Authors: H. Valiense, G.V.O. Fernandes, B. Moura, J. Calasans-Maia, A. Alves, Antonella M. Rossi, J.M. Granjeiro, M. Calasans-Maia
Abstract: The objective of this study was to investigate the bone repair of carbonate apatite (cHA) in comparison to hydroxyapatite (HA, control group), on osseous repair of non-critical size defect in rat calvaria. Spheres (400<ø>500μm) of both materials were synthesized under 37°C (cHA) and 90°C (HA) and were not sintered. Fifteen rats Wistar were submitted to general anesthesia and two perforations (4mm each) were made, one in each parietal bone, for implantation of cHA (left side) and HA spheres (right side). After 1, 3 and 6 weeks, five animals of each group were killed and the two fragments with the biomaterial were collected from the calvaria. The bone blocks with biomaterial were demineralized and 5μm thick semi-serial sections were done for histological analysis. The experimental group of 6 weeks did not show the presence of spheres of both biomaterials and few spheres were observed after 1 and 3 weeks. Histological analysis showed the connective tissue repairing the surgical defect after 1 week and newly formed bone after 3 weeks of surgery. Thus, we concluded both materials are biocompatible, promote osteoconduction and in all studied periods the biomaterials showed to be resorbable.
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Authors: Neusa Motta de Freitas Costa, Bruno Raposo Melo, Rosana Tavares Brito, G.V.O. Fernandes, Vagner Gonçalves Bernardo, Eliene Carvalho da Fonseca, Márcio Baltazar Conz, Gloria Dulce de Almeida Soares, José Mauro Granjeiro
Abstract: The aim of this paper was to evaluate the usefulness of coupling digital image analysis with immunohistochemistry and histomorphometry data to the study of tissue response to hydroxyapatite in a model of critical size bone defect in calvaria of rats. A transosseous defect measuring 8 mm in diameter was performed with a surgical trephine in the parietal bone of 40 rats and divided into two experimental groups according to the treatment: group I (blood clot, control), group II (HA) and killed 1, 3, 6 and 9 months after implantation (n=5/group/period). The skullcaps with overlaying skin were collected and processed for paraffin embedding. The specimens were cut in the laterolateral direction into 5-µm thick semi-serial sections and stained with hematoxylin-eosin for identification and counting of polymorphonuclears cells, mastocytes, and multinucleated giant cells, MNG, or immunolabeled with anti- lysozyme, -factor VIII and –PCNA. Digital images were obtained and analyzed with the ImagePro-Plus® software for cell couting (polymorphonuclears cells, mastocytes, macrophages and MNG) and microvessel density. Image segmentation of anti-PCNA immunostaining was used for cell proliferation analysis. The digital images obtained allowed clear identification of cells of interest by through morphological aspects or immunostaining. Data recording and analysis was facilitated by the use of specific software for image processing and graphical and statistical analysis. It can be concluded that the techniques applied were usefull to identify and count cells, structures and process of interest making easier the effectiveness of hydroxyapatite in the critical size defect in rat calvaria model.
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Authors: G.V.O. Fernandes, Monica Calasans-Maia, F.F. Mitri, Vagner Gonçalves Bernardo, Antonella M. Rossi, G.D.S. Almeida, José Mauro Granjeiro
Abstract: Biomaterials for treatment of bone defects have been studied for a long time. Alloplastic materials, mainly hydroxyapatite (HA), are under intense investigation due to its biocompatibility and osteoconductive properties. The HA can be modified by the incorporation of bivalent cations as Zn2+ known as a positive effectors for bone repair. The purpose of this study was to evaluate comparatively the effectiveness of 5% zinc-containing hydroxyapatite (ZnHA) in the treatment of critical size defect (CSD) in rat’s calvaria. CSD (8mm diameter) created in the skull of forty-five Wistar rats were filled with autogenous bone, HA and ZnHA. Skulls harvested after 30, 90 and 180 days were submitted to histological processing for paraffin embedding. Sections of 5 µm-thick stained with hematoxylin and eosin (HE) allowed histomorphometric analysis. The area of neoformed bone increased (p<0.001) from 30 to 180 days irrespective to treatment groups. ZnHA and the control group showed a large at 180 days but no significant difference compared to HA. Therefore, we concluded that both biomaterials are biocompatibles and osteoconductors, promoting new bone formation and apposition of bone on the surface throughout the periods and the addition of zinc improved the osteogenesis.
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Authors: Monica Calasans-Maia, G.V.O. Fernandes, Antonella M. Rossi, Eliane Pedra Dias, G.D.S. Almeida, F.F. Mitri, José Mauro Granjeiro
Abstract: Hydroxyapatite (HA), widely used as bone graft, can be modified by the incorporation of
bivalent cations (Mg2+ and Zn2+) and its gradual release could favor the bone repair. The purpose of
this research was to evaluate the effect of the HA and zinc-containing hydroxyapatite (Zn-HA) in
the bone repair in rat calvaria in comparison to autogenous bone. Critical size defect in the calvaria
was filled with the graft material and the samples were harvested at the 30, 90 and 180 days. The
light microcopy observations showed the biocompatibility of the graft materials. In the Zn-HA
group the area of neoformed bone was larger than in the HA group, but smaller than in the
autograft. A fibrous connective tissue was more evident around HA granules. It could be conclude
that the presence of zinc ions in HA crystal accelerated the osteogenesis and increased the area of
newly formed bone in relation to HA.
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