant survival was analyzed using a life-table analysis (Wilcoxon [Gehan] test). RESULTS: The overall survival rates of short implants were 99.2% and 98.7% for the implant- and subject-based analyses, respectively. The mean follow-up period was 31 +/- 12.3 months. Two of 532 implants were lost during the observation period. None of the variables studied were statistically associated with implant failure. CONCLUSION: Treatment with short implants can be considered safe and predictable if used under strict clinical protocols.

Clinical outcome of immediately loaded dental implants bioactivated with plasma rich in growth factors: a 5-year retrospective study.

BACKGROUND: The aims of this study were to describe a procedure for the immediate loading of dental implants, evaluate the long-term survival rates of 1,139 immediately loaded dental implants, and analyze the influence of different factors on implant survival. METHODS: A retrospective cohort study design was used. A total of 241 patients received 1,139 immediately loaded implants between 2001 and 2005 in Vitoria, Spain. All implants were placed by one experienced surgeon, and rehabilitations were done by four prosthodontists. Each implant failure was analyzed carefully. The potential influence of demographic, clinical, and surgery-dependent factors and prosthetic variables on implant survival was studied. Implant survival was analyzed using a life-table analysis. RESULTS: The overall survival rates were 99.3%, 96.8%, and 96.9% for the implant-, surgery-, and patient-based analyses, respectively. The mean follow-up period was 28 +/- 15 months. Five of 1,139 implants were lost during the observation period. No variable studied was statistically associated with implant failure. CONCLUSION: Based on these results, the procedure described for the immediate loading of implants can be considered safe and predictable if used under strict clinical protocols.

Establishing cut-off points with clinical relevance for bcl-2, cyclin D1, p16, p21, p27, p53, Sox11 and WT1 expression in glioblastoma - a short report.

PURPOSE: Glioblastoma (GBM) ranks among the most challenging cancers to treat and there is an urgent need for clinically relevant prognostic and diagnostic biomarkers. Here, we set out to investigate the expression of eight proteins (bcl-2, cyclin D1, p16, p21, p27, p53, Sox11 and WT1) in GBM with the specific aim to establish immunohistochemistry cut-off points with clinical relevance. METHODS: Immunohistochemistry (IHC) was used to examine protein expression in 55 surgical GBM specimens using H-scores, and IHC cut-off points were established using the Cutoff Finder web platform. Protein co-expression and its correlation with histopathological features were assessed, and cases were classified according to IDH1 mutation status. Survival curves were determined using Kaplan-Meier analyses. RESULTS: Clinical and molecular parameters found to be correlated with overall survival (OS) were tumor size (r = -0.278; p = 0.048), p53 (r = -0.452; p = 0.001), p16 (r = 0.351; p = 0.012) and Sox11 (r = 0.324; p = 0.020). In addition, we found that tumor size correlated with cyclin D1 (r = -0.282; p = 0.037), p53 (r = 0.269; p = 0.041), Sox11 (r = -0.309; p = 0.022) and WT1 (r = -0.372; p = 0.003). Variables found to be significantly associated with IDH1 mutation status were OS (p < 0.01), age (p < 0.01), cyclin D1 (p = 0.046), p16 (p = 0.019) and Sox11 (p = 0.012). Variables found to be significantly associated with a poor survival were tumor size >5 cm (p < 0.001), bcl-2 score > 40 (p = 0.034), cyclin D1 score ≤ 70 (p = 0.004), p16 score ≤ 130 (p = 0.005), p53 score > 20 (p = 0.003), Sox11 score ≤ 40 (p < 0.001) and WT1 score ≤ 270 (p = 0.02). CONCLUSIONS: Correlations between protein biomarkers and main clinical GBM variables were identified. The establishment of distinct biomarker cut-off points may enable clinicians and pathologists to better weigh their prognostic value.

Novel nanofibrous dressings containing rhEGF and Aloe vera for wound healing applications.

Nanofibrous membranes produced by electrospinning possess a large surface area-to-volume ratio, which mimics the three-dimensional structure of the extracellular matrix. Thus, nanofibrous dressings are a promising alternative for chronic wound healing, since they can replace the natural ECM until it is repaired. Therefore, in this study we have developed a PLGA nanofibrous membrane that contains recombinant human Epidermal Growth Factor (rhEGF) and Aloe vera (AV) extract. Both of them promote wound healing, as EGF is a wound healing mediator and AV stimulates the proliferation and activity of fibroblast. The obtained membranes were composed of uniform and randomly oriented fibers with an average diameter of 356.03±112.05nm, they presented a porosity of 87.92±11.96% and the amount of rhEGF was 9.76±1.75µg/mg. The in vitro viability assay demonstrated that the membranes containing rhEGF and AV improved fibroblast proliferation, revealing the beneficial effect of the combination. Furthermore, these membranes accelerated significantly wound closure and reepithelisation in an in vivo full thickness wound healing assay carried out in db/db mice. Overall, these findings demonstrated the potential of PLGA nanofibers containing rhEGF and AV for the treatment of chronic wounds.

LL37 loaded nanostructured lipid carriers (NLC): A new strategy for the topical treatment of chronic wounds.

The LL37 is a human antimicrobial peptide which not only has a broad spectrum of antimicrobial activity, but it has also been proved to modulate wound healing by participating in angiogenesis, epithelial cell migration and proliferation, and immune response. In this work, LL37 has been encapsulated in nanostructured lipid carriers (NLCs), produced by the melt-emulsification method, in order to improve its effectiveness. The characterisation of the NLC-LL37 showed a mean size of 270nm, a zeta potential of -26mV and an encapsulation efficiency of 96.4%. The cytotoxicity assay performed in Human Foreskin Fibroblasts demonstrated that the NLC-LL37 did not affect cell viability. Moreover, the in vitro bioactivityassay evidenced that the peptide remained active after the encapsulation, since the NLC-LL37 reversed the activation of the macrophages induced by LPS in the same way as the LL37 in solution. In addition, the in vitro antimicrobial assay revealed the NLC-LL37 activity against Escherichia coli. The effectiveness of the nanoparticles was assessed in a full thickness wound model indb/dbmice. The data demonstrated that NLC-LL37 significantly improved healing compared to the same concentration of the LL37 solution in terms of wound closure, reepithelisation grade and restoration of the inflammatory process. Overall, these findings suggest a promising potential of the NLC-LL37 formulation for chronic wound healing.

The topical administration of rhEGF-loaded nanostructured lipid carriers (rhEGF-NLC) improves healing in a porcine full-thickness excisional wound model.

The development of an effective treatment able to reduce the healing time of chronic wounds is a major health care need. In this regard, our research group has recently demonstrated the in vivo effectiveness of the topical administration of rhEGF-loaded lipid nanoparticles in healing-impaired db/db mice. Here we report the effectiveness of rhEGF-NLC (rhEGF loaded nanostructured lipid carriers) in a more relevant preclinical model of wound healing, the porcine full-thickness excisional wound model. The rhEGF-NLC showed a particle size of around 335nm, negative surface charge (-27mV) and a high encapsulation efficiency of 94%. rhEGF plasma levels were almost undetectable, suggesting that no systemic absorption occurred, which may minimise potential side effects and improve treatment safety. In vivo healing experiments carried out in large white pigs demonstrated that 20µg of rhEGF-NLC topically administered twice a week increased the wound closure and percentage of healed wounds by day 25, compared with the same number of intralesional administrations of 75µg free rhEGF and empty NLC. Moreover, rhEGF-NLC improved the wound healing quality expressed in terms of number of arranged microvasculature, fibroblast migration and proliferation, collagen deposition and evolution of the inflammatory response. Overall, these findings demonstrated that topically administered rhEGF-NLC may generate de novo intact skin after full thickness injury in a porcine model, thereby confirming their potential clinical application for the treatment of chronic wounds.

Frequency of obstructive sleep apnea syndrome in dental patients with tooth wear.

STUDY OBJECTIVES: To estimate the frequency of obstructive sleep apnea syndrome (OSAS) in dental patients with tooth wear, and to assess the role of dentists in the identification of patients at risk of OSAS. METHODS: Dental patients with tooth wear and treated with occlusal splint were prospectively recruited to perform sleep study. The severity of tooth wear was established by the treating dentist before patient referral to sleep disorders unit. Sleep questionnaires, anthropometric measurements, and validated respiratory polygraphy were performed. RESULTS: All patients with dental wear were offered a sleepiness analysis. Of 31 recruited patients, 30 (77% males) participated in this study. Patients' mean age was 58.5 ± 10.7 years (range: 35-90 years) and the body mass index was 27.9 ± 3.4 kg/m(2). Tooth wear was mild in 13 patients, moderate in 8 and severe in 9. The mean apnea-hypopnea index (AHI) was 32.4 ± 24.9. AHI < 5 was reported in 2 patients, AHI of 5-29 in 17, and AHI ≥ 30 in 11. A statistically significant association was found between AHI severity and tooth wear severity (Spearman R = 0.505; p = 0.004). CONCLUSIONS: Tooth wear could be a tool to identify those patients at risk of having OSAS. This highlights the importance of dental professionals to identify and refer patients with OSAS.

Infiltration of plasma rich in growth factors enhances in vivo angiogenesis and improves reperfusion and tissue remodeling after severe hind limb ischemia.

PRGF is a platelet concentrate within a plasma suspension that forms an in situ-generated fibrin-matrix delivery system, releasing multiple growth factors and other bioactive molecules that play key roles in tissue regeneration. This study was aimed at exploring the angiogenic and myogenic effects of PRGF on in vitro endothelial cells (HUVEC) and skeletal myoblasts (hSkMb) as well as on in vivo mouse subcutaneously implanted matrigel and on limb muscles after a severe ischemia. Human PRGF was prepared and characterized. Both proliferative and anti-apoptotic responses to PRGF were assessed in vitro in HUVEC and hSkMb. In vivo murine matrigel plug assay was conducted to determine the angiogenic capacity of PRGF, whereas in vivo ischemic hind limb model was carried out to demonstrate PRGF-driven vascular and myogenic regeneration. Primary HUVEC and hSkMb incubated with PRGF showed a dose dependent proliferative and anti-apoptotic effect and the PRGF matrigel plugs triggered an early and significant sustained angiogenesis compared with the control group. Moreover, mice treated with PRGF intramuscular infiltrations displayed a substantial reperfusion enhancement at day 28 associated with a fibrotic tissue reduction. These findings suggest that PRGF-induced angiogenesis is functionally effective at expanding the perfusion capacity of the new vasculature and attenuating the endogenous tissue fibrosis after a severe-induced skeletal muscle ischemia.