Research on implants and osseointegration.

Osseointegration was originally defined as a direct structural and functional connection between ordered living bone and the surface of a load-carrying implant. It is now said that an implant is regarded as osseointegrated when there is no progressive relative movement between the implant and the bone with which it is in direct contact. Although the term osseointegration was initially used with reference to titanium metallic implants, the concept is currently applied to all biomaterials that have the ability to osseointegrate. Biomaterials are closely related to the mechanism of osseointegration; these materials are designed to be implanted or incorporated into the living system with the aims to substitute for, or regenerate, tissues and tissue functions. Objective evaluation of the properties of the different biomaterials and of the factors that influence bone repair in general, and at the bone tissue-implant interface, is essential to the clinical success of an implant. The Biomaterials Laboratory of the Oral Pathology Department of the School of Dentistry at the University of Buenos Aires is devoted to the study and research of the properties and biological effects of biomaterials for dental implants and bone substitutes. This paper summarizes the research work resulting from over 25 years' experience in this field. It includes studies conducted at our laboratory on the local and systemic factors affecting the peri-implant bone healing process, using experimental models developed by our research team. The results of our research on corrosion, focusing on dental implants, as well as our experience in the evaluation of failed dental implants and bone biopsies obtained following maxillary sinus floor augmentation with bone substitutes, are also reported. Research on biomaterials and their interaction with the biological system is a continuing challenge in biomedicine, which aims to achieve optimal biocompatibility and thus contribute to patient health.

The issue of corrosion in dental implants: a review.

Pure titanium or titanium alloys, and to a lesser extent, zirconium, are metals that are often used in direct contact with host tissues. These metallic biomaterials are highly reactive, and on exposure to fluid media or air, quickly develop a layer of titanium dioxide (TiO2) or zirconium dioxide (ZrO2). This layer of dioxide forms a boundary at the interface between the biological medium and the metal structure, determining the degree of biocompatibility and the biological response of the implant. Corrosion is the deterioration a metal undergoes as a result of the surrounding medium (electrochemical attack), which causes the release of ions into the microenvironment. No metal or alloy is entirely inert in vivo. Corrosion phenomena at the interlace are particularly important in the evolution of both dental and orthopedic implants and one of the possible causes of implant failure after initial success. This paper comprises a review of literature and presents results of our laboratory experiments related to the study of corrosion, with special emphasis on dental implants. In situ degradation of a metallic implant is undesirable because it alters the structural integrity of the implant. The issue of corrosion is not limited to a local problem because the particles pmduced as a result could migrate to distant sites, whose evolution would require further studies.

Ploidy studies of pleomorphic adenomas and minor salivary gland carcinomas of the palate.

Ploidy studies of tumors are a diagnostic and prognostic aid. The aim of the present study was to evaluate the DNA content of palate aggressive pleomorphic adenomas (PA) and adenocarcinomas of the salivary glands. Twelve cases of salivary gland tumors of the palate were selected from the archives of the Oral Pathology Department, Faculty of Dentistry, University of Buenos Aires (1966-2001). Six cases corresponded to aggressive pleomorphic adenomas (PA) and the remaining six to adenocarcinomas (AD). Myxoid and epithelial areas of PA were evaluated. The epithelial areas of the most aggressive cases of PA exhibited a high DNA content. The myxoid areas of same cases of PA had a 2C ploidy level. The difference in ploidy values between the myxoid and epithelial areas of PA would suggest the presence of different cell populations. DNA content and the detection of aneuploidy would be prognostic aids in palate salivary gland carcinomas.

Effects of intraosseous implantation of silica-based bioactive glass particles on rat kidney under experimental renal failure.

The aim of the present study is to evaluate the effects of intraosseous implantation of silica-based bioactive glass (BG) particles on rat kidney under experimental renal failure. The animals are assigned to one of the two groups: renal failure (RF) and renal failure + bioactive glass (RF + BG). Particles of melt-derived 45S5 BG are implanted in the marrow of one tibia of each animal in the RF + BG group. The animals are killed 24 h and 14 days postimplantation. The RF + BG group exhibits a statistically significant increase in serum urea 24 h postimplantation. The tibiae of the RF + BG group are resected and embedded in methyl-methacrylate resin. Ground sections are analyzed by light microscopy and energy-dispersive X-ray (EDX) analysis. The presence of silicon, calcium, and phosphorus is evaluated in the BG particles. A 55% reduction in silicon content is observed at 14 days postimplantation as compared with that at 24 h.Light microscopy analysis reveals lesions in kidney parenchyma. Hyperplasia associated with nuclear vacuolization in the tubules and a marked thickening of the basal membrane are observed in the renal cortex of the RF + BG animals killed at 24 h postimplantation, but not in those at 14 days. The present results demonstrate reversible renal cell injury in rats exposed to intraosseous implantation of silica-based BG particles under experimental RF.

Malignant fibrous histiocytoma associated with coxofemoral arthrodesis.

The discovery of biomaterials led to their use in the manufacture of implants for biomedical applications. In vivo, no metal or alloy is completely inert. The potential toxicity of some of the metals most frequently employed in the manufacture of orthopedic implants has been reported. Their carcinogenic potential has been evaluated in experimental animal models. However, few reports have discussed the potential development of malignant tumors associated with prosthetic structures in humans. The present study documents a case of intraosseous sarcoma that developed in the vicinity of a metallic prosthesis 43 months after a coxofemoral arthrodesis with metallic pins and screws. With this report the authors seek to contribute to the understanding of the potential toxicity and risks of using metallic implants. Since metallic implants employed in the rehabilitation of osteo-muscular-articular disorders usually remain in the organism for long periods of time, the need to monitor the metallic structures and the adjacent tissues is extremely relevant.

Meta-analysis of the literature on 1946 cases of minor salivary gland tumors of the palate.

Minor salivary gland tumors are relatively rare and exhibit great diversity in terms of histopathology, localization, biological behavior and classification. The studies of significant case series report controversial data, mainly in terms of the proportion of benign versus malignant tumors and the relative frequency of histological types. Palate tumors are the most frequent, with an incidence of over 50%. The aim of the present study was to perform a meta-analysis to evaluate salivary gland tumors of the palate in terms of the proportion of malignant versus benign tumors, the frequency of the histological types and the data employed for statistical analysis. We analyzed a selection of international publications (1950-1999) of case series of minor salivary gland tumors of the palate, including our own series. The host institutions were classified into 3 categories: A) High Complexity Institutions (HCI), i.e. oncological reference centers and general hospitals that treat cancer patients; B) Medium Complexity Institutions (MCI); C) Low Complexity Institutions (LCI). Based on the main classifications, we joined categories and employed a simplified classification to analyze a total of 1835 cases in the literature and our own series of 111 cases (unpublish data). The results of the meta-analysis of the literature demonstrated that the data employed for statistical analysis depends on the type of host institution. The classification of institutions according to their level of complexity allowed for adequate interpretation of the previously published statistical data. Our interpretation of these studies suggests that the data on the percentage of malignant versus benign tumors and diversity of histological type must be obtained from series of low complexity institutions. LCI data are reliable whereas the HCI data are based on pre-selected cases, rendering the data unreliable.

Effect of titanium dioxide on the oxidative metabolism of alveolar macrophages: an experimental study in rats.

Metallic implants of titanium are used therapeutically in biomedicine because of its excellent biocompatibility. However, no metal or alloy is completely inert. We have previously shown that titanium oxide (TiO(2)) is transported in blood by phagocytic monocytes and deposited in organs such as liver, spleen, and lung 6 months after intraperitoneal injection (ip). Furthermore, it is well known that exposure to metal traces alters the cellular redox status. Thus, the aim of the present study was to determine the presence of titanium in target organs after chronic exposure, assess the potential structural alterations, and evaluate the oxidative metabolism of alveolar macrophages (AM) in the lung. Rats were ip injected with 1.60 g/100 g body wt of TiO(2) in saline solution. Organs (liver, spleen, lung) were processed for histological evaluation. Reactive oxygen species (ROS) in AM obtained by bronchoalveolar lavage (BAL) were evaluated using the nitroblue tetrazolium test and quantitative evaluation by digital image analysis. The histological analysis of organs revealed the presence of titanium in the parenchyma of these organs with no associated tissue damage. Although in lung alveolar macrophages TiO(2) induced a significant rise in ROS generation, it failed to cause tissue alteration. This finding may be attributed to an adaptive response.

Kinetics of MTT-formazan exocytosis in phagocytic and non-phagocytic cells.

MTT is taken up by cells by endocytosis and reduced to formazan in the endosomal/lysosomal compartment. Formazan is deposited intracellularly as blue granules and is later exocytosed as needle-like formazan crystals. The present study involves an analysis of the pattern of exocytosis of MTT in different cell types showing clearcut differences in the response that can be associated to their ability to phagocytose. To further assess the characteristics of the exocytic mechanism of MTT/formazan, different experimental conditions were assayed. When culture medium with decreasing serum concentration was used as a metabolic modulator no variations were observed in the proportion of cells with formazan crystals. Conversely, the markedly sensitivity of phagocytic cells to increasing concentrations of genistein constituted a remarkable difference with non-phagocytic cells. These results must be considered when the modulation of MTT exocytosis is used as a signal of the progress of human diseases.

Analysis of the epidemiological features of oral cancer in the city of Buenos Aires.

Oral cancer comprises 0.6 to 5% of all human malignant tumors. It is accepted in the literature that the clinical evolution of oral cancer has a bad prognosis, i.e. the five year survival rate ranges from 34% to 56%. The aim of the present study was to present a metaanalysis of the most relevant publications on oral cancer in the city of Buenos Aires, including our own series. The publications reviewed herein include the following series: 517 cases (1950-1970), 243 cases (1961-1968), 336 cases (1972-1984), and 274 cases (1992-2000). The clinical end-points evaluated were: age, distribution by sex, tumor site, presence of metastatic adenopathies, and clinical stage. A comparative statistical evaluation of the clinical parameters assessed was performed. Survival was evaluated by the test of Kaplan-Meier. The male/female ratio was 7.1:1 for the 1950-1970 period, 4.3:1 for the 1961-1968 period; 2.3:1 for the 1972-1984 period; and 1.24:1 in our series (1992-2000). The most frequent tumor site (21 to 35% of the cases) was the tongue. At the time of diagnosis, 60-71% of the patients had advanced TNM clinical stages (III and IV). Our follow-up revealed a five-year survival rate after diagnosis of 39%. The overall analysis of all the cases corresponding to the 1950-2000 period revealed that the prevalence of oral cancer in women has risen alarmingly and that the percentage of patients with advanced stages of tumor development continues to be high.

Efficacy of Core Needle Biopsy Technique for Jawbone Diseases.

Core needle biopsy (CNB) has been proven useful for diagnosing bone lesions, although it is not often used for jawbone lesions. The aim of this study was to evaluate the efficacy of the CNB method in a series of cases of intramaxillary lesions. CNB was performed on 85 patients with intraosseous lesions which were grouped according to radiographic appearance as: radiopaque lesions (RO, n=13), radiolucent lesions (RL, n=39) and mixed lesions with both radiolucent and radiopaque areas (RL-RO, n=33). The technique enabled us to obtain several tissue cylinders from each lesion (average 2.5 cylinders), which were processed following routine histopathological technique and H&E stain, plus special techniques when necessary. The histopathological analysis together with clinical data enabled accurate diagnosis (AD) in 81% of the cases and descriptive diagnosis (DD) in 14%. The material obtained in 5% of the cases was not appropriate for study (ND). The difference between successful (AD) and unsuccessful (DD+ND) CNB cases is statistically significant. The highest percentage of successful CBNs was for RO and RLRO lesions (85% and 100% respectively). RL lesions were more difficult because most of them were cystic lesions with fluid content.

La biopsia-punción ósea ( Core needle biopsy, CNB) es un procedimiento de probada utilidad en el diagnóstico de lesiones óseas. Sin embargo, no es una técnica de uso frecuente en las lesiones de los maxilares. La finalidad de este trabajo fue evaluar la eficacia del método de CNB en una serie de casos de lesiones intramaxilares. Se realizaron CNB en 85 pacientes con lesiones intraóseas, las cuales fueron agrupadas según su aspecto radiográfico en lesiones radiopacas ( RO, n=13), lesiones radiolucidas (RL, n=39) y lesiones mixtas con sectores radiolúcidos y radiopacos (RL-RO, n=33). La técnica permitió obtener varios cilindros de tejido de cada lesión ( promedio: 2.5 cilindros) los cuales fueron procesados según técnica histopatológica de rutina con tinción de H&E y técnicas especiales en los casos en que fueron necesarias. El análisis de los cuadros histopatológicos conjuntamente con los datos clínicos, permitió realizar un un diagnóstico de certeza (AD) en el 81% de los casos y un diagnostico descriptivo (DD) en el 14 % . En el 5% de los casos el material obtenido no fue adecuado para su estudio (ND) La diferencia entre los casos de CNB exitosa y no exitosa ( DD+ND) es estadisticamente significativa. El mayor porcentaje de CBN exitosas correspondió a las lesiones RO y RL-RO ( 85% y 100% respectivamente) Las lesiones RL presentaron mayor dificultad debido a que, en su mayoría, eran lesiones quísticas con contenido líquido.

In vitro age dependent response of macrophages to micro and nano titanium dioxide particles.

As a result of corrosion, microparticles (MP) and/or nanoparticles (NP) can be released from the metallic implants surface into the bioenvironment. The biological response to these particles depends not only on the physico-chemical properties of the particles but also on host factors, such as age. Macrophages have attracted wide concern in biomedicine. The aim of this investigation was to study the age related biological response of macrophages to TiO2 -MP and NP in vitro. Alveolar macrophages (AM) obtained from young and senescent rats were cultured and exposed to TiO2 -MP and NP. Cell metabolism, superoxide anion (O2 (-) ) and nitric oxide (NO) generation, and cytokine release (IL-6, TNFα, IL-10) were measured. Cell metabolism was not affected by particle exposure. O2 (-) and NO generation increased in a dose dependent manner. A marked increase on IL-6 release was found in the young-AM subpopulation exposed to TiO2 -MP. Conversely, both particle sizes induced a dose dependent release of TNFα in senescent-AM. Only the highest concentration of TiO2 -particles caused a significant increase in IL-10 release in AM-cultures. These observations lend strong support to the suggestion that cellular response of macrophages to TiO2 -particles is age dependent. The biological effect of the particles would seem to be more deleterious in the senescent age-group.

Exposure to subcutaneously implanted uranium dioxide impairs bone formation.

The introduction of uranium particles into subcutaneous tissue is a risk that affects workers engaged in the extraction, purification, and manufacture of uranium, as well as soldiers who are wounded with uranium shrapnel. The authors evaluated the effect of an internal source of an insoluble form of uranium on bone. Uranium dioxide powder (0.125 gm/kg body weight) was implanted subcutaneously in rats. After 30 days, animals exposed to uranium weighed less than controls. Bone formation activity in endochondral ossification and bone growth were also lower in the experimental animals, as evidenced by histomorphometric and morphometric methods. This is the first study to report bone damage resulting from continuous, nonlethal exposure to an insoluble compound of uranium dioxide over a period of 30 days.

Anatomic variations of the root canal of the rat according to age.

The mesial canal of the first lower molar of the rat has been used as an experimental model in Endodontics. The present study involves a detailed analysis of its structural variations as a function of age and thus contributes to its characterization and adequate use as an experimental model. We evaluated 60 molars of Wistar rats of different body weight: (group 1: 300-399 g, n=22; group 2: 400-499 g, n=16; group 3: 600-700 g, n=22). The samples were radiographed employing a standardized system. Total canal length and volume were measured on enlarged projections of the radiographs. Canal diameter was measured at coronary, middle and apical levels and employed to estimate the area of canal cross-section as an indicator of potential flow or metabolic interchange. From the second age group onwards, we detected radicular hypercementosis that increased canal length and resulted in the formation of a complex apical delta. The canal diameter decreased very significantly in the apical third. The estimated flow expressed in terms of the area of cross-section on the projection decreased from 1.2 mm2 in Group 1 to 0.05 mm2 in Group 2. Group 3 did not differ significantly from Group 2. The present results reveal that the spontaneous, age-related variations in canal anatomy described herein must be considered when performing experimental endodontics in rats.

Impact through time of different sized titanium dioxide particles on biochemical and histopathological parameters.

Due to corrosion, a titanium implant surface can be a potential source for the release of micro (MPs) and nano-sized particles (NPs) into the biological environment. This work sought to evaluate the biokinetics of different sized titanium dioxide particles (TiO2 ) and their potential to cause cell damage. Wistar rats were intraperitoneally injected with 150 nm, 10 nm, or 5nm TiO2 particles. The presence of TiO2 particles was evaluated in histologic sections of the liver, lung, and kidney and in blood cells at 3 and 12 months. Ultrastructural analysis of liver and lung tissue was performed by TEM, deposit concentration in tissues was determined spectroscopically, and oxidative metabolism was assessed by determining oxidative membrane damage, generation of superoxide anion (O2(-)), and enzymatic and non-enzymatic antioxidants. TiO2 particles were observed inside mononuclear blood cells and in organ parenchyma at 3 and 12 months. TiO2 deposits were consistently larger in liver than in lung tissue. Alveolar macrophage O2(-) generation and average particle size correlated negatively (p < 0.05). NPs were more reactive and biopersistent in lung tissue than MPs. Antioxidant activity, particularly in the case of 5 nm particles, failed to compensate for membrane damage in liver cells; the damage was consistent with histological evidence of necrosis.

Subcellular redistribution of NHERF1 in response to dehydroepiandrosterone (DHEA) administration in endometrial glands of Wistar rats.

To understand the regulation of Na(+)/H(+) exchanger regulatory factor (NHERF1) in polycystic ovarian syndrome, we studied the expression of NHERF1 in uterus of Wistar rats injected with (6 mg/kg) of dehydroepiandrosterone (DHEA) for 7 and 20 days. Immunohistochemistry analysis of NHERF1 showed a substantial shift in the intracellular localization of NHERF1 in endometrial glands and areas of luminal epithelium as early as 7 days of DHEA administration. The NHERF1 accumulated in the "Golgi apparatus area" virtually in all the glands in the 7-day protocol, and in the majority of the glands of 20-day protocol. In contrast, NHERF1 is expressed in the apical membrane and slightly in the cytoplasm of the control epithelium. The subcellular redistribution of NHERF1 could affect the sorting of proteins to the apical membrane and the organization of the apical compartment.

Exfoliative cytology and titanium dental implants: a pilot study.

BACKGROUND: Oral exfoliative cytology is a diagnostic method that involves the study of cells exfoliated from the oral mucosa. Ions/particles released from metallic implants can remain in the peri-implant milieu. The aim of the present study is to assess the presence of metal particles in cells exfoliated from peri-implant oral mucosa around titanium dental implants. METHODS: The study comprised 30 patients carrying titanium dental implants, who had neither a metallic prosthesis nor metal restorations in neighboring teeth. Individuals undergoing orthodontic therapy and those who had oral piercing were also excluded from the study. The study sample included patients with and without peri-implantitis. Cytologic samples of the peri-implant area were collected. Samples of the marginal gingiva on the contralateral side of the implant were taken from the same individuals to serve as control. Cytologic analysis was performed using light microscopy. Titanium concentration was determined using inductively coupled plasma-mass spectrophotometry. RESULTS: Metal-like particles were observed inside and outside epithelial cells and macrophages in cytologic smears of peri-implant mucosa of both patients with and without peri-implantitis. No particles were found in the control cytologic samples. The concentration of titanium was higher in the peri-implantitis group compared with the group without peri-implantitis; no traces of titanium were observed in controls. CONCLUSIONS: Regardless of an inflammatory response, ions/particles are released from the surface of the implant into the biologic milieu. Exfoliative cytology is a simple technique that may be used to detect metal particles in cells exfoliated from the peri-implant mucosa.

Oral mucosa tissue response to titanium cover screws.

BACKGROUND: Titanium is the most widely used metal in dental implantology. The release of particles from metal structures into the biologic milieu may be the result of electrochemical processes (corrosion) and/or mechanical disruption during insertion, abutment connection, or removal of failing implants. The aim of the present study is to evaluate tissue response of human oral mucosa adjacent to titanium cover screws. METHODS: One hundred fifty-three biopsies of the supra-implant oral mucosa adjacent to the cover screw of submerged dental implants were analyzed. Histologic studies were performed to analyze epithelial and connective tissue as well as the presence of metal particles, which were identified using microchemical analysis. Langerhans cells, macrophages, and T lymphocytes were studied using immunohistochemical techniques. The surface of the cover screws was evaluated by scanning electron microscopy (SEM). RESULTS: Forty-one percent of mucosa biopsies exhibited metal particles in different layers of the section thickness. Particle number and size varied greatly among specimens. Immunohistochemical study confirmed the presence of macrophages and T lymphocytes associated with the metal particles. Microchemical analysis revealed the presence of titanium in the particles. On SEM analysis, the surface of the screws exhibited depressions and irregularities. CONCLUSIONS: The biologic effects seen in the mucosa in contact with the cover screws might be associated with the presence of titanium or other elements, such as aluminum or vanadium. The potential long-term biologic effects of particles on soft tissues adjacent to metallic devices should be further investigated because these effects might affect the clinical outcome of the implant.

In vivo comparative biokinetics and biocompatibility of titanium and zirconium microparticles.

Titanium and zirconium are biomaterials that present a layer of titanium dioxide (TiO(2)) or zirconium dioxide (ZrO(2)). As a result of corrosion, microparticles can be released into the bioenvironment, and their effect on tissues is seemingly associated with differences in the physicochemical properties of these metals. The aim of this study was to perform a long-term evaluation of the distribution, destination, and potential risk of TiO(2) and ZrO(2) microparticles that might result from the corrosion process. Wistar rats were i.p. injected with an equal dose of either TiO(2) or ZrO(2) suspension. The following end-points were evaluated at 3, 6, and 18 months: (a) the presence of particles in blood cells and liver and lung tissue, (b) Ti and Zr deposit quantitation, (c) oxidant-antioxidant balance in tissues, and (d) O(2)(-) generation in alveolar macrophages. Ti and Zr particles were detected in blood mononuclear cells and in organ parenchyma. At equal doses and times postadministration, Ti content in organs was consistently higher than Zr content. Ti elicited a significant increase in O(2)(-) generation in the lung compared to Zr. The consumption of antioxidant enzymes was greater in the Ti than in the Zr group. The present study shows that the biokinetics of TiO(2) and ZrO(2) depends on particle size, shape, and/or crystal structure.

Expression of NHERF1 in colonic tumors induced by 1,2-dimethylhydrazine in rats is independent of plasma ovarian steroids.

In normal embryonic fibroblasts, the Na(+)/H(+) exchanger regulator factor 1 (NHERF1) stabilizes E-cadherin/ß-catenin binding and the lack of NHERF1 expression promotes cell transformation thus acting as a tumor suppressor gene. We here tested the hypothesis that NHERF1 could act as a tumor suppressor gene in colon cancer as a mediator of estrogens' protective actions in colon carcinogenesis. We studied the expression and localization of NHERF1 and ß-catenin by immunohistochemistry in colonic tumors induced by 1,2 dimethylhidrazine (DMH) in Sprague-Dawley rats. One group of the rats treated with the carcinogen was ovariectomized (OVX) in the middle of the tumor induction, simulating a human menopausal condition. We observed a protective role of estrogens in colon cancer, as non-ovariectomized rats (DMH) had a reduced tumor area compared with the ovariectomized group (DMH + OVX; mean ± SE) 28.98 ± 4.65 vs. 67.58 ± 8.69 (p < 0.00380). Despite the lack of plasma estrogen stimulation, we found abundant expression of NHERF1 in colon tumors from ovariectomized rats. NHERF1 was mainly localized in the cytoplasm of the adenocarcinoma cells and lost the apical localization previously reported in normal colon tissue. We also detected expression of NHERF1 by western blot in the SW48, CACO-2, and HT29 colon cancer cell lines. Non-estrogenic factors in plasma or the tumor microenvironment may regulate NHERF1 expression in transformed colon epithelial cells. Further studies are required to understand the regulation of NHERF1 expression in colon cancer tissue.