E. Maconnachie

The effects of inhibitors of cysteine-proteinases and collagenase on the resorptive activity of isolated osteoclasts

The effects of specific inhibitors of cysteine-proteinases ((Z-Phe-Ala-CHN2: benzyloxycarbonyl-phenyl-alanyl alanyl diazomethane and E-64: trans-epoxy-succinyl-L-leucylamido(4-guanidino)-butane) and collagenase and collagenase ((Cl-1: N-(3-N-benzyloxycarbonyl amino-1-R-carboxypropyl)-L-leucyl-O-methyl-L-tyrosine N-methylamide) have been tested on the osteoclastic resorption of dentine. Chick osteoclasts were cultured in the presence or absence of 12.5 microM Z-Phe-Ala-CHN2, 40 or 60 microM E-64, or 40 or 100 microM Cl-1 for 1 or 2 days. In addition, osteoclasts were cultured on oyster shell calcitostracum with or without 12.5 microM Z-Phe-Ala-CHN2. Specimens were studied by light microscopy to count cells and resorption features and by scanning electron microscopy (SEM) stereophotogrammetry for the measurement of the depths, plan-areas and volumes of resorption pits. The numbers, depths and volumes (but not the plan-areas) of the resorption pits in dentine were significantly reduced by Z-Phe-Ala-CHN2 and E-64. Thus, for a given plan-area, the volumes and the depths of resorption pits were smaller in these experimental groups compared with control dentine specimens. The overall inhibition of resorption was at least 75%. Cl-1 did not have this inhibitory effect on the numbers or sizes of resorption pits in dentine. When the oyster calcitostracum was used as a substrate for the osteoclasts, Z-Phe-Ala-CHN2 did not reduce the numbers or volumes of pits, but increased the plan-areas and prevented the formation of deeper pits.(ABSTRACT TRUNCATED AT 250 WORDS)

Human cranial bone structure and the healing of cranial bone grafts: a study using backscattered electron imaging and confocal microscopy.

SummaryThis study explored the microstructure of human cranial bone at different ages, and the survival, remodelling and modelling of cranial bone grafts. A combination of reflection and fluorescence confocal optical microscopy and scanning electron microscopy in the backscattered electron imaging mode was employed to examine highly polished block faces of plastic-embedded bone fragments as harvested for grafting, or recovered after a period in situ as a graft. The methods enabled remarkably detailed information on bone content, maturation and turnover to be gleaned from tiny scraps of bone. Microfractures in the harvested bone were repaired at the graft site, with welding of old and new bone indicating revascularization. Human cranial bone grafts successfully stimulated bone cell differentiation, supported new bone formation on resorbed and unresorbed surfaces, and underwent bone turnover. The type and organization of new bone reflected the growth rate and maturation of the graft rather than the age of the patient.

Treatment with lithium salts reduces ethanol dehydration shrinkage of glutaraldehyde fixed tissue

SummaryChanges in the area of glutaraldehyde fixed 15 day p.c. mouse embryo limbs were recorded using a Quantimet 720 image analysing computer attached to a light microscope: during a period of treatment with an isotonic salt solution (mostly halides of the alkali or alkaline earth metals); a subsequent wash with distilled water; and dehydration through a 30, 50, 70, 80, 90, and 100% ethanol series. Pretreatment with NaCl, KCl, RbCl had no significant effect. Treatment with LiCl, LiNO3, LiF (0.03 M), CsF and CsCl caused an increase (relative to Na, K or Rb treated samples) in the specimen volume during dehydration, which persisted in 100% ethanol. Li treated samples showed the largest post-critical-point-drying (CPD) volumes, followed by Cs treated tissue. Pretreatment with Be, Mg, Ca, Sr and Ba chlorides caused shrinkage and the 100% ethanol and post-CPD volumes of these samples were all lower than those treated with the monovalent cation containing salts.

SEM study of surface alterations of bioactive glasses and glass--ceramics in a bony implantation bed.

Back scattered electron (BSE) images of glasses and glass-ceramics with different degrees of bone-bonding behaviour have been investigated. After the implantation of probes in the femur of rats changes of the surface material, induced by leaching and corrosion (material response) and of the tissue in the drill hole (host response), were studied. The results demonstrate advantages of using BSE images and give further information for understanding the sequential changes in the implant material which occur mainly in bone-bonding materials. The density of the leached zone was not homogeneous and it therefore might be possible that preferential leaching of the glass matrix contributes to the mineralization of extracellular matrix. These findings could be of use for further development of surface reactive materials.