Olga Roda

Synergism between melatonin and atorvastatin against endothelial cell damage induced by lipopolysaccharide

Abstract:  The beneficial effects of atorvastatin are based on both cholesterol‐dependent and independent mechanisms. The latter probably include the ability of the estatin to enhance the expression of endothelial nitric oxide synthase (eNOS) and to cause a vasodilatation. In turn, the antioxidant and anti‐inflammatory actions of melatonin are related to its vascular protection. In the present study, we investigated the efficacy of the combination of melatonin plus atorvastatin against endothelial cell damage induced by inflammation and oxidative stress injury. Human umbilical vein endothelial cells (HUVEC) were cultured with bacterial lipopolysaccharide (LPS) in the presence or absence of melatonin and/or atorvastatin. LPS inhibited eNOS mRNA and protein expression, which was reversed by atorvastatin and, to a lesser extent, by melatonin. Together, melatonin + atorvastatin induced higher eNOS protein expression than either compound alone. Melatonin, but not atorvastatin, reduced free radical generation, lipid peroxidation, and interleukin‐6 levels induced by LPS. In the presence of atorvastatin, the effects of melatonin were maintained or even improved. These data suggest that melatonin improves the beneficial effects of atorvastatin and reduces its side effects in endothelial cells during inflammation and under conditions of oxidative stress.

In vitro characterization of a nanostructured fibrin agarose bio-artificial nerve substitute

Neural tissue engineering is focused on the design of novel biocompatible substitutes to repair peripheral nerve injuries. In this paper we describe a nanostructured fibrin–agarose bioartificial nerve substitute (NFABNS), based on nanostructured fibrin–agarose hydrogels (FAHs) with human adipose‐derived mesenchymal stem cells (HADMSCs). These NFABNSs were mechanically characterized and HADMSCs behaviour was evaluated using histological and ultrastructural techniques. Mechanical characterization showed that the NFABNSs were resistant, flexible and elastic, with a high deformation capability. Histological analyses carried out in vitro during 16 days revealed that the number of HADMSCs decreased over time, with a significant increase after 16 days. HADMSCs formed cell clusters and degraded the surrounding scaffold during this time; additionally, HADMSCs showed active cell proliferation and cytoskeletal remodelling, with a progressive synthesis of extracellular matrix molecules. Finally, this study demonstrated that it is possible to generate biologically active and mechanically stable tissue‐like substitutes with specific dimensions, based on the use of HADMSCs, FAHs and a nanostructure technique. However, in vivo analyses are needed to demonstrate their potential usefulness in peripheral nerve repair. Copyright

Development of the human shoulder joint during the embryonic and early fetal stages: anatomical considerations for clinical practice

Although several studies have been published regarding the morphology and anatomical variations of the human shoulder joint, most have dealt with adult individuals. Those looking into the development of the joint have been focused on specific structures or have observed specimens in advanced gestational stages. The goal of this paper is to perform a complete analysis of the embryonic and early fetal development of the elements in the shoulder joint, and to clarify some contradictory data in the literature. In our study, serial sections of 32 human embryos (Carnegie stages 16–23) and 26 fetuses (9–13 weeks) were analyzed. The chondrogenic anlagen of the humerus and the medial border of the scapula can be observed from as early as Carnegie stage 17, whereas that of the rest of the scapula appears at stage 18. The osteogenic process begins in week 10 for the humeral head and week 11 for the scapula. At stage 19 the interzone becomes apparent, which will form the glenohumeral joint. In the next stage the glenohumeral joint will begin delaminating and exhibiting a looser central band. Denser lateral bands will join the humeral head (caput humeri) and the margins of the articular surface of the scapula, thus forming the glenoid labrum, which can be fully appreciated by stage 22. In 24‐mm embryos (stage 21) we can observe, for the first time, the long head of the biceps tendon (which is already inserted in the glenoid labrum by week 9), and the intertubercular sulcus, whose depth is apparent since week 12. Regarding ligamentous structures, the coracohumeral ligament is observed at the end of Carnegie stage 23, whereas the primitive glenohumeral ligament already appeared in week 10. The results of this study provide a detailed description of the morphogenesis, origin and chronological order of appearance of the main intrinsic structures of the human shoulder joint during late embryonic and early fetal development. We expect these results to help explain several functional aspects of the shoulder joint, and to clarify some contradictory data in the literature regarding this complex anatomical and biomechanical structure, helping future researchers in their efforts.

Generation of a bioengineered autologous bone substitute for palate repair: an in vivo study in laboratory animals

We carried out an in vivo study to evaluate the potential usefulness of a novel bioengineered bone substitute for the repair of palate defects in laboratory rabbits, using tissue‐engineering methods. Our results showed that the use of a bioengineered bone substitute was associated with more symmetrical palate growth as compared to the controls, and the length and height of the palate were very similar on both sides of the palate, with differences from negative controls 4 months after artificial bone grafting for bone length. The histological analysis revealed that the regenerated bone was well organized and expressed osteocalcin. In contrast, bone corresponding to control animals without tissue grafting was immature, with areas of osteoid tissue and remodelling, as determined by MMP‐14 expression. These results suggest that bone substitutes may be a useful strategy to induce the formation of a well‐structured palate bone, which could prevent the growth alterations found in cleft palate patients. This opens a door to a future clinical application of these bone substitutes. Copyright

Histological and immunohistochemical study of an unusual type of gallbladder duplication

Gallbladder duplication is a rare congenital anomaly, with an incidence of 1 in 3,800 autopsies. The correct diagnosis and treatment of this type of entity is important in clinical practice, because it may cause some clinical and surgical problems. In this report, we present the clinical case of a 28-year-old female with abdominal pain. Ultrasound of the upper abdomen showed a distended gallbladder with the presence of a septum that could suggest a congenital anomaly of the extrahepatic biliary system. During surgery, a distended and inflamed gallbladder with a lithiasis was found. In addition, a complete septum and double cystic duct were observed. The gross and histopathological evaluation of the surgical specimen allowed us to confirm the diagnosis of a Y- shaped type gallbladder duplication according to Boydens classification. In conclusion, in presence of an atypical imaging of the gallbladder, diagnosis of this group of congenital anomalies should be considered in order to adequately plan surgical intervention if necessary.

Lysophosphatidic Acid Pretreatment Prevents Micromolar Atorvastatin-Induced Endothelial Cell Death and Ensures the Beneficial Effects of High-Concentration Statin Therapy on Endothelial Gene Expression

Because of the pleiotropic effects of statins, it may potentially be used as a locoregional adjuvant in vascular revascularization, tissue engineering, and regenerative procedures. Electron probe X-ray microanalyses and oligonucleotide microarrays were used to identify the global effects of micromolar concentrations of atorvastatin on the gene expression and cell viability of endothelial cells in different states of lysophosphatidic acid (LPA)-induced activation. Treatment with 1-μM atorvastatin for 24 hours significantly reduced the viability of human vascular endothelial cells (HUVECs). However, the same treatment of LPA-preactivated HUVECs produced elevated cell viability levels and an optimal vascular gene expression profile, including endothelial nitric oxide synthase overexpression, endothelin-1 repression, an anti-inflammatory genetic pattern, and upregulation of molecules involved in maintaining the endothelial barrier (vascular endothelial cadherin, claudin 5, tight junction protein 1, integrin β4). The atorvastatin treatment also produced a repression of microRNA 21 and genes involved in cell proliferation and neointimal formation (vascular endothelial growth factor [VEGF] A, VEGF receptor 1, VEGFC). Results obtained suggest that micromolar atorvastatin therapy can enhance global endothelial function, but its effects on cell viability vary according to the baseline state of cell activation (preactivated, postactivated, or not activated). Preactivation with LPA protects HUVECs against atorvastatin-induced apoptosis and delivers optimal levels of cell viability and functionality.

Anatomic study of pedicled bipolar teres major transfer for irreparable posterosuperior rotator cuff tears

BACKGROUND Treatment of rotator cuff (RC) tears has not included bipolar muscle-tendon transfers to date. The objective of this study was to verify the feasibility of pedicled bipolar teres major (TM) transfer over and under the long head of the triceps brachii (LHT) and compare its versatility with monopolar transfer in a model of supraspinatus (SS) tears in cadavers. METHODS In 6 shoulders of cryopreserved cadavers, we re-created complete SS tears, conducting monopolar and bipolar TM transfers over and under LHT. We compared the morphology of the SS and TM, defect coverage, angle between the transferred TM and major SS axis, and axillary nerve overlap with each technique. RESULTS The TM and SS were morphologically similar. Defect coverage was significantly lower with monopolar transfer (12 ± 4 mm) than with bipolar transfer (39 ± 9 mm under the LHT, P = .003, and 38 ± 8 mm over the LHT, P = .004). The bipolar transfer course over the LHT was the nearest to the SS axis (39° ± 11°, P = .005). We found a greater axillary nerve overlap with bipolar transfer under the LHT (27 ± 8 mm) than with bipolar transfer over the LHT (1 ± 2 mm, P = .005) or monopolar transfer (0 mm, P < .001). CONCLUSION Bipolar TM transfer is possible without neurovascular pedicle interference, obtaining greater RC defect coverage and the closest path to the SS axis when conducted over the LHT compared with monopolar or bipolar transfer under the LHT. Accordingly, it can be considered an alternative option for the treatment of posterosuperior RC defects.

Predicting successful prosthetic rehabilitation in major lower-limb amputation patients: a 15-year retrospective cohort study

OBJECTIVE To determine and compare specific factors that could be associated and predictive with successful prosthetic rehabilitation in major lower-limb amputations. METHODS A 15-year long (2000-2014) retrospective observational cohort study was conducted. Two different criteria were used to define successful prosthetic rehabilitation: (1) the ability to walk at least 45m, regardless of assistive devices; and (2) walking >45m without other ambulatory aids than one cane (if required). Age, gender, comorbidities, cause and level of amputation, stump characteristics, ulcers in the preserved limb, and time between surgery and physical therapy were examined as predictors of successful prosthetic rehabilitation. RESULTS A total of 169 patients (61.60±15.9 years) were included. Regarding walking ability with or without walking aids, the presence of ulcers in the preserved limb was individually associated with failed prosthetic rehabilitation (p<0.001), while being male (OR=0.21; 95%CI=0.06-0.80) and transtibial level of amputation (OR=6.73; 95%CI=1.92-23.64) were identified as independent predictors of failure and success, respectively. Regarding the criterion of successful rehabilitation, a shorter time until rehabilitation was individually associated with improved walking ability (p<0.013), while failure could be predicted by comorbidities (OR=0.48; 95%CI=0.29-0.78) and age groups of 65-75 years old (OR=0.19; 95%CI=0.05-0.78) and over 75 years old (OR=0.19; 95%CI=0.04-0.91). CONCLUSIONS Regarding walking ability with or without walking aids, male gender and transtibial level of amputation are independently associated with failure and success respectively, whereas older age and comorbidities can predict failed prosthetic rehabilitation when assistive walking devices are considered. Future prospective cohort studies are needed to confirm these findings.