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Biomedical subjects

C M Stanford

Publications and source records attributed to C M Stanford.

33 records · Page 2Linked to original sources
PubMed1995

Rapidly forming apatitic mineral in an osteoblastic cell line (UMR 106-01 BSP).

This study evaluated a rapid biomineralization phenomenon exhibited by an osteoblastic cell line, UMR 106-01 BSP, when treated with either organic phosphates [beta-glycerophosphate (beta-GP), Ser-P, or Thr-P], inorganic phosphate (P(i)), or calcium. In a dose-dependent manner, these agents (2-10 mM) stimulated confluent cultures to deposit mineral in the cell layer (ED50 of approximately 4.6 mM for beta-GP (30 +/- 2 nmol Ca2+/microgram DNA) and approximately 3.8 mM (29 +/- 2 nmol Ca2+/microgram DNA) for P(i)) with a plateau in mineral formation by 20 h (ET50 approximately 12-15 h). beta-GP or P(i) treatment yielded mineral crystals having an x-ray diffraction pattern similar to normal human bone. Alizarin red-S histology demonstrated calcium mineral deposition in the extracellular matrix and what appeared to be intracellular paranuclear staining. Electron microscopy revealed small, needle-like crystals associated with fibrillar, extracellular matrix deposits and intracellular spherical structures. Mineral formation was inhibited by levamisole (ED50 approximately 250 microM), pyrophosphate (ED50 approximately 1-10 microM), actinomycin C1 (500 ng/ml), cycloheximide (50 micrograms/ml), or brefeldin A (1 microgram/ml). These results indicate that UMR 106-01 BSP cells form a bio-apatitic mineralized matrix upon addition of supplemental phosphate. This process involves alkaline phosphatase activity, ongoing RNA and protein synthesis, as well as Golgi-mediated processing and secretion.

Alkaline Phosphatase↗
PubMed1995

Proliferative and phenotypic responses of bone-like cells to mechanical deformation.

Limited in vivo and in vitro experiments suggest that bone and bone-like cells respond to mechanical signals in a trigger-like rather than a dose-response fashion; i.e., they fail to respond until they have been stimulated with some given number of cycles of loading, and then once they respond, additional cycles produce little or no effect. To explore this notion, rat calvaria-derived osteoblast-like cells and the cell line MC3T3-E1 were plated at a high cell density (5,000 cells/mm2) on silicone membranes coated with type-I collagen and were allowed to attach for 24 hours. The membranes then were exposed to vacuum pressure (-1 kPa, 0.5 Hz) on a daily basis, and cultures were assayed every 2 days for 2 weeks. The proliferation of nontransformed cells increased 7-fold with as few as four daily cycles but not with one cycle per day. Furthermore, 1,800 cycles of vacuum did not result in a greater response than four cycles per day. We observed inverse phenotypic responses: the expression of osteocalcin was depressed compared with controls in the cultures of osteoblast-like cells that were strained with as few as four cycles per day. Alkaline phosphatase activity was depressed in the cultures of both the osteoblast-like cells and the MC3T3-E1 cells exposed to low vacuum pressures (-1 kPa) with four daily cycles of vacuum pressure. Increasing the vacuum magnitude did not affect the occurrence of a "trigger response" between one and four cycles of vacuum application.(ABSTRACT TRUNCATED AT 250 WORDS)

3T3 Cells↗
PubMed1995

Cellular deformation reversibly depresses RT-PCR detectable levels of bone-related mRNA.

Osteoblastic cells respond to mechanical stimuli with alterations in proliferation and/or phenotypic expression. In some cases, these responses occur within only a few applications of stimuli (i.e. 'cycle-dependent trigger response') rather than in a dose-dependent manner. To explore potential mechanisms of the cycle dependent trigger response, we raised the following questions: (1) Does strain of bone cells alter gene expression; if so, how quickly does it occur and how long does it last? (2) Are alterations in message level strain magnitude dependent? (3) Are alterations in steady-state message levels cycle dependent? Cultures were evaluated for osteocalcin mRNA one week following a daily stretch application at four stretch magnitudes and four cycle numbers and compared to nonstretched controls. Steady state mRNA message was ascertained prior to and at 10, 20, 30, 60, 120, 180, and 240 min following initiation of stretch. Following mRNA isolation, first strand cDNA synthesis was performed and fluorometrically quantitated. A reverse transcriptase based PCR (RT-PCR) approach allowed assessment of osteocalcin mRNA levels from microcultures (50,000 cells per 10 microliters culture or 5000 cells mm2) of rat calvarial osteoblasts. Optimized PCR was performed using primers to the bone specific protein, osteocalcin (OC) and two 'housekeeping' genes, beta-actin and GAP-DH. PCR products were separated on 4% agarose gels and band intensities digitized with relative quantitation based on internal standards in each gel. The lowest magnitude of stretch (- 1 KPa) at 1800 cycles per day reproducibly depressed message for osteocalcin, but not beta-actin when assayed immediately following the cessation of strain application. By three hours following the initiation of stretch, message levels returned to control values. At the time of stretch cessation, the 1800 cycle stretch regimen diminished (p < 0.0001) steady-state osteocalcin message independently of the four stretch magnitudes. Stretch for 300 cycles failed to depress (p = 0.05) osteocalcin message cultures at any time, but 600 cycles depressed message by 30 min. By one and two hours, cultures stretch 600, 900, and 1800 cycles showed similar levels of message depression. Four hours following the initiation of stretch, message levels returning to nonstrained levels in all groups. We conclude that alterations in cell response to strain are in part mediated by gene expression, that alterations last 3-4 h in this system, and that the message mechanism itself exhibits a trigger-response dependency to cycle number.

Actins↗
PubMed1994

Characterizations of titanium implant surfaces. III.

There are several reports in the literature concerning the similarities and the differences between the oxide on cpTi and Ti-6A1-4V alloy; however, their biological sequelae are not entirely known. In this work, a series of surface characterization techniques were used in conjunction with short term in vitro biological assays to assess the effects of materials selection (cpTi and Ti alloy) on osteoblast-like cell responses. Surface analysis indicated that with the exception of oxide thickness, there were no significant differences in surface characteristics between the two implant materials. These results were reflected in the biological studies, where the levels of cell attachment and adaptation of the attached cells to the titanium surfaces were similar. These results are in general agreement with previous in vivo studies and continue to indicate that cpTi and Ti alloy are suitable, biologically compatible materials for fabrication of dental implants.

Animals↗
PubMed1994

How connective tissues temporally process mechanical stimuli.

Connective tissues have the capacity to adapt to a changing mechanical environment. Considerable experimental evidence suggests, however, that tissues respond to only a small portion of mechanical loads they experience and thus ignore the majority of the load they experience. Evidence suggests that tissues 'temporally process' these external mechanical signals in specific ways: 1) they respond in a trigger-like manner after a relatively few events or cycles of loading; 2) they respond only to some window of strain magnitude; 3) they exhibit a refractory period after a response; 4) they have a memory for previous stimuli. These characteristics do not necessarily mirror distinct cellular phenomena but rather reflect typical features of experimental and theoretical models. The manner in which tissues temporally process mechanical signals can be explained by energy transfer over short periods of time from the matrix to the second messenger systems. Much of the energy input into connective tissues during deformation is recovered, and relatively little is 'lost' and thus potentially transferable to the cells. Since evolutionary principles suggest that a pathway of response needs to be defined by biologically optimal use of energy, it is logical that cells would have evolved mechanisms to respond to a small finite level of integrated stimuli in order to rapidly respond (fitness) especially in times of environmental duress.

Adaptation, Physiological↗
PubMed1994

Bone cell expression on titanium surfaces is altered by sterilization treatments.

Phenotypic responses of rat calvarial osteoblast-like cells (RCOB) were evaluated on commercially pure titanium (cpTi) surfaces when cultured at high density (5100 cells/mm2). These surfaces were prepared to three different clinically relevant surface preparations (1-micron, 600-grit, and 50-microns-grit sand-blast), followed by sterilization with either ultraviolet light, ethylene oxide, argon plasma-cleaning, or routine clinical autoclaving. Osteocalcin and alkaline phosphatase, but not collagen expression, were significantly affected by surface roughness when these surfaces were altered by argon plasma-cleaning. In general, plasma-cleaned cpTi surfaces demonstrated an inverse relationship between surface roughness and phenotypic markers for a bone-like response. On a per-cell basis, levels of the bone-specific protein, osteocalcin, and the enzymatic activity of alkaline phosphatase were highest on the smooth 1-micron polished surface and lowest on the roughest surfaces for the plasma-cleaned cpTi. Detectable bone cell expression can be altered by clinically relevant surfaces prepared by standard dental implant preparation techniques.

Alkaline Phosphatase↗
PubMed1992

Short-term plasma-cleaning treatments enhance in vitro osteoblast attachment to titanium.

The purpose of this research was to characterize the in vitro cellular behavior of osteoblast-like cells on titanium surfaces prepared with argon plasma-cleaning (PC) treatments for various lengths of time. The highest levels of cell attachment were observed for surfaces which had been plasma-treated for one min. Surface analyses indicated that although PC treatments dramatically improved surface wettability, the presence of inorganic contaminants was observed with longer treatment times and may have interfered with cell attachment. Further work is suggested to investigate the longer-term phenotypic expression of osteoblasts when grown on implant surfaces.

Analysis of Variance↗
PubMed1991

The concept of osseointegration and bone matrix expression.

Osseointegration has been defined as the direct structural and functional connection between ordered, living bone and the surface of a load-carrying implant. To date, this concept has been described by descriptive histological and ultrastructural criteria but not by biochemical means. This review evaluates the basic science work performed on this concept and then applies the concept to the principle of osseous healing. Specific studies are cited where alterations in the healing response are due to clinical management of implant placement and how studies of surface properties may lead to further insights on implant design and prognosis. In addition, a review of bone expression as a function of in vitro stress applications is given. This is followed by an indepth review of the collagens and noncollagenous proteins, described to date, within isolated bone matrix. It is this collagenous matrix (especially type I) that is described as being close to and oriented with a glycoprotein component next to the implant surface. In turn, the large family of noncollagenous proteins are important in mediating bone proliferation, matrix accumulation, orientation, mineralization, and turnover. This section is followed by a discussion of specific growth factors as they may relate to osseous healing around an implant.

Animals↗
PubMed1991

In vitro periodontal ligament fibroblast attachment to plasma-cleaned titanium surfaces.

The objective of this research was to characterize the in vitro cellular behavior of fibroblast-like cells derived from rat periodontal ligament on commercially pure titanium surfaces which were sterilized by a variety of treatments. Following standard surface preparation protocols, the Ti specimens were sterilized by either steam autoclaving, exposure to ethylene oxide gas, exposure to ultraviolet light, or plasma-cleaning in argon for either one min or five min. Fibroblast-like cells in serum-supplemented media were incubated on the various Ti specimens for up to two h. In general, the levels of cell attachment for plasma-cleaned surfaces were significantly higher than those for steam-sterilized surfaces, but were significantly lower than the attachment levels for both the ultraviolet-treated surfaces and the tissue culture plastic control. The duration of plasma cleaning itself did not have a significant effect on the percentage of cell attachment at any time period. SEM evaluations indicated that by two h, the cellular morphology was different on the variously treated specimens. These studies indicate that the method of sterilization following implant surface preparation can affect the initial in vitro biological events of cell attachment and spreading.

Animals↗
PubMed1987

Radiopacity of light-cured posterior composite resins.

Radiographic images of teeth and restorations were used to evaluate the radiopacity of 11 light-cured posterior composite resins. The radiopacity of these composite resins provided enough variation on radiographs so that clinicians distinguished the images of the restoration from adjacent tooth structure.

Aluminum↗
PubMed1984

Effects of backing reflectance and mold size on polymerization of photo-activated composite resin.

The in vitro depth of cure of a visible-light-activated microfilled composite, as influenced by backing reflectance and mold size, was investigated using microhardness measurements. More extensive polymerization was obtained with higher values of backing reflectance. Mold size larger than the cross-sectional area of the photo-activating beam also increased the degree of polymerization. Backing reflectance and mold size are influential factors and need to be defined in depth-of-cure evaluations if in vitro measurements are to be valid predictors of clinical performance.

Chemical Phenomena↗
PubMedDate not supplied

In situ evaluation of sodium hexametaphosphate-containing dentifrices.

The purpose of this investigator-blinded, five-treatment, crossover human intraoral study was to evaluate the effects of two experimental dentifrice formulations containing either stannous fluoride (SnF(2)) or sodium fluoride (NaF) packaged with sodium hexametaphosphate in a dual-phase delivery system on demineralization-remineralization using an in situ model system. The experimental dentifrice formulations' ability to alter demineralization-remineralization was compared to a series of three controls: SnF(2)-positive control, NaF-positive control and no-fluoride placebo-negative control. The single-section crown model, developed at the University of Iowa, was used to assess the fluoride efficacy of two experimental products versus the placebo containing no fluoride and positive controls. The results of the current in situ study suggest a clinical level of anticaries activity for the experimental SnF(2) and NaF dentifrice formulations that was as good as either of the positive controls, when evaluated using polarized light microscopy. This supports the conclusion that the use of the sodium hexametaphosphate ingredient does not interfere with the normal fluoride activity of these toothpastes. In addition, the experimental SnF(2) product was numerically better than both the NaF and placebo controls at preventing demineralization of sound root surfaces.

Adult↗
PubMedDate not supplied

Osteoblastic cell attachment to hydroxyapatite-coated implant surfaces in vitro.

Hydroxyapatite (HA) used as a coating for implants can exhibit varying levels of interaction with the biologic environment. The crystallinity of the HA-based coating has been shown to control the rate of dissolution and appears to play a role in the initial cellular interaction with the implant surfaces. An osteoblastic cell attachment assay was employed to examine the cell attachment to untreated and pretreated (pH 5.2, 24 hours) titanium and HA coatings of low (50%), medium (75%), and high (90%) crystallinity. A slightly higher percentage of cell attachment (%CA) was found on untreated and pretreated HA surfaces as compared to the titanium surface. No significant difference could be found in the %CA between the 3 levels of crystallinity. However, higher levels of %CA were observed on pretreated HA surfaces than on untreated HA surfaces (t test, P < .05). Elevated calcium and phosphate levels in culture medium did not have any effect on cell attachment. Scanning electron microscopic examinations revealed surface degradation of the HA coating following pretreatment in the simulated inflammatory media (pH 5.2, 24 hours). The results suggest that the altered surface topography may influence the initial cell attachment to HA surfaces.

Analysis of Variance↗