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Biomedical subjects

G Giavaresi

Publications and source records attributed to G Giavaresi.

At least 19 recordsLinked to original sources

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

Laser Doppler evaluation of microcirculation behaviour during an ischaemia-reperfusion injury.

Laser Doppler flowmetry (LDF) was tested in an experimental ischaemic model on rat limbs to evaluate the degree of ischaemia and to find a possible correlation with values obtained with this device and prognosis. Under general anaesthesia, 40 Wistar rats were submitted to 4 h and 30 min of ischaemia of the left hind limb. Ten rats formed the control group (group 1). Two enzymes, native superoxide dismutase (SOD) and SOD modified with polyethylene glycol, were employed in 15 rats each (groups 2 and 3). Data were collected by means of LDF both in the sole and muscles before ischaemia (steady state), during ischaemia and at the beginning of reperfusion, and only in the sole after 1 h of reperfusion. A range of predictive (95%) perfusion values (PU) for limb healing or necrosis was identified at the beginning of reperfusion. During ischaemia, PU changed from 0 to 10, both in the sole and in the muscle. A three-factor ANOVA (site, group, time) did not show interaction of these factors with PU (F = 1.655; p = 0.195), even if every single effect was significant (p < 0.0005). A two-factor ANOVA (group, time) showed a significant interaction of these factors with PU (F = 4.079; p = 0.019). The logistic regression between the reperfusion PU of each site and the survival of the limb was observed at the beginning and after 1 h of reperfusion in the sole only. Furthermore, a correlation between sole and muscle PU at the steady state and at the beginning of the reperfusion period was observed. The results showed the effectiveness of LDF, which can be considered a quite reliable tool to evaluate the degree of ischaemia and to have a good correlation with prognosis in this kind of experiments.

Animals

In vitro osteoinduction of demineralized bone.

Among numerous available materials for osseous repair and reconstruction, those presenting osteoinductive characteristics and promoting bone regeneration are preferable. Fresh autologous bone is one of the most effective, but it has some disadvantages and risks. Demineralized bone matrix (DBM) is considered to be a valid alternative, because it seems to show osteogenic potential, ascribed to the presence of bone morphogenetic proteins. In addition it can be prepared without difficulty and preserved without losing osteoinductive properties. The aim of the study was to evaluate the osteoinductive ability of xenogenic DBM, by testing DBM powder obtained from rabbit long bones, in cell culture of murine fibroblasts, alone or associated with electromagnetic field (EMF), that are known to exhibit biologic effects on cells: in particular they are used in orthopedics to improve bone formation. At the end of experiment, alkaline phosphatase (ALP) activity, calcium levels and cell proliferation and morphology were evaluated. A statistically significant stimulation of ALP activity and cell proliferation and a morphological change of fibroblasts were found. The results obtained show how DBM and EMF have different effects on cells, and that together they have synergic action toward bone induction.

Animals

pO2 measurement in an experimental model of patellar tendon autograft pro-anterior cruciate ligament.

Thirty-four sheep were submitted to surgery substituting the native ACL with the central third of the patellar tendon, ten enter this study. The purpose was to find a possible relationship between tissue pO2 and healing processes considering also the biomechanical and histomorphological aspects of the grafts. Four of them were sacrificed under general anaesthesia after 6 months, and six after 1 year in order to perform tissue pO2 measurement and an analysis of microvessel density on specimens of the normal ACL and the graft. Our data showed higher pO2 values of the autografts after 6 months. After 1 year the data was comparable to those of native ACL. This was confirmed by a microvessel count of the histological specimens and the data was in relationship to biomechanical and histomorphological analysis. Tissue pO2 can be observed and recorded in "in vivo" ACL, and patellar tendon used as graft, with no injury to their integrity. The monitoring system might be considered as an experimental tool for indirect controls of the anterior cruciate substitutes.

Animals

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

Experimental evaluation of a resorbable intramedullary plug for cemented total hip replacement.

In order to evaluate degradation kinetics and biocompatibility of a resorbable poly(D,L-lactic acid) (PDLLA) plug for total cemented hip prostheses, an experimental in vitro and in vivo study was carried out. Degradation rate studies were performed in Ringer solution and after in vivo plug implantation in the femoral medullar cavity of rabbits. In vitro biocompatibility was evaluated in murine fibroblast cell cultures. PDLLA plugs showed faster degradation kinetics in vivo than in vitro. Histological evaluations showed that PDLLA completely disappeared in vivo 26 weeks after implantation. Fibrous tissue in the medullar cavity was observed at 13 weeks, but no histological changes were observed after 26 weeks. Also, the in vitro tests showed good biocompatibility of the biomaterial. Our results show the possibility of considering this resorbable plug for clinical situations instead of the traditionally used plugs [polyethylene, poly(methyl methacrylate) or cancellous bone] due to its biocompatibility, degradation properties and simplicity of use.

Animals

Biomaterials for orthopedic surgery in osteoporotic bone: a comparative study in osteopenic rats.

UNLABELLED: To evaluate orthopedic devices in pathological bone, an experimental study was performed by implanting Titanium (Ti) and Hydroxyapatite (HA) rods in normal and osteopenic bone. Twenty-four rats were used: 12 were left intact ( CONTROL: C) while the other 12 were ovariectomized (OVX). After 4 months all the animals were submitted to the implant of Ti or HA in the left femoral condyle (Ti-C, HA-C, Ti-OVX, HA-OVX). Two months later the animals were sacrificed for histomorphometric, ultrastructural and microanalytic studies. Our results show a significant difference between the Affinity Index (A.I.) of HA-C and Ti-C (77.0 +/- 7.4 vs 61.2 +/- 9.7) (p < 0.05). No significant differences were observed between the osteointegration of Ti-C and Ti-OVX (61.2 +/- 9.7 vs 48.2 +/- 6.7). Significant differences also exist between the osteointegration of HA-C and HA-OVX (77.0 +/- 7.4 vs 57.6 +/- 11.5) (p < 0.01). Microanalysis shows some modifications in Sulphur (S) concentration at the bone/biomaterial interface of the Ti-OVX group. Therefore our results confirmed the importance of biomaterials characteristics and of bone quality in osteointegration processes.

Animals

Bioabsorbable polyphosphazene matrices as systems for calcitonin controlled release.

To provide a suitable delivery system for the calcitonin controlled release 80 mg bioabsorbable polyphosphazene matrices were obtained with entrapped 50 or 250 micrograms calcitonin. The in vitro behaviour demonstrated a release burst for about 24 hours, followed by a period of slow release of the peptide lasting for weeks. Matrices containing 250 micrograms calcitonin were implanted under general anaesthesia in osteoporotic female rats, while a group of animals (control group) received unloaded matrices. After thirty days a second batch of matrices was implanted in both groups to prolong the period of treatment until two months. The explanted matrices were histologically evaluated together with the surrounding tissues, and the dosage of the residual calcitonin was also performed. Results demonstrated the good biocompatibility of the system and the complete release of the calcitonin from the matrices 30 days after implantation. The therapeutic effect, after sixty days of treatment was confirmed by the better densitometric values observed in the femoral bone of treated animals than in controls.

Animals

Pericellular pO2 as an alternative method to test cytotoxicity.

Pericellular pO2 (ppcO2) was compared with the release of cytoplasmatic enzyme lactate dehydrogenase (LDH) and mitochondrial metabolic function (tetrazolium salt reduction, MTT) as an alternative method to evaluate cytotoxicity. L-929 cells were seeded and incubated with 3 different control materials to test their cytotoxicity effects by these methods. High density polyethylene and copper were respectively used as negative and positive controls, while a 0.9% NaCl solution was as reagent control and extraction vehicle. PpcO2 was measured by a rod polarographic Clark-type probe connected to Licox pO2 computer (GMS mbH, Kiel, Germany). One-way ANOVA test showed significant differences among groups regarding every methods (p < 0.0005). The comparison of the mean variation coefficients of three methods showed no significant differences. In addition, a significant correlation was found between ppcO2 and MTT (r = 0.621; p < 0.001) as well as ppcO2 and LDH data (r = 0.474; p < 0.001). In conclusion, ppcO2 may be considered a reliable, safely and alternative method to test cytotoxicity.

Animals

[The osseointegration process: a still open chapter in experimental and clinical research].

The osseointegration of biomaterials for dental and orthopaedic use is a still open topic in experimental and clinical research. Some aspects of this complex mechanism are not completely research. Some aspects of this complex mechanism are not completely explained yet. These reasons make the process of osseointegration a topical subject both in clinical and experimental view points. The authors describe how this complex event has not been studied with standard investigation and report also most of present research lines on the basis of their experience. A particular attention is paid to some factors related to both biomaterial and implant site, which can influence significantly the process of osseointegration.

Animals

[Experimental osteoporosis caused by estrogen deficiency in rats. Role of a lactose, L-arginine and L-lysine combination in the prevention and therapy].

The role of a lactose, L-arginine and L-lysine association in the treatment of osteoporosis was evaluated in an in vivo model. Fifty Sprague Dawley female rats, 10 months age, were divided into 5 groups: 10 non-ovariectomized animals sacrificed at the beginning of the study (I group); 10 ovariectomized animals (OVX) without treatment and sacrificed at three months (II group); 10 OVX animals without treatment sacrificed at 6 months; 10 OVX animals treated starting from the I post-operative day with the association for three months (IV group, prevention) and 10 OVX animals treated like those of the IV group starting three months after surgery (V group, therapy). Animals were sacrificed in general anesthesia according to the scheduled times for radiologic, densitometric, histologic and mineralogic investigations. Preliminary results, which will need further researches, seem to show a progressive reduction of bone mass after ovariectomy in 10 months female rat and that the pharmacological association given as therapy (V group) may be able to lessen the reduction of bone mineral content.

Animals

Pathophysiological and histomorphological evaluation of polyacryloylmorpholine vs polyethylene glycol modified superoxide dismutase in a rat model of ischaemia/reperfusion injury.

Twenty Wistar rats were divided into two groups. Both underwent acute ischaemia followed by reperfusion of the left hind limb. The first group was a control group while the second was treated with PAcM-SOD. The survival percentage of the limb after 10 days was 30% for the first group and 70% for the second. Neither linear regression nor correlation were found between groups as far as the survival percentage of the limb after 10 days and reperfusion pmO2 data were concerned. After ten days the histomorphological analysis was significant regarding the fibre diameter and the percentage of central located nuclei in the specimens of PAcM-SOD treated limbs compared to normal limbs, but not when compared to the muscular fibres of the control group. Comparing these results with others obtained with native SOD and monomethoxypoly(ethylene glycol) modified SOD (mPEG-SOD) used in the same experimental model, we can conclude that the clinical and morphological results were better using mPEG-SOD, and that PAcM-SOD does have a protective effect on ischaemic muscle damage, although it is not as effective as mPEG-SOD in preventing ischaemia/reperfusion injury.

Acrylic Resins

Comparison of hemodialysis versus hemoperfusion in the clearance of high-dose methotrexate in pigs.

To evaluate the different capabilities of hemo-perfusion (HP) and hemodialysis (HD) treatment in resolving the side effects during high-dose methotrexate (MTX) therapy, an experimental comparative study was done. Under general anesthesia, 12 female Large-White pigs, 35 +/- 5 kg body weight, underwent a bilateral renal vessel ligature to create an acute renal failure and avoid the physiologic elimination of MTX. An MTX i.v. infusion was performed using a dose of 12.5 g/m2 and for a specific time according to its pharmacokinetics. The animals were divided into 2 groups of 6 animals each and connected to the extracorporeal circuit; the first group was submitted to a 1-h HP with an anion exchange resin (Dow 1X-2, Dow Chemical) and the second group to 1-h HD with capillary polysulfone fibers (F6, Fresenius). Blood samples were taken at established intervals for hematological and biochemical evaluations. The results demonstrate a higher capability of HP treatment (P < 0.001) related to HD in MTX removal.

Animals

In vitro evaluation of heparin adsorption during haemoperfusion with Dowex 1 x 2 anion exchange resin.

Haemoperfusion treatments have found clinical applications in various pathologies, even though problems due to the insufficient biocompatibility of sorbents have often limited its employment. Among these problems, there is that of heparin adsorption which can determine coagulation disorders or the necessity of a dosage correction during the extra corporeal circulation. The aim of the study was to test the influence of different methods of coating on heparin adsorption from an anion exchange resin coated with poly-hydroxyethyl-methacrylate. Eight haemoperfusion cartridges were prepared using 8 g of resin for each (4 cartridges contained resin coated by spray-coating technique and 4 coated by phase-separation one). Haemoperfusion was performed by using 600 IU of heparin (25,000 IU) corresponding to 0.12 ml in 500 ml of swine blood and a closed circuit. Blood flow rate was maintained at 15 ml/min and the circuit pressure was 70 torr during haemoperfusion. Blood samples were withdrawn at timed intervals before and after the column in order to evaluate blood heparin levels. The results demonstrated the superiority of the phase-separation technique that does not permit any interactions between adsorbent substance and drug. As a result of this, the adjustment of the dosage may not be necessary during further in vivo experiments.

Adsorption

Biopolymeric modification of superoxide dismutase (mPEG-SOD) to prevent muscular ischemia-reperfusion damage.

The efficacy of preventing ischemia-reperfusion damage by employing native or modified (mPEG-SOD) superoxide dismutase in an experimental animal model of acute ischemia of the left hindlimb was tested. Four hours and thirty minutes complete warm ischemia was induced in the left hindlimb of 43 Wistar rats, by clamping the femoral artery and monitoring its efficacy with Laser Doppler flowmetry. After ten days, a significative difference (p = 0.004) of the survival leg rate was found in the group treated with mPEG-SOD (86.6%) compared with the control group (30%). Histomorphological and ultrastructural analysis were performed at different time intervals confirming what the clinical course had already pointed out. These results show that SOD in its modified form, despite the lower dosage, can provide good protection against ischemia/reperfusion injury of muscles.

Animals

[In vivo experimental research in vascular surgery. Methodology and current Italian law].

Authors underscore the importance of research in vascular surgery progress and approach some basic aspects on the correct conduction of an experimental research in vascular surgery fields and in particular on synthetic prosthesis of different diameter. Some important aspects of the current Italian law on animal experimentation are shown because they could cause new changes either on methodology or animal selection. Finally some experimental models adopted in the department of experimental surgery of the Rizzoli Orthopaedic Institutes on vascular prosthesis are described.

Animal Welfare

Biological and synthetic conduits in peripheral nerve repair: a comparative experimental study.

Two different types of conduits, one biological, obtained with homologous glutaraldehyde preserved vein segments and the other synthetic bioabsorbable, made with Poly [L-lactide-co-6-caprolactone], were evaluated as guides for nerve repair in alternative to autologous grafts in an experimental animal model. Under general anesthetic, the ischiatic nerve of a number Wistar rats was transected to create a 1 cm gap, which was then repaired by means of the conduits or autologous grafts. Controls were performed at 1, 3 and 6 months; nerve regeneration was effective with both conduits, but the count of myelinated axons showed a significant difference between the synthetic and biological tubes (p < 0.001). The Poly [L-lactide-co-6-caprolactone] guide was still intact 30 days after implant; progressive signs of degradation were present at 90 and 180 days. These results show that the synthetic conduits are better than those obtained with preserved vein segments and might be considered in alternative to autologous grafts in peripheral nerve reconstruction.

Animals