Celia Clemente

Bone regeneration in rabbit calvaria with novel monetite granules

The aim of this study was to evaluate whether local application of monetite granules would induce bone regeneration in critical size defects on rabbits calvaria. Novel monetite granules were synthesized by thermal conversion of preset brushite cement. Twelve female New Zealand rabbits were used for this study. Two identical 10-mm-diameter bicortical cranial defects were created in each animal. One of the defects was grafted with monetite granules while the contralateral was left unfilled as negative control. Animals were sacrificed at 4 and 8 weeks after surgery, and biopsies were taken for histological and histomorphometrical evaluation under light microscopy. Wilcoxon test was used for statistical analysis. The histological observations showed signs of graft resorption as newly formed bone tissue grew surrounding and penetrating the monetite granules. Histomorphometric evaluation showed that the augmented bone volume as well as the augmented mineral tissue was higher in the defects treated with monetite granules (p < 0.05) 8 weeks after the intervention. In this animal model, local application of the novel monetite granules in bone defects enhances bone healing significantly.

Vasoactive intestinal peptide enhances growth and angiogenesis of human experimental prostate cancer in a xenograft model

We show that vasoactive intestinal peptide (VIP) exerts trophic and proangiogenic activities in experimental prostate cancer in vivo. Nude mice were subcutaneously injected with Matrigel impregnated with LNCaP prostate cancer cells. Cell treatment with 100 nM VIP for 1h before xenograft resulted in increased tumor growth after 8 and, more remarkably, 15 days of injection. The same occurred with the mRNA expression of the main angiogenic factor, vascular endothelial growth factor (VEGF), as shown by real-time RT-PCR quantification. The proangiogenic activity of VIP was further established by showing increases of hemoglobin levels, Masson trichromic staining, and immunohistochemical CD34 staining in tumors excised 15 days after subcutaneous injection of VIP-treated cells as compared to control conditions. All these parameters indicate that VIP increases vessel formation. This xenograft model is a useful tool to study in vivo the effects of VIP-related peptides in tumor growth and development of blood supply as well as their therapeutical potential in prostate cancer.

Effects of Local Melatonin Application on Implant Osseointegration

PURPOSE The aim of this study was to assess the effect of local melatonin administration on bone osseointegration around implants in rabbit tibiae. MATERIAL AND METHODS Ten female, 6-month-old New Zealand rabbits were randomly divided into two groups: the experimental group, where five rabbits were treated with local application of melatonin (3 mg) to implant sites when placed into the rabbit tibia, and the control group, those who where without additive materials. Four weeks later, animals were sacrificed; tibiae were dissected from soft tissues and fixed in buffered formaldehyde, and then included in methacrylate. Histological sections were performed to be studied under light microscopy and analyzed morphometrically to evaluate the amount of bone to implant contact (BIC), trabecular area density, and cortical area density. One-way analysis of variance test was used for statistical evaluation. p < .05 was considered to be significant. RESULTS Histological evaluation showed more trabecular reaction in the melatonin group. Morphometrical analysis showed a statistically significant increase in trabecular BIC in the melatonin group when compared with the control group (24.61% ± 2.87 vs 13.62% ± 1.44; p < .01). Cortical BIC was decreased in the melatonin group, without statistical significance (71.08 ± 3.63 vs 76.28 ± 2.57; p = 0.31). Trabecular area density was increased significantly in the melatonin group (8.68 ± 1.61 vs 4.02 ± 0.36; p < .05). Cortical area density was decreased significantly in the melatonin group (91.31 ± 1.6 vs 95.7 ± 0.5; p < .05). CONCLUSION Within the limitation of this animal study, local melatonin application at the time of implant placement might induce more trabecular bone at implant contact and higher trabecular area density.

G‐proteins and β‐adrenergic stimulation of adenylate cyclase activity in the diabetic rat prostate

The consequences of experimental diabetes on membrane lipids, β‐adrenergic stimulation of adenylate cyclase activity, and G‐protein levels in the prostate gland are not defined.

Fracture bone healing and biodegradation of AZ31 implant in rats

The ideal temporary implant should offer enough mechanical support to allow healing of the fracture and then biodegrade and be resorbed by metabolic mechanisms without causing any toxic effect. The aim of this research has been to simultaneously study in situ bone healing and the biodegradation of AZ31 Mg alloy as an osteosynthesis material. The in vivo study was carried out in AZ31 implants with and without Mg-fluoride coating inserted in un-fractured and fractured femurs of Wistar rats for long experimentation time, from 1 to 13 months, by means of computed tomography, histological and histomorphometric analysis. Tomography analysis showed the bone healing and biodegradation of AZ31 implants. The fracture is healed in 100% of the animals, and AZ31 maintains its mechanical integrity throughout the healing process. Biodegradation was monitored, quantifying the evolution of gas over time by 3D composition of tomography images. In all the studied groups, gas pockets disappear with time as a result of the diffusion process through soft tissues. Histomorphometric studies reveal that after 13 months the 46.32% of AZ31 alloy has been resorbed. The resorption of the coated and uncoated AZ31 implants inserted in fractured femurs after 1, 9 and 13 months does not have statistically significant differences. There is a balance between the biodegradation of AZ31 and bone healing which allows the use of AZ31 to be proposed as an osteosynthesis material.

Analysis of metallic traces from the biodegradation of endomedullary AZ31 alloy temporary implants in rat organs after long implantation times

AZ31 alloy has been tested as a biodegradable material in the form of endomedullary implants in female Wistar rat femurs. In order to evaluate the accumulation of potentially toxic elements from the biodegradation of the implant, magnesium (Mg), aluminium (Al), zinc (Zn), manganese (Mn) and fluorine (F) levels have been measured in different organs such as kidneys, liver, lungs, spleen and brain. Several factors that may influence accumulation have been taken into account: how long the implant has been in place, whether or not the bone is fractured, and the presence of an MgF2 protective coating on the implant. The main conclusions and the clinical relevance of the study have been that AZ31 endomedullary implants have a degradation rate of about 60% after 13 months, which is fully compatible with fracture consolidation. Neither bone fracture nor an MgF2 coating seems to influence the accumulation of trace elements in the studied organs. Aluminium is the only alloying element in this study that requires special attention. The increase in Al recovered from the sampled organs represents 3.95% of the amount contained in the AZ31 implant. Al accumulates in a statistically significant way in all the organs except the brain. All of this suggests that in long-term tests AZ31 may be a suitable material for osteosynthesis.

Effects of Pygeum africanum extract (Tadenan®) on vasoactive intestinal peptide receptors, G proteins, and adenylyl cyclase in rat ventral prostate

Tadenan® (a Pygeum africanum extract) is a drug used in the treatment of benign prostatic hyperplasia. Its effects on prostate fibroblast proliferation and bladder function after partial outlet obstruction have been demonstrated in various pharmacological studies. However, its effects at the molecular level are poorly documented.

Effect of flutamide-induced androgen-receptor blockade on adenylate cyclase activation through G-protein coupled receptors in rat prostate

The effect of the antiandrogen flutamide on the prostatic vasoactive intestinal peptide (VIP) receptor/effector system was studied in rats. Rats were s.c. injected with a daily dose of flutamide (15 mg/kg B.W.) or vehicle for 14 days. Drug treatment resulted in histological evidence of gland involution and increased plasma membrane fluidity as estimated by fluorescence spectroscopy. The number of VIP receptors and the stimulatory effect of VIP on adenylate cyclase activity in prostatic membranes decreased in flutamide-treated rats. However, the pattern of forskolin stimulation of the enzyme activity was not modified by this drug. Androgen-receptor blockade by flutamide also decreased the prostatic levels of alpha(s,) alpha(i1/2), and alpha(i3/0) G-protein subunits, as estimated by an immunological procedure. Whereas apoptotic DNA fragmentation was evidenced in prostate from 3-day castrated animals, a heterogeneous electrophoretic pattern was observed after flutamide treatment. Thus, androgen-receptor blockade by flutamide results in an important impairment of the components of the VIP receptor/effector system in rat prostate as well as in a modification of their coupling extent, which is presumably due to differences observed in plasma membrane fluidity. These results represent a crosstalk in the prostate between two mechanisms of signal transduction involved in cell proliferation.

Osseointegration of TI6Al4V dental implants modified by thermal oxidation in osteoporotic rabbits

BackgroundIn this work, the effect of the heat treatment on Ti6Al4V implants and topical administration of growth hormone to address a better osseointegration in osteoporotic patients has been analysed.Methods The osseointegration process of Ti6Al4V implants modified by oxidation treatment at 700 °C for 1 h and the influence of local administration of growth hormone (GH) in osteoporotic female rabbits after 15 and 30 days of implantation have been studied. Bone response was analysed through densitometric and histomorphometric studies. Characterization of the surface was provided by scanning electron microscopy.ResultsThe oxidation treatment promotes the formation of an oxide scale grown on the Ti6Al4V implants that alters the nanoroughness of the surface. Bone mineral density (BMD) increases from 0.347 ± 0.014 (commercial) to 0.383 ± 0.012 g cm−2 (modified), and bone-to-implant contact (BIC) goes from 48.01 ± 14.78 (commercial) to 55.37 ± 15.31 (modified) after 30 days of implantation.ConclusionsThe oxidation treatment on the Ti6Al4V dental implants enhances the early bone formation at the longest periods of implantation. No significant differences in the BMD and BIC results in healthy and osteoporotic rabbits were revealed with respect to the local administration of GH in the implantation site.

Local Application of Growth Hormone to Enhance Osseointegration in Osteoporotic Bones: A Morphometric and Densitometric Study

PURPOSE The aim of this study was to assess the effect of local application of growth hormone on osseointegration of dental implants inserted in osteoporotic bones. MATERIALS AND METHODS Twenty female New Zealand rabbits were used in this study. Ten were ovariectomized and fed a low-calcium diet for 6 weeks, and the others remained intact. A titanium implant was inserted into each tibia, in both groups. In half of the rabbits, 2 IU of growth hormone was placed into the ostectomy prior to the implant insertion. Two weeks after implant surgery, all animals were sacrificed. Tibiae were dissected from soft tissues, and included in methacrylate to be studied under light microscopy. Bone-to-implant contact (BIC) and bone mineral density (BMD) were measured by morphometric and densitometric analysis, respectively. Multifactorial analysis of variance (ANOVA) was used for statistical evaluation. P < .05 was considered to be significant. RESULTS Ovariectomy induced less BIC and BMD in regions closer to the implant compared with the control group. Local application of growth hormone was able to increase the BIC in the ovariectomized group, with statistically significant differences with respect to the control group (P < .01). Regarding the BMD, no statistically significant differences were found. CONCLUSION Within the limitations of this experimental study, local application of 2 IU of recombinant human growth hormone at the moment of titanium implant insertion in rabbit tibiae significantly enhanced the BIC around titanium implants 15 days after the implantation in this experimental osteoporotic animal model, without affecting the BMD.