An Important Role of Macrophages for Wound Margin Regeneration in a Murine Flap Model

BACKGROUND: Macrophages have been known to have diverse roles either after tissue damage or during the wound healing process; however, their roles in flap wound healing are poorly understood. In this study, we aimed to evaluate how macrophages contribute to the flap wound regeneration.METHODS: A murine model of a pedicled flap was generated, and the time-course of the wound healing process was determined. Especially, the interface between the flap and the residual tissue was histopathologically evaluated. Using clodronate liposome, a macrophage-depleting agent, the functional role of macrophages in flap wound healing was investigated. Coculture of human keratinocyte cell line HaCaT and monocytic cell line THP-1 was performed to unveil relationship between the two cell types.RESULTS: Macrophage depletion significantly impaired flap wound healing process showing increased necrotic area after clodronate liposome administration. Interestingly, microscopic evaluation revealed that epithelial remodeling between the flap tissue and residual normal tissue did not occurred under the lack of macrophage infiltration. Coculture and scratch wound healing assays indicated that macrophages significantly affected the migration of keratinocytes.CONCLUSION: Macrophages play a critical role in the flap wound regeneration. Especially, epithelial remodeling at the flap margin is dependent on proper macrophage infiltration. These results implicate to support the cellular mechanisms of impaired flap wound healing.

Experimental evidence of pharmacological management of anchorage in Orthodontics: A systematic review

Introduction: Orthodontic anchorage is one of the most challenging aspects of Orthodontics. Preventing undesired movement of teeth could result in safer and less complicated orthodontic treatment. Recently, several reviews have been published about the effects of different molecules on bone physiology and the clinical side effects in Orthodontics. However, the effects of local application of these substances on the rate of orthodontic tooth movement have not been assessed.Objectives: The aim of this research was to analyze the scientific evidence published in the literature about the effects of different molecules on orthodontic anchorage.Methods: The literature was systematically reviewed using PubMed/Medline, Scopus and Cochrane databases from 2000 up to July 31st, 2014. Articles were independently selected by two different researchers based on previously established inclusion and exclusion criteria, with a concordance Kappa index of 0.86. The methodological quality of the reviewed papers was performed.Results: Search strategy identified 270 articles. Twenty-five of them were selected after application of inclusion/exclusion criteria, and only 11 qualified for final analysis. Molecules involved in orthodontic anchorage were divided into three main groups: osteoprotegerin (OPG), bisphosphonates (BPs) and other molecules (OMs).Conclusions: Different drugs are able to alter the bone remodeling cycle, influencing osteoclast function and, therefore, tooth movement. Thus, they could be used in order to provide maximal anchorage while preventing undesired movements. OPG was found the most effective molecule in blocking the action of osteoclasts, thereby reducing undesired movements.

Introdução: a ancoragem ortodôntica é um dos aspectos mais desafiadores da Ortodontia. A prevenção de movimentos dentários indesejados poderia resultar em um tratamento ortodôntico mais seguro e menos complexo. Recentemente, foram publicadas várias revisões de literatura sobre os efeitos de diferentes substâncias na fisiologia do tecido ósseo e os efeitos colaterais clínicos na Ortodontia. Porém, os efeitos da aplicação local dessas substâncias no grau de movimentação dentária ortodôntica não foram avaliados.Objetivos: o objetivo da presente pesquisa foi analisar a evidência científica publicada na literatura sobre os efeitos de diferentes substâncias na ancoragem ortodôntica.Métodos: a literatura foi sistematicamente revisada utilizando-se as bases de dados PubMed/Medline, Scopus e Cochrane, de 2000 a 31 de julho de 2014. Os artigos foram selecionados, de maneira independente, por dois pesquisadores diferentes, tendo como base critérios de inclusão e exclusão previamente estabelecidos, com um índice Kappa de concordância de 0,86. A qualidade metodológica dos artigos revisados foi analisada.Resultados: a estratégia de pesquisa identificou 270 artigos; 25 artigos foram selecionados após a aplicação dos critérios de inclusão e exclusão, mas apenas 11 foram qualificados para a análise final. As substâncias envolvidas na ancoragem ortodôntica foram divididas em três grupos principais: osteoprotegerina (OPG), bisfosfonatos (BFs) e outras substâncias (OSs).Conclusões: diferentes substâncias são capazes de alterar o ciclo de remodelação óssea, influenciando na função dos osteoclastos e, portanto, na movimentação dentária. Sendo assim, essas substâncias podem ser utilizadas para promover o máximo de ancoragem e prevenir movimentos indesejados. A OPG foi a substância mais eficaz no bloqueio da ação dos osteoclastos, reduzindo os movimentos indesejados.

Adjuvant Effect of IV Clodronate on the Delay of Bone Metastasis in High-Risk Prostate Cancer Patients: A Prospective Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: High-risk prostate cancer patients undergoing treatment often experience biochemical recurrence. The use of bisphosphonates as an adjuvant treatment delays skeletal events, yet whether or not bisphosphonates also delay metastastic development remains to be determined. MATERIALS AND METHODS: A total of 140 high-risk prostate cancer patients who were undergoing definitive treatment and who had clinically organ-confined disease and who suffered from biochemical recurrence were administered intravenous (IV) clodronate. The patients were treated with a radical retropubic prostatectomy (RP) or curative radiotherapy (RTx). Upon androgen deprivation therapy initiation, tri-monthly IV clodronate was added to the treatment to prevent bone demineralization. Twenty-six out of 60 operated cases and 45 out of 80 irradiated cases received bisphosphonate. The length of time until the first bone metastasis was recorded and analyzed. RESULTS: No statistical difference was found for the type of primary treatment (RP or RTx) on the time to the first bone metastasis (95% confidence interval [CI], 0.40 to 2.43; p=0.98). However, there was a clear advantage favoring the group that received bisphosphonate (p<0.001). The addition of bisphosphonate delayed the appearance of the first bone metastasis by seven-fold (95% CI, 3.1 to 15.4; p<0.001). CONCLUSION: Treatment with tri-monthly IV clodronate delayed the time to the first bone metastasis in high-risk prostate cancer patients who were experiencing an increase in the prostate specific antigen level after definitive treatment.

The investigation of macrophage infiltration in the early phase of ischemic acute renal failure in mice

BACKGROUND/AIMS: Inflammation plays a key role in ischemic acute renal failure (ARF). The present study investigated the infiltration of macrophages in the early phase of ischemic ARF in mice. METHODS: Ischemic ARF was induced by renal clamping for 22 min, while the control mice underwent sham surgery (no clamping). The serum creatinine and blood urea nitrogen (BUN) levels were measured in the control and post-ischemia mice. Immunofluorescence staining was used to measure the number of CD 11b-positive cells in the kidney tissue sections to determine the amount of post-ischemic macrophage infiltration. Lipo-Cl2MBP (clodronate) for macrophages depletion was injected via a tail vein 5 d before ischemia induction and again 2 d before ischemia induction. RESULTS: The study found that the post-ischemia mice had higher levels of serum creatinine and BUN at 16 and 24 h compared to the controls. Immunofluorescence staining showed there were more macrophages in the post-ischemic tissue at 2, 8, 16 and 24 h compared to the control tissue, and that most of these macrophages were located in the outer medulla. The mice treated with clodronate prior to ischemia induction were found to have lower levels of serum creatinine compared to those mice that weren't treated with clodronate. CONCLUSIONS: There was significant infiltration of macrophages from the early phase of ischemic ARF, and this peaked at 16-24 h. Macrophage depletion using clodronate was protective against ischemic ARF.

Macrophage Depletion by Clodronate Liposomes Suppresses Neointimal Formation After Carotid Artery Injury in Apolipoprotein E-Deficient Mice

BACKGROUND AND OBJECTIVES: Clodronate liposomes deplete phagocytic cells, thereby suppressing inflammation after vascular injury. We compared the effect of clodronate liposomes on macrophage depletion and neointimal formation in apolipoprotein E-deficient mice [ApoE (-) mice]. MATERIALS AND METHODS: ApoE (-) mice were randomly assigned to the clodronate liposomes group (Clodronate Group, n=7) and the vehicle liposomes group (Control Group, n=7). Clodronate (0.1 mL/10 g) was injected via the tail vein starting 2 days (d-2) before left common carotid artery injury. RESULTS: The percentage of blood monocytes was subsequently decreased after clodronate injection (14.0+/-7.4% at baseline, 6.8+/-4.9% at 24 hours and 0.7+/-0.3% at 1 week after the clodronate liposome injection). The percentage of macrophages in the plaque area was significantly lower in the clodronate group at week 2 (32.0+/-6.5 vs. 68.7+/-7.6%, respectively, p<0.05) and at week 4 (37.3+/-8.5 vs. 62.6+/-9.4%, respectively, p<0.05). The interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha concentrations were significantly decreased in the clodronate group at week 4 (12.3+/-2.5 vs. 22.9+/-3.5 pg/mL, respectively, p<0.05 for IL-6 and 16.6+/-2.2 vs. 43.6+/-6.1 pg/mL, respectively, p<0.05 for TNF-alpha). The plaque volume was significantly greater in the control group at week 2 (0.345+/-0.063 vs. 0.153+/-0.053 mm2, respectively, p<0.05) and at week 4 (0.320+/-0.027 vs. 0.167+/-0.070 mm2, respectively, p<0.05). CONCLUSION: Intravenous administration of clodronate liposomes depleted monocytes and macrophages, and so this reduced the inflammatory markers and neointimal formation in ApoE (-) mice.

Experimental study for thrombocytopenic purpura therapy by targeting macrophages in liver and spleen / 中国实验血液学杂志

The study purpose was to explore whether dichloromethylene diphosphonate (Cl(2)MDP)-loaded gelatin particles can induce the depletion of macrophage in reticuloendothelial system of liver and spleen or can depress the immunity of macrophage in SD rat models of immune thrombocytopenic purpura (ITP) to treat the ITP rats. New Zealand rabbits were immunized with platelets of SD rats to prepare rabbit anti-rat platelet serum, and the serum was intravenously injected into SD rats to produce the ITP model. In experimental ITP models, 150 microl of anti-platelet serum was intravenously injected into SD rats per 24 hours. The platelet counts maintained pathological level and were persistently less than 50 x 10(9)/L in the models during experiment process. The MTT test of macrophage RAW264.7 was carried out by means of Cl(2)MDP-loaded gelatin particles in vitro. After intravenous injection of a group dose of Cl(2)MDP-gelatin particles, the platelet counts of the rats were measured at the time of 4 hours, 24 hours, 48 hours, 72 hours and 96 hours, respectively, and bleeding times were detected in 24 hours. The results showed that Cl(2)MDP-loaded gelatin particles increased the platelet counts of ITP models to mean of 180 x 10(9)/L, a physiological level in 24 hours after injection, and kept this platelet level through whole process of 120 hours. Furthermore, rats pre-treated with Cl(2)MDP-loaded gelatin particles avoided the decrease of platelet counts significantly when they were injected anti-platelet serum. It is concluded that Cl(2)MDP-loaded gelatin particles restrain multiplication of macrophage RAW264.7, and promptly, effectively restore platelet counts of ITP models to physiological level in a dose dependent manner. So, the targeting therapy of drug-loaded gelatin particles offers a new idea and approach to treat ITP, and this strategy is worthy of further studies.

Uso do clodronato endovenoso na calcinose difusa em uma criança com síndrome de superposição esclerose sistêmica e dermatomiosite / Use of clodronate in extensive calcinosis in a child with systemic sclerosis and dermatomyositis overlap

Os autores descrevem o caso de uma garota negra, com diagnóstico de síndrome de superposição dermatomiosite e esclerose sistêmica, que desenvolveu calcinose difusa, complicada por infecções secundárias e significativa limitação funcional de membros. Tratamento com colchicina, diltiazem e alendronato sódico não se mostrou eficaz no controle da calcinose, requerendo uso endovenoso bimestral de clodronato, que contribuiu para significativa melhora na cicatrização das úlceras cutâneas e na qualidade de vida.

Revisão: há espaço para o uso dos bisfosfonatos na osteodistrofia? / Should bisphosnates be used to treat osteodystrophy?

Os bisfosfonatos são drogas que inibem a reabsorção óssea e são utilizados clinicamente para o controle das hipercalcemias, calcificações ectópicas e osteoporose. Pacientes com insuficiência renal crônica em diálise e transplantados poderiam se beneficiar do uso dessas drogas. Entretanto, por serem drogas de excreção renal, exigem cautela e conhecimento da sua farmacologia. Nos pacientes em diálise, começam a ser empregados para o controle do hiperparatiroidismo, melhorando a hipercalcemia e permitindo o uso de calcitriol. Em transplantados, vêm sendo utilizados para controlar a perda de massa óssea no pós-transplante, causada por persistência do hiperparatiroidismo ou pelo uso das drogas imunossupressoras, principalmente os corticóides.

Tratamiento de trombocitopenias inmunes en un modelo experimental / Treatment of immune thrombocytopenias in an experimental model

La púrpura trombocitopénica inmune (PTI) es un desorden hematológico caracterizado por un aumento del consumo de plaquetas. La destrucción plaquetaria es mediada por el sistema retículo-endotelial (SRE), en especial macrófagos hepáticos y esplénicos. Nosotros demostramos que la eliminación específica de estas células utilizando clodronato encapsulado en liposomas (lipclod) induce un aumento del recuento plaquetario. Además, encontramos que la trombocitopenia puede puede ser revertida por el lipclod sin comprometer totalmente la integridad del SRE al cabo de 18h de tratamiento. Los tiempos de sangría de los animales tratados fueron similares a los de los de los animales controles, sugiriendo que la hemostasia está bien controlada en estos animales. Estos datos indican que el uso de lip-clod merece ser considerado como una estrategia de tratamiento para aquellos trombocitopénicos donde se procura elevar rápidamente el recuento plaquetario.

Bisfosfonato e movimentaçäo dentária induzida: avaliaçäo microscópica de seus efeitos / Biophosphonates and induced tooth movement: microscopical evaluation of its effects

Os biofosfonatos säo eficientes bloqueadores das reabsorçöes e, por isso, säo utilizados no controle das doenças metabólicas ósseas. Os mecanismos pelos quais essas drogas interferem nos processos de reabsorçäo ainda näo estäo totalmente esclarecidos, mas incluem a inibiçäo da funçäo e a alteraçäo da morfologia dos clastos e precursores, além da atuaçäo como citotóxicos sobre os macrófagos e os osteoblastos. Num modelo experimental de movimentaçäo dentária induzida, procurou-se verificar, microscopicamente, a influência do clodronato sobre a morfologia do osso alveolar, do ligamento periodontal e do cemento dos primeiros molares superiores de 45 ratas com 90 dias de vida, nos tempos experimentais de 24 horas, 3, 5, 7, 10 e 21 dias de movimento. Os resultados microscópicos revelaram que a administraçäo do clodronato näo interferiu em nenhuma das situaçöes referidas, inclusive durante o movimento dentário induzido, nas diversas fases analisadas. A presença localizada mais intensa de infiltrado inflamatório crônico sugere uma influência dos bisfosfonatos nos processos inflamatórios

Experiencia en el uso del clodronato en tastasis oseas del cancer de la mama / Experience in using clodronato in breast cancer`s osseus methastasis

Los bifosfonatos son compuestos de estructura similar a los pirofosfatos, con un atomo de C uniendo los fosfatos. Son efectivos en el tratamiento de metastasis oseas por via oral o endovenosa, si bien no presentan accion antitumoral. Se presenta la evolucion de 5 pacientes que recibieron clodronato por via oral. En todas ellas se obtuvo mejoria del dolor oseo, con una mayor y mejor sobrevida, sin aparicion de fracturas, compresiones ni otras manifestaciones de recurrencia que necesitaran radio o quimioterapia. Dos casos llevan mas de dos años de tratamiento. El efecto del clodronato es mas favorable cuando se administra tempranamente, con la aparición de las metástasis óseas

use of bisphosphonates in skeletal disease due to breast cancer

The variability of different breast cancers in the susceptibility to metastatic bone disease is poorly understood, but may depend in part upon gene expression of PTHrP and the vitamin D receptor. In contrast, much more is known of the manner which metastatic breast disease affects bone remodelling to induce osteolytic bone disease. Mechanisms include a generalised increase in activation frequency at sites close to metastatic tissue, an imbalance between the amount of bone formed and that resorbed within resorption cavities, and uncoupling of bone formation from bone resorption. The greatest morbidity from metastatic bone disease arises from osteolytic disease and gives rise to hypercalcemia, bone pain and fractures. Since osteolysis is primarily mediated by the activation of osteoclasts, there has been a great deal of interest in the use of agents which primarily affect bone pain and fractures. Since osteolysis is primarily mediated by the activation of osteoclasts, there has been a great deal of interest in the use of agents which primarily affect bone metabolism to alter the natural history of metastatic bone disease. Non-steroidal anti-inflammatory agents and cytotoxic agents are capable of inducing responses in bone, but are limited by their toxicity when effective doses are utillized. The use of calcitonin in the long term suppression of osteolysis has also been disappointing. The bisphosphonates are, however, capable of inducing sustained decreases in osteoclast activity and numbers in patients with osteolytic bone disease. There are now several studies which have examined the effects of the bisphosphonates on skeletal morbidity in breast cancer. Both clodronate and pamidronate decrease the incidence of hypercalcemia, bone pain and pathological fractures, but do not significantly alter mortality. Given, however, the unchanging survival in patients with metastatic bone disease, significant improvements in the quality of remaining life is an important therapeutic effect

Clodronate in hypercalcemia and osteolytic bone disease due to breast cancer

Progressive osteolysis is a significant cause for morbidity in patients with neoplasia affecting the skeleton. It gives rise to fractures, bone pain and hypercalcaemia. The mechanism for osteolysis is principally mediated by the activation of bone resorbing cells. The bisphosphonates are specific inhibitors of osteoclast mediated bone resorption and have been widely used in the management of osteolysis. Clodronate is one of the bisphosphonates that may be given by mouth or by parenteral injections. Both formulations lower serum calcium in the vast majority of affected patients due to the inhibition of bone resorption. Moreover, the agent also has significant effects on bone pain. There is increasing evidence that the long-term use of Clodronate decreases the incidence of intercurrent hypercalcaemia, bone pain and fracture and thereby improves the quality of life of affected patients