Sterilization of allograft bone: is 25 kGy the gold standard for gamma irradiation?

For several decades, a dose of 25 kGy of gamma irradiation has been recommended for terminal sterilization of medical products, including bone allografts. Practically, the application of a given gamma dose varies from tissue bank to tissue bank. While many banks use 25 kGy, some have adopted a higher dose, while some choose lower doses, and others do not use irradiation for terminal sterilization. A revolution in quality control in the tissue banking industry has occurred in line with development of quality assurance standards. These have resulted in significant reductions in the risk of contamination by microorganisms of final graft products. In light of these developments, there is sufficient rationale to re-establish a new standard dose, sufficient enough to sterilize allograft bone, while minimizing the adverse effects of gamma radiation on tissue properties. Using valid modifications, several authors have applied ISO standards to establish a radiation dose for bone allografts that is specific to systems employed in bone banking. These standards, and their verification, suggest that the actual dose could be significantly reduced from 25 kGy, while maintaining a valid sterility assurance level (SAL) of 10−6. The current paper reviews the methods that have been used to develop radiation doses for terminal sterilization of medical products, and the current trend for selection of a specific dose for tissue banks.

The effect of parasitization by Trichogramma australicum on Helicoverpa armigera host eggs and embryos

Histological investigations of the pathology of Helicoverpa armigera (Hiibner) eggs after attack by the egg parasitoid, Trichogramma australicum (Girault), indicate that the developing embryo is immediately killed by envenomation. Soon afterward the histological staining characteristics of parasitized host embryos change and the embryonic germ band dissociates into a mass of individual rounded cells. Hosts attacked by females sterilized by gamma-irradiation showed the same pathological effects as normally parasitized hosts, indicating that host degeneration is due to female venom rather than factors derived from the parasitoid embryo or larva. Cell death also occurred in older host embryos although tissue breakdown was delayed. These findings have allowed us to determine not just that the host dies but what happens to the cells and tissues, i.e., their physical appearance, the time course of their degeneration, and that the process is retarded in older hosts. These processes can possibly be emulated in artificial diets. (C) 2003 Elsevier Inc. All rights reserved.

In vitro bioactivity of MOEP grafted ePTFE membranes for craniofacial applications

The bioactivity of three methacryloyloxyethyl phosphate (MOEP) grafted expanded polytetrafluoroethylene (ePTFE) membranes with varying surface coverage as well as unmodified ePTFE was investigated through a series of in vitro tests: calcium phosphate (CaP) growth in simulated body fluid (SBF), serum protein adsorption, and a morphology and attachment study of human osteoblast-like SaOS-2 cells. The graft copolymers were prepared by means of gamma irradiation induced grafting and displayed various surface morphologies and wettabilities depending on the grafting conditions used. Unmodified ePTFE did not induce nucleation of Cal? minerals, whereas all the grafted membranes revealed the growth of Cal? minerals after 7 days immersion in SBF. The sample with lowest surface grafting yield (24% coverage), a smooth graft morphology and relatively high hydrophobicity (theta(adv) = 120 degrees, theta(rec) = 80 degrees) showed carbonated hydroxyapatite growth covering the surface. On the other hand, the samples with high surface grafting yield (76% and 100%), a globular graft morphology and hydrophilic surfaces (theta(adv) = 60 degrees and 80 degrees, theta(rec) = 25 degrees and 15 degrees, respectively) exhibited irregular growth of non-apatitic Cap minerals. Irreversibly adsorbed protein measured after a 1 h immersion in serum solution was quantified by the amount of nitrogen on the surface using XPS, as well as by weight increase. All grafted membranes adsorbed 3-6 times more protein than the unmodified membrane. The sample with the highest surface coverage adsorbed the most protein. Osteoblast-like SaOS-2 cells cultured for 3 h revealed significantly higher levels of cell attachment on all grafted membranes compared to unmodified ePTFE. Although the morphology of the cells was heterogeneous, in general, the higher grafted surfaces showed a much better cell morphology than both the low surface-grafted and the control unmodified sample. The suite of in vitro tests confirms that a judicious choice of grafted monomer such as the phosphate-containing methacrylate monomer (MOEP) significantly improves the bioactivity of ePTFE in vitro. (c) 2005 Elsevier Ltd. All rights reserved.

Polymeric grafting of acrylic acid onto poly(3-hydroxybutyrate-co-3-hydroxyvalerate): Surface functionalization for tissue engineering applications

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) melt processed disks and solvent cast films were modified by graft co-polyinerization with acrylic acid (AAc) in methanol solution at ambient temperature using gamma irradiation (dose rate of 4.5 kGy/h). To assess the presence of carboxylic acid groups on the surface, reaction with pentafluorophenol was performed prior to X-ray photoelectron spectroscopy analysis. The grafting yield for all samples increased with monomer concentration (2-15%), and for the solvent cast films, it also increased with dose (2-9 kGy). However, the grafting yield of the melt processed disks was largely independent of the radiation dose (2-8 kGy). Toluidine blue was used to stain the modified materials facilitating, visual information about the extent of carboxylic acid functionalization and depth penetration of the grafted copolymer. Covalent linking of glucosamine to the functionalized surface was achieved using carbodimide chemistry verifying that the modified substrates are suitable for biomolecule attachment.

Methacryloxyethyl phosphate-grafted expanded polytetrafluoroethylene membranes for biomedical applications

Expanded polytetrafluoroethylene (ePTFE) membranes were modified by graft copolymerization with methacryloxyethyl phosphate (MOEP) in methanol and 2-butanone (methyl ethyl ketone (MEK)) at ambient temperature using gamma irradiation. The effect of dose rate (0.46 and 4.6 kGyh(-1)), monomer concentration (1-40 %) and solvent were studied and the modified membranes were characterized by weight increase, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). XPS was used to determine the % degree of surface coverage using the C-F (ePTFE membrane) and the C-C (MOEP graft copolymer) peaks. Grafting yield, as well as surface coverage, were found to increase with increasing monomer concentration and were significantly higher for samples grafted in MEK than in methanol solution. SEM images showed distinctly different surface morphologies for the membranes grafted in methanol (smooth) and MEK (globular), hence indicating phase separation of the homopolymer in MEK. We propose that in our system, the non-solvent properties of MEK for the homopolymer play a more important role than solvent chain transfer reactions in determining grafting outcomes. (c) 2005 Society of Chemical Industry.

The effects of ionizing radiation on the reproductive capacity of adult Penaeus (Marsupenaeus) japonicus (Bate)

The reproductive capacity of adult Penaeus (Marsupenaeus) japonicus (Bate) was assessed after exposure to ionizing gamma radiation from a cobalt-60 source. Males and females were each exposed to 0, 10 and 20 Gray (Gy) of ionizing radiation (IR) and reciprocally crossed to give nine mating combinations. Fecundity and hatch rate of resulting spawnings were used as measures of reproductive capacity. IR significantly (P