Biomedical subjects

R Giardino

Publications and source records attributed to R Giardino.

At least 37 records · Page 2Linked to original sources

PubMed2005

Prosthetic devices shaped as tubular chambers for the treatment of large diaphyseal defects by guided bone regeneration.

Guided tissue regeneration is based on the hypothesis that the different tissues have unequal abilities to penetrate a wounded area during the healing process. The use of a device acting as a chamber allows the growth of a particular tissue and prevents the ingrowth of other tissues which impair the healing process. At the same time the chamber protects and maintains in situ the intrinsic growth factors so that they may perform their specific activity. Guided tissue regeneration currently plays a well-recognized role mostly in dentistry and peripheral nerve surgery but interesting perspectives have also opened up in orthopedics. Considering the possibility of using guided bone regeneration in the repair of diaphyseal bone defects, this updated survey highlights some critical points and pathways related to the state-of-the-art of this promising procedure, focusing particularly on the properties of the material to make the tubular chamber, the use of osteopromotive factors and the most appropriate animal model to be used for the experimental evaluation.

Animals↗

PubMed2005

Shock wave therapy as an innovative technology in skeletal disorders: study on transmembrane current in stimulated osteoblast-like cells.

Extracorporeal shock wave treatment (ESWT) is successfully used in various musculoskeletal disorders and pathologies. Despite the increasing use of this kind of therapy, some aspects of its mechanism of action are still unclear. In vitro bone cell behavior under ESWT were previously investigated by the present author and MG63 osteoblast-like cells showed an enhancement in proliferation and in the osteoblast differentiation after therapy with a low-energy flux density. The aim of the present study was to evaluate the effect of ESWT on the permeabilization of cell membrane. We characterized physiological changes in the MG63 associated with ESWT generated by an ESW device and patch clamp recording was performed to study ion channels. Experiments were carried out using the whole-cell recording configuration of the patch-clamp technique and the ionic current measurements were performed on cell samples of ESW treated and control groups. The patch-clamp technique showed the effect of ESWT on the amplitude of transmembrane currents. The treatment with ESW enhanced the transmembrane current as well the voltage dependence of Ca-activated and K channels that mediate these currents: the differences between treated cells and control at 80mV were over 1000 pA (p<0.05). These modifications of ion channels activity positively influence cell proliferation (MTT test, p<0.0001) without interfering with the normal synthesis activity of stimulated osteoblasts.

Cell Count↗

PubMed2005

Soft tissue response to a new austenitic stainless steel with a negligible nickel content.

This study evaluates the soft tissue response to a new austenitic stainless steel with a low nickel content (P558) in comparison with a conventional stainless steel (SSt)and a titanium alloy (Ti6Al4V). Previous findings showed its in vitro biocompatibility by culturing P558 with healthy and osteoporotic osteoblasts and its in vivo effectiveness as bone implant material. Regarding its use as a material in osteosynthesis,P558 biocompatibility when implanted in soft tissues, as subcutis and muscle, was assessed. Disks and rods of these metals were implanted in rat subcutis and in rabbit muscle, respectively. Four and twelve weeks post surgery implants with surrounding tissue were retrieved for histologic and histomorphometric analysis: fibrous capsule thickness and new vessel formation were measured. Around all implanted materials, light microscopy highlighted a reactive and fibrous capsule formation coupled with ongoing neoangiogenesis both in rats and in rabbits. Histomorphometric measurements revealed a stronger inflammatory response,in terms of capsule thickness,surrounding SSt implants (9.8% Ni content) both in rat subcutis and in rabbit muscle independently of shape and site of implantation. A progressive decrease in capsule thickness around P558 (<0.02% Ni content) and Ti6Al4V, respectively, was seen. Regarding new vessel density, the data showed a different response dependent on the site of implantation. However,in the light of the previous and present studies, P558 is a good material, instead of titanium alloys, in orthopedic research.

Animals↗

PubMed2005

Cell dynamics in the correct control of bone metabolism using natural treatments.

The present study was undertaken in order to assess the efficacy of a commercial product containing calcium and silicon (Osteosil-Calcium) on cell metabolism. MG-63 osteblast-like cells were cultured in the presence of three different drug concentrations (10, 5 and 2.5 microg/mL). Either serum-free culture and standard culture with serum were investigated. Morpho-functional tests (MTT and ALP), scanning electron microscopy (SEM), microanalysis (EDAX) and time-lapse video microscopy were performed. Cell actin cytoskeletal modification with fluorescence phalloidin staining was also tested. Our data show the in vitro functional efficacy of Osteosil-Calcium on MG63 cell viability and ALP production. This study demonstrates its positive effect on the metabolism of the single cell and suggests wider uses of this drug in health protection and or in Regenerative Medicine therapies which are currently applied to the elderly.

Analysis of Variance↗

PubMed2004

A new austenitic stainless steel with a negligible amount of nickel: an in vitro study in view of its clinical application in osteoporotic bone.

A biomaterial named P558 is a new austenitic stainless steel (SS) with a negligible amount of Ni (<0.20%). In previous in vitro and in vivo studies it was compared with conventional SS and Ti6Al4V and shown to be a promising material in orthopedics. Because osteoporosis is a type of pathology very often encountered in implanted patients and can be studied with in vitro models, the purpose of the present study was to evaluate P558 in vitro through comparison of normal (nOB) with osteopenic (oOB) bone-derived primary rat osteoblasts. Osteoblasts were cultured directly on P558 and polystyrene as controls for 72 h. Osteoblast proliferation, adhesion, and activity (ALP, OC, TGF-beta1, and IL-6) were evaluated at 24 and 72 h. Results demonstrated that the growth of nOB and oOB cultured on P558 was not affected negatively when compared to control. Cells on P558 did not show any alteration in terms of adhesion, proliferation, and metabolic marker production in nOB and oOB cultures, and a significant increase in ALP, OC, and TGF-beta1 production was observed. SEM images revealed no alteration in cell morphology. The current findings demonstrate that P558 promotes osteoblast proliferation, activation, and differentiation not only in normal bone, but also in osteopenic bone-derived osteoblasts.

Biocompatible Materials↗

PubMed2004

Osteointegration of bioactive glass-coated and uncoated zirconia in osteopenic bone: an in vivo experimental study.

In elderly and osteoporotic patients an age-related loss of osteoinductivity could be the biological cause of implant failure regardless of the high quality of the implanted device. yttria stabilized tetragonal zirconia (YSTZ), either coated with the bioactive glass named RKKP bioglaze (RKKP) or uncoated, was implanted in the distal femurs of sham-operated and ovariectomized female rats. Animals were sacrificed at 30 and 60 days. Histomorphometry and microhardness tests were performed to assess osteointegration rate as well as bone quality around the implants. Significant decreases (p < 0.0005) in trabecular bone volume, BV/TV (41%), trabecular bone surface BS/TV (33%), trabecular thickness Tb.Th (20%), and trabecular number Tb.N (32%), together with a significant increase in trabecular separation Tb.Sp (184%), were found for the osteopenic rats compared with the sham-operated rats. At both experimental times the RKKP coating ensured a better osteointegration rate with higher AI values than the uncoated YSTZ, even when osteopenic rats were used (48% at 30 days and 12% at 60 days). No differences were observed at the bone-biomaterial interfaces for either material when comparing sham-operated with osteopenic rats. The present results demonstrate that the RKKP bioactive glass used as a coating ensures a high osteointegration rate even in osteoporotic bone, which is already visible from postoperative day 30 and is still apparent on day 60.

Animals↗

PubMed2004

Early detachment of titanium particles from various different surfaces of endosseous dental implants.

Titanium (Ti) endosseous dental screws with different surfaces (smooth titanium--STi, titanium plasma-sprayed-TPS, alumina oxide sandblasted and acid-etched--Al-SLA, zirconium oxide sandblasted and acid etched--Zr-SLA) were implanted in femura and tibiae of sheep to investigate the biological evolution of the peri-implant tissues and detachment of Ti debris from the implant surfaces in early healing. Implants were not loaded. Sections of the screws and the peri-implant tissues obtained by sawing and grinding were analysed by light microscopy immediately after implantation (time 0) and after 14 days. All samples showed new bone trabeculae and vascularised medullary spaces in those areas where gaps between the implants and host bone were visible. In contrast, no osteogenesis was induced in the areas where the implants were initially positioned in close contact with the host bone. Chips of the pre-existing bone inducing new peri-implant neo-osteogenesis were surrounded by new bone trabeculae. The threads of some screws appeared to be deformed where the host bone showed fractures. Ti granules of 3-60 microm were detectable only in the peri-implant tissues of TPS implants both immediately after surgery and after 14 days, thus suggesting that this phenomenon may be related to the friction of the TPS coating during surgical insertion.

Animals↗

PubMed2004

Histomorphometric bone modifications induced by growth hormone treatment in a rabbit model of short bowel syndrome.

The effects of recombinant human growth hormone (rhGH) on cancellous and cortical bone were investigated in an experimental rabbit model of short bowel syndrome (SBS). Eighteen young male New Zealand rabbits, 2.0 +/- 0.2 kg b.w., were divided into three groups: an SBS Group submitted to a 70% midjejunoileal enterectomy and reanastomosis; an SBS-GH Group undergoing the same surgery and receiving 0.4 mg/kg/day rhGH for 28 days; a Control Group which was sham-operated. Thirty-five days after surgery, all the animals were pharmacologically euthanised and their femurs and L5 vertebrae were used for densitometric and histomorphometric studies. Vertebral and femoral densitometric results showed that the SBS Group presented significantly (P<0.01) lower values (10-25%) than the Control and SBS-GH Groups. Significant differences in the cancellous histomorphometric parameters, namely the trabecular bone area (-7% to 46%), trabecular thickness (-21% to 34%) and trabecular separation (17-32%), were observed between the SBS Group and the other groups. Both the SBS and SBS-GH Groups showed significantly (P<0.05) higher values than the Control Group in terms of cross-sectional area (approximately 24%), cortical area (approximately 20%), and periosteal perimeter (approximately 9%), while medullary area (41%) and endocortical perimeter (18%) were significantly higher (P<0.05) in SBS Group than those of Control Group. The current findings are encouraging and suggest that GH administration in SBS animal model used may improve skeletal tissue remodelling.

Animals↗

PubMed2004

New polymers for drug delivery systems in orthopaedics: in vivo biocompatibility evaluation.

The use of biodegradable polymers for drug delivery systems excluded the need for a second operation to remove the carrier. However, the development of an avascular fibrous capsule, reducing drug release, has raised concern about these polymers in terms of tissue-implant reaction. Five novel polymers were evaluated in vivo after implantation in the rat dorsal subcutis and compared to the reference polycaprolactone (PCL). Poly(cyclohexyl-sebacate) (PCS), poly(L-lactide-b-1,5-dioxepan-2-one-b-L-lactide) (PLLA-PDXO-PLLA), two 3-hydroxybutyrate-co-3-hydroxyvalerate copolymers (D400G and D600G), and a poly(organo)phosphazene (POS-PheOEt:Imidazole) specimens were histologically evaluated in terms of the inflammatory tissue thickness and vascular density at 4 and 12 weeks from surgery. The highest values of inflammatory tissue thickness were observed in D600G (P < 0.01), PCS (P < 0.001) and PLLA-PDXO-PLLA (P < 0.001) at 4 weeks, while POP-PheOEt:Imidazole showed the lowest value of inflammatory tissue thickness (P < 0.05) at 12 weeks. D400G, D600G, PLLA-PDXO-PPLA and POP-PheOEt:Imidazole showed higher (P < 0.001) values of vascular density near the implants in comparison to PCL at 4 weeks. Finally, D400G and D600G increased their vessel densities while POP-PheOEt:Imidazole and the synthetic polyester PLLA-PDXO-PLLA presented similar vessel density values during experimental times. These different behaviours to improve neoangiogenesis without severe inflammatory tissue-responses could be further investigated with drugs in order to obtain time-programmable drug delivery systems for musculoskeletal therapy.

Animals↗

PubMed2004

Adsorption of cationic antibacterial on collagen-coated titanium implant devices.

Two different cationic antimicrobial molecules, chlorhexidine (CH) and poly(hexamethylenebiguanide) (PH), were adsorbed from aqueous solution to titanium implant devices surface-modified by the covalent coupling of collagen on a polyanionic acrylic acid overlayer. Results show that more antimicrobial was adsorbed on surface modified implants as compared to control titanium devices. Moreover, the kinetic of release was affected by the interaction between the polyanionic overlayer and the cationic antimicrobial, leading to slower kinetic of release in the case of CH and stable adsorption in the case of polycationic PH . These data indicate that biochemically modified collagen coated surfaces could be endowed also by antimicrobial properties, in the spirit of present researches on multifunctional implant surfaces.

Adsorption↗

PubMed2004

Oral low-dose cyclophosphamide in metastatic hormone refractory prostate cancer (MHRPC).

The chemotherapeutic approach to hormone-refractory metastatic prostate cancer (MHRPC) for a long time included only estramustine. Then, attempts have been made with other various agents as cyclophosphamide, vinblastine, etoposide, taxanes and carboplatinum. Although the new drugs and combinations have increased the response rate of MHRPC, they have had no impact on the natural history of MHRPC, which is about 1 year as median time of survival. After an occasional observation of prolonged response in a patient with MHRPC treated with a very well tolerated oral low-dose of cyclophosphamide, from February 1996 to October 2002, seven more patients with MHRPC and progressive disease were consecutively recruited. Response to treatment was evaluated by conventional radiological procedures and/or serial serum PSA measurements. The decline of PSA value was considered to assess the response consistent with the response guidelines from the prostate specific antigen-working group. All eight studied patients continuously received oral low dose cyclophosphamide until progression or the occurrence of significant toxicity. So far three patients (37.5%) progressed (PD), two (25%) showed PR and the three remaining SD. Response rate was 25%, and clinical benefit occurred in 62.5% of the studied patients. In the five patients with clinical benefit on cyclophosphamide median duration of clinical benefit, PR and SD were 9, 24+ and 8 months, respectively. In these five patients median overall survival times from cyclophosphamide and from the first regimen of chemotherapy were 17 and 33+ months respectively, while in the three patients with PD they were 4 and 13 months. The same interval times in patients with > or =50% decline of serum PSA were 29 and 50.5 months, while in those with <50% decline of the same marker, they were 13 and 32 months, respectively. Grade 2 or 3 neutropenia were observed in all the studied patients. In four (50%) of them pulmonary and urinary infections that were easily cured by the common antibiotics occurred. These data suggest that the metronomic use of cyclophosphamide, given alone, has similar or higher activity with lower toxicity than when administered with other active drugs. So it can be an useful option before or after the use of other single or combined potentially active chemotherapeutic agents.

Administration, Oral↗

PubMed2004

Osteoporosis and biomaterial osteointegration.

Biomaterial osteointegration depends not only on the properties of the implanted material but also on the characteristics and regenerative capability of the host bone. For this reason, researchers involved in biomaterial evaluation now place great importance on the various pathologies often present in orthopaedic patients which can negatively affect the success of surgical implants. Osteoporosis is undoubtedly one of the most frequently encountered of such diseases. Models reproducing the osteoporotic condition can be useful to understand the influence of the pathology on cell behaviour, bone regeneration and osteointegration processes, thus increasing our basic knowledge and allowing the development of surgical techniques and implant biomaterials more suitable for use in the surgical treatment of fractures in osteoporotic patients. The present paper is a literature review and, after a short description of how the presence of osteoporosis could influence bone regenerative processes, the results of the main studies on biomaterial biocompatibility and osteointegration both in vitro and in vivo in the presence of osteoporotic condition are reported. Both cell cultures and animal models are able to demonstrate the different response of bone to biomaterials by comparing healthy and pathological conditions. The use of pathological bone-derived cells and pathological animals is therefore recommended to test candidate orthopaedic materials.

Animals↗

PubMed2004

Different diagnostic techniques for the assessment of cortical bone on osteoporotic animals.

The aim of this study was to assess the capability of ultrasonography and densitometry to predict the mechanical competence of cortical bone in healthy and osteopenic rats, respectively. Thirty 10-month-old Sprague-Dawley retired breeder female rats were used and randomized into three groups of 10 animals each. A group underwent bilateral ovariectomy by dorsal approach (Ovx), another group underwent a simulated ovariectomy (Sham-Ovx), and the last group served as a sham-aged control group (Control). Sixteen weeks after surgery, the animals were euthanized and the femurs of each rat excised for ultrasonographic and densitometric measurements, and mechanical analyses. The Ovx Group had a significantly decreased amplitude dependent speed of sound (AD-SoS-about 7-8%) when compared to the other groups (p<0.0005). For Ovx animals compared with Sham-Ovx and Control rats, significant decreases in densitometric data were observed (6-13%), as well as significant decreases in femoral Max. Load (about 18%) and flexural rigidity (about 30%). The best correlation (R2=0.55, p<0.0005) found was between SoS and femoral shaft bone mineral density (SBMD). The regression coefficient R2 increased when power-law fits were used, particularly from 0.34 (p<0.001) to 0.36 (p<0.0005) in the correlation between SoS and Max. Load and from 0.21 (p<0.05) to 0.25 (p<0.01) in the correlation between SBMD and Max. Load. The ability of QUS or DXA to accurately predict the actual mechanical characteristics of bone, and in particular bone elasticity, remained relatively poor and the improvement of the power-law model did not describe exhaustively the relationships between the variables tested. The DXA and QUS capability to discriminate between ovariectomized and non-ovariectomized rats did not improve when tested together.

Absorptiometry, Photon↗

PubMed2004

Analysis of 1H-NMR relaxation time distributions in L1 to L6 rat lumbar vertebrae.

A better knowledge of the NMR relaxation behavior of bone tissue can improve the definition of imaging protocols to detect bone diseases like osteoporosis. The six rat lumbar vertebrae, from L1 to L6, were analyzed by means of both transverse (T(2)) and longitudinal (T(1)) relaxation of (1)H nuclei at 20 MHz and 30 degrees C. Distributions of relaxation times, computed using the multiexponential inversion software uniform penalty inversion, extend over decades for both T(2) and T(1) relaxation. In all samples, the free induction decay (FID) from an inversion-recovery (IR) T(1) measurement shows an approximately Gaussian (solid-like) component, exp[-1/2(t/T(GC))2], with T(GC) approximately 12 micros (GC for Gaussian component) and a liquid-like component (LLC) with initially simple-exponential decay. Averaging and smoothing procedures are adopted to obtain the ratio alpha between GC and LLC signals and to get separate T(1) distributions for GC and LLC. Distributions of T(1) for LLC show peaks centered at 300-500 ms and shoulders going down to 10 ms, whereas distributions of T(1) for GC are single broad peaks centered at roughly 100 ms. The T(2) distributions by Carr-Purcell-Meiboom-Gill at 600 micros echo spacing are very broad and extend from 1 ms to hundreds of ms. This long echo spacing does not allow one to see a peak in the region of hundreds of micros, which is better seen by single spin-echo T(2) measurements. Results of the relaxation analysis were then compared with densitometric data. From the study, a clear picture of the intratrabecular and intertrabecular (1)H signals emerges. In particular, the GC is presumed to be due to (1)H in collagen, LLC due to all the fluids in the bone including water and fat, and the very short T(2) peak due to the intratrabecular water. Overall, indications of some trends in composition and in pore-space distributions going from L1 to L6 appeared. Published results on rat vertebrae obtained by fitting the curves by discrete two-component models for both T(2) and T(1) are consistent with our results and can be better interpreted in light of the shown distributions of relaxation times.

Animals↗

PubMed2004

Fibroin hydrogels for biomedical applications: preparation, characterization and in vitro cell culture studies.

Silk fibroin hydrogels prepared either by treating a 2% (w/v) silk fibroin aqeuous solution at 4 degrees C (thermgel) or by adding 30% (v/v) of glycerol (glygel), were characterized by using Environmental Scanning Electron Microscopy (ESEM), Fourier Transform Infrared Spectroscopy (FT-IR), Differential Scanning Calorimetry (DSC), Thermogravimetrical Analysis (TGA) and molecular weight determination. The preparation procedure affected morphology and molecular weight of hydrogels, with no or negligible differences being displayed by FT-IR and DSC analyses. While thermgel presented a well uniform porous structure, the morphology of glygel appeared to be non-porous and heterogeneous. Glygel presented lower water content and lower degradation temperatures, associated with the presence of glycerol but likely also to less-organized protein structures. Cytoxicity tests with human osteoblast-like cells indicated that both gels were not cytoxic, while cell cultures pointed out a faster cell proliferation on glygel and a higher cell activation and differentiation on thermgel. These gels could be used as scaffolds able to promote in situ bone regeneration.

Animals↗

PubMed2004

Biomedical coatings to improve the tissue-biomaterial interface.

One of the most important factors determining the degree of tissue interaction of an implanted device is the property of its surface. Thus, great importance is given to chemical and morphological characteristics of biomaterial surfaces to improve biocompatibility, cell migration, proliferation and differentiation, mechanical stability and endogenous tissue ingrowth. In order to obtain new and healing stimulating properties, it is possible to apply a coating or more generally a surface treatment to the surface of a prosthetic device. One of the most versatile methods for coating is thermal spray technology. This paper considers the principle of thermal spray processes and their application in the biomedical field, namely the coatings used for orthopedic prostheses and dental implants. Among thermal spray processes, plasma spray as well as High Velocity Oxygen Fuel (HVOF) processes will be particularly considered and their most important aspects will be illustrated.

Bone Substitutes↗

PubMed2004

In vitro models to test orthopedic biomaterials in view of their clinical application in osteoporotic bone.

The development of in vitro cell cultures, in association with in vivo experimentation, greatly improved the characterization of biomaterials for orthopedic devices before their clinical use. In recent years an increasing interest has arisen in the use of both pathological osteoblast cultures and animal models to perform in vitro and in vivo tests on biomaterial behavior. A growing number of prostheses, in fact, are implanted in osteoporotic patients, due to the increasing age of the population. Moreover, the presence of osteoporosis may affect bone-biomaterial osteointegration in these patients. The present paper is a literature review and, after a short description of in vitro studies for characterization of osteoblasts derived from osteoporotic bone, the results of in vitro studies on biomaterial biocompatibility and osteointegration rate in the presence of osteoporotic bone derived osteoblast cultures are reported. Pathological cell culture models are able to demonstrate the different behavior of osteoblasts in response to biomaterials, when comparing normal and pathological conditions.

Animals↗

PubMed2004

A biomimetic gelatin-calcium phosphate bone cement.

The interest in new bone substitutes is rapidly increasing in the field of orthopedic surgery. A variety of calcium phosphate bone cement has been developed and different additives have been used to improve their biocompatibility and bioactivity. Following a biomimetic strategy aimed at reproducing bone characteristics, this study investigates the biological properties of a new gelatin enriched calcium phosphate cement (GEL-CP) that exhibits improved mechanical properties with respect to cement prepared without gelatin (C-CP). Human osteoblast MG63 were cultured on the surfaces of GEL-CP and were compared to cells cultured on C-CP samples, and on polystyrene of plate culture as control (C). Cell attachment, proliferation and differentiation were evaluated up to 21 days. SEM revealed that osteoblasts grown on GEL-CP showed a normal morphology and biological tests demonstrated very good rate of proliferation and viability in every experimental time. The presence of gelatin stimulated alkaline phosphatase activity, collagen and transforming growth factor 31 production. The data indicate that the new cement GEL-CP favors osteoblast proliferation, activation of their metabolism and differentiation. The remarkable improvement of the setting properties of the calcium phosphate cement due to the presence of gelatin suggest that the biomimetic composite material could be successfully applied as bone substitute.

Alkaline Phosphatase↗