Effects of caffeic and chlorogenic acids on the rat skeletal system.

OBJECTIVE: Caffeic acid, predominantly as esters linked to quinic acid (chlorogenic acids), is a phenolic acid present at high levels in coffee. The aim of the study was to investigate effects of caffeic and chlorogenic acids on the skeletal system of female rats with normal estrogen levels and estrogen-deficient. MATERIALS AND METHODS: Caffeic acid (5 and 50 mg/kg p.o. daily) and chlorogenic acid (100 mg/kg p.o. daily) were administered for 4 weeks to non-ovariectomized and bilaterally ovariectomized mature Wistar rats, and their effects were compared with appropriate controls. Moreover, estradiol (0.2 mg/kg p.o. daily) was administered to ovariectomized rats. Bone turnover markers, mass, mineralization and mechanical properties were examined. RESULTS: Although caffeic acid at a low dose exerted some unfavorable effects on the skeletal system, at high doses, caffeic and chlorogenic acids slightly increased mineralization in the tibia and improved mechanical properties of the femoral diaphysis (compact bone). Unlike estradiol, they did not counteract the worsening of the tibial metaphysis bone strength (cancellous bone) and increases in osteocalcin concentration induced by estrogen deficiency. CONCLUSIONS: High doses of the phenolic acids slightly favorably affected the rat skeletal system independently of the estrogen status.

Impact of inter- and intramolecular interactions on the physical stability of indomethacin dispersed in acetylated saccharides.

Differential scanning calorimetry (DSC), broadband dielectric (BDS), and Fourier transform infrared (FTIR) spectroscopies as well as theoretical computations were applied to investigate inter- and intramolecular interactions between the active pharmaceutical ingredient (API) indomethacin (IMC) and a series of acetylated saccharides. It was found that solid dispersions formed by modified glucose and IMC are the least physically stable of all studied samples. Dielectric measurements showed that this finding is related to neither the global nor local mobility, as the two were fairly similar. On the other hand, combined studies with the use of density functional theory (DFT) and FTIR methods indicated that, in contrast to acetylated glucose, modified disaccharides (maltose and sucrose) interact strongly with indomethacin. As a result, internal H-bonds between IMC molecules become very weak or are eventually broken. Simultaneously, strong H-bonds between the matrix and API are formed. This observation was used to explain the physical stability of the investigated solid dispersions. Finally, solubility measurements revealed that the solubility of IMC can be enhanced by the use of acetylated carbohydrates, although the observed improvement is marginal due to strong interactions.

Enhancement of the physical stability of amorphous indomethacin by mixing it with octaacetylmaltose. inter and intra molecular studies.

PURPOSE: To demonstrate a very effective and easy way of stabilization of amorphous indomethacin (IMC) by preparing binary mixtures with octaacetylmaltose (acMAL). In order to understand the origin of increased stability of amorphous system inter- and intramolecular interactions between IMC and acMAL were studied. METHODS: The amorphous IMC, acMAL and binary mixtures (IMC-acMAL) with different weight ratios were analyzed by using Dielectric Spectroscopy (DS), Differential Scanning Calorimetry (DSC), Raman Spectroscopy, X-ray Diffraction (XRD), Infrared Spectroscopy (FTIR) and Quantitative Structure-Activity Relationship (QSAR). RESULTS: Our studies have revealed that indomethacin mixed with acetylated saccharide forms homogeneous mixture. Interestingly, even a small amount of modified maltose prevents from recrystallization of amorphous indomethacin. FTIR measurements and QSAR calculations have shown that octaacetylmaltose significantly affects the concentration of indomethacin dimers. Moreover, with increasing the amount of acMAL in the amorphous solid dispersion molecular interactions between matrix and API become more dominant than IMC-IMC ones. Structural investigations with the use of X-ray diffraction technique have demonstrated that binary mixture of indomethacin with acMAL does not recrystallize upon storage at room temperature for more than 1.5 year. Finally, it was shown that acMAL can be used to improve solubility of IMC. CONCLUSIONS: Acetylated derivative of maltose might be very effective agent to improve physical stability of amorphous indomethacin as well as to enhance its solubility. Intermolecular interactions between modified carbohydrate and IMC are likely to be responsible for increased stability effect in the glassy state.

A new way of stabilization of furosemide upon cryogenic grinding by using acylated saccharides matrices. The role of hydrogen bonds in decomposition mechanism.

Recently it was reported that upon mechanical milling of pure furosemide significant chemical degradation occurs (Adrjanowicz et al. Pharm. Res.2011, 28, 3220-3236). In this paper, we present a novel way of chemical stabilization amorphous furosemide against decomposing that occur during mechanical treatment by preparing binary mixtures with acylated saccharides. To get some insight into the mechanism of chemical degradation of furosemide induced by cryomilling, experimental investigations supported by density functional theory (DFT) computations were carried out. This included detailed studies on molecular dynamics and physical properties of cryoground samples. The main thrust of our paper is that we have shown that furosemide cryomilled with acylated saccharides forms chemically and physically stable homogeneous mixtures with only one glass transition temperature, Tg. Finally, solubility measurements have demonstrated that furosemide cryomilled with acylated saccharides (glucose, maltose and sucrose) is much more soluble with respect to the crystalline form of this active pharmaceutical ingredient (API).

Effect of concurrent administration of alendronate sodium and retinol on development of changes in histomorphometric parameters of bones induced by ovariectomy in rats.

Retinol is a commonly used vitamin, especially by elderly people. Alendronate sodium, an aminobisphosphonate, is a potent antiresorptive drug used in the treatment of osteoporosis in postmenopausal women. Frequently, alendronate sodium and retinol are used concurrently. There are no reports on the interaction between alendronate sodium and retinol. The aim of the present study was to investigate the effect of concurrent administration of alendronate sodium and retinol on bone remodeling in ovariectomized rats. The histomorphometric parameters of long bones were studied. The experiments were carried out on 3-month-old Wistar rats, divided into 7 groups: I (C) - sham operated control rats, II (OVX) - ovariectomized control rats, III (OVX + ALN) - ovariectomized rats + alendronate sodium (3 mg/kg po), IV (OVX + R-1) - ovariectomized rats + retinol (700 IU/kg po), V (OVX + R-2) - ovariectomized rats + retinol (3500 IU/kg po), VI (OVX + ALN + R-1) - ovariectomized rats + alendronate sodium (3 mg/kg po) + retinol (700 IU/kg po), VII (OVX + ALN + R-2) - ovariectomized rats + alendronate sodium (3 mg/kg po) + retinol (3500 IU/kg po). The drugs were administered to the rats daily by oral gavage (alendronate sodium in the morning, retinol in the afternoon) for 28 days. Body mass gain, bone mass, mineral content in the tibia, femur and L-4 vertebra, histomorphometric parameters of the right tibia (width of osteoid, periosteal and endosteal transverse growth, area of the transverse cross section of the bone marrow cavity and the cortical bone) and the right femur (width of epiphyseal and metaphyseal trabeculae, width of epiphyseal cartilage) were studied. Bilateral ovariectomy induced osteopenic skeletal changes in mature female rats. Alendronate sodium administered at a dose of 3 mg/kg po daily inhibited the development of changes induced by ovariectomy in the skeletal system of rats. Retinol, especially administered at the dose of 3500 IU/kg daily, intensified the changes in the osseous system caused by estrogen deficiency in rats. Retinol administered concurrently with alendronate sodium attenuated the antiresorptive effect of alendronate sodium on the skeletal system in ovariectomized rats.

Effects of doxycycline on the development of bone damage caused by prednisolone in rats.

The aim of the present study was to investigate the effects of doxycycline on the development of bone damage caused by prednisolone in rats. The experiments were carried out on male WAG rats (200-260 g), divided into 2 control and 6 experimental groups receiving prednisolone (5 mg/kg im daily) or/and doxycycline (100 mg/kg po daily) for 2 or 4 weeks. The animals were sacrificed on the 15th or 29th day of the experiment and the following characteristics were examined: mass, length, mechanical properties, mineral and calcium content in the tibia and femur, width of endosteal and periosteal osteoid, endosteal and periosteal transverse growth, transverse cross-section area of the diaphysis and of the marrow cavity in the tibia, width of epiphyseal cartilage and width of trabeculae in the femur. Prednisolone caused features of osteopenia (inhibition of bone formation and intensification of bone resorption), which were stronger after 4 weeks of the experiment. Doxycycline administered alone for 2 or 4 weeks intensified the processes of bone formation and resorption. Doxycycline to some degree attenuated the influence of prednisolone on rat bones.

Effects of pamidronate on the development of changes in bone mechanical properties and bone structure caused by the administration of prednisolone in rats.

The purpose of the study was to investigate the influence of pamidronate on mechanical properties, growth, and structural changes in bones of rats in which experimental osteopenia was induced by administration of prednisolone. The experiment was carried out on male WAG rats divided into three groups: I. Control, II. Prednisolone (5 mg/kg im daily) and III. Disodium pamidronate (3 mg/kg sc daily) + prednisolone (5 mg/kg im daily). After three weeks of the experiment, the animals were sacrificed and their femoral and tibial bones were prepared. The administration of prednisolone resulted in morphological and metabolic changes in the osseous system, characteristic of osteopenia. The increased osteopsathyrosis was noted, manifesting in lowered resistance to fractures and lesser deformability in comparison with the control group. The administration of pamidronate resulted in the reduction of the destructive action of prednisolone on bones.

The effects of pamidronate on mechanical properties, growth and structural changes in rat bones.

Although a number of properties of bisphosphonates have been recognized which influence the metabolic process in bones, particularly those concerning the remodelling processes, the influence of this new group of drugs on the mechanical properties of bones remains an open issue. In order to clarify this problem, the present study concentrated on the influence of a new generation bisphosphonate, i.e. pamidronate upon the mechanical properties, growth, and morphological changes in the femoral and tibial bones in rats. The experiments carried out concerned pamidronate administration to male Wistar rats in doses of 3 mg/kg of body mass subcutaneously, for the period of 3 or 6 weeks. The total changes in the osseous tissue after pamindronate administration indicate the drug to foster the development of osteopetrosis in rats, the prominent sings of the disease being mainly deformations of epiphysis, decreased bone growth, increased thickness of epiphysial cartilage and bone trabeculae, as well as lowered resistance to fractures and decreased susceptibility to deformations.