Release of lithium from 3D printed polycaprolactone scaffolds regulates macrophage and osteoclast response.

The immunomodulatory effects of lithium have been reported across a range of models and contexts. Lithium appears to have a positive effect on osteogenesis in vivo, while in vitro outcomes throughout the literature are varied. Tissue engineering approaches have rarely targeted local lithium delivery within a regenerative setting. We hypothesized that part of the positive effects of lithium in vivo may be due to an immunomodulatory effect manifesting in a local environment. To achieve a sustained lithium release from scaffold constructs, we blended lithium carbonate, a soluble salt of lithium, with the biomaterial polymer polycaprolactone (PCL). We printed constructs of PCL alone, and with 5% (5Li) and 10% (10Li) lithium carbonate. Mechanical testing revealed that mechanical properties were largely retained with lithium carbonate incorporation, and we measured a consistent release of the ion over a 7 day period. The efficacy of our construct system was then assessed using a primary mouse macrophage culture, and a differentiated osteoclast culture. We found that the lithium released from constructs had a great effect on macrophage polarization, resulting in pronounced upregulation of immunomodulatory (M2) genes, and a decrease in pro-inflammatory (M1) genes. This was reflected in cytokine expression, and illustrated through immunofluorescent staining. Osteoclast activity was greatly suppressed by the lithium incorporation, with a marked effect on gene expression and actin ring formation. Our work demonstrated an effective system for local lithium delivery, confirmed the pronounced effects that lithium has on macrophage and osteoclast response, and sets the stage for further innovations in ion release for targeted tissue engineering.

Lithium entrapped chitosan nanoparticles to reduce toxicity and increase cellular uptake of lithium.

Lithium carbonate is an effective drug against bipolar disorders. Direct use of lithium carbonate has been reported to result in lithium toxication and pulmonary complications. With chitosan micro and nanoparticles gaining attention for their protein absorption, drug targeting and improved dissolution rate of sparingly water-soluble drugs, this work has focused on chitosan loaded Li as a possible alternative to Li alone for cellular uptake. Well standardized ionic gelation technique employed in this study resulted in Li loaded chitosan nanoparticles with hydrodynamic diameter below 300 nm and zeta potential of + 30 mV and oval morphology. Through various techniques electrostatic interaction as well as Claritin dependent endocytic pathway is suggested as facilitating 1.3 times increase in cell proliferation in lithium carbonate loaded chitosan nanoparticles treated PC12 cells. A controlled Li release to the extent of less than 50% in 48 h from the nanoparticle was observed. This observation has very high significance as it ensures that the lithium toxicity can be avoided. These results indicated that chitosan is a promising carrier for lithium carbonate and may improve its therapeutic efficacy and also overcome toxicity during its use in the treatment of neuropsychiatric disorders.

Desenvolvimento, caracterização e desempenho biológico de um novo biomaterial de liberação controlada à base de carbonato de lítio aplicado ao reparo ósseo / Development, characterization and biological performance of a new biomaterial of controlled release based on lithium carbonate applied to bone repair

Neste trabalho desenhou-se bases de desenvolvimento, caracterização e avaliação dos aspectos biológicos relacionados ao Sistema BoneLithium, idealizado a partir da associação de partículas de carbonato de lítio dispersas em matriz gel de carbopol®, com capacidade de atuar como um sistema liberador de fármacos. Metodologicamente este estudo se dividiu em quatro partes: Na primeira delas, o objetivo central foi o desenvolvimento e a caracterização do biomaterial através da manipulação farmacológica. Na segunda etapa, avaliou-se a reação tecidual em subcutâneo de ratos, na terceira a influência das partículas de lítio liberadas pelo Sistema BoneLithium no reparo ósseo através de modelos experimentais utilizando coelhos, e por ultimo, a capacidade de cicatrização de defeitos ósseos criados cirurgicamente em calvária de ratos, tratados com o biomaterial e diferentes opções de enxertos ósseos com o objetivo de comparar a eficiência do Sistema BoneLithium aos protocolos pré-existentes. Experimentalmente, avaliou-se a reação tecidual onde se utilizou 15 ratos machos divididos aleatatoriamente em 5 grupos onde implantouse no subcutâneo tubos de butterfly contendo o biomaterial por períodos de preservação recomendados pela norma ADA 10993 para teste de reação tecidual. Os resultados demonstram que o Sistema BoneLithium apresenta reação tecidual normal. Para a avaliação do comportamento biológico do Sistema BoneLithium foram utilizados coelhos brancos adultos da raça New Zealand nos quais defeitos ósseos bilaterais de 1 cm de diâmetro foram confeccionados cirurgicamente na calvária e foram Tratados com o Sistema Bone Lithium do (lado Direito) e somente o Gel de Carbopol (lado esquerdo)/Coágulo sanguíneo (controle). A Histomorfometria demonstrou comportamento favorável ao reparo ósseo e adicionalmente através de Microtomografia Computadorizada (CT SKYSCAN), foi possível constatar diferenças significativas considerando p> 0.05 (ANOVA, Tukey) para o processo de reparo ósseo. A avaliação da performance do Sistema BoneLithium utilizando ratos Wistar nos quais foram criados defeitos críticos no centro da calvária e tratados com diferentes modalidades de enxertos ósseos (controle, autógeno, osso de banco (Unioss®, Marília Brasil), Bio-Oss® e associações com o Sistema BoneLithium. A histomorfometria mostrou diferenças significativas considerando p> 0.05 (ANOVA, Tukey) para avaliação de tecido conjuntivo pré-osteogênico e tecido ósseo neoformado, e quando avaliado qualitativamente por tomografia computadorizada de feixe cônico (I cat Cone Beam FOV 0.05 Xoran Tecnology, LLC, EUA e E-vol, CDT, Brasil), observaram-se áreas de neoformação óssea compatíveis com hiperdensidade óssea em toda a extensão do defeito quando apuradas em analises de paridade em escala Hounsfield. Dessa forma, conclui-se que no contexto deste estudo é possível concluir que Sistema BoneLithium representa uma alternativa com potencial viabilidade clínica e necessita seguimento de aplicação em novas metodologias.(AU)

In this work, bases for the development, characterization and evaluation of the biological aspects related to the BoneLithium System were designed, based on the association of lithium carbonate particles dispersed in carbopol® gel matrix, capable of acting as a drug-releasing system. Methodologically this study was divided in four parts: In the first one, the central objective was the development and characterization of the biomaterial through the pharmacological manipulation. In the second step, the tissue reaction was evaluated in subcutaneous of rats, in the third the influence of the lithium particles released by BoneLithium System in the bone repair through experimental models using rabbits, and finally, the capacity of healing of bone defects created surgically in Calvaria of rats, treated with the biomaterial and different options of bone grafts with the objective to compare the efficiency of the BoneLithium System to the preexisting protocols. Experimentally, the tissue reaction was evaluated in which 15 male rats were randomly divided into 5 groups, where butterfly tubes containing the biomaterial were implanted in the subcutaneous tubes for preservation periods recommended by the ADA 10993 standard for biocompatibility test. The results demonstrate that the BoneLithium System is tissue reaction positive. To evaluate the biological behavior of the BoneLithium System, adult New Zealand white rabbits were used in which bilateral bone defects of 1 cm in diameter were surgically made on calvaria and treated with the Bone Lithium System (right side) and only Gel Of Carbopol (left side) / blood clot (control). Histomorphometry showed a favorable behavior to bone repair and, in addition, through Computerized Microtomography (CT SKYSCAN), it was possible to verify significant differences considering p> 0.05 (ANOVA, Tukey) for the bone repair process. The evaluation of the performance of the BoneLithium System using Wistar rats in which critical defects were created at the calvarial center and treated with different bone graft modalities (control, autogenous, bone bank (Unioss®, Marília Brazil), Bio-Oss® and associations (ANOVA, Tukey) for evaluation of pre osteogenic connective tissue and neoformed bone tissue, and when assessed qualitatively by cone beam computed tomography (I cat - Cone Beam - FOV 0.05 - Xoran Tecnology, LLC, USA and E-vol, CDT, Brazil), areas of bone neoformation compatible with bone hyperdensity throughout the extent of the defect were ascertained in Hounsfield scale parity analyzes, It is concluded that in the context of this study it is possible to conclude that the BoneLithium System represents an alternative with potential clinical feasibility And requires follow-up of application in new methodologies.(AU)

Effects of Nanosized Lithium Carbonate Particles on the Functional Activity of Macrophages During Development of Hepatocarcinoma 29.

The functional activity of macrophages in response to injection of nanosized lithium carbonate particles after initiation of hepatocarcinoma 29 in male CBA mice was evaluated by the production of NO, arginase activity, and absorption of zymosan granules. In intact animals, NO production by peritoneal macrophages increased by 4 times and arginase activity 3.1 times in response to a single injection of nanosized particles into the hip muscle. The level of NO production by macrophages remained high after 4 and 5 injections, while arginase activity returned to normal. The level of phagocytic peritoneal macrophages increased by 1.4 times after 5 injections of the particles. The level of NO production by macrophages gradually increased in animals with hepatocarcinoma developing in the hip muscle: by 1.6 times on day 3, 3.2 times on day 7, and by 2.6 times on day 13 in comparison with the corresponding parameters in intact animals. The increase of NO production by peritoneal macrophages after tumor process initiation was not paralleled by changes in arginase activity and absorption of zymosan granules. The results indicated that injection of nanosized lithium carbonate particles after inoculation of hepatocarcinoma 29 cells in the right hip muscle tissue was inessential for the function of peritoneal macrophages by the studied parameters.

Dosimetric evaluation of lithium carbonate (Li2CO3) as a dosemeter for gamma-radiation dose measurements.

This work reports the possibility of using lithium carbonate as a dosimetric material for gamma-radiation measurements. Carboxi-radical ions, CO(2)(-) and CO(3)(-), arise from the gamma irradiation of Li(2)CO(3), and these radical ions can be quantified by electron paramagnetic resonance (EPR) spectrometry. The EPR-signal response of gamma-irradiated lithium carbonate has been investigated to determine some dosimetric characteristics such as: peak-to-peak signal intensity versus gamma dose received, zero-dose response, signal fading, signal repeatability, batch homogeneity, dose rate effect and stability at different environmental conditions. Using the conventional peak-to-peak method of stable ion radicals, it is concluded that lithium carbonate could be used as a gamma dosemeter in the range of 3-100 Gy.

Lithium carbonate in the management of cannabis withdrawal in humans: an open-label study.

Cannabis is the most widely used illicit substance in the world. Estimates suggest that approximately 10-20% of cannabis users meet criteria for cannabis dependence and a significant proportion experience withdrawal discomfort on cessation of use. To date, there has been an absence of any clinically validated treatments to manage withdrawal. The current study is an open-label trial exploring the utility of lithium carbonate for the management of cannabis withdrawal symptoms in treatment seeking adult humans. In total, 20 participants were recruited to the study (19 men). All met DSM-IV cannabis-dependence criteria and had been smoking cannabis daily or almost daily for a mean 9 years. Participants were admitted to an inpatient detoxification facility and prescribed lithium 500 mg b.d. for 7 days. Cannabis withdrawal was assessed daily with the Marijuana Withdrawal Checklist (MWC). Two participants were withdrawn from the trial because of possible adverse effects. Sixty percent of participants completed the 7-day treatment program. Follow-up was conducted at a mean of 107 days following treatment. The mean percentage of days abstinent in the period between treatment cessation and follow-up was 87.57%. Twenty-nine percent of participants (n=5) reported continuous abstinence that was biochemically verified at follow-up. Agreement between self-reported cannabis use and urinalysis at follow-up was moderate (kappa=0.47). Significant reductions in symptoms of depression and anxiety and cannabis-related problems were also reported. This study provides evidence for the potential clinical utility and safety of lithium in the management of cannabis withdrawal. A randomised, placebo-controlled trial is recommended.

Room temperature ICl-induced dehydration/iodination of 1-acyl-5-hydroxy-4,5-dihydro-1H-pyrazoles. a selective route to substituted 1-acyl-4-iodo-1H-pyrazoles.

A number of new functionally substituted 1-acyl-5-hydroxy-4,5-dihydro-1H-pyrazoles have been prepared in moderate to excellent yields from the corresponding 2-alkyn-1-ones. The resulting dihydropyrazoles undergo dehydration and iodination in the presence of ICl and Li2CO3 at room temperature to provide 1-acyl-4-iodo-1H-pyrazoles.