Biomechanical and histological comparison of two suture configurations for soft tissue grafts: speedtrap™ versus krackow stitch.

To compare 2 different graft preparation techniques to determine biomechanical strength and resultant tissue trauma evaluated by histology. Twelve common flexors of the finger's tendons were prepared with either tubulization (SpeedTrap™) or transtendon stiches (Orthocord™). The stiffness, resistance and energy at maximum load were tested for biomechanical assessment in both groups. After load testing, Samples were stained with hematoxylin and eosin (HE) to evaluate histological damage. We observe that the time to prepare tendons with SpeedTrap™ was 8.3 times faster (1:25 min) than traditional ones (15:02 min). In all cases, the mean values for SpeedTrap™ were higher in terms of strength, stiffness and energy at maximum load than for traditional suture but without significant difference (p > 0.05). The Krackow stitch produces greater structural damage to the collagen fibers while SpeedTrap™ maintains better organized arrangement of the fibers after tubulization preparation. With the results obtained, we can conclude that the tubulization technique allows faster graft preparation with less structural damage to the manipulated tissue without altering the biomechanical resistance provided by the transtendon suture technique.

Combined use of novel chitosan-grafted N-hydroxyethyl acrylamide polyurethane and human dermal fibroblasts as a construct for in vitro-engineered skin.

A rich plethora of information about grafted chitosan (CS) for medical use has been reported. The capability of CS-grafted poly(N-hydroxyethyl acrylamide) (CS-g-PHEAA) to support human dermal fibroblasts (HDFs) in vitro has been proven. However, CS-grafted copolymers lack good stiffness and the characteristic microstructure of a cellular matrix. In addition, whether CS-g-PHEAA can be used to prepare a scaffold with a suitable morphology and mechanical properties for skin tissue engineering (STE) is unclear. This study aimed to show for the first time that step-growth polymerizations can be used to obtain polyurethane (PU) platforms of CS-g-PHEAA, which can also have enhanced microhardness and be suitable for in vitro cell culture. The PU prepolymers were prepared from grafted CS, polyethylene glycol, and 1,6-hexamethylene diisocyanate. The results proved that a poly(saccharide-urethane) [(CS-g-PHEAA)-PU] could be successfully synthesized with a more suitable microarchitecture, thermal properties, and topology than CS-PU for the dynamic culturing of fibroblasts. Cytotoxicity, proliferation, histological and immunophenotype assessments revealed significantly higher biocompatibility and cell proliferation of the derivative concerning the controls. Cells cultured on (CS-g-PHEAA)-PU displayed a quiescent state compared to those cultured on CS-PU, which showed an activated phenotype. These findings may be critical factors in future studies establishing wound dressing models.

Association of high-risk human papillomavirus with ocular surface squamous neoplasia: a case-control study in Mexico.

OBJECTIVE: To investigate the association of high-risk hu-man papilloma virus (HR-HPV) and other risk factors with ocular surface squamous cell neoplasia (OSSN). MATERIALS AND METHODS: We obtained DNA from 22 fresh frozen OSSN tissues and 22 pterygia as controls, we used a broad-spectrum HPV DNA amplification short PCR fragment to identify HPV infection in all specimens and then genotyped HPV by a reverse hybridization line probe assay. We also obtained demographic, sun exposure, and tobacco consump-tion information. RESULTS: HR-HPV frequency was 40.9% in the OSSN group and 4.5% in the pterygia group (p=0.009). After covariate adjustment, OSSN was associated with HR-HPV (OR=16.3, 95%CI=1.2,218.1, p=0.03) and sunburn (OR=10.8, 95%CI=1.8,86.0, p=0.02). CONCLUSIONS: Ocular surface squamous cell neoplasia is a multifactorial disease. The strong association between HR-HPV and OSSN, suggests that HR-HPV could play an etiological role in OSSN development.

Association of high-risk human papillomavirus with ocular surface squamous neoplasia: a case-control study in Mexico / Asociación del virus del papiloma humano de alto riesgo con la neoplasia escamosa de la superficie ocular: estudio de casos y controles en México

Abstract: Objective: To investigate the association of high-risk human papilloma virus (HR-HPV) and other risk factors with ocular surface squamous cell neoplasia (OSSN). Materials and methods: We obtained DNA from 22 fresh frozen OSSN tissues and 22 pterygia as controls, we used a broad-spectrum HPV DNA amplification short PCR fragment to identify HPV infection in all specimens and then genotyped HPV by a reverse hybridization line probe assay. We also obtained demographic, sun exposure, and tobacco consumption information. Results: HR-HPV frequency was 40.9% in the OSSN group and 4.5% in the pterygia group (p=0.009). After covariate adjustment, OSSN was associated with HR-HPV (OR=16.3, 95%CI=1.2,218.1, p=0.03) and sunburn (OR=10.8, 95%CI=1.8,86.0, p=0.02). Conclusions: Ocular surface squamous cell neoplasia is a multifactorial disease. The strong association between HR-HPV and OSSN, suggests that HR-HPV could play an etiological role in OSSN development.

Resumen: Objetivo: Investigar la asociación del virus del papiloma humano de alto riesgo (VPH-AR), así como de otros factores, con neoplasia escamosa de la superficie ocular (NESO). Material y métodos: Se obtuvieron 22 especímenes de tejido fresco de NESO y 22 de pterigión como controles; se utilizó una técnica molecular altamente sensible para identificar la infección por VPH en todos los especímenes, así como la genotipificación del VPH. También se obtuvo información demográfica sobre exposición a la luz solar y tabaquismo. Resultados: La frecuencia de infección por VPH-AR fue de 40.9% en el grupo de NESO y de 4.5% en el grupo control (p=0.009). Después de ajustar por covariables, NESO se asoció con el VPH-AR (OR=16.3, IC95%=1.2,218.1, p=0.03) y el eritema solar (OR=10.8, IC95%=1.8,86.0, p=0.02). Conclusiones: La neoplasia escamosa de superficie ocular en una neoplasia multifactorial. Los presentes resultados sugieren que el VPH-AR podría tener un papel etiológico en el desarrollo de NESO.

A novel model to culture cells from giant cell tumor of bone using three-dimensional (3D) polycaprolactone scaffold.

Two-dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing GCTB cells on a polycaprolactone (PCL)-printed scaffold. We also evaluated the viability of the cultured cells and their adherence to the PCL scaffold at day 14 days using immunofluorescence analysis with calcein, vinculin, and phalloidin. Using the histological technique with hematoxylin and eosin staining, we observed all the histological components of GCTB in this 3D model. Immunohistochemical assays with cathepsin K, p63, and receptor activator of nuclear factor (NF)-κB ligand (RANKL) yielded positive results in this construct, which allowed us to confirm that the seeded cells maintained the expression of GCTB markers. Based on these findings, we concluded that the PCL scaffold is an efficient model to culture GCTB cells, and the cell viability and adherence to the scaffold can be preserved for up to 14 days. Moreover, this model can also be used in subsequent studies to assess in vitro cell-cell interactions and antineoplastic efficacy of certain agents to establish a treatment against GCTB.

Gamma radiation-induced grafting of poly(2-aminoethyl methacrylate) onto chitosan: A comprehensive study of a polyurethane scaffold intended for skin tissue engineering.

A novel brush-like poly(2-aminoethyl methacrylate) (PAEMA) was grafted onto chitosan (CS) through gamma radiation-induced polymerization. The copolymer (CS-g-PAEMA) was used to prepare a sodium acetate leached poly(urethane-urea) scaffold. The above derivatives were developed, synthesized, and characterized to meet the specific characteristics of biomaterials. The results revealed that this method is an easy and successful route for grafting PAEMA onto CS. The feasibility of preparing a CS-g-PAEMA polyurethane foam was confirmed by mechanical, morphometric, spectroscopic, and cytotoxic studies. The scaffold showed high biocompatibility both in vitro and in vivo. The first experiment proved that CS-based polyurethane efficiently allows the dynamic culturing of human fibroblast cells. Additionally, an in vivo study in a murine model indicated a complete integration of the scaffold to surrounding subcutaneous tissue as supported by the histological and histochemical assessments. The aforementioned results support the use of CS-g-PAEMA poly(saccharide-urethane) as a model of in vitro-engineered skin.

Gota tofácea en la columna lumbar causando radiculopatía / Tophaceous gout in the lumbar spine causing radiculopathy

Los depósitos de tofos en columna lumbar son una condición rara. Presentamos el caso de un varón de 44 años con dolor lumbar y radiculopatía. Las radiografías mostraron espondilolistesis lítica de L5, la resonancia magnética (RM) imagen hipointensa en T1 y heterogénea en T2 localizado en el espacio interespinoso L4-L5 y en articulación facetaria izquierda que invade el neuroforamen izquierdo. El ultrasonido de rodilla izquierda (RI) evidenció «doble contorno» del cóndilo femoral medial. Se realizó laminectomía descompresiva y artrodesis de L5-S1, el estudio histopatológico reportó material amorfo con reacción de células gigantes multinucleadas de tipo a cuerpo extraño

Tophaceous deposits in lumbar spine is considered a rare condition. We report the case of a 44-year-old patient with low back pain and radiculopathy. Radiographs revealed lytic spondylolisthesis in L5. Magnetic resonance imaging showed hypointense signal on T1 and a heterogeneous signal on T2 located in the L4-L5 interspinous space and in the left facet joint that invades left neuroforamen. The left knee ultrasound showed «double contour» of the medial femoral condyle. Decompressive laminectomy with arthrodesis at the level of L5-S1 was performed. The histological examination revealed amorphous material with a foreign body giant cell reaction

Tophaceous Gout in the Lumbar Spine Causing Radiculopathy. / Gota tofácea en la columna lumbar causando radiculopatía.

Tophaceous deposits in lumbar spine is considered a rare condition. We report the case of a 44-year-old patient with low back pain and radiculopathy. Radiographs revealed lytic spondylolisthesis in L5. Magnetic resonance imaging showed hypointense signal on T1 and a heterogeneous signal on T2 located in the L4-L5 interspinous space and in the left facet joint that invades left neuroforamen. The left knee ultrasound showed «double contour¼ of the medial femoral condyle. Decompressive laminectomy with arthrodesis at the level of L5-S1 was performed. The histological examination revealed amorphous material with a foreign body giant cell reaction.

Co-culture of dedifferentiated and primary human chondrocytes obtained from cadaveric donor enhance the histological quality of repair tissue: an in-vivo animal study.

To compare the quality of the repair tissue in three-dimensional co-culture of human chondrocytes implanted in an in vivo model. Six cadaveric and five live human donors were included. Osteochondral biopsies from the donor knees were harvested for chondrocyte isolation. Fifty percent of cadaveric chondrocytes were expanded until passage-2 (P2) while the remaining cells were cryopreserved in passage-0 (P0). Fresh primary chondrocytes (P0f) obtained from live human donors were co-cultured. Three-dimensional constructs were prepared with a monolayer of passage-2 chondrocytes, collagen membrane (Geistlich Bio-Gide®), and pellet of non-co-cultured (P2) or co-cultured chondrocytes (P2 + P0c, P2 + P0f). Constructs were implanted in the subcutaneous tissue of athymic mice and left for 3 months growth. Safranin-O and Alcian blue staining were used to glycosaminoglycan content assessment. Aggrecan and type-II collagen were evaluated by immunohistochemistry. New-formed tissue quality was evaluated with an adaptation of the modified O'Driscoll score. Histological quality of non-co-cultured group was 4.37 (SD ±4.71), while co-cultured groups had a mean score of 8.71 (SD ±3.98) for the fresh primary chondrocytes and 9.57 (SD ±1.27) in the cryopreserved chondrocytes. In immunohistochemistry, Co-culture groups were strongly stained for type-II and aggrecan not seen in the non-co-cultured group. It is possible to isolate viable chondrocytes from cadaveric human donors in samples processed in the first 48-h of dead. There is non-significant difference between the numbers of chondrocytes isolated from live or cadaveric donors. Cryopreservation of cadaveric primary chondrocytes does not alter the capability to form cartilage like tissue. Co-culture of primary and passaged chondrocytes enhances the histological quality of new-formed tissue compared to non-co-cultured cells.

Multicentric Spinal Cord Glioblastoma.

A 40-year-old man was referred to our center with a 4-month history of progressive quadriparesis. Previous brain imaging revealed no abnormality. Magnetic resonance imaging (MRI) with contrast of the cervicothoracic spine revealed 2 heterogeneously enhancing lesions at the level of C3-C5 and T2-T4 (A); T1- and T2-weighted MRI showed hypointense and hyperintense lesions, respectively, which is characteristic of spinal cord astrocytoma (B and C). Intraoperative biopsies supported the diagnosis of glioblastoma (hypercellularity, vascular proliferation, and tumor cell palisading around necrosis) (D). Though resection was attempted, surgery was aborted due to hemodynamic instability and signal changes in neuromonitoring. The patient stabilized in the intensive care unit and experienced slight improvement in upper extremity strength. After discharge without further complications, the patient was expected to return for further adjuvant therapy/surgical planning but unfortunately passed away 1.5 months after his operation, most likely to rapid disease progression.

Hepatic expression of ghrelin and adiponectin and their receptors in patients with nonalcoholic fatty liver disease.

BACKGROUND & AIM: Adiponectin and ghrelin are hormones that participate in hepatic lipid metabolism, and their expression in liver tissue could have important implications for nonalcoholic fatty liver disease (NAFLD). The aim of this study was to evaluate the hepatic expression of ghrelin, adiponectin, AdipoR, and IL-6 in patients with NAFLD and normal liver. METHODS: We studied patients with clinical-pathological diagnosis of NAFLD or a normal liver. Patients were classified according to their diagnosis into three groups: normal liver, nonalcoholic hepatic steatosis, and nonalcoholic steatohepatitis (NASH). Adiponectin, AdipoR1, AdipoR2, IL-6, and ghrelin mRNA levels were assessed in biopsies by reverse transcriptase-polymerase chain reaction. RESULTS: Of the 21 patients, three had a normal liver biopsy, 14 had nonalcoholic steatosis, and four had NASH. Patients with NAFLD exhibited significantly higher HOMA-IR and triglyceride concentration (both P<0.05). There was a nonsignificant trend towards higher ghrelin expression in patients with NASH > nonalcoholic steatosis > normal liver. Patients with NASH had significantly higher mRNA adiponectin levels and lower IL-6 levels than did those with a normal liver (P<0.05). AdipoR expression did not differ significantly between groups. CONCLUSION: Adiponectin overexpression was observed in patients with NASH. The role of hepatic ghrelin in NAFLD requires further research.

Effect of Helicobacter pylori infection on gastric ghrelin expression and body weight.

BACKGROUND & AIM: Ghrelin is a peptide mainly produced by gastric tissue playing an important role in energy homeostasis. It has been suggested that inflammatory and atrophic events induced by Helicobacter pylori (H. pylori) infection in gastric mucosa compromise the survival of the ghrelin-producing cells. The aim of this study was to investigate the effect of H. pylori infection on gastric ghrelin expression and body weight. METHODS: Consecutive patients referred for upper endoscopy were invited to participate. Patients with H. pylori infection (determined by histology) were defined as cases and patients without infection as controls. The density of colonization was classified in mild, moderate, or severe infection. Body mass index (BMI) was calculated. Ghrelin-immunoreactive cells were quantified in gastric biopsies by immunohistochemical staining. RESULTS: We studied 189 cases (92 males, 97 females) and 94 controls (55 males, 39 females). Cases were older (48.16 +/- 16.44 vs. 42.88 +/- 17.04 years, p < 0.05) and exhibited a lower percentage of ghrelin-immunoreactive cells (2.13% vs. 10.43%, p < 0.05) than controls. The prevalence of obesity was significantly lower than normal-weight among all cases, independently of the severity of infection (mild infection, 17.6% vs. 47.3%, p = 0.001; moderate-severe infection, 10.4% vs. 50%, p = 0.001). Univariate analysis showed a non-significant trend suggesting a protective effect of H. pylori against obesity. Nevertheless, BMI did not differ significantly between cases and controls. CONCLUSION: Chronic H. pylori infection contributes to a lower percentage of gastric ghrelin-immunoreactive cells but has no effect on the body weight of infected patients.

Hepatitis C virus infection and non-Hodgkin's lymphoma: a review and case report of nine patient.

The role of hepatitis C virus (HCV) is well established in the development of chronic hepatitis, cirrhosis and hepatic carcinoma, as well as in mixed type II cryoglobulinemia, membranoproliferative glomerulonephritis(MPGN) and porphyria cutanea tarda (PCT). Increasing evidence has been reported of a close association of HCV infection with autoimmune and hematological processes, mainly cytopenias and lymphoproliferative disorders such as B cell non-Hodgkin's lymphoma. We describe the demographic, clinical and histopathological findings of nine patients from the Mexican population with non-Hodgkin's lymphoma and HCV infection.

Low serum levels of ghrelin are associated with gallstone disease.

AIM: To explore the role of ghrelin in gallstone disease. METHODS: We carried out a cross-sectional study in 150 subjects, 38 with gallstones (cases) and 112 controls. We also did a real-time PCR-RT study in twenty gallbladder samples each. Body mass index (BMI), serum insulin, ghrelin, and serum lipids were measured. Logistic regression analyses (univariate and multivariate) were conducted to estimate the probability of gallstone disease associated with serum ghrelin concentrations. RESULTS: Cases were statistically different from controls in gender distribution (P = 0.01), age (53 vs 44 yr, P = 0.002), BMI (28 vs 25; P = 0.004), and glucose (5.26 vs 4.98 mmol/L; P = 0.05). The prevalence of ghrelin serum levels above the third tercile was lower in subjects without metabolic syndrome (P < 0.05). In a multivariate model, we found a protective effect, when ghrelin values were higher than the median value (OR = 0.27, 95%CI 0.09-0.82, P = 0.02). Twenty (20%) gallbladder specimens expressed ghrelin mRNA. CONCLUSION: Serum ghrelin concentrations are associated with a protective effect of GD.

The Amerindian's genes in the Mexican population are associated with development of gallstone disease.

BACKGROUND: It has been suggested that genes related to Amerindian ancestry account for the high prevalence of gallstone disease (GD) observed in Mexican-Americans. The HLA-B39 is an allele found in higher frequency in Amerindians whereas HLA-B15 is rarely found. The aim of this study was to test the hypothesis that gallstone susceptibility genes are more prevalent in Mexicans with recent Amerindian ancestry. METHODS: We carried out a prospective case-controlled study. Subjects were divided into those who had stones visible on gallbladder ultrasound (cases), and those whose ultrasounds were negative for gallstones (controls). Body mass index (BMI) was calculated, and serum lipids and lipoprotein, and glucose levels were measured. Class I HLA (HLA-B) typing was performed by PCR amplification of genomic DNA. RESULTS: Of the 1,101 subjects, 146 were classified as subjects with GD (cases) and 955 as subjects without GD (controls). Mean age of the cases was 53.5 +/- 12.5 yr versus 44.78 +/- 12.0 yr for the controls, p= 0.001. A family history of GD was observed in 48% of the cases versus 28.4% of the controls, p= 0.001. HLA-B39 was more frequently increased in GD subjects (0.162), compared with controls (0.063), p= 0.008. The odds ratio of having HLA-B39 was 2.8 and 95% (CI 95%= 1.3-6.3) for GD; HLA-B15 was more frequently increased in controls than in cases. CONCLUSIONS: The most prevalent HLA alleles detected in these GD cases are characteristic of Amerindian populations, supporting the role of genetics in the high prevalence of the development of GD in Mexican mestizos.

[Obesity and steatohepatitis. Histologic aspects]. / Obesidad y esteatohepatitis. Aspectos histológicos.

The traditional concept of adipose tissue as a passive reservoir for energy storage is no longer valid because it has been demonstrated that adipose tissue is a complex, essential, and highly active metabolic and endocrine organ that not only responds to afferent signals from traditional hormone systems and the central nervous system (CNS), but also expresses and secretes factors with important endocrine functions. These factors include leptin and other cytokines. Adipose tissue is also a major site for metabolism of sex steroids and glucocorticoids. The important endocrine function of adipose tissue is emphasized by adverse metabolic consequences of both adipose tissue excess and deficiency. Adipose tissue excess, particularly in visceral compartment, is associated with insulin resistance, hyperglycemia, dyslipidemia, hypertension, and prothrombotic and proinflammatory states. Liver is one of the principal targets of lipid-associated damage by mechanisms that involve apoptosis activation by source of tumoral necrosis factor-alpha and caspase activation and liberation of oxygen-reactive species by oxidative stress and enzymatic chains such as P450, CYP2E1, and CYP3A4, resulting in a continuum involving non alcohol-related fatty liver, non-alcoholic steatohepatitis with or without fibrosis, and liver cirrhosis. This work presents an overview of endocrine functions of adipose tissue and its influence on mechanisms of liver damage.

Prospective study of factors affecting postoperative pain and symptom persistence after stapled rectal mucosectomy for hemorrhoids: a need for preservation of squamous epithelium.

PURPOSE The purpose of the study was to determine the variables associated with postoperative pain and the clinical response of patients with uncomplicated hemorrhoidal disease treated with stapled rectal mucosectomy in the medium term. METHODS Patients with Grade II to IV, uncomplicated hemorrhoidal disease who underwent stapled rectal mucosectomy were prospectively included. The basal characteristics of the population were evaluated and level of stapling and placement of hemostatic suture determined. Histologically, the type of resected epithelium and presence of muscle fibers was evaluated. Postoperative pain was evaluated by means of a visual analog scale. Complications and clinical response were evaluated. RESULTS One hundred patients with a mean age of 43.9 years were included. Only columnar epithelium was resected in 48, transitional epithelium in 47, and squamous epithelium in 5 patients. Smooth muscle fragments were found in 55 patients, and, in 12 of these, fibers from the external muscular layer of rectum were also seen. Follow-up was 12.6 +/- 3.4 (range, 7-24) months. A total of 79 patients were completely asymptomatic at the end of follow-up. Resected squamous epithelium was associated with a higher postoperative pain level in the multivariate analysis (coefficient beta = 1.16 (95 percent confidence interval, 0.08-2.24); P = 0.035). CONCLUSIONS Rectal mucosectomy with stapler is an effective method for the treatment of uncomplicated prolapsing hemorrhoidal disease. Intensity of postoperative pain was associated with the type of resected epithelium. This suggests that low transection of hemorrhoids must be avoided.