Biomedical subjects

L Di Silvio

Publications and source records attributed to L Di Silvio.

At least 19 recordsLinked to original sources

PubMed2007

In vitro osteoblastic response to 30 vol% hydroxyapatite-polyethylene composite.

Hydroxyapatite-reinforced high-density polyethylene (HA-HDPE) composite, as a bone replacement material, has successfully been used clinically as middle ear prostheses and orbital floor implants. The aim of this study was to examine its in vitro biocompatibility in order to develop a further application, that is, as skull reconstruction implants. Human osteoblast cells isolated from femoral heads and crania were used to determine the biological response of the composites. HA-HDPE composites (30 vol %) with two grades of HA filler that had different surface morphologies were selected for this in vitro assessment. The results showed that HA-HDPE composite was bioactive and supported osteoblast attachment, proliferation, and differentiation. The composite with rough-surfaced HA filler demonstrated slightly better cellular response than the composite with smooth-surfaced HA filler. Although osteoblastic cells derived from skull showed an overall slower response compared with those from femoral heads, these in vitro results show that HA-HDPE composite potentially could be used as a skull implant.

Bone Substitutes↗

PubMed2006

Chemical-physical characterization and in vitro preliminary biological assessment of hyaluronic acid benzyl ester-hydroxyapatite composite.

HYAFF11 is a biocompatible, biodegradable benzyl ester of hyaluronic acid. However, in order to use it for orthopedic application, its mechanical performance needs to be improved. In this study, a novel composite based on HYAFF11 polymer matrix reinforced with hydroxylapatite (HA) has been developed. Its advantage is having a similar component of the mineral phase of bone resulting in favorable osteoconductive properties. The present study has examined the compressive mechanical and surface chemical-physical properties of the novel HYAFF11-HA composite. Preliminary biological investigations, including pH and cytotoxicity studies of the material extracts, have also been performed using an in vitro primary human osteoblast-like cell model. Moreover, protein, especially fibronectin adsorption has been investigated following incubation in culture medium and human plasma. The results show a grainy surface topography composed mainly of C, P, and Ca, with a Ca/P atomic ratio indicating HA on the composite surface. Mechanical analysis shows an improvement of the compressive properties of HYAFF11 matrix, both in the dry and swollen states, with values in the range of that of spongy bone. No cytotoxic effects and no inhibition of cell proliferation have been observed in the presence of the material extracts with pH values within acceptable ranges for cell vitality. Protein studies reveal a similar pattern, but a higher amount of fibronectin following incubation in human plasma when compared with culture medium. The results show that the novel HYAFF11-HA composite shows a great potential for application in orthopedic fields, especially as vertebral trabecular bone substitute.

Biocompatible Materials↗

PubMed2005

Bioresorbable glass fibres facilitate peripheral nerve regeneration.

This is a proof of principle report showing that fibres of Bioglass 45S5 can form a biocompatible scaffold to guide regrowing peripheral axons in vivo. We demonstrate that cultured rat Schwann cells and fibroblasts grow on Bioglass fibres in vitro using SEM and immunohistochemistry, and provide qualitative and quantitative evidence of axonal regeneration through a Silastic conduit filled with Bioglass fibres in vivo (across a 0.5 cm interstump gap in the sciatic nerves of adult rats). Axonal regrowth at 4 weeks is indistinguishable from that which occurs across an autograft. Bioglass fibres are not only biocompatible and bioresorbable, which are absolute requirements of successful devices, but are also amenable to bioengineering, and therefore have the potential for use in the most challenging clinical cases, where there are long inter-stump gaps to be bridged.

Absorbable Implants↗

PubMed2005

Scandia--a potential biomaterial?

Biocompatibility is a pre-requisite for all biomaterials used for medical application. During the last two decades significant advances have been made in the development of novel materials and selection and use of these materials has been directly dependent upon their biocompatibility. Several materials containing calcium or titanium cations demonstrate biocompatibility and are routinely used in various forms within the human body. Due to its position in the periodic table, scandium in the form of its oxide scandia (Sc(2)O(3)) was studied as the first stage of a wider exploration of the biocompatibility of ceramics. A commercial human osteoblast-like cell line (HOS TE 85) was used to study the biocompatibility of both sintered and abraded scandia surfaces. Scanning electron microscopy was used to examine cell adhesion, the MTT assay was used to measure cell metabolic function and the alamarBlue for the assessment of proliferation. Although the results are only preliminary findings, qualitative observations showed that both sintered and abraded surfaces favoured cell adhesion to the same extent. Quantitatively, a significant increase in cell proliferation was observed on Sc(2)O(3) compared to Thermanox, tissue culture control. Furthermore, Sc(2)O(3) has been shown to be non-toxic, able to be maintain cell viability and support cell growth and proliferation.

Biocompatible Materials↗

PubMed2005

The chemical constitution and biocompatibility of accelerated Portland cement for endodontic use.

AIM: To evaluate the biocompatibility of mineral trioxide aggregate and accelerated Portland cement and their eluants by assessing cell metabolic function and proliferation. METHODOLOGY: The chemical constitution of grey and white Portland cement, grey and white mineral trioxide aggregate (MTA) and accelerated Portland cement produced by excluding gypsum from the manufacturing process (Aalborg White) was determined using both energy dispersive analysis with X-ray and X-ray diffraction analysis. Biocompatibility of the materials was assessed using a direct test method where cell proliferation was measured quantitatively using Alamar Blue dye and an indirect test method where cells were grown on material elutions and cell proliferation was assessed using methyltetrazolium assay as recommended by the International standard guidelines, ISO 10993-Part 5 for in vitro testing. RESULTS: The chemical constitution of all the materials tested was similar. Indirect studies of the eluants showed an increase in cell activity after 24 h compared with the control in culture medium (P<0.05). Direct cell contact with the cements resulted in a fall in cell viability for all time points studied (P<0.001). CONCLUSIONS: Biocompatibility testing of the cement eluants showed the presence of no toxic leachables from the grey or white MTA, and that the addition of bismuth oxide to the accelerated Portland cement did not interfere with biocompatibility. The new accelerated Portland cement showed similar results. Cell growth was poor when seeded in direct contact with the test cements. However, the elution made up of calcium hydroxide produced during the hydration reaction was shown to induce cell proliferation.

Aluminum Compounds↗

PubMed2004

Porosity variation in hydroxyapatite and osteoblast morphology: a scanning electron microscopy study.

The biocompatibility of hydroxyapatite has been demonstrated by previous studies, with enhancement of osteointegration through the use of porous hydroxyapatite (pHA). Emphasis has been focused on the use of coralline hydroxyapatite or the introduction of macroporosity into synthetic hydroxyapatite. The current study investigates the role of macro- and microporosities in synthetic phase-pure porous hydroxyapatite on the morphological aspects of human osteoblast-like cells using scanning electron microscopy. Cells were seeded on four different types of porous hydroxyapatite (HA1, HA2, HA3 and HA4) and examined following 1, 2, 14 and 30 days of incubation in vitro. The results indicated that the cells had an affinity to micropores through filopodia extensions, at initial stage of attachment. Cellular proliferation and colonization was evident on all materials with cells forming cellular bridges across the macropores at day 14 with cellular canopy formation covering entire macropores observed by day 30. This study demonstrates that while the introduction of microporosity has no evident effect on cellular morphology at later time points, it seems to play a role in initial cellular anchorage and attachment.

Bone and Bones↗

PubMed2004

An ultrastructural study of cellular response to variation in porosity in phase-pure hydroxyapatite.

Hydroxyapatite has been shown to be biocompatible and bioactive. Incorporation of porosity has been shown to enhance osteointegration; however, difficulty in controlling the extent and type of porosity has limited investigation into determining the role of both macro- and microporosity. The current investigation reports on the synthesis of four types of phase-pure hydroxyapatite with varying levels of porosity (HA1-HA4), and with defined levels of macro- and microporosities. Transmission electron microscopy was used to evaluate qualitatively the effect of these two parameters on cell-material interactions following a 30-day incubation period. Biological mineralization was observed within vesicles and the needle-like minerals were confirmed as hydroxyapatite using X-ray microanalysis. This demonstrated the suitability of primary human osteoblast-like cells as a tool to assess the extent of mineralization. Furthermore, internalization of hydroxyapatite particles was observed. Our findings show that the variation in macro- and microporosity does not affect the extent of cell-material interaction, with collagen synthesis evident in all samples.

Cell Communication↗

PubMed2004

The fundamentals of tissue engineering: new scaffolds.

The ability to regenerate new bone for skeletal use is a major clinical need. In this study, two novel porous calcium phosphate materials pure HA and biphasic HA/beta-Tricalcium phosphate (HA/beta -TCP) were evaluated as potential scaffolds for cell-seeded bone substitutes using human osteoblast-like cells (HOS) and primary human mesenchymal stem cells (hMSCs). A high rate of proliferation was observed on both scaffolds. A greater increase in alkaline phosphatase (ALP- an indicator of osteoblast differentiation) was observed on HA/beta -TCP compared to HA. This observation indicates that HA/TCP may play a role in inducing osteoblastic differentiation. Although further evaluation is required both materials show potential as innovative synthetic substitutes for tissue engineered scaffolds.

Animals↗

PubMed2003

Initial in vitro interaction of osteoblasts with nano-porous alumina.

In the present study we have used a characterised primary human cell culture model to investigate cellular interactions with nano-porous alumina. This material, prepared by anodisation, is being developed as a coating on titanium alloy implants. The structure of the alumina, as determined by X-ray diffraction and transmission electron microscopy, was amorphous. When studying cell/material interactions we used both biochemical and morphological parameters. Cell viability, proliferation and phenotype were assessed by measurement of redox reactions in the cells, cellular DNA, tritiated thymidine ([3H]-TdR) incorporation and alkaline phosphatase (ALP) production. Results showed a normal osteoblastic growth pattern with increasing cell numbers during the first 2 weeks. A peak in cell proliferation was seen on day 3, after which cell growth decreased, followed by an increase in ALP production, thus indicating that the osteoblastic phenotype was retained on the alumina. Cell adhesion was observed, the osteoblast-like cells having a flattened morphology with filipodia attached to the pores of the material. SDS-PAGE and western blot measurements showed that the nano-porous alumina was able to adsorb fibronectin. Trace amounts of aluminium ions were measured in the surrounding medium, but no adverse effect on cell activity was observed.

Adsorption↗

PubMed2003

The effect of varying percentage hydroxyapatite in poly(ethylmethacrylate) bone cement on human osteoblast-like cells.

Poly(ethylmethacrylate) (PEMA) bone cement has been developed, and the cements mechanical properties are improved by the incorporation of particulate fillers, such as hydroxyapatite (HA). In this in vitro study, human osteoblast-like (HOB) cells were used to examine the effect on cellular behavior of the addition of HA to PEMA using a plain PEMA control. Thymidine uptake ((3)H-TdR) and total DNA were used to assess cell growth and proliferation. Confocal laser scanning microscopy (CLSM) was used to study focal contacts and actin cytoskeletal organisation. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to assess cell morphology and cellular ultrastucture. The early time points showed preferential anchorage to the HA exposed on the cement surface, but no difference in adhesion or proliferation. These results have been attributed to increases in residual monomer with HA incorporation, as shown by proton nuclear magnetic resonance (H(1)-NMR) spectra.

Journal Article↗

PubMed2003

Enhanced HAPEX topography: comparison of osteoblast response to established cement.

The use of poly(methylmethacrylate) PMMA cement by Charnley in the 1960s revolutionized orthopaedic medicine. Since this time, however, little has changed. The development of bioactive composites, such as HAPEX (a composite of 40% vol hydroxyapatite (HA) in a polyethylene matrix) have potential in orthopaedic applications. The composite has been shown to allow direct bone bonding in vivo, and in vitro studies have shown preferential attachment to HA exposed on the composite surface. In vitro study has also shown that altering the topography HAPEX can enhance osteoblast response. This study uses microscopical investigation of osteoblast cytoskeleton, and biochemical measurement of proliferation (by thymidine incorporation) and phenotype (by alkaline phosphatase activity) to compare primary human osteoblast (HOB) activity on HAPEX and PMMA cement. The study shows large increases in HOB response to the new generation material compared to PMMA, the current implant standard.

Journal Article↗

PubMed2002

New acrylic bone cements conjugated to vitamin E: curing parameters, properties, and biocompatibility.

Acrylic bone cement formulations with antioxidant character were prepared by incorporation of a methacrylic monomer derived from vitamin E (MVE). Increasing concentrations of this monomer provided decreasing peak temperature values, ranging from 62 to 36 degrees C, and increasing setting time with values between 17 and 25 min. Mechanical properties were evaluated by compression and tension tests. Compressive strength of the new formulations were superior to 70 MPa in all cases. The cement containing 25 wt % MVE, however, showed a significant decrease in tensile properties. Biocompatibility of the new formulations was studied in vitro. The analysis of the effect of leachables from cements into the media showed continued cell proliferation and cell viability with a significant increase for the cement containing 15 wt % MVE. This formulation also showed a significant increase in cellular proliferation over a period of 7 days as indicated by the Alamar Blue test. The cells were able to differentiate and express phenotypical markers in presence of all materials. A significant increase in alkaline phosphatase activity was observed on the cements prepared in presence of 15-25 wt % MVE compared with PMMA. Morphological assessment showed that the human osteoblast (HOB) cells were able to adhere, retain their morphology, and proliferate on all the cements.

Acrylates↗

PubMed2002

Osteoblast behaviour on HA/PE composite surfaces with different HA volumes.

A hydroxyapatite (HA) reinforced polyethylene (PE) composite (designated HAPEX), with high mechanical specification and a bioactive HA phase, has been optimised as a bone analogue material. Manufacturing conditions and machining of the materials were carefully controlled to give a reproducible material surface roughness with minimal batch variation. The effect of surface composition was examined in vitro using primary human osteoblasts (HOB). HOBs were cultured in direct contact with the test materials containing 20% and 40% vol. HA. The results showed that 40% HA/PE enhanced cellular activity by increasing proliferation rate and differentiation compared to the 20% vol. HA composite. The cytoskeletal organisation of the cells was also examined and HOBs cultured on 40% HA/PE were flatter and had an enhanced rate of cytoskeletal organisation and an increase in focal contact points compared to the 20% HA/PE.

Cell Differentiation↗

PubMed2002

Increasing hydroxyapatite incorporation into poly(methylmethacrylate) cement increases osteoblast adhesion and response.

Poly(methylmethacrylate) (PMMA) is the current standard for cement held prostheses. It forms a strong bond with the implant, but the bond between the cement and the bone is considered to be weak, with fibroblastic cells observed at the implant site, rather than direct bone contact, a contributing factor leading to implant failure. Incorporation of hydroxyapatite (HA) increases the biological response to the cement from tissue around the implant site, thus giving increased bone apposition. In this study, PMMA discs with 0, 4.6 and 8.8 vol%. HA were examined. Primary human osteoblast-like cells (HOBs) were used for the biological evaluation of the response to the cements in vitro. Morphology was observed using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Measurement of tritiated thymidine (3H-TdR) incorporation and alkaline phosphatase (ALP) activity were used to assess proliferation and differentiation. A synergy between increasing focal contact formation, cytoskeletal organisation, cell proliferation and expression of phenotype was observed with increasing HA volume. Preferential anchorage of HOBs to HA rather than PMMA was a prominent observation.

Bone Cements↗

PubMed2002

Initial attachment of osteoblasts to an optimised HAPEX topography.

The interactions between an implant material and the surrounding tissue are of a complicated nature, and the initial attachment of cells to the surface is important in determining the implant success. HAPEX has been developed as a second-generation orthopaedic biomaterial, with both mechanical and biological characteristics that make it suitable for bone augmentation. Further optimisation of the material is being continued to increase the attachment of osteoblasts coupled with improving mechanical characteristics, so it may be used in load bearing applications. It has been previously observed that polishing followed by roughening the surface of HAPEX enhances osteoblast proliferation and phenotype. This article discusses the recruitment of primary human osteoblast cells onto the optimised surface, by examining morphology and cytoskeletal changes using scanning electron microscopy and confocal laser scanning microscopy. The results show that the cells attach in greater numbers to the optimised surface, and develop notably faster, than cells on machined HAPEX.

Arthroplasty, Replacement, Hip↗

PubMed2002

In vitro adhesion and biocompatability of osteoblast-like cells to poly(methylmethacrylate) and poly(ethylmethacrylate) bone cements.

A bone cement, poly(ethylmethacrylate)/n-butylmethacrylate (PEMA/nBMA) has been developed with lower exotherm and monomer leaching compared to the traditional poly(methylmethacrylate)/methylmethacrylate (PMMA/MMA) cement. This study compares the in vitro biological response to the cements using primary human osteoblast-like cells (HOB). Cell attachment was qualified by immunolocalization of vinculin and actin cytoskeleton, showing more organization on PEMA/nBMA compared to PMMA/MMA. Proliferation was assessed using tritiated thymidine incorporation, and phenotype expression determined by measuring alkaline phosphatase (ALP) activity. An increase in proliferation and ALP activity was observed on PEMA/nBMA compared to PMMA/MMA. The results confirm the biocompatability of PEMA/nBMA, and an enhanced cell attachment and expression of differentiated cell phenotype.

Journal Article↗

PubMed2001

Initial interaction of osteoblasts with the surface of a hydroxyapatite-poly(methylmethacrylate) cement.

Failure of the bone/cement interface in cemented joint prostheses is a contributor to implant loosening. The introduction of a bioactive phase, such as hydroxyapatite (HA), to cement may enhance fixation by encouraging direct bone apposition rather than encapsulation of the implant by fibrous tissue. The effect of poly(methylmethacrylate) (PMMA) bone cement (incorporating 17.5% HA wt.) on bioactivity has been investigated using primary human osteoblast-like cells (HOB). A significantly higher cell proliferation and differentiation was seen on the PMMA/HA cement compared to the PMMA cement alone, with retention of phenotype up to 21 days of culture on both materials.

Bone Cements↗

PubMed2001

Effects of carrier release kinetics on bone morphogenetic protein-2-induced periodontal regeneration in vivo.

BACKGROUND: Bone morphogenetic proteins (BMPs) have shown considerable promise as a therapeutic agent to enhance periodontal regeneration although the optimal characteristics of a suitable release system are not known. AIM: The aim of this study was to compare the effects of slow and fast degrading gelatin carriers on BMP-2-induced periodontal healing. METHOD: Recombinant human bone morphogenetic protein-2 (rhBMP-2) was incorporated into gelatin and subsequently differentially cross-linked to produce slow and fast release carrier systems. Release kinetics were confirmed in vitro, by measuring release of 125I-growth hormone from similar gelatin plugs. Effects of BMP were evaluated in surgically created rat periodontal fenestration defects which were processed for histology 10 days post-operatively. The rats were divided into 4 groups and the control defects were treated with either slow or fast degrading gelatin (CONs or CONf respectively), whilst test groups were treated with 1.25 microg rhBMP-2 in the slow or fast degrading gelatin (BMPs or BMPf respectively). RESULTS: BMPf greatly increased bone formation compared with the control (CONf) (1.67 +/- 0.65 versus 0.34 +/- 0.11 x 10(-4) m2), but no significant differences were observed with BMPs and CONs. In contrast, new cementum formation was significantly greater in the BMPs group compared with all other groups (p<0.05). CONCLUSION: Release kinetics of BMP may have important effects on the outcome of BMP-induced periodontal regeneration. New bone formation may be affected by rapid-release kinetics although further investigation is necessary to confirm this. In contrast, new cementum formation is promoted by slow release of BMP.

Alveolar Bone Loss↗