Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats.

Cells from fetal or neonatal skeleton can synthesize bone-like tissue in vitro. In contrast, formation of bone-like tissue in vitro by cells derived from adult animals has rarely been reported and has not been achieved using cells from bone marrow. We have explored development of bone-like tissue in vitro by bone marrow stromal cells. Marrow stromal cells obtained from 40-43-day-old Wistar rats were grown in primary culture for 7 days and then subcultured for 20-30 days. Cells were cultured in either alpha-minimal essential medium containing 15% fetal bovine serum, antibiotics, and 50 micrograms/ml ascorbic acid, or the above medium supplemented with either 10 mM Na-beta-glycerophosphate, 10(-8) M dexamethasone, or a combination of both. Cultures were examined using phase-contrast microscopy, undemineralized and demineralized tissue histology, histochemistry (for alkaline phosphatase activity), immunohistochemistry (for collagen type, osteonectin, and bone Gla-protein), scanning and transmission electron microscopy, energy dispersive X-ray microanalysis, and X-ray diffraction. Collagenous, mineralized nodules exhibiting morphological and ultrastructural characteristics similar to bone were formed in the cultures, but only in the presence of both beta-glycerophosphate and dexamethasone. Cells associated with the nodules exhibited alkaline phosphatase activity. The matrix of the nodules was composed predominantly of type-I collagen and both osteonectin and Gla-protein were present. X-ray microanalysis showed the presence of Ca and P, and X-ray diffraction indicated the mineral to be hydroxyapatite. The nodules were also examined for bone morphogenetic protein-like activity. Paired diffusion chambers containing partly demineralized nodules and fetal muscle were implanted intraperitonealy in rats. Induction of cartilage in relation to muscle was observed histologically after 40 days in the chambers. This finding provided further support for the bone-like nature of the nodules. The observations show that bone-like tissue can be synthesized in vitro by cells cultured from young-adult bone marrow, provided that the medium contains both beta-glycerophosphate and, particularly, dexamethasone.

Evaluation of shear strength at the cement-endodontic post interface.

The strength characteristics of the cement-implant interface were evaluated for smooth-tapered, threaded, and porous-surfaced endodontic implants with the use of different cements. Specifically, tensile and torsional shear strengths were measured for zinc phosphate, polycarboxylate, glass-ionomer, silicophosphate, and AH-26 cements. The results indicated superior shear strength characteristics for threaded endodontic implants on axial loading. However, this strength was diminished when torsional forces were applied. Porous-surfaced endodontic implants showed strong resistance to both axial and torsional loading.

Evaluation of the retention of endodontic implants.

The study investigated the retentive strength of endodontic implants measured by forced removal (pull-out or push-out tests) as a function of implant design and cement type. Smooth-tapered, threaded, and an innovative porous-surfaced implant were evaluated. Specimens were cemented in single-rooted human teeth with five different cements: zinc phosphate, polycarboxylate, glass ionomer, silicophosphate, or AH-26. The results indicated superior retention for the threaded and porous-surfaced implants, and stronger retention with glass-ionomer and AH-26 cements.

Threaded versus porous-surfaced designs for implant stabilization in bone-endodontic implant model.

An endodontic implant model system was used to compare the effect of implant design on stabilization in bone. Specifically a porous-surfaced design was compared to conventional threaded and smooth-tapered endodontic implant designs. All implants were placed in immediate function thereby assessing the effect of early limited movement on the fixation achieved. A total of eighty-three endodontic implants were inserted in the mandibles of six adult mongrel dogs. Animals were sacrificed immediately after implantation and after 3, 6, and 12 months. Implants were evaluated by clinical and radiographic examination and after animal sacrifice by pull-out tests of the implant from the tissues, SEM examination of the pulled-out implants and, finally, histology. The pull-out test results indicated increasing shear strength with implantation time for the porous-surfaced implants in contrast to the gradual loss of fixation for the threaded implants and the continuous low shear strength for the smooth implants. Histological studies and SEM examination indicated the reason for these changes. Smooth implants became encapsulated by fibrous connective tissue from early post-implantation time periods. Threaded implants, although initially mechanically interlocked with bone, developed a fibrous connective tissue capsule that gradually thickened with time until, by 6 months, little mechanical interlock of bone and implant was present. It was assumed that this fibrous capsule thickening was caused by implant movement. The porous-surfaced implants, however, became stabilized by bone ingrowth and showed more extensive bone formation within the surface pores with time. It is concluded that for implants that are made functional immediately after implantation, as in this study, porous-surfaced implants can become strongly fixed by bone ingrowth, in contrast to conventional threaded or smooth-surfaced designs, thus presenting a more favourable long term prognosis.

A histological assessment of the initial healing response adjacent to porous-surfaced, titanium alloy dental implants in dogs.

We report here the results of a histological assessment of the initial healing response following implantation into the dog mandible of a porous-surfaced, titanium alloy endosseous dental implant. Two implants were placed in edentulous areas on each side of the mandible of each dog and covered with a full-thickness mucoperiosteal flap. The implant sites on one side of the mandible were allowed to head for four weeks, while those on the other side were allowed to head for eight weeks before the animals were killed. Histological specimens were obtained and assessed both qualitatively and by computer-assisted morphometry. All but one of the 24 implants were well-tolerated and healed with a variable ingrowth of bone into the porous-surface geometry. The histomorphometric measurements revealed that bone ingrowth had reached a plateau by four weeks of initial healing.

Observations on the effect of movement on bone ingrowth into porous-surfaced implants.

Although porous-surfaced orthopedic implants have been designed for fixation by bone ingrowth, there is clinical evidence that this does not always occur. Initial implant movement relative to host bone can result in attachment by a nonmineralized fibrous connective tissue layer. The ranges of movement that result in either bone or fibrous connective tissue fixation are observed in dogs in two independent studies. Experimentally, bone ingrowth can occur in the presence of some movement, albeit very small (up to 28 mu), while excess movement (150 mu or more) can result in attachment by mature connective tissue ingrowth.

A scanning electron microscopic study of the odontoblast process in human coronal dentine.

It is generally accepted that the odontoblast process occupies the dentinal tubules only in the inner part of the dentine, extending approx 0.7 mm from the pulp in both animals and man. Twenty-three premolars, molars and third molars from subjects aged 11-24 yr, all caries-free or only slightly decayed, were processed immediately after extraction by one of four methods. (I) Teeth were split in liquid nitrogen and then fixed, dehydrated in ascending ethanol, and dried by critical point drying (CPD). (II) Teeth were fractured by use of a mallet and a chisel and then prepared as in Method I. (III) The root was cut off using a diamond disk or a mallet and a chisel and the crown was then fixed, fractured in liquid nitrogen, dehydrated and CPD. (IV) As Method III but following fixation the teeth were freeze-dried. All specimens were examined in the scanning electron microscope (SEM). In specimens prepared by Methods II, III or IV the odontoblast process was limited to the inner third of crown dentine. However in all the specimens prepared by Method I the odontoblast process extended to the dentine-enamel junction.