Biomedical subjects

Milena Fini

Publications and source records attributed to Milena Fini.

At least 37 records · Page 2Linked to original sources

PubMed2003

Human osteopenic bone-derived osteoblasts: essential amino acids treatment effects.

The development of in vitro cell culture methods has made it possible to study bone cell metabolism and growth and obtain a deeper insight into the pathophysiology of common orthopedic diseases such as osteoporosis. After analyzing the effect of two essential amino acids, L-arginine (Arg) and L-lysine (Lys), in previous in vitro and in vivo studies, the present authors investigated the administration of Arg and Lys in osteoblasts derived from human osteopenic bone. After isolation, osteoblasts were cultured in DMEM supplemented with either Arg (0.625 mg/ml/day, Arg Group) or Lys (0.587 mg/ml/day, Lys Group), or both of them (Arg-Lys Group), whereas the Control Group was sham-treated. After 7 days the following parameters were tested in all groups: MTT proliferation test, Alkaline Phosphatase (ALP), Nitric Oxide (NO), Calcium (Ca), Phosphorus (P), Osteocalcin (OC), C-Terminal Procollagen type I (PICP), Interleukin-6 (IL-6), Transforming Growth Factor-beta 1 (TGF-beta 1), Platelet Derived Growth Factor (PDGF) and Insulin-Like Growth Factor-I (IGF-I). Results were compared with those obtained from human healthy bone to verify the effect of the amino acids on osteoblasts derived from pathological tissue. In addition, a comparison was also made with the results obtained from rat osteopenic bone to assess reliability of the in vitro model. The current results support previous findings and indicate that Arg and Lys stimulation has a positive effect on osteoblast proliferation, activation and differentiation. Therefore, administration of these amino acids may be useful in clinical treatment and prevention of osteoporosis.

Aged↗

PubMed2003

Osteoblasts cultured from osteoporotic bone: a comparative investigation on human and animal-derived cells.

In vitro studies on pathophysiology and innovative treatments of many orthopaedic diseases, based on the investigations of cells from pathologic skeletal tissues, greatly improve basic knowledge of osteoporosis. Primary osteoblast (OB) cultures derived from osteopenic bone from different species (human, rat, sheep) were compared to assess the differences that should be taken into account when performing in vitro biocompatibiliy tests or investigating pharmacological and physical treatments. Primary OB were isolated from osteopenic patients and animals by well-established methods and their metabolism was assessed with or without 1,25(OH)2D3. The greatest significant differences were observed between rat and human cells both under basal conditions and after 1,25(OH)2D3 stimulation. In addition, the response to 1,25(OH)2D3 stimulation of OBs from osteopenic rats was significantly different from that of human and sheep OB cultures, in terms of NO, OC, IL-6, and TGF-beta1. Cells derived from osteopenic sheep behaved much more similarly to those from humans, except for a significant difference in terms of TGF-beta1 observed both under basal conditions and after stimulation.

Animals↗

PubMed2003

Primary osteoblasts response to shock wave therapy using different parameters.

Over the past decade extracorporeal shock-wave therapy (ESWT) has been increasingly applied to orthopaedic and musculoskeletal pathologies, the aim of this study was to assess how the energy density of the shock waves and the number of impulses affect viability, differentiation and synthetic activity of osteoblasts. Primary sheep osteoblasts cultures were treated with ESWT with an electro-hydraulic shock wave generator by selecting three different energy levels (14-21-28 kV corresponding at 0.15-0.31-0.40 mJ/mm2) and two different total numbers of impulses (500, 1000) for each level. At the end of treatment, cell counts and viability were recorded. Cells were then cultivated for 48 hours starting from a concentration of 1 x 10(4) cells/ml. The biological activity and viability were evaluated at 24 and 48 hours after treatment. No cytodestructive effects were observed in Group A, while a cytodestructive effect of ESWT was seen in cultures receiving the highest energy treatments. The different shock wave treatment induced differences in MTT assays after 24 and 48 hours, in particular the highest level showed a detrimental effect on cell respiration at both experimental times as compared to the Control Group and the protein metabolism was generally depressed by ESWT with impulses at the highest energy level. After 24 hours such effect further increased with the growing number of impulses. The lowest energy level appeared to significantly improve the metabolic parameter in primary cell cultures as compared to controls when 500 impulses were selected. The current study has demonstrated that one of the most important aspects to be considered is not the total number of impulses used but the energy level of the shock waves, thus confirming that ESWT has a dose-dependent effect on cells.

Animals↗

PubMed2003

Effect of extracorporeal shock wave therapy on osteoblastlike cells.

Extracorporeal shock wave therapy has been used increasingly in musculoskeletal disorders although its biologic mechanisms are not understood completely. The current study evaluated the effects of extracorporeal shock wave therapy on human osteoblastlike cells by using an electrohydraulic shock wave generator and comparing three energy levels. (Group A, 14 kV and 0.15 mJ/mm2; Group B, 21 kV and 0.31 mJ/mm2; Group C, 28 kV and 0.40 mJ/mm2; Control Group, no energy) and two total impulses (500, 1000) for each level. At the end of treatment, a reduction by approximately 76% was observed in Group C cell number versus basal value when compared with the other groups. Viability, biochemical activity, and gene expression of cultured cells were evaluated 24 and 48 hours after treatment. The viability test showed a decrease in Group C viability of approximately 54% at both culture times as compared with the other groups. Significant increases in nitric oxide, osteocalcin, and transforming growth factor-beta1 production ranging from 10% to 35% were found in Group A. All treated groups had lower C-terminal procollagen Type I values than the Control Group, but important increases were observed between 24 and 48 hours in all groups except Group C. This particular finding reveals that osteoblast differentiation in Group A is enhanced strongly during the first 24 hours after exposure leading after another 24 hours to an increase in C-terminal procollagen Type I production and consequently in bone matrix deposition. The current study showed that one of the most important aspects to be considered is not the total number of impulses used, but the energy level of the shock waves, therefore confirming that extracorporeal shock wave therapy has a dose-dependent initial destructive effect on cells when the selected energy is higher than 21 kV.

Cell Survival↗

PubMed2003

Hemodynamic indices versus gastric tonometric measurements for prognosis of hemorrhagic shock: a porcine model.

The aim of the study reported here was to assess the prognostic value of gastric tonometry and its implications in the initial phases of hemorrhagic shock. Hemorrhagic shock was induced by use of femoral arterial bleeding in 12 hybrid swine under general anesthesia. Approximately 30% of the circulating blood volume was removed, until mean arterial pressure of 45 mmHg was reached. The shock conditions were observed over a limited period (90 min) by comparing traditional hemodynamic parameters with gastric tonometric measurements and tissue oxygenation. After a shock period of 90 min without pharmacologic treatment, blood was collected in acid-citrate dextrose-treated bags and was reinfused via the right femoral vein. At the end of the experiment, seven animals had good hemodynamic recovery on reinfusion (group A), whereas values in five animals deceased in the same phase (group B). Hemodynamic and gastric tonometric results were compared between survivors and nonsurvivors. Intravascular volume restoration and reduction of systemic vascular resistance (SVR) enabled the animals of group A to maintain standard ventricular kinetics and recover in terms of splanchnic regional flow. In addition, increase in intramucosal gastric pH (pHi), decrease in the pH-gap (pHa-Hi), and progressive restoration in gastric wall tissue oxygenation (PtO2) also were observed. These results suggest that useful diagnostic and therapeutic indications can be obtained by acquisition of simple hemodynamic measurements at the beginning of the shock period. On the basis of results of statistical analysis, only mean arterial pressure and SVR were good indicators of shock development, whereas pHi was not a significant factor in this experimental model.

Animals↗

PubMed2002

Osteointegration of hydroxyapatite-coated and uncoated titanium screws in long-term ovariectomized sheep.

This study was designed to evaluate the osteointegration of HA-coated and uncoated titanium screws in the cortical bone of long-term (24 months) ovariectomized sheep (OVX group) compared to sham-aged sheep (Control group). At 12 weeks after implantation, the screws were tested biomechanically (extraction torque) and histomorphometrically (affinity index: AI) in both femoral and tibial diaphyses. Cancellous bone status was assessed by iliac crest biopsy. BMD of the L5 vertebra and a histomorphological study of the femoral and tibial shafts were performed to acquire data on cortical bone. A significant difference was found between the OVX and Control groups for BMD (p<0.0005), and a significant reduction in the cancellous bone area was observed in the OVX group. Femoral and tibial cortical bone parameters showed significant differences between the groups. The type of material selected (femurs: p<0.0005; tibiae: p<0.0005) and ovariectomy (femurs: p<0.005; tibiae: p<0.005) had a significant effect on the extraction torque. AI results were related to the presence or absence of ovariectomy (p<0.05) and strictly depended on the material implanted in the femur and tibia (p<0.0005). In conclusion, at implantation OVX sheep showed a significant loss of trabecular and cortical bone versus sham-aged sheep. The biomechanical and histomorphological results achieved suggest employing HA-coated screws in the presence of osteopenic cortical bone.

Animals↗

PubMed2002

The effect of water in inorganic microsponges of calcium phosphates on the porosity and permeability of composites made with polymethylmethacrylate.

Bone-substitute compounds were obtained by mixing pre-polymerised powders of polymethylmethacrylate and liquid methylmethacrylate monomer with two aqueous dispersions of alpha-tricalcium phosphate (alpha-TCP) grains and calcium-deficient hydroxyapatite (CDHA) powders. The final composites appeared to be porous. The total open porosity was a function of the amount of water added. The water, which was the pore-forming agent, vapourised after the polymerisation process, leaving behind empty spaces in the polymeric matrix. The inorganic powders placed inside the polymeric matrix were shown to act as local microsponges. The water capacity of these microsponges can be determined by a centrifugation step carried out on aqueous dispersion of alpha-TCP and/or CDHA powders that occur before any reaction with the organic compound. The relationship between the total open porosity of the composites and the amount of water trapped inside the inorganic agglomerates proved to be almost linear. The effect of the chemical composition of the powder on the total open porosity is not too great, provided that the two kinds of pellets are prepared with the same amount of water. Both the permeability and shape of the pores proved to be a function of the total open porosity. An increase of the latter parameter produces an increase in permeability and a decrease in tortuosity. Osteoconductivity and osteoinductivity of the final composites were tested in vivo by implantation in rabbits. Formation of new trabecular bone was observed inside the pores where the inorganic powders had been placed.

Animals↗

PubMed2002

A bone substitute composed of polymethylmethacrylate and alpha-tricalcium phosphate: results in terms of osteoblast function and bone tissue formation.

The biological properties of a composite polymeric matrix (PMMA + alpha-TCP) made of polymethylmethacrylate (PMMA) and alfa-tricalciumphosphate (alpha-TCP) was tested by means of in vitro and in vivo investigations. PMMA was used as a comparative material. Osteoblast cultures (MG 63) demonstrated that PMMA + alpha-TCP significantly and positively affected osteoblast viability, synthetic activity and interleukin-6 level as compared to PMMA. At 12 weeks, the PMMA + alpha-TCP implants in rabbit bone successfully osteointegrated in trabecular and cortical tissue (affinity index: 57.14+/-8.84% and 68.31+/-6.18%, respectively). The newly formed bone after tetracycline labelling was histologically observed inside PMMA + alpha-TCP porosity. The microhardness test at the bone-PMMA + alpha-TCP interface showed a significantly higher rate of newly formed bone mineralization compared with PMMA (+83.5% and +58.5%, respectively), but differences still existed between newly formed and pre-existing normal bone. It is herein hypothesized that the present positive results may be ascribed to the porous macroarchitecture of PMMA + alpha-TCP and the presence of the bioactive ceramic material that could have a synergic effect and be responsible for the improvement of (a) the material colonization by bone cells, (b) osteoblast activity, (c) osteoinduction and osteoconduction processes, (d) bone remodelling.

Bone Cements↗

PubMed2002

The effect of pulsed electromagnetic fields on the osteointegration of hydroxyapatite implants in cancellous bone: a morphologic and microstructural in vivo study.

Effects of pulsed electromagnetic fields (PEMFs, 75 Hz. 1.6 mT) were investigated in 12 rabbits after placing hydroxyapatite (HA) implants in their femoral condyles. Six animals were stimulated with PEMFs for three consecutive weeks, 6 h/day, while the remaining animals were sham-treated (Control Group). Rabbits were sacrificed at 3 and 6 weeks (after a 3-week non-stimulation period) for histomorphometric analysis and microhardness testing (at 200, 500, 1,000, 2,000 microm from the implant) around the implants. Histomorphometric analysis did not highlight any significant changes. On the contrary, there were statistically significant differences between the effects produced by PEMFs and Control Groups (F = 149.70, p < 0.0005) on the Affinity Index results, as well as by the experimental time of 6 and 3 weeks (F = 17.12, p = 0.001) on the same results. In PEMF-stimulated animals the microhardness (HV) values measured in trabecular bone at a distance of 200 and 500 microm from the implants, were significantly higher with respect to controls. At 6 weeks, HV values at the bone-implant interface in PEMF-stimulated animals were not significantly different with respect to normal bone, while they remained significantly lower in control animals. Both morphological and structural results demonstrated a positive therapeutic effect of PEMFs in accelerating HA osteointegration in trabecular bone.

Animals↗

PubMed2002

Controlling the lateral distribution and alignment of human gingival fibroblasts by micropatterned polysaccharide surfaces.

Adhesion and alignment of primary human gingival fibroblast to Petri dishes were controlled by proper modification of the surface chemistry of the solid substrate. In particular, air plasma (glow-discharge) treatment, coupled to a masking technique, was used to pattern cell-adhesive areas on a cell-adhesion-resistant alginate coated surface. Fibroblasts were successfully confined to the air plasma-treated areas, which show a much greater growth rate and cell density as compared to the original alginate coated surface. Beside the implications for the study of the mechanisms of bio-adhesion at hydrophilic surfaces, these systems can be of interest for a number of applications in dental surgery and materials.

Alginates↗

PubMed2002

The role of different chemical modifications of superoxide dismutase in preventing a prolonged muscular ischemia/ reperfusion injury.

It is well know that a long period of ischemia followed by reperfusion can create an irreversible tissue damage, also due to the excessive generation of oxygen-derived free radicals. A possibility for avoiding this syndrome is represented by the use of free radical scavengers, such as the superoxide dismutase (SOD). The current authors compared the results achieved through different modifications of this enzyme in an experimental rat hind limb model of ischemia/reperfusion. 60 rats that had a 4 hour and 30 minute ischemia of the left hind limb were divided into four groups of 15 each and treated using a physiological solution (control group), native SOD, monomethoxypolyethylene-glycol-SOD (mPEG-SOD) or poly(acryloilmorpholine)-SOD (PAcM-SOD). The outcomes obtained in terms of limb survival (p < 0.05), as well as histomorphologic studies (p < 0.0005), revealed a superior capacity of mPEG-SOD when compared with the other three substances.

Acrylic Resins↗

PubMed2002

Human osteoblast cultures from osteoporotic and healthy bone: biochemical markers and cytokine expression in basal conditions and in response to 1,25(OH)2D3.

The development of methods to culture bone cells has enhanced in vitro studies and allowed researchers to investigate bone cell metabolism in healthy tissue and in various different bone diseases. Greater knowledge of cultures of pathologic bone tissue-derived osteoblasts may be helpful in performing in vitro experiments that test biomaterials and therapies to be used in the orthopedic field, since this kind of approach better reflects the conditions of clinical relevance to many patients. In the present study primary cultures of human osteoblastic cells were isolated from donors with osteoporosis (HOB, Human Osteopenic Bone) and their respective controls (HNB, Human Normal Bone). They were then characterized in baseline conditions and after stimulation with 10(-9) M 1,25(OH2)D3. Specific biochemical markers of bone cells and cytokines involved in bone turnover were evaluated to assess cell metabolism and any possible differences between osteoblasts derived from healthy and osteopenic bone tissue. Under baseline conditions, HNB and HOB in vitro cultures showed some differences in proliferation (MTT test), PICP, OC and IL-6. The HNB response to 1,25-(OH2)D3 stimulation differed significantly from that of the HOB cultures but only with regard to the MTT test, and ALP and PICP levels; the other selected parameters showed a similar behavior for both cultures. The current findings should be taken into account when cultures derived from human bone are used for in vitro experiments.

Alkaline Phosphatase↗

PubMed2002

Enhanced guided bone regeneration with a resorbable chamber containing demineralized bone matrix.

BACKGROUND: The effectiveness of a nonporous poly-DL-lactide tubular chamber in guiding bone regeneration through a long bone defect had already been assessed in an experimental model using the rabbit radius. The injection of bone marrow stem cells into the chamber had proven to enhance bone regeneration. METHODS: The present study reports on the development of the above research project in a subsequent stage. Demineralized bone matrix (DBM) obtained by milling New Zealand rabbit femoral and tibial diaphyses was placed into a tubular chamber. A 10-mm defect was bilaterally created in the radii of 10 rabbits. On the left side (chamber side) the defect was treated by means of a poly-DL-lactide chamber filled with DBM, whereas DBM alone was used on the right side (control). RESULTS: Controls were performed at 3 and 6 months by radiographs and histomorphometry and demonstrated better bone growth on the chamber side versus the control side. A comparison with the results previously obtained by stem cell injection into the chamber revealed significant acceleration of bone regrowth in the first 3 months because of the addition of DBM to the chamber. However, no significant difference was found between the two sides after 6 months. CONCLUSION: These results have confirmed the effectiveness of the chamber as a container for the factors promoting bone regeneration, probably because the osteogenetic activity is maintained in situ.

Absorbable Implants↗

PubMed2002

Characterization of bone defect repair in young and aged rat femur induced by xenogenic demineralized bone matrix.

BACKGROUND: The osteoinductive effect of some biomaterials could be affected by those systemic conditions typical of old age. The aim of the present paper was to assess the effects of age on the healing of bone defects treated with demineralized bone matrix (DBM). METHODS: The study was conducted in young (3 month old) and aged (18 month old) rats to assess the efficacy of DBM in the treatment of osseous defects in bone with limited repair capacities. A standard bone defect was created in the distal femoral condyles of male Wistar rats: the left condyle was filled with rabbit DBM granules, while the right condyle was left empty (control). Histological and microhardness analyses were performed at 30 and 45 days after implant surgery. RESULTS: After implantation of xenogenic DBM, bone healing areas of the aged and young groups showed a significant increase in the formation of newly mineralized bone relative to controls. Measurements of trabecular thickness on day 45 revealed no differences between newly formed and preexisting bone in the young group, while control values were lower. Microhardness measurements demonstrated that newly mineralized bone, either induced by DBM or not, and preexisting bone were comparable in terms of trabecular hardness after 45 days. CONCLUSIONS: In conclusion, xenogenic DBM seems to be effective in bone defect healing, since it increases mineralized tissue volume. In both DBM-filled and empty sites, age seems to have a detrimental effect on the volume of new bone formation but no influence on bone maturation.

Aging↗

PubMedDate not supplied

Titanium alloy osseointegration in cancellous and cortical bone of ovariectomized animals: histomorphometric and bone hardness measurements.

PURPOSE: Histomorphometry and microhardness measurements were performed after Ti6AI4V implantation in cancellous and cortical bone of healthy and ovariectomized animals to determine characterization of the bone-biomaterial interface in osteopenic bone. MATERIALS AND METHODS: Nine ovariectomized and 9 sham-aged rats were used. Four months later, nails were implanted in the distal femurs, and the animals were sacrificed after 8 weeks. Moreover, 3 ovariectomized and 3 sham-aged sheep were used. Twenty-four months later, screws were implanted in the tibial diaphyses and the animals were sacrificed after 12 weeks. RESULTS: Histomorphometry showed the development of osteopenia in both trabecular and cortical bone, and revealed a significant decrease in the osseointegration rate in osteopenic versus sham-aged animals for both trabecular (Affinity Index: -18.6%, P < .001) and cortical bone (Affinity Index: -23.5%, P < .005; Bone Ingrowth: -9%, P <.05). At the interfaces of the sham-aged animals in both trabecular and cortical bone, a decrease of bone microhardness was observed in comparison with preexisting bone (trabecular: -9.8%, P < .0005; cortical: -19.3%, P < .0005). In case of osteopenia, this decrease was even more extensive (trabecular: -15.5%, P < .0005; cortical: -24.7%, P < .0005). DISCUSSION: The present data suggest that bone formation around Ti6AI4V was not associated with complete bone maturation, even in healthy animals. In case of osteopenia, both bone formation and maturation were delayed. CONCLUSION: These results apparently demonstrate the utility of investigating biomaterials in osteopenic bone and the importance of careful evaluation of the healing rate and bone maturation degree around implanted biomaterials.

Alloys↗

PubMedDate not supplied

Surface chemistry effects of topographic modification of titanium dental implant surfaces: 1. Surface analysis.

PURPOSE: To analyze the surface composition of 34 different commercially available titanium dental implants. MATERIALS AND METHODS: Surface composition was evaluated by x-ray photoelectron spectroscopy (XPS). Samples were divided into 4 groups, depending on their surface topography (machined, sandblasted, acid etched, or plasma sprayed). RESULTS: Statistical analysis of the data showed a clear relationship between surface composition and topography, which can be easily accounted for by the chemical effects of the surface treatment performed. On average, acid-etched and plasma-sprayed surfaces had higher titanium and lower carbon concentration than machined surfaces. DISCUSSION AND CONCLUSION: Current studies aimed at the evaluation of implants with different topography should not implicitly assume that topography is the only variable controlling the biologic response. Rather, when comparing different topographies, it should be taken into account that surface chemistry may be a variable as well.

Acid Etching, Dental↗

PubMedDate not supplied

Surface chemistry effects of topographic modification of titanium dental implant surfaces: 2. In vitro experiments.

PURPOSE: To determine, in vitro, cytotoxicity and cell adhesion on 3 different implant surfaces. MATERIALS AND METHODS: All samples had machined surfaces, but they were subjected to different cleaning procedures, which produced 3 different surface chemistries. One of the samples was "as-produced" from the machining tools. The other samples were subjected to partial and total cleaning routines. Cytotoxicity was evaluated using mouse fibroblast cultures, and cell adhesion was evaluated with osteoblast-like SaOS-2 cells. RESULTS: The "as-produced" sample showed a pronounced surface contamination by lubricating oils. For partially and totally cleaned samples, an increasing amount of titanium and a decreasing carbon/titanium ratio was observed as cleaning became more complete. DISCUSSION: Differences in surface chemistry such as those normally found on titanium implant surfaces (see part 1 of this series) can lead to those same effects which, in in vitro experiments, are normally accounted for in terms of surface topography alone. CONCLUSION: Effects related to surface chemistry can operate over and above surface topography, making it impossible, without proper characterization, to make definite statements about the role of topography alone.

Animals↗

PubMedDate not supplied

Surface analysis and effects on interfacial bone microhardness of collagen-coated titanium implants: a rabbit model.

PURPOSE: The aim of this study was to evaluate the surface chemistry and the microhardness at the implant-bone interface using a recently developed collagen-coated titanium implant in a short-term rabbit model. MATERIALS AND METHODS: Surface chemistry was evaluated by x-ray photoelectron spectroscopy (XPS), while in vivo studies involved 4-week implants mid-diaphysis in the lateral femurs of adult male rabbits. After conventional embedding and evaluation of histologic sections, the resinembedded blocks containing the implanted screws were used to measure bone hardness by means of an indentation test. RESULTS: Decomposition of the C1s peak obtained by XPS analysis confirmed that surface-immobilized collagen retained all the molecular features of the control, nonimmobilized reference. As to microhardness measurement, newly formed bone at the collagen-coated-implant/bone interface was significantly harder than bone at the interface of the uncoated control implant and bone. DISCUSSION: These results suggested that collagen coating significantly improves bone maturation and mineralization at the interface in comparison with uncoated commercially pure titanium. Surface modification of titanium implants by collagen coating has recently been discussed as a promising approach to the biochemical modification of implant surfaces. The present results support previous histologic findings and demonstrated that the biomolecular layer linked over the titanium implant can increase the bone healing rate, at least in this animal model. CONCLUSIONS: The present microhardness measurement at the bone-implant interface showed that collagen coating can significantly improve bone maturation and mineralization at the interface in comparison with uncoated commercially pure titanium, confirming and substantiating previous findings by histomorphometric measurements from the same model.

Animals↗