Camptothecin: solubility, in-vitro drug release, and effect on human red blood cells and sperm cold preservation.

BACKGROUND: Camptothecin (CPT) is an anticancer drug, and is not employed in the clinic because of its high hydrophobicity and low active form stability. CPT may also have potential for use in cold preservation. OBJECTIVE: To overcome these drawbacks, CPT solubility variations in the presence of cyclodextrins (CDs) and polyethylene glycol (PEG) were evaluated by Higuchi solubility experiments. MATERIALS AND METHODS: CPT was encapsulated in different cyclodextrins and polyethylene glycol using a co-evaporation method. The CPT interactions with CDs and PEG 6000 were investigated by Fourier-transformed infrared spectroscopy (FT-IR), and X-ray powder diffraction (XRPD). Then, CPT complexes were evaluated for in-vitro drug release. To evaluate the potential anticancer efficacy of the CPT complexes system, in-vitro cytotoxicity studies on human red blood cells were carried out using UV assay. The impact of the CPT complex systems on sperm motility protection during cold preservation at 4 degree C was studied using CASA. RESULTS: The dissolution profile of these preparations shows the improvement of the dissolution of the CPT following a fickien diffusion. The CPT solubility and stability improvement were the cause of the cytotoxicity on the red blood cells test. However, CPT alone, encapsulated, dispersed, and chemically modified protected spermatozoids during cold preservation. CONCLUSION: We confirm the interest in CPT encapsulated and dispersed in anticancer treatments. We also found that CPT encapsulated or dispersed could protect sperm against oxidative damage and improve the membrane integrity of human sperm. Consequently, CPT encapsulated our dispersed could eventually be beneficial for infertility therapy. Doi: 10.54680/fr23210110712.

Drug release, stability and efficiency of vitamin e loaded in liposomes for bovine sperm protection in cryopreservation medium.

BACKGROUND: It is known that a considerable number of drugs in clinical use or under development are water-insoluble drugs with poor bioavailability. The liposomal delivery system has drawn attention as one of the noteworthy approaches to increase both dissolution and absorption because of its biocompatibility and ability to encapsulate hydrophobic molecules in the lipid domain. However, several drawbacks have been reported, the most common is liposome structural instability. OBJECTIVE: To encapsulate alpha tocopherol into liposomes, to determine the new formulation stability and to study the drug-release of alpha tocopherol into the sperm cryopreservation medium. MATERIALS AND METHODS: The liposomes prepared by an ethanol injection method were characterized for size stability, alpha tocopherol release and sperm motility tests. RESULTS: The prepared unilamellar vesicles had both narrow size distribution (around 99 nm) and a good physical and chemical stability at 4°C during 12 months. The liposomes did not release the vitamin E immediately, but retained the protectant for 24 hours, probably due to the rigidity of the liposomal fence which was reinforced by adding cholesterol. Then, all vitamin E molecules were released by 48 hours. Release was potentially by Fickian diffusion probably by the creation of mini-ducts due to both agitation and fence hydration. Moreover, semen motility treated with vitamin E liposome preparations was significantly improved compared to all other treatments (including commonly used sperm conservation media). CONCLUSION: The stable vitamin E liposomes formulated in this work are a promising alternative for semen cryopreservation protection.

Encapsulation Nanotechnology in Sperm Cryopreservation: Systems Preparation Methods and Antioxidants Enhanced Delivery.

Sperm cryopreservation promotes the storage and transport of germplasm for its use in artificial insemination (AI) and other advanced reproductive technologies. However, sperm cryopreservation causes several stresses including thermal shock, osmotic damage, and ice crystal formation, thereby reducing sperm quality. Supplementing cryoprotectant media with antioxidants has been reported to be positive in different species. It has been widely suggested to combine antioxidants with nanotechnology, to maximize therapeutic activity and to minimize undesirable side effects. In this review, we discuss the role of different antioxidants in sperm cryopreservation and their improved therapeutic effect through their formulation using nanotechnology. In addition, we report the different nano-systems preparation methods present in literature. Whilst the use of nanotechnology in animal production is still in its infancy, encouraging results from nutrition, biocidal, remedial, and reproductive studies are driving further investigations.

Development, validation and method stability study of a LC-MS method to quantify leuprolide (Hormone analog) in human plasma.

A rapid, highly sensitive and simple high-performance liquid chromatographic-tandem mass spectrometric (LC-MS) assay is developed and validated for the quantification of leuprolide: a Gonadotropin Releasing Hormone analog (GnRH) in human plasma. Moreover, various parameters of the method stability are determined. After the addition of stable isotope (internal standard), the leuprolide was extracted from human plasma by a C18 solid phase micro extraction (MEPS) cartridge and directly injected into LC-MS/MS system. Chromatographic separation was achieved using a Waters Atlantis HILIC, C18, 150 × 2.1 mm, 5 µ column. Mobile phase was a mixture of acetate buffer (pH 3) and acetonitrile (25/75). Drug detection was performed by MS using electrospray ionization in positive mode. Multiple reaction monitoring (MRM) with a tandem mass spectrometer was used to detect the analytes. Precursor to product ion transitions of: m/z 605.5 â†’ m/z 110.2 and m/z 609.1 â†’ m/z 249.1 were used to quantify leuprolide and leuprolide-13C6-15N, respectively. Sample analysis time was 3 min for each injection. The assay exhibited a linear dynamic range of 0.0500-40 ng/ml for each analyte with a lower limit of quantification (LLOQ) of: 0.0500 ng/ml. Furthermore, a complete analytic validation was carried out, including tests on: The specificity, precision, accuracy, matrix effect and stability under different storage conditions. Importantly, the obtained results established: an acceptable precision and accuracy for concentration over standard curve range. Nevertheless, it is to emphasize the simplicity, rapidity and also the high precision and accuracy of this novel LC-MS method, offering useful information about solution stability. Finally, this work is a good alternative to quantify Leuprolide concentration in human blood, especially on human clinic trials step.

Rosmarinus officinalis Essential Oil Preloaded in ß-Cyclodextrin: Effect on Ram Spermatozoa Motility, Membrane Integrity and Oxidative Status During 4°C Storage.

BACKGROUND: Rosmarinus officinalis essential oil (Rom) has been reported recently to be of interest for use in sperm cryopreservation. However, related to its lipophilic characteristics, encapsulation in cyclodextrin could enhance Rom positive effects by increasing its solubility in sperm extenders. OBJECTIVE: To compare the effect of Rom preloaded in hydroxypropyl-ß-cyclodextrin (Rom-cd) to Rom alone (Rom) on ram sperm conserved at 4°C. MATERIALS AND METHODS: Ram epididymal sperm was collected from six testes. Each collected sperm was split into four equal aliquots. The control aliquot was diluted with Tris extender (Tris + citric acid + fructose + penicillin), two aliquots were treated with Rom at 0.5µl ml-1 and 1µl ml-1 respectively, and one aliquot treated with Rom-cd at 1µl ml-1. All sperm aliquots were analyzed for motility after 0, 2, 4, 24 and 48 h of storage at 4°C using a Computer Aided Semen Analysis (CASA). Membrane integrity and oxidative stress status were measured after 48 h of storage. RESULTS: The results indicated that motility parameters were best preserved in the extender containing Rom-cd compared to the groups treated by Rom without cyclodextrin. Rom alone resulted to higher sperm motility than the control group. Lower oxidative stress and more cell membrane protection were observed in Rom treated samples, especially when using Rom-cd. CONCLUSION: The ability of Rom to protect ram sperm against cryopreservation damages was improved after encapsulation in hydroxypropyl-ß-cyclodextrin (HPßCD).

Protective effect of proteins derived from Calotropis procera latex against acute inflammation in rat.

The non-dialysable proteins present in the latex of plant Calotropis procera possess anti-inflammatory and analgesic properties. The aim of this study was to evaluate the effect of latex proteins (LP) on the level of inflammatory mediators, oxidative stress markers and tissue histology in the rat model of carrageenan-induced acute inflammation. This study also aimed at evaluating the anti-inflammatory efficacy of LP against different mediators and comparing it with their respective antagonists. Paw inflammation was induced by subplantar injection of carrageenan, and the effect of LP was evaluated on oedema volume, level of TNF-α, PGE(2), myeloperoxidase, nitric oxide, reduced glutathione, thiobarbituric acid-reactive substances and tissue histology at the time of peak inflammation. Paw inflammation was also induced by histamine, serotonin, bradykinin and PGE(2), and the inhibitory effect of LP against these mediators was compared with their respective antagonists at the time of peak effect. Treatment with LP produced a dose-dependent inhibition of oedema formation, and its anti-inflammatory effect against carrageenan-induced paw inflammation was accompanied by reduction in the levels of inflammatory mediators, oxidative stress markers and normalization of tissue architecture. LP also produced a dose-dependent inhibition of oedema formation induced by different inflammatory mediators, and its efficacy was comparable to their respective antagonists and more pronounced than that of diclofenac. Thus, our study shows that LP has a potential to be used for the treatment of various inflammatory conditions where the role of these mediators is well established.

Fast dissolving cyclodextrin complex of piroxicam in solid dispersion part I: influence of ß-CD and HPß-CD on the dissolution rate of piroxicam.

Sublingual drug delivery is an interesting route for drug having significant hepatic first-pass metabolism or requiring rapid pharmacological effect as for patients suffering from swallowing difficulties, nausea or vomiting. Sublingual absorption could however be limited by the kinetic of drug dissolution. This study evaluated influences of cyclodextrins (ß-CD or HP-ß-CD) and their different inclusion process (spray-drying or freeze-drying) on the drug dissolution kinetic of solid dispersions in poly(ethylene glycol) (PEG, Mw 6000Da) of piroxicam, used as poor hydrosoluble drug model. A secondary objective was to determine influences of drug dispersion process in PEG (evaporation or melting methods) on the drug dissolution kinetic of piroxicam. Piroxicam solid dispersions containing or not cyclodextrins were characterized by different scanning calorimetry (DSC), Thermogravometry analyser (TGA) and Fourier transform-infrared spectroscopy (FT-IR) spectroscopy. In vitro drug dissolution study of these solid dispersions was then performed. The results demonstrated the high potential and interest of solid dispersions of drug previously included in cyclodextrins for sublingual delivery of hydrophobic drugs. This study also showed the advantages of evaporation method on the melting ones during drug dispersion in PEG. Indeed, drug complexation with cyclodextrins as dispersion by melting prevented the presence in solid dispersions of drug in crystalline form which can represent up to 63%. Moreover, dispersion in PEG by evaporation method gave more porous drug delivery system than with melting methods. This allowed complete (limited at most at 80-90% with melting methods) and quick drug dissolution without rebound effect like with melting ones.

Constrictive pericarditis presenting as chylothorax.

Chylothorax is a rare clinical condition that can be attributed to a damaged thoracic duct. The condition is suggested by aspiration of milky white fluid from the pleural cavity and is commonly associated with either malignant diseases or trauma (e.g. cardiothoracic surgery). We present the case of a 15-year-old boy with chylothorax, whose effusion was due to constrictive pericarditis. The definitive treatment of chylothorax involves identification and management of the underlying pathology. We suggest that when dealing with cases of chylothorax, constrictive pericarditis should be considered among the causes.