José Cavalheiro

Granules of osteopatite and glass-reinforced hydroxyapatite implanted in rabbit tibiae

Granules of a modified hydroxyapatite, glass-reinforced hydroxyapatite composite and commercial hydroxyapatite were implanted in rabbit tibiae. Histological studies were carried out after 1 and 2 months implantation periods using light and fluorescence microscopy. A much higher percentage of bone contact was developed for both the glass-reinforced hydroxyapatite composite and the modified hydroxyapatite when compared to commercial hydroxyapatite (89–91% versus 66%) after 2 months implantation. The mechanism of bone formation and growth around implants is discussed in terms of the influence of elements incorporated into these novel materials which are commonly found in bone tissues, such as Na, K and Mg, and the presence of a soluble β-tricalcium phosphate phase in the microstructure of the composite.

Acute and Long-Term Effects of Hyperthermia in B16-F10 Melanoma Cells

Objective Hyperthermia uses exogenous heat induction as a cancer therapy. This work addresses the acute and long-term effects of hyperthermia in the highly metastatic melanoma cell line B16-F10. Materials and Methods Melanoma cells were submitted to one heat treatment, 45°C for 30 min, and thereafter were kept at 37°C for an additional period of 14 days. Cultures maintained at 37°C were used as control. Cultures were assessed for the heat shock reaction. Results Immediately after the heat shock, cells began a process of fast degradation, and, in the first 24 h, cultures showed decreased viability, alterations in cell morphology and F-actin cytoskeleton organization, significant reduction in the number of adherent cells, most of them in a process of late apoptosis, and an altered gene expression profile. A follow-up of two weeks after heat exposure showed that viability and number of adherent cells remained very low, with a high percentage of early apoptotic cells. Still, heat-treated cultures maintained a low but relatively constant population of cells in S and G2/M phases for a long period after heat exposure, evidencing the presence of metabolically active cells. Conclusion The melanoma cell line B16-F10 is susceptible to one hyperthermia treatment at 45°C, with significant induced acute and long-term effects. However, a low but apparently stable percentage of metabolically active cells survived long after heat exposure.

An in vitro and in vivo investigation of the biological behavior of a ferrimagnetic cement for highly focalized thermotherapy

The cancer treatment by local hyperthermia, using a high frequency electromagnetic field is an extensively studied subject. For this propose it was developed a ferrimagnetic cement (FC) to be injected directly into the tumor. In this study it was determined the FC injectability, its capability to generate heat when placed within a magnetic field and its interaction with a modified simulated body fluid using SEM/EDS and XRD. The FC biological response was assessed by the intramuscular implantation in rats and histological analysis of the surrounding tissues. The results suggest that FC can be injected directly into the tumor, its temperature can be increased when exposed to a magnetic field and the surface of the immersed samples quickly becomes coated with precipitate denoting its ionic change with the surrounding medium. The histological analysis revealed a transient local inflammatory reaction, similar to the control material, only slightly more abundant during the first weeks, with a gradual decrease over the implantation time. Based on these results, we concluded that FC might be useful for highly focalized thermotherapy, with a good potential for clinical use.

Highly focalised thermotherapy using a ferrimagnetic cement in the treatment of a melanoma mouse model by low temperature hyperthermia

Purpose: Evaluation of the effectiveness of highly focalised thermotherapy (HFT) in a melanoma mouse model, using a ferrimagnetic cement (FC) and repeated low hyperthermia treatments. Materials and methods: A melanoma mouse model was induced with B16F10 cells in C57BL6 mice. The FC, injected into the tumour, was used as the magnetic vehicle for HFT. FC location within the tumour was assessed by radiography and its capability to generate heat, when exposed to an external high frequency magnetic field (HFMF), monitored by thermal camera. The HFT treatment consisted of three HFMF exposures, with 48-h intervals, each one lasting 30 min, with a 5–6°C tumour temperature increase. At the end of the experiment, FC samples were characterised by scanning electron microscopy (SEM) and energy dispersion spectroscopy (EDS). The presence of iron contents was analysed in the tumour, lungs, liver and spleen. Histological evaluation and immunohistochemical staining for caspase-3 were performed. Tumour growth was monitored during the experiment. Results: Surface analysis showed FC stabilisation within the tumour, and iron was absent. The thermal camera confirmed the localised temperature increase in the tumour. HFT treatments inhibited the tumour growth by ∼70% compared to controls. This was due to cell destruction by necrosis and apoptosis. Conclusions: The HFT, using the FC, proved to be a minimally invasive technique that statistically inhibited tumour growth. Results suggested that this methodology seems to be a promising technique for the treatment of solid tumours, allowing repeated low hyperthermia treatments, which can be easier and less traumatic than other hyperthermia techniques.

GUIDED BONE REGENERATION USING OSTEOPATITE GRANULES AND POLYTETRAFLUOROETHYLENE MEMBRANES

Granules of a modified hydroxyapatite, Osteopatite®, were implanted in the right posterior tibiae of adult rabbits. We studied the extent of bone regeneration in bone holes. In the right tibiae, that were filled with granules of this biomaterial covered with a polytetrafluoro-ethylene (PTFE) membrane using, as a control, uncovered granules. In the left tibia, an empty hole was covered with PTFE membrane and a second hole was left empty to be used as a control. A histomorphometric study was carried out using light microscopy, four and eight weeks after the surgery. The covered granules presented a higher percentage of bone contact than the uncovered ones, and it was also possible to observe a better bone tissue organization, mainly produced by the immobilization action of the PTFE membrane. Empty bone defects covered with PTFE membranes, two months after implantation, presented large areas of Haversian bone and direct bone contact to the PTFE membrane.

Silicon and iron levels in tissues of animals treated with a “ferrimagnetic ceramic” with oncotherapeutic potential (anti-tumor) value

The stability in a biological environment of an injectable cement with oncotherapeutic potential--consisting of a glass powder of SiO2 (35.6%), CaO (42.4%), P2O5 (17%), Na2O (5%) and 30% of its weight of Fe3O4 dissolved in (NH4)2HPO4 plus NH4H2PO4--was evaluated referring to the release of silicon and iron. The experimental model was the rat, and organs (liver, kidney, spleen, lung, heart, and brain) of the implanted and control animals were collected for quantification of these elements by electrothermal atomization atomic absorption spectrometry methods. In most of the analysed organs no significant difference in the contents of silicon and iron between the implanted and the control animals was found.

Modified Hydroxyapatite Granules and Plaster as Binder: In Vitro and Clinical Tests

ABSTRACT Hydroxyapatite (HA), produced by chemical precipitation is normally used as a bone graft. Hydroxyapatite modified (HAM), allows the fabrication of granules of a different material with HA structure. The aqueous medium that involves HAM particles has higher pH than the particles of HA, improving the conditions of formation of new bone. In this work the chemical and physical properties of this new HAM granules were studied. A clinical test in dental alveolus in 10 patients, using plaster as binder was carried out. Microradiographic results and histological cuts, retrieved from experimental surgery in dogs, showed the useful behaviour of the association HAM/plaster.