Comparison of calcitonin, alendronate and fluorophosphate effects on ovariectomized rat bone.

The effects of calcitonin, alendronate and fluorophosphate preventive treatment on ovariectomized rat femur were studied by comparing densitometric, mechanical, mineralogical and histomorphometric data. Sixty retired breeder female Sprague-Dawley rats, aged 10 months, were randomly divided into six groups. A group (baseline) was euthanized at the beginning of the study as a baseline group; four groups were ovariectomized and one was sham-operated (sham) and considered as a sham-aged group. A group of ovariectomized rats was used as a sham-therapy control (OVX) and received only deionized drinking water, while the other three received: a) salmon calcitonin (SCN) at a dose of 2 IU/kg/d s.c. (OVX + SCN); b) alendronate sodium salt (ALN) at a dose of 6 microg/kg/d administered by gavage (OVX + ALN); and c) L-glutamine monofluorophosphate (G-MFP) and calcium at a rate of 1:30 F/Ca at a dose of 0.21 mg F/6.30 mg Ca per kg/d by gavage (OVX + MFP). Significant increases (P < 0.05) of about 15 and 27% in femoral proximal epiphysis bone mineral density (BMD) of the OVX + ALN group were observed versus healthy groups and the OVX group, respectively. The OVX + ALN group also showed significant increases in femoral mid-diaphysis BMD when compared to OVX (18%, P < 0.001), OVX + SCN (14%, P < 0.05) and OVX + MFP (18%, P < 0.001) groups. In the OVX + MFP group, the three-point bending test demonstrated significant increases (P < 0.05) in maximal load of 21 and 22% when compared to the OVX and OVX + SCN groups, respectively. Also, stiffness data showed significant increases of the OVX + MFP (17%) and sham (14%) groups in comparison with the OVX group. A decrease in Mg (42%, P < 0.05), and increases in Ca (15%, P < 0.0001) and PO4 (8%, P < 0.005) content were found by comparing OVX + MFP and OVX groups. Trabecular bone volume results showed significant increases by comparing OVX + ALN and OVX groups (12.20%, P < 0.0005), as well as control groups. Tested agents were able to reduce the bone loss due to estrogen deficiency, but this did not always produce an increase in strength of the treated bone. Alendronate treatment prevented a decrease in bone mineral density and maintained bone mechanical properties after ovariectomy without impairment of bone mineralization in aged rats.

Calcitonin release system in the treatment of experimental osteoporosis. Histomorphometric evaluation.

Bioabsorbable polyphosphazene matrices charged with 250 microg calcitonin were evaluated as calcitonin delivery systems for the controlled release of this drug in an in vivo experimental model. Matrices were implanted under general anesthesia in osteopenic female rats, while a group of osteopenic animals (control group) received unloaded matrices. After 30 days a second series of matrices, loaded and unloaded was, respectively, implanted in both groups. In a third group of osteopenic female rats (positive control), 10 IU calcitonin were injected daily for two months. A baseline group and a sham operated group of animals were also included in the study. The explanted matrices were histologically evaluated together with the surrounding tissues, and bone histomorphometry was performed on undecalcified sections from femurs. The results showed the good biocompatibility of the matrices. Bone histomorphometry revealed that bone architecture in the treatment group was improved and results were not different from those observed after calcitonin injection. These data confirm a good in vivo behavior of the system, as well as a therapeutic effect of the released calcitonin in the osteopenic condition.

Transplantation of chondrocytes seeded on a hyaluronan derivative (hyaff-11) into cartilage defects in rabbits.

Different methods have been used to improve chondrocyte transplantation for the repair of articular cartilage defects. Several groups of biomaterials have been proposed as support for in vitro cell growth and for in vivo implantation. Here. we describe a new approach investigating the healing of rabbit cartilage by means of autologous chondrocytes seeded on a hyaluronan derivative referred to as Hyaff-11. Full thickness defects were created bilaterally in the weight-bearing surface of the medial femoral condyle of both femora of New Zealand male rabbits. The wounds were then repaired using both chondrocytes seeded on the biomaterial and biomaterial alone. Controls were similarly treated but received either no treatment or implants of the delivery substance. Histologic samples from in and around the defect sites were examined 1, 3 and 6 months after surgery and were scored from 0 to 16. Statistically significant differences in the quality of the regenerated tissue were found between the grafts carried out with biomaterial carrying chondrocyte cells compared to the biomaterial alone or controls. This study demonstrates the efficacy of this hyaluronan-based scaffold for autologous chondrocytes transplantation.

[Words and action of medical instruction: surgical lectures of Antonio Scarpa]. / Parola e gesto nell'insegnamento medico: le lezioni chirurgiche di Antonio Scarpa.

Antonio Scarpa (1752-1832) was chairman of anatomy and surgery at the university of Pavia for thirty years since 1783. The study of the manuscripts containing his surgical lectures, collected by his pupils, is the basis for the knowledge of the method of teaching in its twofold dimension of the theoretical matter and of the practical demonstration. Starting from the analysis of one of these manuscripts, entitled "Lezioni di operazioni chirurgiche del Cav. Prof. Antonio Scarpa" the value of these collections of notes is discussed and their significance as a textbook for the students is emphasized. The survey of every single lecture demonstrates that the theoretical exposition immediately preceded the operative procedure on the corpse. Finally the qualities expected by Scarpa in the teachers and the students for a satisfactory education in surgery are considered. Fulfilling his mastership Scarpa struggled to convey to the pupils the method learned when he himself was a student. He was therefore considered the head of the anatomy and surgery school in Pavia.

rhVEGF and experimental rat skin flaps: systemic or local administration and morphological characteristics.

Skin flap survival is a significant problem in skin surgery; in particular, inadequate arterial or venous blood supply results in necrosis of the distalmost portion. The aim of this study was to evaluate the ability of Vascular Endothelial Growth Factor (VEGF) of modifying the morphological features of skin flaps. Bilateral epigastric skin flaps were raised in 16 Wistar male rats. The epigastric artery and vein of the left flaps were clamped and then injected with rhVEGF (8 rats) or saline (8 rats). The right flaps were not clamped and received rhVEGF or saline systemically. The rats were euthanized on the seventh day and flap skin samples collected. Tissue fragments were subject to immunohistochemical (rhVEGF, VEGFr, VIII factor, CD34 antibodies), ultrastructural and morphostructural investigations. The results showed that rhVEGF improved the condition of flaps and that systemic administration was effective in promoting the development of an adequate vascular network.

Discriminant capacity of quantitative ultrasound versus dual X-ray absorptiometry to determine cancellous bone loss in ovariectomized rats.

The capacity of dual x-ray absorptiometry and quantitative ultrasound to discriminate bone loss and to predict the mechanical and microarchitectural properties of cancellous bone in an animal model of osteopenia was evaluated. Thirty-five female Sprague-Dawley rats (10 months old) were randomized into three groups: baseline group, 10 rats killed at the beginning of the study; ovx group, 15 rats ovariectomized; and sham group, 10 rats sham operated. At the beginning and end of the study, all the animals underwent osteosonography to record the proximal tail (C3 vertebra) bone speed of sound. Sixteen weeks after surgery, the animals were euthanized and the L5-6 lumbar vertebrae of each rat were excised for densitometric, biomechanical (compression test), and histomorphometric studies. Significant differences were found among the groups for final speed of sound (p = 0.01). The L5 bone mineral density of the ovx group decreased by 12.1% (p = 0.049) and 12.6% (p = 0.035) compared, respectively, with baseline and sham groups. The biomechanical parameters of the ovx group decreased by 15-47% compared with the other groups, showing significant differences between the ovx and sham groups both for maximal stress (p = 0.026) and elastic modulus (p = 0.013). Histomorphometric parameters of the ovx group showed significant decreases in comparison with other groups. Logistic regression analysis showed that dual X-ray absorptiometry and quantitative ultrasound discriminate ovariectomized and healthy rats with a similar capacity, classifying correctly all rats used in the model in a range of 61-70%. This similar capacity seems to derive from two different capacities to detect bone changes. Dual X-ray absorptiometry, depending on bone mineralization and density, is able to detect modifications in bone stiffness and strength, confirmed also by the correlation with biomechanical data. On the contrary, quantitative ultrasound seems to depend more on cancellous bone microarchitecural changes because it is correlated to histomorphometric parameters.

The effect of osteopenia on the osteointegration of different biomaterials: histomorphometric study in rats.

The osteointegration of Hydroxyapatite (HA), Titanium (Ti-6Al-4V: Ti), Zirconia (ZrO2), Alumina (Al2O3) and 2 biological glasses (AP40 and RKKP) was comparatively investigated in normal and osteopenic rats by means of histomorphometry. Thirty-six Sprague Dawley female rats were left intact (Group C) while 36 were ovariectomized (Group OVX). Group C and OVX were further divided into 6 subgroups. After 16 weeks all animals were submitted to the femoral implant of nails made of the above-mentioned materials. Eight weeks after implantation the animals were euthanized, the femurs were harvested for histomorphometric analysis. The data showed that: (1) all the tested materials were biocompatible in vitro; (2) no significant differences existed in Affinity Index (AI) of Group C; and (3) results from paired comparison applied to the AI showed significant differences among the Groups C and OVX. The AI did not significantly change among intact groups, while it significantly decreased when some materials were implanted in OVX subgroups (AP40, ZrO2 and Ti-6Al-4V: p < 0.0005, p < 0.05 and p < 0.01). It is confirmed that bone mineral density is a strong predictor of the osteointegration of an orthopedic implant and that the use of pathological animal models is necessary to completely characterize biomaterials.

Bone response to orthodontic loading of endosseous implants in the rabbit calvaria: early continuous distalizing forces.

The purpose of this experimental study was to evaluate the effect of early orthodontic loading on the stability and bone-implant interface of titanium implants in a rabbit model. Twenty-four short threaded titanium fixtures were inserted in the calvarial mid-sagittal suture of 10 rabbits. Two weeks following insertion, 20 implants (test group) were subjected to continuous distalization forces of 150 g for a period of 8 weeks. The remaining four implants (control group) were left unloaded for the same follow-up interval. Clinically, all implants except for one test fixture were stable, and exhibited no mobility or displacement throughout the experimental loading period. Histologically, all stable implants were well-integrated into bone. No differences could be found between the pressure and tension surfaces of the test implants relative to bone quality and density within a range of 1000 microns from the fixture surface. Similarly, qualitative differences were not observed between the apical and coronal portions of test fixtures. Morphometrically, a mean percentage bone-to-implant contact of 76.00 +/- 18.73 per cent was found at the test pressure sides, 75.00 +/- 11.54 per cent at the test tension sides, and 68.00 +/- 15.55 per cent at the control unloaded surfaces. No statistically significant differences in the percentage of bone-to-metal contact length fraction were found between test pressure surfaces, test tension surfaces, and unloaded control surfaces. Marginal bone resorption around the implant collar or immediately beneath it was found in roughly the same percentage of analysed sites in the test and control fixtures. In contrast, slight bone apposition was demonstrated at the implant collar of the test pressure surfaces, while no apposition or resorption were observed in the test tension zones. This study suggests that short endosseous implants can be used as anchoring units for orthodontic tooth movement early in the post-insertion healing period.

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.

[Trauma of the peripheral nervous system: experimental assessments with guided tissue regeneration]. / Lesioni traumatiche del Sistema Nervoso Periferico: valutazioni sperimentali sulla rigenerazione tessutale guidata.

Guided tissue regeneration open interesting perspectives in reconstructive surgery of peripheral nerves. Artificial conduits for nerve repair can be obtained with biodegradable polymers. Lactic and caproic acid copolimers and poliphosphazenes are biocompatible materials and have a slow reabsorption rate. Two types of conduits obtained with Poli[L-lactide-co-6-caprolactone] and poli [bis (etilalanate) phosphazene] were evaluAted as guides for nerve regeneration in an experimental model on two groups of six Wistar rats. Under general anesthesia and with microsurgical technique, the ischiatic nerve was bilaterally isolated. On the right side a segment of the nerve was removed to create a 10 mm gap. The defect was then repaired using the conduit. On the controlateral limb after the creation of the same defect, the nerve continuity was restored using as an autograft the segment removed from the right side. Control were performed at 30, 90, 180 days and consisted in histological and electron microscopy investigations. They showed the gradual degradation of both the conduits without signs of local toxicity. The regeneration of the nerve fibers in the lumen was not significantly different from that observed in the autologous grafts. Both the conduits may be considered effective for guided nerve regeneration, but polyphosphazenes allow the possibility of use the polymer as a carrier for neurite-promoting factors.

Role of intramarrow penetration in osseous repair: a pilot study in the rabbit calvaria.

BACKGROUND: The use of intramarrow penetration in combination with osseous reconstructive surgery has been recommended in various periodontal and oral surgical applications. The purpose of this pilot study was to evaluate the effect of intramarrow penetration on the rate of bone neogenesis in protected spaces created on the calvarial bone using occlusive titanium domes in 16 adult white rabbits. METHODS: In each rabbit, 2 standardized circular 1 mm deep slits were prepared in the bone, one on each side of the midline using a trephine. Within the perimeter of one of the slits, the external cortical surface of the skull bone was mechanically perforated using a carbide round bur (experimental site), while the bone surface within the other slit (control site) was left intact. Subsequently, 2 prefabricated titanium domes with an inner diameter of 4.8 mm and an inner height of 3.0 mm were anchored in the prepared slits. The animals were divided into 4 groups of 4 rabbits each and were sacrificed at 10, 21, 42, and 60 days. The specimens were assessed for the percentage bone fill and density of the newly formed bone within the boundaries of the domes. RESULTS: Histomorphometric analysis showed significantly increased bone neogenesis within the experimental domes at 10, 21, 42, and 60 days. A mean total area of 29% newly formed bone was demonstrated in the experimental sites at 21 days, while negligible bone formation (9%) occurred with the control domes at the same evaluation interval. Bone density was consistently higher in experimental domes at all healing intervals. CONCLUSIONS: Within the limits of this study, the results demonstrate that intramarrow penetration accelerates initial bone neogenesis and results in increased bone fill and density, suggesting that its use can be beneficial in bone regenerative procedures.

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.

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.

Resorbable device for fracture fixation: in vivo degradation and mechanical behaviour.

The availability of a high molecular weight poly(L-lactide), with supposed good mechanical properties, encouraged us to carry out an in vivo experimental study to evaluate the degradation rate and accompanying loss of strength during interaction with bone. Small cylindrical PLLA rods were manufactured by compression moulding. Thirty-six male New Zealand rabbits (b.w. 3 kg), randomly divided into six groups of six animals each, were submitted to implantation of the rods under general anaesthesia (phi 3 x 7 mm rods in the lateral condylic region of the right femur and phi 3 x 50 mm rods in the intracondylic channel of the left femur). The animals were then sacrificed at the end of each experimental time (4, 8, 12, 24, 52, 64 weeks), hence the femurs were explanted. Histology showed no inflammatory cells with fragmentation or resorption of bone trabeculae. Mechanical studies showed a decrease in mechanical properties and flexional stiffness, and the absence of evident material resorption. Finally, the properties of this newly developed resorbable device, when confirmed, seem to justify future clinical introduction of this synthetic material.

[In vivo experimental research in vascular surgery. Methodology and current Italian law]. / La ricerca sperimentale in vivo in chirurgia vascolare. Metodologia ed attuale normativa italiana.

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.

The first Billroth II gastric resection as reported by an Italian medical journal.

A report on the first Billroth II surgical intervention for gastric cancer is entirely reprinted. The description is original and is taken from a letter published by a young Italian physician. Eugenio Casati, who was present in the operative theatre of the Second Surgical Clinic of the University of Vienna on January 15, 1885, while he was on a scientific journey in Europe. A biographical profile of the writer is provided. The origin of the surgical procedure for gastric resection known as Billroth II is also examined, and its significance in the early surgery of the alimentary tract is highlighted. A short account by Billroth's pupil, Viktor von Hacker, on the same intervention also allows comparison and a better knowledge of Billroth's surgical strategy in performing one of the starting points of modern gastric surgery.

Preparation of a new nerve guide from a poly(L-lactide-co-6-caprolactone).

A copolymer of L-lactide and 6-caprolactone (50:50, w/w) was synthesized and characterized. The thermal behaviour of this material did not show any crystallinity for several months; only after more than 1 yr of aging at room temperature and, particularly, in the in vitro degradation tests did it partially crystallize. The values of tensile strength, percent elongation at break and elastic modulus were, respectively, 25 MPa, 490% and 3 MPa. Transparent, elastic nerve guides having inner diameter of 1.3 mm and wall thickness of 175 microns were prepared.