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Biomedical subjects

D Zaffe

Publications and source records attributed to D Zaffe.

At least 19 recordsLinked to original sources

Histological study on sinus lift grafting by Fisiograft and Bio-Oss.

The work aims to provide a histological investigation of Fisiograft, a PLA/PGA copolymer, used as filler for bone defects in humans. The study was performed on biopsies of sinus lifts where Bio-Oss and Fisiograft gel were applied as graft material. Bone regeneration was satisfactory in all sinus lifts, even when Fisiograft was applied alone. Due to remarkable osteoclast activity, Bio-Oss granules were cleared from the majority of biopsy cores. At histology, Fisiograft gel appeared as globes enveloped by fibroblasts, displaying an epithelial-like cell appearance. Due to its solubility in solvents, undegraded Fisiograft (recorded for 7 months or more) did not stain whereas degraded Fisiograft stained positive. The loose connective tissue, that surrounded Fisiograft and bone contained isolated mastocytes. Bone grew inside the loose connective and often reached the surface of Fisiograft by intervening cells. The results seem to indicate that Fisiograft may be considered both a polymer useful for fastening bone substitutes inside a defect and in addition a material capable of prompting bone regeneration, with or without the use of a bone substitute. In addition to space-former and space-maintainer functions, Fisiograft shows potential bone stimulation function, which may be labelled as osteopromotive capability.

Autoradiography↗

Release of elements from retrieved maxillofacial plates and screws.

Vitallium appliances and surrounding tissues were investigated to evaluate the release and accumulation of elements. Four microplates, sixteen screws and surrounding tissues were removed from three patients presenting inflammation 4 to 6 years after surgery and were submitted to SEM and X-ray microprobe analysis. Histology was performed on paraffin or PMMA sections of tissues.A continuous release of elements from metallic appliances into soft tissues was observed. Cobalt, chromium, and nickel were detected in soft and boney tissues in close proximity to the appliance. Aluminium, as a component of screw coatings, accumulated in soft tissues, and a remarkable amount of aluminium was detected in the dense lamella of lamellar bone. The results suggest that coatings containing aluminium should be avoided and the time these appliances are allowed to remain in patients should be shortened. Further studies on element release and the fate of aluminium in bone are warranted.

Aged↗

PLGA microspheres for oral osteopenia treatment: preliminary "in vitro"/"in vivo" evaluation.

The aim of this work was to prepare and to evaluate "in vitro"/"in vivo" microspheres based on poly(D,L-lactide-co-glycolide) copolymers containing ipriflavone, for the local treatment of oral bone loss. The first objective was the preparation and "in vitro" characterization of ipriflavone loaded microspheres, by emulsion/solvent evaporation method. Process parameters such as drug:polymer weight ratio, and molecular weight of copolymers, were also investigated. The second objective was to elaborate a suitable animal model of mandibular osteoporosis, to evaluate the efficacy of these microparticulate drug delivery systems. "In vivo" experiments were carried out on female rats, in which oral osteopenia was induced by gonadectomy and molar avulsion. Morphometric analysis of mandibular segment were carried out to quantify the development of oral osteopenia and the efficacy of drug loaded microspheres. Results showed that ipriflavone loaded PLGA microspheres can be successfully obtained with good "in vitro" characteristics, utilizing the emulsification/solvent evaporation method. "In vivo" experiments revealed that local administration of microspheres produced only mild inflammation on the injection site. Morphometric analyses showed, at the level of the third molar, a slight increase in spongy and total bone mass on rat jaw treated with microspheres with respect to control. Control animals exhibited a scarce degree of osteopenia demonstrating that this animal model is not suitable for this purpose.

Animals↗

Protective effect of melanocortin peptides in rat myocardial ischemia.

The influence of the melanocortin peptide ACTH-(1-24) (adrenocorticotropin) on the consequences of short-term coronary ischemia (5 min) followed by reperfusion, and the effect of the long-acting melanocortin [Nle(4),D-Phe(7)]alpha-melanocyte-stimulating hormone (NDP-MSH) on the damage induced by a permanent coronary occlusion, were investigated in anesthetized rats. Ischemia was produced by ligature of the left anterior descending coronary artery. Reperfusion-induced arrhythmias [ventricular tachycardia (VT), ventricular fibrillation (VF)] and survival rate within the 5 min following reperfusion, blood levels of free radicals detected 2 min after reperfusion by electron spin resonance spectrometry, and amount of healthy myocardial tissue, measured 72 h after permanent coronary occlusion on immunohistologically stained serial sections, were evaluated. Postischemic reperfusion induced VT in all saline-treated rats, and VF and death in a high percentage of animals (87%). In rats treated i.v. (2.5 min after coronary occlusion) with ACTH-(1-24) (0.16-0.48 mg/kg) there was a significantly dose-dependent reduction in the incidence of arrhythmias and lethality. Ischemia/reperfusion caused a large increase in free radical blood levels; treatment with ACTH-(1-24) (0.48 mg/kg i.v.) almost completely prevented this increase. In rats subjected to permanent coronary occlusion, the amount of healthy myocardial tissue was much reduced in saline-treated rats, while in rats treated s.c. with NDP-MSH (0.27 mg/kg every 12 h) it was significantly higher. The present data demonstrate, for the first time, an unforeseen property of melanocortin peptides, i.e., their ability to significantly reduce both heart ischemia/reperfusion injury and size of the ischemic area induced by permanent coronary occlusion.

Animals↗

[Metallic elements in tissues surrounding internal rigid fixation (IRF) devices].

BACKGROUND: Plates and other devices made by several alloys have been introduced to reach the stability of bone fractured fragments. Elements constituting alloys could be detected especially in organs, yet also in local tissues. Aim of the present study is the analysis of tissues surrounding IRF devices analyzing the morphology of released particles and studying the behavior of adjacent tissues to check metallic elements diffusion. METHODS: Biopsies were retrieved from 18 patients, aged 20 to 76 years. The patients received IRF by plates, screws and grids from 4 months to 9 years. They were divided into five groups according to the local phlogistic degree. Ordinary light microscopy, scanning electron microscopy and X-ray microprobe analysis (EDS system) was used to perform morphological investigations and identification of metal particles and elements. RESULTS: Metal particles or elements arising from plates, screws or grid may undergo tissular diffusion and cellular uptake. Not only Chromium, Iron or Aluminium but also Titanium may be easily released from devices and engulfed in tissues. In particular Titanium diffusion is evident in fibrous tissue surrounding IRF devices. Aluminium appears to be particularly accumulated in a persistent way in fibrous tissues and shows a characteristic embedding pattern in lamellar bone. CONCLUSIONS: The degree of local phlogosis appears to be strictly correlated to metallosis. Chromium, Iron, Aluminium and also Titanium, even if at different degree, give rise to phlogistic effects. Metallosis and phlogosis can produce a cascade process in which they are both the cause and the effect at the same time. The abundant release of Titanium, which does not normally produce clinical phlogosis as i.e. Aluminium, should be worthy of further investigations on its cellular effects.

Adult↗

Osteocyte-osteoclast morphological relationships and the putative role of osteocytes in bone remodeling.

An osteocyte lacunae differential count under the light microscope (LM) (1-lacunae with live osteocytes, 2-empty lacunae and lacunae with degenerating osteocytes) was carried out outside the reversal lines of osteonic lamellar bone from various mammals and man to evaluate the possibility of osteocyte survival where osteoclast resorption had occurred. The polarized light microscope (PLM) was used to establish the curvature of bony lamellae outside the convexity of reversal lines: concave lamellae indicate osteocytes reabsorbed on their vascular side where they radiate long vascular dendrites; convex lamellae indicate bone resorption on the osteocyte mineral side, radiating short dendrites. In all samples it was found that: a) about 60% of osteocytes outside the reversal lines were live; b) the percentage of alive osteocytes close to reversal lines is higher when they are attacked on their mineral side. The present data support our view that surviving osteocytes, particularly those attacked from their mineral side, might intervene in the final phase of bone resorption (osteoclast inhibition?). The fact that under the transmission electron microscope (TEM) intercellular contacts were never observed between osteocytes and osteoclasts indicates that if a modulation should occur between these two cellular types it could take place by a paracrine route only. The putative role of the cells of the osteogenic system, particularly osteocytes, in the bone remodeling cycle is also discussed.

Journal Article↗

Neuroprotective effect of gamma-hydroxybutyrate in transient global cerebral ischemia in the rat.

The effect of gamma-hydroxybutyrate on the histological and behavioral consequences of transient brain ischemia was studied in the four vessel occlusion rat model. In saline-treated animals, 30 min ischemia caused a massive loss of neurons in the hippocampal CA1 subfield (normal neurons: 14%, 5%, 23% and 30% on the 3rd, 10th, 15th and 65th day after ischemia, respectively). gamma-Hydroxybutyrate - 300 mg/kg intraperitoneally (i.p.) 30 min before or 10 min after arteries occlusion, followed by 100 mg/kg i.p. twice daily for the following 10 days - afforded a highly significant protection (normal neurons on the 3rd, 10th, 15th and 65th day after ischemia: 88% and 91%, 80% and 80%, 91% and 90%, 72% and 71% in rats receiving the first dose before or after arteries occlusion, respectively). The ischemia-induced sensory-motor impairment was significantly attenuated in rats receiving the first dose of gamma-hydroxybutyrate before arteries occlusion. Finally, the ischemia-induced impairment in spatial learning and memory, evaluated starting 27 days after the ischemic episode, was significantly attenuated by gamma-hydroxybutyrate, either injected first at 30 min before or 10 min after arteries occlusion. Lower doses of gamma-hydroxybutyrate had no significant effect. In conclusion, these results indicate that gamma-hydroxybutyrate provides significant protection against both histological and behavioral consequences of transient global cerebral ischemia in rats.

Animals↗

[Clinical evaluation, radiologic and histologic analysis in mandibular alveolar distraction procedures. Preliminary study].

BACKGROUND: Alveolar distraction osteogenesis is a process to form new alveolar bone to correct alveolar deformities in ridge height and width. Aim of this work is to study the bone processes to optimize the implantoprosthetic rehabilitation. METHODS: Alveolar distraction osteogenesis was applied in 7 patients with ridge deformities to obtain the desired ridge augmentation. Clinical and radiological evaluations were performed during the following 12 weeks, before implant insertion. Biopsies at 40, 60 and 88 days were studied after general, specific and histochemical staining of slides; microradiographs were analyzed to evaluate the trabecular bone volume. RESULTS: Forty days after the end of distraction, soft callus shows the start of ossification. Sixty days after the end of distraction, soft callus was widely converted into a network of trabecular woven bone; osteogenic activities were low; trabecular bone volume was about 50%. Eighty-eight days after the end of distraction bone amount appeared reduced, with a more ordered structure, further reduction of bone formation activity, whereas osteoclast erosion was active. CONCLUSIONS: Results show an almost steady-state bone deposition processes 60 days after the end of distraction and a regress with longer time. The results suggest the possibility of an early implant insertion to avoid bone loss due to mechanical unloading.

Adolescent↗

Neovascularized bone grafts: experimental investigation.

Vascularized bone grafts are standardized procedures in reconstructive surgery but there are some disadvantages: donor site morbidity, limited number of "natural" donor sites, and complex technique. In this study, we test the possibility of creating a "neovascularized" bone graft utilizing a vascular implantation procedure in a rabbit model. Sixteen New Zealand adult white rabbits were used. In each animal, two iliac crest bone grafts (7 x 7 x 10 mm) were harvested. Vascular implantation of the right superficial femoral vessels was performed in one of the two grafts, which was wrapped in a silicone envelope to avoid neovascularization from the surrounding tissues and positioned in a subcutaneous pocket in the right medial thigh. On the left side, the bone block, wrapped in the silicone envelope, was buried subcutaneously without vascular implantation. The operated animals were divided into two groups: Group I included eight rabbits explanted 4 weeks postoperatively and Group II included eight rabbits explanted 8 weeks postoperatively. Tetracycline injection was performed 72 hours preexplantation to evaluate new bone formation. Selective colloidal ink injection in the axial artery was performed to investigate the neovascularization before inclusion in poly-methyl-methacrylate (PMMA). Histological examination was performed in all explanted specimens comparatively. Histological examination 8 weeks after surgery showed a marked neovascularization, with normal bone cells. Tetracycline labeling showed new bone formation with a normal pattern. In all nonvascularized specimens, no viable cells or neovascularization and no bone formation were found. The vascular implantation procedure can induce a good neovascularization with new bone formation in a small bone graft. The possibility of neovascularization induction by the simple vascular implantation procedure has several clinical implications in reconstructive surgery.

Animals↗

Effects of essential amino acids and lactose on bony fractures and defects in rabbits: a preliminary histomorphometric study.

An experimental study was performed in order to test the possibility of improving bone repair with the administration of a drug (Calciofix, Farmaceutici Damor SpA, Naples, Italy) containing essential amino acids and lactose. Fifty rabbits were submitted to an open transversal fracture of the left fibula and to a right femoral condyle defect. They were left untreated or treated daily with the drug orally and were divided into subgroups depending on the experimental time: 15, 30, 40, 50, 60 days. Histomorphometric data showed a significantly faster healing rate in treated animals compared with untreated ones. Firstly, on day 30 there was a significantly larger amount of cartilage in the control bone callus (P < 0.01). On day 50 a significant difference existed between trabecular and lacunar percentages in the two subgroups (P < 0.0005). At 60 days no significant differences were observed, but bony trabeculae had become more oriented parallel to the long axis of the bone in treated animals. Secondly, after 15 days the defect area was significantly smaller in treated animals than in the untreated ones (P < 0.01). At 30 and 40 days, respectively, significant differences existed between the two subgroups in connective tissue and mature bone percentages (P < 0.01 and P < 0.001). Our results seem to demonstrate that the drug significantly accelerates the rate of bone formation in fractures and bone defects in rabbits.

Amino Acids↗

Histomorphometric study of bone reactions during orthodontic tooth movement in rats.

The biological response to orthodontic tooth movement has generally focused on reactions within the periodontal ligament (PDL), whereas less attention has been paid to the behavior of neighboring bone. The purpose of the study was to describe the influence of orthodontic force on bone surrounding the displaced tooth and the adjacent, untreated teeth. Bone changes in relation to treatment time and different sites were investigated. A mesial tipping of the left maxillary first molar was obtained from 54 adult male Wistar rats. Oxytetracycline was injected subcutaneously 48 h before killing, which took place after 4, 7, or 14 days. The maxilla was fixed in paraformaldehyde and embedded undecalcified in methylmethacrylate. A set of thick horizontal sections was taken from the cervical, intermediate, and apical levels of the roots. The sections were microradiographed and analyzed microscopically under bright-field and fluorescent illumination. Bone fraction and PDL width was measured using a Zeiss Videoplan device equipped with an overlay system. New bone formation was detected by oxytetracycline labels. The analysis showed a consistent, significant decrease of the alveolar bone fraction around both displaced and adjacent teeth at all treatment times. Apposition, indicated by the tetracycline uptake, was found on the periosteal side of the treated hemimaxilla and, after 14 days, also on the surface toward which the tooth was moving and around the adjacent teeth. These results suggest that a time rather than a space relationship exists between bone resorption and formation and that the whole hemimaxilla reacts to the mechanical challenge, resembling the regional acceleratory phenomenon (RAP) observed in other circumstances.

Adaptation, Physiological↗

Guided bone regeneration using titanium grids: report of 10 cases.

In order to ensure an adequate space where new bone can be formed in guided bone regeneration (GBR), most surgeons fill bone defects with biomaterials. In this work we evaluated new bone regeneration in 10 patients using only a blood clot protected with titanium grids and non-resorbable membranes, without any filling material. A manual measurement of the size of the bone defect, using a plastic probe, was performed at 2 surgical steps. After 5 months of treatment, a biopsy was taken from each patient, fixed and embedded in PMMA, examined microradiographically and morphologically to evaluate the newly-formed bone. Our results showed a good repair of the defects by bone regeneration (about 85% overall), high mineral density of new bone around the implants after 5 months, and steady state deposition processes. These results in GBR, without filling material, appear very promising for implantology and reconstructive odontostomatology practice.

Adult↗

Induction and pharmacological treatment of oral osteopenia in rats.

BACKGROUND: Osteoporosis is a major public health problem, responsible for a great number of fractures, associated with devastating costs to society. In addition, oral bone loss has an enormous impact on the health quality of life of patients, affecting up to 90% of elderly individuals. The aim of this work was to elaborate an animal model of mandibular and maxillary osteoporosis in which to evaluate bone loss and possible prevention by pharmacological treatment. METHODS: Six Sprague-Dawley rats were gonadectomized and treated with clodronate (male) or 17 beta-estradiol (female) for two months. Six gonadectomized and six sham-operated rats of both sexes were treated with placebo. The mandible and maxilla, fixed and methacrylate embedded, were serially sectioned, microradiographed and processed for histomorphometric analysis. RESULTS: Gonadectomy did not modify the amount of compact and trabecular bone in mandibles of rats of either sex, treated or not with clodronate or estrogens, compared to sham-operated rats. Compared to sham-operated rats, a 10-25% increase of bone porosity was found in the maxilla of ovariectomized rats, either receiving estrogens or not, while in male rats no difference among groups could be evidenced. CONCLUSIONS: The conclusion is drawn that rats, due to their peculiar masticatory habits yielding huge loads on oral bones, do not represent a suitable experimental model for studying oral bone loss related to skeletal osteoporosis. In order to worsen oral osteopenia it would be mandatory to combine gonadectomy with a mechanical unloading (i.e. after molar extraction) of mandibular or maxillary bone.

Animals↗

Morphology of peri-implant regenerated bone, in sheep's tibia, by means of guided tissue regeneration.

BACKGROUND: No data on the resistance to functional loads of bone regenerated by means of guided tissue regeneration, can be found. In this paper the new-formed bone is analyzed, in cases that see the surgical creation of defects in testing animals. The morphological aspects of perimplant bone, regenerated by means of bone substitutes or not, were evaluated. METHODS: A coronal defect was created by means of a counter-sink bur. The following step provided for osseointegrated fixtures, with a machined or blasted (TiOblast) surface, to be installed. Some bone defects were filled with Bio-Oss, and covered with resorbable membrane (Vycril), others were left unfilled. The animals were sacrificed after different periods of time (24 and 45 days). Some thick sections (200 microns) underwent then microradiography and were examined with a microscope, under transmitted and polarized light. Other sections, thin (5 microns), were coloured with toluidine blue, Gomori's Stain and Solochrome Cyanine/Congo red. RESULTS: Data confirm the excellent behaviour of bovine bone and of TiO blasted fixtures. The experimental results bring into evidence that, in cases of Guided Bone Regeneration (GBR) by means of membrane, the use of fixtures with a blasted surface (TiOblast), and of Bio-Oss as bone substitute, made osseointegration easier. CONCLUSIONS: Results, moreover, suggest that in case of implant surgery with GBR, in order to allow the maturation of regenerated bone, it's healing time must be prolonged.

Animals↗

Immunohistochemical detection of cell-cycle associated markers on paraffin embedded and formalin fixed needle biopsies of prostate cancer: correlation of p120 protein expression with AgNOR, PCNA/cyclin, Ki-67/MIB1 proliferation-scores and Gleason gradings.

Paraffin embedded and formalin fixed needle biopsies of prostate cancer (PC) were used to immunocytochemically detect the p120 nucleolar protein in relation to the Gleason histological gradings (GHG), the labelling indices of proliferating nuclear immunocytochemical markers (PCNA/Cyclin, Ki-67/MIB1) and the argyrophilic nucleolar region (AgNOR) rate. The twenty-six cases of PC (6 from large histological samples and 20 from needle biopsies) were equally distributed into low (< or = 6) or high (> or = 7) GHG groups. The p120 nucleolar protein immunocytochemical reaction was randomly expressed in large histological sections but uniformly distributed without gaps in needle biopsy sections. Only on the latter were quantitative values of PCNA/Cyclin (23.2 in low and 45.3 in high GHG), Ki-67/MIB1 (13.8 in low and 43.3 in high GHG) and AgNOR (5.0 in low and 7.5 in high GHG) related to those of p120 nucleolar protein (0.8 in low and 3.8 in high GHG). The values of all these cell cycle markers increased from low to high GHG of PC, all four reaching high statistical significance between the two groups (ANOVA-two tailed p < 0.0001). The PCNA/Cyclin index showed a higher positivity than the Ki-67/MIB1 index in PC with low GHG but not in PC with high GHG. In conclusion, paraffin embedded and formalin fixed PC needle biopsies exhibit a higher diagnostic PCNA/Cyclin than Ki-67/MIB1 index for cases presenting differentiated features, whereas p120 nucleolar protein detection seems to be a suitable marker of poorer outcome of PC.

Aged↗

A reabsorbable tubular chamber for the treatment of large diaphyseal bone defects. Experimental study in rabbits.

Twenty-five rabbits were used to test the healing of diaphyseal defects by means of a reabsorbable tubular chamber. In 20 animals a 10 mm gap was created in the left radius and bridged with a poly-L-lactide chamber (experimental side). The same defect was created on the right radius, remaining untreated (control side). In the remaining 5 animals, the right radius was left intact and they were used only for biomechanical tests at 12 months. X-rays and histomorphometry showed no bone repair at the control side while the inner surface of the chamber was progressively lined by neoformed bone (75% of the normal cortical thickness). Biomechanical data demonstrated that elastic modulus and ultimate stress were not significantly different in both experimental and intact radii. The chamber, by preventing the passage of soft tissues and creating a close space for intrinsic repair factors, seems to be a suitable device for the repair of long bone defects.

Analysis of Variance↗

Morphostructural relationships between bone and implant: comparative analyses by optical microscopy and microradiography.

This study assessed the relationship between the surfaces of a hollow cylindrical titanium implant and the consequent bone organization around it. A human bone maxillary specimen was taken that enclosed an implant which had lasted 28 months under prosthetic loading. The implant was removed because of fracture that prevented the implant from being used as a supporting element for new prosthetic aims. The bone block-section was fixed and embedded, several sections were obtained, and each section was analyzed using three different histologic analyses: optical microscopy under ordinary light and polarized light, and microradiography. Comparative analyses were required on every single section to provide a complete morphostructural analysis of the peri-implant bone. Data retrieved by this research showed the presence of woven bone in the bone layer facing the external implant surface several months after its surgical fitting and functionalization. This finding demonstrates the importance of woven bone, first as holding sheath in the immediate postsurgical stage, and second as a mold for the subsequent centripetal accrementition of secondary lamellar bone.

Dental Implantation, Endosseous↗

[Osteo-conductive materials: animal experiments and instrumental analysis II].

BACKGROUND: In daily dental surgery practice, when it is not possible to obtain-enough autogenous bone graft to fill bone defects, bone substitutes can be useful for bone regeneration. MATERIALS AND METHODS: After a preliminary microscopic examination of histologic samples obtained from semithin sections of the tibias of rats showing no major differences in the results both for the single material and implant time, further studies on five different osteoconductive materials used in oral clinical practice were carried out with microradiography, SEM and X-ray microanalysis. In experimental animals (rats) bone defects 3 mm large were surgically induced in both tibias: one of the defects was filled by bone substitutes and on the other side the bone defect healed naturally. RESULTS AND CONCLUSIONS: The findings obtained were studied in depth, and allowed to conclude that the materials tested are suitable for the purpose for which they are marketed. Non-demineralized bovine bone showed the best results with a high degree of osteointegration. However, the results also show that not all the materials tested offer healing advantages over controls.

Animals↗