Hipofosfatasemia crónica persistente de causa no determinada y su repercusión músculo-esquelética / Persistent hypophosphatasemia and its musculoskeletal repercussion

La fosfatasa alcalina baja o hipofosfatasemia, ya sea debida a causas genéticas (hipofosfatasia) o secundarias, presenta correlato clínico. Nuestro objetivo es estimar la prevalencia de hipofosfatasemia crónica persistente y describir sus hallazgos osteometabólicos. Se realizó una búsqueda electrónica de afiliados adultos al Hospital Italiano de Buenos Aires, entre 2013 y 2017, con al menos 2 determinaciones de fosfatasa alcalina igual a 30 UI/l o menor y ninguna mayor de 30 UI/l (rango de referencia 30-100 UI/l). Se excluyeron aquellos con causas secundarias diagnosticadas y se analizaron los correlatos clínico y bioquímico. Se detectó hipofosfatasemia crónica persistente en 78 de 105.925, 0,07% (0,06-0,09) de los afiliados. Solo uno fue excluido por tener causa secundaria. Eran 61,1% mujeres de 44 (34-56) años, fosfatasa alcalina 24 (20-27) UI/L, fosfatemia 4,1 (3,8-4,6) mg/dl. Se observaron osteoartritis, calcificaciones vasculares y fracturas, menos frecuentemente litiasis renal, calcificación del ligamento longitudinal común anterior, pérdida dental y convulsiones. El 63,6% tenían al menos una de las características clínico-radiológicas evaluadas, pero en solo 5,2% fue mencionado el diagnóstico de hipofosfatasemia en la historia clínica. La densitometría evidenció algún grado de afección (osteopenia u osteoporosis) en 76,2%. Se constataron 19 fracturas, con predominio en radio. La prevalencia de hipofosfatasemia fue similar a lo previamente reportado. El reconocimiento fue bajo; sin embargo, se observaron variadas manifestaciones músculo-esqueléticas, similares a las descriptas en la hipofosfatasia del adulto, por lo cual ­ante una hipofosfatasemia sin causa secundaria­ se sugiere considerar este diagnóstico. (AU)

Low alkaline phosphatase (ALP) or hypophosphatasemia either due to genetic (hypophosphatasia) or secondary causes, presents a clinical correlate. Our objectives are to estimate the prevalence of persistent hypophosphatasemia and to describe the clinical findings. We performed a search using the electronic medical records of the members of the Hospital Italiano de Buenos Aires health care system, between 2013 and 2017. Adult members with ≥ 2 ALP ≤ 30 IU/l, no ALP >30 IU/l (normal range 30-100 UI/l) and without diagnosed secondary causes were analyzed. Persistent hypophosphatasemia was detected in 78 of 105.925, 0.07% (0.06-0.09) of members. Only one was excluded due to a secondary cause, 61.1% were women, 44 (34-56) year-old, ALP 24 (20-27) IU/l and phosphatemia 4.1 (3.8-4.6) mg/dl. Osteoarthritis, vascular calcifications and fractures were detected, and nephrolithiasis, DISH (Diffuse idiopathic skeletal hyperostosis), tooth loss, and seizures were less frequently observed. At least one of the mentioned characteristics were present in 63.6 %, but only 5.2% had hypophosphatasemia registered in their clinical record. Densitometry showed osteopenia or osteoporosis in 76.2%. There were 19 fractures, most of them in radius. The prevalence of hypophosphatasemia was similar to what has been previously reported. Hypophosphatasemia finding in medical records was low, but far from being asymptomatic, clinical manifestations were observed. In the presence of hypophosphatasemia without a secondary cause, adult hypophosphatasia should be uspected. (AU)

Fosfatasa alcalina ósea sérica elevada no mediada por metástasis en un paciente con cáncer renal / Elevated bone alkaline phosphatase not mediated by metastasis in a patient with renal cancer

Presentamos un paciente de 63 años con cáncer renal y aumento de fosfatasa alcalina sérica de tipo óseo de acuerdo con su reactividad con anticuerpos monoclonales específicos. Se descartaron las causas conocidas de aumento de la isoenzima, incluyendo metástasis óseas. Los niveles enzimáticos cayeron abruptamente con la remoción del tumor, por lo que consideramos a este último como su origen. Diversas isoenzimas de fosfatasa alcalina pueden ser producidas y secretadas por tumores como manifestación paraneoplásica. El conocimiento de esto puede, en ocasiones, orientarnos hacia la presencia de una neoplasia oculta. Además, los cambios en los niveles séricos de esas isoenzimas pueden ser indicadores de respuesta al tratamiento o de recidiva tumoral. (AU)

A 63-year old man was seen in the outpatient clinic because of renal cancer and elevation in bone alkaline phosphatase measured by monoclonal antibodies assay. Known causes of bone isoenzyme augmentation, including bone metastases, were ruled out. The tumoral origin of the isoenzyme was diagnosed because after removal of the tumor the enzymatic levels fell sharply. Several alkaline phosphatase isoenzymes can be produced and secreted by tumors as a paraneoplasic manifestation and their elevation could be a manifestation of an occult neoplasia. Furthermore the monitoring of their blood levels can be useful means of treatment response and a tool to monitoring recurrence if a sharp decrease after removal of the tumor is observed. (AU)

TGF-β1 on induced osteogenic differentiation of human dermal fibroblast

PURPOSE: To evaluate the role of transforming growth factor beta 1 (TGF-β1) on the induced osteogenic differentiation of human dermal fibroblasts. METHODS: We performed four groups with cultured dermal fibroblasts according to the culture medium: CONTROL (DMEM culture medium); TGF-β1 (DMEM culture medium with 10 ng/ml of TGF-β1); OSTEOG (DMEM culture medium with 0.5 µg/ml of ascorbic acid, 10 mmol/l of β-glycerophosphate and 10 nmol/L of dexamethasone); and OSTEOG/TGF-β1 (osteogenic medium with 10 ng/ml of TGF-β1). Alkaline phosphatase (ALP) activity and the amount of osteocalcin (OC) in the supernatant, as well as the capability to form calcium phosphate deposits, were analysed for 28 days RESULTS: There were significant differences (p<0.05) between CONTROL and TGF-β1 groups in comparison with OSTEOG and OSTEOG/TGF-β1 groups in the ALP activity and OC amount. Although, both osteogenic groups had the same behavior with regard the expression curve during the experimental time, the OSTEOG/TGF-β1 group achieved significantly higher ALP and OC levels and showed no significant difference in the levels of mineralized deposits and in comparison with the levels found in the OSTEOG group. CONCLUSION: The addition of transforming growth factor beta 1 to the osteogenic culture medium increased the activity of alkaline phosphatase and the amount of osteocalcin, but TGF-β1 did not alter the presence of mineralized calcium phosphate deposits. .

Rol de la fosfatasa alcalina salival en el diagnóstico de las enfermedades periodontales / The role of alcaline phosphatase on the diagnosis of periodontal diseases

Uno de los tejidos más afectados en esta enfermedad es el tejido óseo que recubre a la pieza dentaria (hueso alveolar), que como consecuencia de la progresión de la enfermedad, se va produciendo una reabsorción y destrucción total del mismo. Sería de utilidadpoder contar con algún criterio diagnóstico, como son los marcadores biológicos, que permita detectar la posible agresión a los tejidos. La FAL ósea es una enzima directamente relacionada con el metabolismo osteológico de nuestro organismo, particularmente cuando se produce el remodelamiento (reabsorción-neoformación) del mismo. El propósito de esta revisión bibliográfica es determinar el rol de la fosfatasa alcalina salival como marcador bioquímico de la enfermedad periodontal.

Addition of bone morphogenetic protein type 2 to ascorbate and β-glycerophosphate supplementation did not enhance osteogenic differentiation of human adipose-derived stem cells

Bone morphogenetic protein type 2 (BMP-2) is a potent local factor, which promotes bone formation and has been used as an osteogenic supplement for mesenchymal stem cells. OBJECTIVES: This study evaluated the effect of a recombinant BMP-2 as well as the endogenous BMP-4 and BMP-7 in the osteogenic differentiation of adipose-derived stem cells (ASCs) in medium supplemented with ascorbate and β-glycerophosphate. MATERIAL AND METHODS: Human ASCs were treated with osteogenic medium in the presence (ASCs+OM+BMP-2) or absence (ASCs+OM) of BMP-2. The alkaline phosphatase (ALP) activity was determined and the extracellular matrix mineralization was evaluated by Von Kossa staining and calcium quantification. The expressions of BMP-4, BMP-7, Smad1, Smad4, and phosphorylated Smad1/5/8 were analyzed by western blotting. Relative mRNA expressions of Smad1, BMP receptor type II (BMPR-II), osteonectin, and osteocalcin were evaluated by qPCR. Results: ASCs+OM demonstrated the highest expression of BMP-4 and BMP-7 at days 21 and 7, respectively, the highest levels of BMPR-II mRNA expression at day 28, and the highest levels of Smad1 mRNA at days 14 and 28. ASCs+OM+BMP-2 demonstrated the highest levels of Smad1 mRNA expression at days 1, 7, and 21, the highest expression of Smad1 at day 7, the highest expression of Smad4 at day 14, the highest ALP activity at days 14 and 21, and expression of phosphorylated Smad1/5/8 at day 7. ASCs+OM and ASCs+OM+BMP2 showed similar ALP activity at days 7 and 28, similar osteonectin and osteocalcin mRNA expression at all time periods, and similar calcium depositions at all time periods. CONCLUSIONS: We concluded that human ASCs expressed endogenous BMP-4 and BMP-7. Moreover, the supplementation of ASCs with BMP-2 did not increase the level of osteogenic markers in the initial (ALP activity), intermediate (osteonectin and osteocalcin), or final (calcium deposition) phases, suggesting that the exogenous addition of BMP-2 did not improve the in vitro osteogenesis process of human ASCs.

Avaliação da influência do fluoreto no reparo ósseo em alvéolo dental de ratos: análise morfométrica e perfil de atividade das MMP-2 e 9 / Evaluation of the fluoride influence in alveolar bone repair in rats: morphometric analysis and MMP-2 activity

O flúor (F), na forma de fluoreto de sódio (NaF), é um elemento presente no cotidiano de quase a totalidade da população brasileira, sendo usado na água de abastecimento (Brasil) e também como terapêutico para osteoporose (comumente na Europa). Assim sendo, é importante o entendimento do efeito do flúor no reparo ósseo. Neste estudo foram utilizados 4 grupos com ratos Wistar de 80 dias (n=200), os quais receberam água de beber contendo 5, 15 e 50 ppm de flúor e água deionizada (grupo controle) durante todo experimento. Esses animais tiveram o incisivo superior direito extraído. Os animais foram eutanasiados O hora, 7, 14, 21 e 30 dias após a extração, sendo coletado sangue (análise da concentração de flúor), a região do alvéolo dental para análise microscópica (análise histológica e morfometria) e extração de proteínas referentes ao reparo ósseo (metaloproteinases de matriz - MMPs 2 e 9). A análise de concentração de flúor no plasma sangüíneo mostrou maior presença desse elemento no grupo tratado com 50 ppm de F nos períodos de 21 e 30 dias. A análise histológica detectou osso neoformado em todos os animais após 30 dias, porém o grupo de 50 ppm de F apresentou menor formação óssea que os outros grupos. A análise morfométrica mostrou um aumento na densidade de volume de osso neoformado, entre 7 e 30 dias, nos grupos controle, 5, 15 e 50 ppm de F, com concomitante diminuição na densidade de volume de tecido conjuntivo e coágulo sangüíneo. A atividade da MMP-2 foi mais acentuada no período inicial, enquanto a MMP-9 teve maior atividade no período final. Concluiu-se que o F, em altas concentrações, pode retardar o reparo alveolar diminuindo a formação de novo tecido ósseo, associado a um atraso na remissão do coágulo.

The fluorine (F), in the sodium fluoride (NaF) form, is an element daily present for most people, being used in water supply (Brazil) and also as therapy for osteoporosis (common in Europe). In such case it is important the understanding of the effect of fluoride in physiologic repair in bone tissue. This study used 4 groups of rats for 80 days (n = 200), which received drinking water containing 5, 15 and 50 ppm of fluoride, and deionized water (control group) throughout experimento These animais had the right upper incisor extracted. The animais were euthanized O hours, 7, 14, 21 and 30 days after extraction, and blood collected (analysis of the concentration of fluorine), the region of the dental alveolus for microscopic exa'mination (histological analysis and morphometry) and extraction of proteins for the bone repair (the matrix metalloproteinases - MMP 2 and 9). The analysis of concentration of fluoride in blood plasma showed greater presence of that element in the group treated with 50 ppm of F in periods of 21 and 30 days. Histological analysis detected neoformed bone in ali animais after 30 days, but the group of 50 ppm F showed lower bone formation than the other groups. Morphometric analysis showed an increase in the volume density of neoformed bone, between 7 and 30 days in control groups, 5, 15 and 50 ppm F, with a concomitant decrease in the volume density of connective tissue and blood dot. The activity of MMP-2 was more pronounced in the initial period, while MMP-9 had higher activity in the 30 days. It was concluded that the F, in high concentrations, may delay the tooth socket repair reducing the formation of new bone tissue, associated with a delay in remission of blood clot.

effects of zinc supplementation on serum zinc, alkaline phosphatase activity and fracture healing of bones

To determine the effect of zinc supplementation on callus formation, serum zinc and alkaline phosphatase activity in humans. This randomized, double-blind, placebo controlled clinical trial was conducted on 60 patients with traumatic bone fracture referred to Shohada Hospital of Tabriz, Iran from August to December 2007. Subjects were randomly divided into 2 groups: cases [n=30], receiving one capsule of zinc sulfate consists of 50 mg zinc each day and the controls [n=30], receiving placebo for 60 days. Individual and clinical information was determined by a questionnaire: nutritional intake by 3 days food records at the beginning and the end of trial. Serum zinc and alkaline phosphatase was measured by atomic absorption spectroscopy, and by enzymatic method. Callus formation during fracture healing was evaluated by radiography of the bone. There was no significant difference in physical activity, gender, age, type of fractures, and nutrient intake, between the 2 groups. The administration of zinc caused a significant elevation of serum zinc and alkaline phosphatase activity. Assessment of bone x- rays showed a significant progress in callus formation in cases compared to the controls. This study shows that zinc supplementation can stimulate fracture healing, however, it needs further study

Utilidad de la fosfatasa alcalina urinaria como marcador precoz de lesión tubular renal / Utility of urinary alkaline phosphatase as early marker of renal tubular failure

El objetivo de este estudio fue determinar la actividad de la fosfatasa alcalina urinaria (FALur) para evaluar precozmente lesión tubular y su utilidad diagnóstica. Los pacientes estudiados fueron: 20 Controles (C), 11 hipertensos (HTA), 23 diabéticos (DBT) y 34 con insuficiencia renal de diverso origen (IRDO). Se realizaron las determinaciones de: creatinina, clearence de creatinina (Jaffé cinético), FAL sérica y urinaria (cinético DGKC), microalbuminuria (inmunoturbidimétrico), proteiunuria (turbidimétrico), uroproteinograma, SDS-PAGE (al 12,5%) e isoenzimograma de FAL. La FAL sérica hallada fue normal, sin diferencia entre grupos y sin relación con el aumento de la FALur. El valor de corte recomendado para FALur fue de 8 UI/L. La FALur estuvo elevada en HTA e IRDO y normal en individuos con DBT. Los aumentos de FALur en IRDO se relacionaron con la lesión tubular estructural y en pacientes con HTA podrían relacionarse con alteración tubular precoz. Se propone la determinación de FALur para la detección temprana de lesión tubular ante falla renal establecida o en individuos con riesgo de desarrollarla, y se establece su utilidad en pacientes: - con DBT y HTA para seguimiento (junto a microalbuminuria y clearence de creatinina), - internados en riesgo de insuficiencia renal aguda: para orientar tratamientos, - con insuficiencia renal crónica: como indicador de lesión y pronóstico.

The objective of this study was to determine the activity of urinary Alkaline Phosphatase (ALPur) to evaluate early tubular failure and its diagnostic usefulness. The patients studied were: 20 Controls (C), 11 with Hipertensión (HTA), 23 Diabetic (DBT) and 34 with renal Insufficiency of diverse origin (IRDO). The creatinine, creatinine clearence (kinetic Jaffé) serum and urinary ALP (kinetic DGKC), microalbuminuria (Immunoturbidimetric), proteiunuria (Turbidimetric), uroproteinogram, SDS-PAGE (12.5%) and ALP isoenzymes determinations were made. The results indicate that serum ALP was normal, without difference between groups, and no relation with the increase in ALPur. Recommended cut-off value of ALPur was 8 UI/L. ALPur was elevated in HTA and IRDO, and normal in DBT. Increases in ALPur in IRDO were related to the structural tubular injury, and those in HTA could be related to early tubular alteration. Determination of ALPur is proposed for early detection of tubular injury, before renal failure is established or when there is risk of developing it, establishing its usefulness in: - DBT and HTA patients: screening (together with microalbuminuria and creatinine clearence). - Hospitalized patients in risk of acute renal insufficiency: in order to orient treatments. - Patients with chronic renal insufficiency: as an indicator of injury and prognosis.

Study of the new human molehydatiform alkaline phosphatase

Alkaline Phosphatase [EC: 3.1.3.1] is synthesized by kidney, liver, bone, Intestine and placenta. This enzyme is a glycoprotein and dimmer 4 Zn+2 and Mn+2 in each dimmer. It hydrolyzes mono ester phosphate to organic compound and phosphate_in alkaline medium. The purpose of this research is to compare this enzyme with placental alkaline phosphatase. Human Molehydatiform was purified by folds of precipitation of bybutanol, acetone, Amoniumm sulphate, Sephadex G200, affinity chromatography and preparative electrophoresis. Human Molehydatiform was purified 611.8 times. We obtained specific activity, optimum temperature and optimum Ph equaling to 611.8 U/mg, 40 centigrade degrees and 10.4 respectively. Purified Human Molehydatiform Alkalie phosphatase is different from Human placental Alkaline phosphatase due to optimum pH and optimum temperature

Determinación de fosfatasas en hongos de praderas / Phosphatases determination in praire fungi

Se realizó un estudio para determinar la actividad de la fosfatasa ácida y alcalina en 180 cepas fúngicas aisladas desde suelo rizósferico y la rizósfera de tres plantas forrajeras (Dactilys glomerata, Lolium perenne y Trifollium repens) cultivadas en una pradera en rotación y una pradera permanente. Las fosfatasas fueron determinadas por el método desarrollado por Tabatabai & Bremner (1969) y se leyeron en un espectrofotómetro a 400 nm. Los resultados obtenidos se sometieron a un análisis de varianza (ANDEVA). En las cepas fúngicas ensayadas se determinó actividad para fosfatasa ácida y alcalina. Los mayores valores tanto para fosfatasa ácida (139.68 mg/mL*g micelio) y alcalina (127.12 mg/mL*g micelio) los registró la cepa 14-2R de Penicillium chrysogenum aislada de la rizófera de L. perenne cultivado en pradera permanente. Entre las mejores cepas evaluadas de la pradera permanente existió una mejor concordancia entre los valores determinados para fosfatasa ácida y alcalina.