Guided bone regeneration (GBR) is employed to encourage the formation of new bone in osseous defects by restricting the infiltration of soft tissues. While a variety of membranes have been evaluated for this surgical procedure, the non-resorbable material of choice is currently expanded polytetrafluoroethylene (e-PTFE). A new alkali-cellulose membrane produced by a biotechnological process has been developed as an alternative to e-PTFE for GBR. In this study, the biocompatibility of this novel alkali-cellulose membrane and e-PTFE was compared using tissue culture and an in vivo GBR model. In vitro both materials supported the attachment, migration and differentiation of osteoblast-like cells in culture for up to 3 weeks. The in vivo model was based upon full-thickness transcortical bone defects in the mandibular rami of Sprague-Dawley rats. The right rami were used as controls, contralateral defects being covered bucally and lingually with either e-PTFE or alkali-cellulose membranes. Pathological and histomorphometric analysis was undertaken at 4 and 10 weeks post-implantation. Bone regeneration associated with alkali-cellulose membranes was predominantly endochondral in type in contrast to e-PTFE which induced direct bone formation (intramembranous ossification). The amount of new bone formed in defects was similar for both types of membrane, but alkali-cellulose membranes induced significantly greater inflammatory response; characterized by lymphocytes, macrophages and multinucleated giant cells. Degradation and possible exposure of individual cellulose fibres may account for the poor performance of alkali-cellulose membranes in vivo. This animal and in vitro study indicates that when choosing a non-resorbable membrane for GBR, e-PTFE membranes are likely to perform better than those produced from alkali-cellulose.

The plasma copolymerization of acrylic acid and methyl vinyl ketone with 1,7-octadiene is reported. A range of surfaces including hydrocarbon plasma polymer, carbonyl-containing plasma copolymer and carboxyl-containing plasma copolymers have been prepared and characterized by means of X-ray photoelectron spectroscopy. The stability of these surfaces in water has been assessed at 37°C. The influence of their functional group chemistries on the attachment and spreading of osteoblast-like (ROS 17/2.8) and bone marrow stromal cells (BMSC) has been explored. The data reported show a strong correlation between the carboxyl group and the attachment and spreading of both ROS cells and BMSC. This relationship has been explored in some detail with the ROS cells. Actin-staining by direct immunofluorescence was used to visualize changes in cytoskeleton of ROS cells with substratum chemistry. As the concentration of the carboxyl groups increased cell number and cell spreading were notably enhanced. As few as 5 carboxyls per 100 carbons were sufficient to support good attachment and with cells showing well-defined polygonal cell morphology. Although cells attached to a hydrocarbon plasma polymer surface these cells had failed to spread. The attachment and spreading of BMSC were compared on a carboxyl-containing surface and on a hydrocarbon surface. There was a more marked difference in the number of cells that had attached to these two surfaces (cf. ROS cells). The spreading on the carboxyl surface was much more typical of BMSC. Results from an extended culture using primary cultures of bone marrow cells (BMC) are reported. These cells were not trypsinized and appear to be less sensitive to differences in surface chemistry (cf. BMSC). Even so, the results at 10 days indicate much greater biosynthetic activity on the carboxyl-containing surface.

The in vitro biocompatibility of a group of ionomeric cements (ICs) was evaluated with respect to their ion release properties. These ICs were made from a defined series of glasses with the general formula 1.5SiO2⋅0.5P2O5⋅Al2O3⋅(1.0-Z)CaO⋅0.75CaF2 where Z was the mole fraction (ranging from 0–0.1) of an alkali metal oxide, either sodium or potassium or a mixture of both. For these alkali metal ICs, the amount of sodium released was directly related to the sodium content of the constituent glass. Similarly, the amount of potassium released was directly related to the potassium content. There was no correlation between the aluminum content of the glass and the aluminum ion release. Increasing the monovalent cation concentration, however, produced ICs with increased fluoride release. The biocompatibility of the ICs, as assessed by in vitro cell growth and viability measurements, was inversely proportional to aluminum ion release. Fluoride ion release, although important in terms of in vitro biocompatibility, would appear to be less important than aluminum ion release in determining the overall biocompatibility of the ICs studied. © 1998 Kluwer Academic Publishers

The first comparative study of cell attachment to self-assembled monolayers (SAMs) and plasma-deposited films is reported. Osteoblast-like cells attached extensively to acid-terminated alkyl thiol SAMs and to a plasma copolymer of acrylic acid and octa 1,7-diene (acid-PCP). However, they attached poorly to methyl:terminated SAMs and a plasma polymer of octa-1,7-diene (OD-PP).

The purpose of this study was: (a) to examine the effect of plasma-gas composition on plasma polymer oxygen/carbon (O/C) ratio, functional group composition and stability in water, and then (b) to examine cell attachment to surfaces containing different concentrations of O/C and functional groups. Oxygen-functionalised surfaces were deposited by means of the plasma copolymerisation of acrylic acid/1,7-octadiene. The use of a diluent hydrocarbon allowed the deposition of surfaces with a range of O/C concentrations. Plasma copolymer surfaces were characterised by X-ray photoelectron spectroscopy (XPS). Changes in functional group composition with % acrylic acid monomer and the non-dispersive and dispersive parts of the surface energy of these plasma copolymers were measured. The solubility of the plasma copolymers was assessed by means of XPS. The degree of attachment of ROS 17/2.8 osteoblast-like cells to plasma copolymer surfaces deemed to be ‘stable’ in aqueous medium was measured. Tissue culture polystyrene (TCPS) was included as a control. Attachment was found to be greatest to the plasma copolymer surface with an O/C of 0.11. This surface had a carboxylic acid concentration of ca. 3%. Attachment did not correlate with increased surface wettability (i.e. the non-dispersive component of the surface energy).

Using antisera to regions of human parathyroid hormone-related protein (PTHrP) the saccus vasculosus (SV) of the sea bream (Sparus aurata) has been shown to contain immunoreactive PTHrP. By immunohistochemistry (IHC) the epithelial coronet cells in fixed and wax-embedded SV tissue reacted with antisera to the prepro region of human PTHrP (−13 to +2), the N-terminus PTHrP (1–16), and the midmolecule PTHrP (50–69). Sodium dodecyl sulfate–polyacrylamide gel electrophoresis of saccus extracts and incubation media contained two major proteins of 14.3 and 15 kDa. By Western blotting these two proteins both reacted with the three antisera used for IHC, suggesting that they are immunochemically similar to human PTHrP (1–84). Ultrastructurally the coronet cells ofSparussaccus vasculosus resembled coronet cells described for other teleosts, with an abundant smooth endoplasmic reticulum (SER) which was more highly organized in the coronets. IHC at EM level showed reaction mainly with the membranes of the SER. These results suggest thatS. auratasaccus vasculosus may produce a PTHrP-like molecule similar to human PTHrP.

The in vivo response of two defined groups of set ionomeric cements (ICs), were evaluated following implantation in the midshaft of three week old Wistar rat femora for four weeks. New bone formation was associated with all the IC implants, the amount of new bone increasing with increasing sodium or calcium fluoride content of the basic glass component. Previous work has shown that there is a link between glass composition and ion release, fluoride ion release increasing as the sodium or fluoride content of the glass increases. It thus appears that in the series studied improved bone formation associated with the ICs was mediated by increased fluoride ion release.

Glass ionomer cements (GICs) are composite materials with the potential for use as improved bone substitutes and cements. The hydrophilic nature of the GIC matrix may confer the ability to release therapeutic agents after surgical implantation which would aid the development of GICs for wider biomedical application. Acrylic and GIC were loaded (5% w/w> with either a model dye or high molecular weight proteins and eluted in vitro over 84 days to study simulated drug release. Serum proteins were also adsorbed on to the surface of acrylic and two different GICs and desorption measured over six days. GIC was a suitable matrix for simple dye and protein release, protein release being greater from the GIC than from the acrylic cement. Selective desorption from the two different GICs studied was noted indicating GICs may be formulated to release specific drugs or proteins.

Using antibodies to the amino-terminal region of human parathyroid hormone-related protein (PTHrP) we have demonstrated PTHrP immunoreactivity in pituitaries and plasma of the sea bream (Sparus aurata). Pituitary cells at two distinct locations contained immunodetectable PTHrP; an anterior group in the rostral pars distalis which also contained immunoreactive thyroid stimulating hormone (TSH), and a posterior group lying at the border of the pars intermedia and proximal pars distalis between cells which stained with antibody to human corticotrophin-like intermediate lobe peptide. By Western blot analysis pituitary extracts contained two immunoreactive isoforms of PTHrP, one of 29 kDa and the other of 26 kDa. Media of pituitaries incubated for up to 14 days in Krebs-Ringer bicarbonate also had several isoforms of immunodetectable PTHrP, two of them corresponding to the 29- and 26-kDa molecular forms but there were in addition both larger and smaller molecules. The concentration of PTHrP in sea bream plasma was comparable with levels observed in human subjects with humoral hypercalcaemia of malignancy. There was no reaction between pituitary cells or pituitary extracts and antibody to human parathyroid hormone. Thus sea bream pituitary contains immunoreactive PTHrP, which appears to be released into medium during in vitro incubation and which may be a significant source of plasma immunoreactive PTHrP in vivo.