Mônica Diuana Calasans-Maia

Effect of Surface Modifications on Early Bone Healing Around Plateau Root Form Implants: An Experimental Study in Rabbits

PURPOSE The objective of the present study was to evaluate the biomechanical fixation and bone-to-implant contact (BIC) of plateau root form implants of varied surfaces. MATERIALS AND METHODS Plateau root form implants, 3.5 mm in diameter, 8 mm in length, with 4 surfaces (n = 16 each)--machined, alumina-blasted/acid-etched, alumina-blasted/acid-etched plus nanothickness bioceramic coating, and plasma-sprayed calcium-phosphate--were used. They were bilaterally placed at the distal femur of 16 New Zealand rabbits and remained in place for 2 and 4 weeks in vivo. After euthanizing the rabbits, the implants were subjected to torque to interface fracture and were subsequently processed as nondecalcified approximately 30-microm-thickness slides for histomorphologic analysis and BIC determination. Statistical analysis was performed using analysis of variance at the 95% level of significance, considering implantation time and implant surface as independent variables and the torque-to-interface fracture and BIC as dependent variables. RESULTS The torque-to-interface fracture was significantly affected by the implant surface (P < .001) but was not affected by the implantation time (P > .20). The implantation time and implant surface had significant effects on the BIC (P < .04 and P < .001, respectively). The greatest torque-to-interface fracture and BIC was observed for the plasma-sprayed calcium-phosphate. CONCLUSION The implant surface significantly influenced early bone healing around plateau root form implants.

The rabbit as an animal model for experimental surgery

The white New Zealand rabbit (Oryctolagus cuniculus) is frequently used as a model for in vivo studies. However, information on precautions when using this animal as an experimental model is limited. This review of the literature covers the gamut from the selection of the animal model all the way to its death, and describes procedures for transporting, raising, breeding, housing, administering anesthesia and handling so as to rationalize the utilization of this species while exploiting its unique characteristics. Based upon the literature and our own experience with white New Zealand rabbits, we conclude that the rabbit is an adequate model for experimental surgery.

Early osseointegration driven by the surface chemistry and wettability of dental implants

Objective The objective of this study was to investigate the impact of two different commercially available dental implants on osseointegration. The surfaces were sandblasting and acid etching (Group 1) and sandblasting and acid etching, then maintained in an isotonic solution of 0.9% sodium chloride (Group 2). Material and Methods X-ray photoelectron spectroscopy (XPS) was employed for surface chemistry analysis. Surface morphology and topography was investigated by scanning electron microscopy (SEM) and confocal microscopy (CM), respectively. Contact angle analysis (CAA) was employed for wetting evaluation. Bone-implant-contact (BIC) and bone area fraction occupied (BAFO) analysis were performed on thin sections (30 μm) 14 and 28 days after the installation of 10 implants from each group (n=20) in rabbits’ tibias. Statistical analysis was performed by ANOVA at the 95% level of significance considering implantation time and implant surface as independent variables. Results Group 2 showed 3-fold less carbon on the surface and a markedly enhanced hydrophilicity compared to Group 1 but a similar surface roughness (p>0.05). BIC and BAFO levels in Group 2 at 14 days were similar to those in Group 1 at 28 days. After 28 days of installation, BIC and BAFO measurements of Group 2 were approximately 1.5-fold greater than in Group 1 (p<0.05). Conclusion The surface chemistry and wettability implants of Group 2 accelerate osseointegration and increase the area of the bone-to-implant interface when compared to those of Group 1.

A randomized controlled clinical trial to evaluate a new xenograft for alveolar socket preservation

OBJECTIVE The aim of this clinical trial was to compare the effect of Bio-Oss(®) and a new bovine xenograft (Osseus(®) ) in alveolar sockets after a 24-week healing period. MATERIALS AND METHODS A total of 20 adult volunteers ages 30-60 were subjected to single tooth extraction. A tooth extraction was performed at the baseline. All sites were randomly allocated to two test groups (TG1: grafted using a new bovine xenograft, Osseus(®) , and TG2: grafted using commercially available bovine xenograft-Bio-Oss(®) ). Six months later, a sample of the grafted area was obtained and implants were inserted in the same site. Histological sections were examined focusing on the presence of fibrous connective tissue (CT), and newly formed bone in direct contact with the graft. The HE-stained sections were subjected to histomorphometrical evaluation using Image Pro-Plus(®) software (Release 7.0). The definitive crown was placed 3 months later. RESULTS Upon completion of the study, no patients were removed from the study and all inserted implants (10 in each group) were eventually integrated. After 6 months, in the TG1, the mean value of new bone formation was 33.7 (± 7.1), for CT was 32.3 (± 8.9) and for the remaining biomaterial was 10.7 (± 16.2). In the TG2, the mean value of new bone formation was 19.3 (± 22.6), of the CT was 49.9 (± 14.1) and of the remaining biomaterial was 22.6 (± 7.9). CONCLUSIONS No statistically significant difference was observed between TG1 and TG2 after 6 months (P > 0.05), and both biomaterials afforded a more favorable implant position.

In vitro and in vivo evaluation of strontium-containing nanostructured carbonated hydroxyapatite/sodium alginate for sinus lift in rabbits

Various synthetic bone substitutes have been developed to reconstruct bone defects. One of the most prevalent ceramics in bone treatment is hydroxyapatite (HA) that is a useful material as bone substitute, however, with a low rate of biodegradation. Its structure allows isomorphic cationic and anionic substitutions to be easily introduced, which can alter the crystallinity, morphology, biocompatibility, and osteoconductivity. The objective of this study was to investigate the in vitro and in vivo biological responses to strontium-containing nanostructured carbonated HA/sodium alginate (SrCHA) spheres (425<ϕ <600 μm) that were used for sinus lifts in rabbits using nanostructured carbonated HA/sodium alginate (CHA) as a reference. Cytocompatibility was determined using a multiparametric assay after exposing murine preosteoblasts to the extracts of these materials. Twelve male and female rabbits underwent bilateral sinus lift procedures and were divided into two groups (CHA or SrCHA) and in two experimental periods (4 and 12 weeks), for microscopic and histomorphometric analyses. The in vitro test revealed the overall viability of the cells exposed to the CHA and SrCHA extracts; thus, these extracts were considered cytocompatible, which was confirmed by three different parameters in the in vitro tests. The histological analysis showed chronic inflammation with a prevalence of macrophages around the CHA spheres after 4 weeks, and this inflammation decreased after 12 weeks. Bone formation was observed in both groups, and smaller quantities of SrCHA spheres were observed after 12 weeks, indicating greater bioresorption of SrCHA than CHA. SrCHA spheres are biocompatible and osteoconductive and undergo bioresorption earlier than CHA spheres.

Effect of Hydroxyapatite and Zinc-Containing Hydroxyapatite on Osseous Repair of Critical Size Defect in the Rat Calvaria

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.

Short-term in vivo evaluation of zinc-containing calcium phosphate using a normalized procedure

The effect of zinc-substituted calcium phosphate (CaP) on bone osteogenesis was evaluated using an in vivo normalized ISO 10993-6 protocol. Zinc-containing hydroxyapatite (ZnHA) powder with 0.3% by wt zinc (experimental group) and stoichiometric hydroxyapatite (control group) were shaped into cylindrical implants (2×6 mm) and were sintered at 1000 °C. Thermal treatment transformed the ZnHA cylinder into a biphasic implant that was composed of Zn-substituted HA and Zn-substituted β-tricalcium phosphate (ZnHA/βZnTCP); the hydroxyapatite cylinder was a highly crystalline and poorly soluble HA implant. In vivo tests were performed in New Zealand White rabbits by implanting two cylinders of ZnHA/βZnTCP in the left tibia and two cylinders of HA in the right tibia for 7, 14 and 28 days. Incorporation of 0.3% by wt zinc into CaP increased the rate of Zn release to the biological medium. Microfluorescence analyses (μXRF-SR) using synchrotron radiation suggested that some of the Zn released from the biomaterial was incorporated into new bone near the implanted region. In contrast with previous studies, histomorphometric analysis did not show significant differences between the newly formed bone around ZnHA/βZnTCP and HA due to the dissolution profile of Zn-doped CaP. Despite the great potential of Zn-containing CaP matrices for future use in bone regeneration, additional in vivo studies must be conducted to explain the mobility of zinc at the CaP surface and its interactions with a biological medium.

Histomorphometric Analysis of Bone Repair in Critical Size Defect in Rats Calvaria Treated with Hydroxyapatite and Zinc-Containing Hydroxyapatite 5%

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.

Cytocompatibility and biocompatibility of nanostructured carbonated hydroxyapatite spheres for bone repair

ABSTRACT Objective The aim of this study was to investigate the in vitro and in vivo biological responses to nanostructured carbonated hydroxyapatite/calcium alginate (CHA) microspheres used for alveolar bone repair, compared to sintered hydroxyapatite (HA). Material and Methods The maxillary central incisors of 45 Wistar rats were extracted, and the dental sockets were filled with HA, CHA, and blood clot (control group) (n=5/period/group). After 7, 21 and 42 days, the samples of bone with the biomaterials were obtained for histological and histomorphometric analysis, and the plasma levels of RANKL and OPG were determined via immunoassay. Statistical analysis was performed by Two-Way ANOVA with post-hoc Tukey test at 95% level of significance. Results The CHA and HA microspheres were cytocompatible with both human and murine cells on an in vitro assay. Histological analysis showed the time-dependent increase of newly formed bone in control group characterized by an intense osteoblast activity. In HA and CHA groups, the presence of a slight granulation reaction around the spheres was observed after seven days, which was reduced by the 42nd day. A considerable amount of newly formed bone was observed surrounding the CHA spheres and the biomaterials particles at 42-day time point compared with HA. Histomorphometric analysis showed a significant increase of newly formed bone in CHA group compared with HA after 21 and 42 days from surgery, moreover, CHA showed almost 2-fold greater biosorption than HA at 42 days (two-way ANOVA, p<0.05) indicating greater biosorption. An increase in the RANKL/OPG ratio was observed in the CHA group on the 7th day. Conclusion CHA spheres were osteoconductive and presented earlier biosorption, inducing early increases in the levels of proteins involved in resorption.

Stimulatory Effect on Osseous Repair of Zinc-Substituted Hydroxyapatite: Histological Study in Rabbit’s Tibia

The study was carried out aiming the evaluation of the effect of hydroxyapatite (HA) and zinc-substituted hydroxyapatite (ZnHA) on osseous repair of rabbit’s tibia. For the study, 15 adult animals, weighing around 2.5 and 3.0 Kg, sourced by the Fluminense Federal University Animal Lab were acquired. Two perforations were created in each tibia and filled with cylinders (2x6 mm) of HA (group 1, right) or ZnHA (group 2, left). The animals were killed after 7, 14, and 28 days for evaluating the histological aspects of the interface site and bone repair. No sign of inflammatory reaction surrounding the cylinders area were observed, neither giant cells. Osteogenesis was evidently accelerated in all healing periods for the ZnHA group in regards to the HA group. Randomly dispersed areas of neoformed bone among wide areas of fibrous connective tissue was observed in the HA group. We conclude that both biomaterials are biocompatible, but zinc-containg HA enhanced and accelerated the osteogenesis in relation to HA.