Biomedical subjects

S Keila

Publications and source records attributed to S Keila.

15 recordsLinked to original sources

PubMed2006

Prostaglandin E2 (PGE2) increases the number of rat bone marrow osteogenic stromal cells (BMSC) via binding the EP4 receptor, activating sphingosine kinase and inhibiting caspase activity.

Prostaglandin E(2) (PGE(2)) is bone-anabolic, i.e. stimulates bone formation and increases bone mass. In this study, we explored possible intracellular mechanisms of its increase of osteogenic cells in rat bone marrow. Adherent rat bone marrow cells were counted after 12-48 h or cultured for 21 days and mineralized nodules were counted. Also, apoptosis of marrow cells was measured after in vivo PGE(2) injection. PGE(2) (100 nM) increased 2-3 fold the number of adherent BMSC, an effect which was mediated via binding the EP(4) receptor since it was mimicked by forskolin and 11-deoxy-prostaglandin E(1) (PGE(1)) and was blocked by DDA and L-161982 (EP(4) antagonist). PGE(2) stimulated sphingosine kinase (SPK) activity since its effects were blocked by DMS (SPK inhibitor) and mimicked by SPP (SPK product). PGE(2) reduced the activity of caspase-3 and -8 in BMSC and their inhibitors increased BMSC number and nodule formation. In vivo, PGE(2) prevented the increase in the apoptosis of bone marrow cells caused by indomethacin. We propose that PGE(2) exerts an anti-apoptotic effect on BMSC, thereby increasing their number and subsequent osteoblastic differentiation. Such an effect could explain how PGE(2) stimulates bone formation in vivo.

Animals↗

PubMed2004

A selective EP4 receptor antagonist abrogates the stimulation of osteoblast recruitment from bone marrow stromal cells by prostaglandin E2 in vivo and in vitro.

Recent evidence indicates that systemic administration of PGE2 increases bone formation and bone mass via activation of the EP4 receptor. Previously, we demonstrated that osteoblastic recruitment from rat bone marrow stromal cells (BMSC) is a major mechanism for the anabolic effect of PGE2. In this study, we used a selective EP4 antagonist to test if the stimulation of osteoblast differentiation from rat BMSC in vitro and in vivo involves the EP4 receptor. In vitro, PGE2 (100 nM) increased nodule formation and alkaline phosphatase (ALP) activity in cultures of rat BMSC 1.5- to 2-fold. These effects were abolished by the EP4 antagonist at 10(-6) M but not 10(-9) M. Furthermore, PGE2 increased the number of surviving adherent BMSC by approximately 225% and the EP4 antagonist prevented this effect as well. The antagonist had no effect on basal levels of nodule formation and adherent cell number. In vivo, daily systemic administration of PGE2 at 6 mg/kg for 2 weeks increased cancellous bone area (by approximately 50%) and increased nodule formation (measured as mineralized area) in ex vivo stromal cultures by approximately 50%. Pre-administration of the EP4 antagonist at 10 mg/kg abrogated both the increase in bone mass as well as the increase in nodule formation. These data indicate that PGE2 stimulates osteoblastic commitment of BMSC via activation of the EP4 receptor.

Alkaline Phosphatase↗

PubMed2004

In vitro effects of enamel matrix proteins on rat bone marrow cells and gingival fibroblasts.

Emdogain (EMD), a formulation of Enamel Matrix Proteins (EMP), is used clinically for periodontal regeneration, where it stimulates cementum formation and promotes gingival healing. In this study, we investigated the in vitro effects of EMD on rat bone marrow stromal cells (BMSC) and gingival fibroblasts (GF). EMD (at 25 micro g/mL) increased the osteogenic capacity of bone marrow, as evidenced by approximately three-fold increase in BMSC cell number and approximately two-fold increase in alkaline phosphatase (ALP) activity and mineralized nodule formation. The presence of EMD in the initial stages (first 48 hrs) of the culture was crucial for this effect. In contrast, EMD did not induce osteoblastic differentiation of GF (evidenced by lack of mineralization or ALP activity) but increased up to two-fold both their number and the amount of matrix produced. These in vitro data on BMSC and GF could explain the promotive effect of EMD on bone formation and connective tissue regeneration, respectively.

Alkaline Phosphatase↗

PubMed2002

Accuracy of a new apex locator: an in vitro study.

AIM: The purpose of this study was to test in an in vitro model the accuracy of a Bingo 1020 electronic apex locator, to compare the results to those of a well known apex locator, Root ZX, as well as to those of the radiographic method of tooth length determination. METHODOLOGY: A total of 120 extracted teeth, preserved in Thymol solution and kept refrigerated, was used for the study. The experiment was performed on single-rooted teeth and on one-root canal, chosen randomly, in multirooted teeth. The teeth were randomly divided into 12 groups of 10 teeth each. After access preparation, the actual length (AL) was measured. The teeth were embedded in an alginate model specially developed for testing apex locators. Electronic tooth length measurements (EL) were carried out prior to root canal preparation using the two electronic apex locators (EAL) - Root ZX and Bingo 1020; three measurements were taken and an average computed. After the third measurement, the file was left in the root canal and a periapical radiograph was taken. The radiographic length (RL) was recorded by measuring the file length from the coronal reference point to the tip of the file. Each root canal was then prepared to a no. 40 K-file diameter using a standardized technique; saline was used for irrigation. Upon completion of the root canal preparation, EL measurements were taken by each EAL in dry conditions and with different irrigation solutions. Each measurement was repeated three times. The RL was recorded according to the last EL measurement. Results were subjected to statistical analysis. RESULTS: In all parameters tested, a significant statistical difference was found between Bingo 1020 and the Root ZX. Measurements obtained using the Bingo 1020 were consistently closer to the AL (0.08 mm) than those obtained using the Root ZX. Both EALs measured the tooth length with great accuracy and a positive correlation of 0.76 (P = 0.00) existed between the two devices. No significant difference was found between the two apex locators when measurements were taken with the different irrigants (P = 0.34) and the content of the root canal did not affect the accuracy of the measurements. Lengths obtained by calculations from the radiographs were longer than the AL as well as the length obtained by both EALs (P = 0.00). CONCLUSIONS: The Bingo 1020 proved to be as reliable as Root ZX and was user friendly. Under the experimental conditions, electronic measurements were more reliable than radiographs in the process of root length determination.

Analysis of Variance↗

PubMed2002

Inhibition of cyclo-oxigenase-2 activity does not abolish the anabolic effect of prostaglandin E2 in vivo or in vitro.

It was previously reported that the expression of cyclo-oxigenase-2 (COX-2) is induced by prostaglandin E(2) (PGE(2)) in vitro in an osteogenic cell line and organ culture, suggesting an autoamplification mechanism. In this study, we first tested whether this phenomenon also occurs in bone tissue in vivo and found that a single anabolic dose of PGE(2) (5 mg/kg) induced (between 30 and 120 min) in rat tibiae, an increase in the mRNA level of COX-2 (2.5- to 9-fold) but not that of COX-1. Secondly, to test whether COX-2 activity in generating endogenous prostaglandins (PGs) is required for the in vivo anabolic properties of PGE(2), young male rats were injected daily with either vehicle (8% ethanol) or 5 mg/kg PGE(2) for 21 days. PGE(2)-injected rats received, 45 min prior to PGE(2), either dimethyl sulphoxide (as vehicle) or one of two doses of NS-398, a selective COX-2 inhibitor: a low dose (3 mg/kg) or a high dose (10 mg/kg). PGE(2) increased bone formation (measured as cancellous mineralizing surface, mineral apposition rate and bone formation rate) and bone mass (measured as cancellous bone area and surface and cortical width). None of these increases was suppressed by pre-administration of NS-398. In contrast, the high dose of NS-398 effectively suppressed an increase in rat hind-paw volume induced by a local carrageenan injection. Furthermore, since COX-2 inactivation may affect PG receptor expression, we found that pre-administration of NS-398 did not abolish the induction in EP(4) receptor mRNA levels, caused by PGE(2) in rat bone tissue. For in vitro testing, rat femoral bone marrow stromal cell cultures were initiated and were incubated in the absence or presence of PGE(2) at 100 nM (as an inducer) and with increasing concentrations of NS-398 (10(-8) M to 10(-5) M) for 21 days, after which time mineralized (Von-Kossa positive) nodules were counted. PGE(2) increased nodule formation as previously reported; however, NS-398 reduced nodule formation in both control and PGE(2)-treated cultures to the same extent. We conclude that while the level of COX-2 mRNA is increased in vivo by administration of PGE(2), inhibition of its activity (i.e. generation of endogenous PGs) does not abolish the anabolic effect of PGE(2).

Animals↗

PubMed2001

Systemic prostaglandin E2 increases cancellous bone formation and mass in aging rats and stimulates their bone marrow osteogenic capacity in vivo and in vitro.

Prostaglandin E(2) (PGE(2)) has been shown to exert a bone anabolic effect in young and adult rats. In this study we tested whether it possesses a similar effect on bone formation and bone mass in aging rats. Fifteen-month-old rats were injected daily with either PGE(2) at 5 mg/kg or vehicle for 14 days. PGE(2) treatment stimulated the rate of cancellous bone formation (a approximately 5.5-fold increase in bone formation rate), measured by the incorporation of calcein into bone-forming surfaces at the tibial proximal metaphysis. This effect resulted in increased cancellous bone area (+54%) at the same site. Since PGE(2) treatment resulted in a much higher proportion of bone surface undergoing bone formation and thus lined with osteoblasts, we tested the hypothesis that PGE(2) stimulates osteoblast differentiation from bone marrow precursor cells both in vivo and in vitro. We found that ex vivo cultures of bone marrow stromal cells from rats injected for 2 weeks with PGE(2) at 5 mg/kg per day yielded more ( approximately 4-fold) mineralized nodules and exhibited a greater (by 30-40%) alkaline phosphatase activity compared with cultures from vehicle-injected rats, attesting to a stimulation of osteoblastic differentiation by PGE(2). We also compared the osteogenic capacity of bone marrow from aging (15-month-old) versus young (5-week-old) rats and its regulation by PGE(2) in vitro. Bone marrow stromal cell cultures from aging rats exhibited a greatly diminished osteogenic capacity, reflected in reduced nodule formation ( approximately 6% of young animals) and lower alkaline phosphatase activity ( approximately 60% of young animals). However, these parameters could be stimulated in both groups of animals by incubation with 10-100 nM PGE(2). The magnitude of this stimulation was greater in cultures from aging rats (+550% vs +70% in nodule formation of aging compared with young rats). In conclusion, we demonstrate here that PGE(2) exerts a bone anabolic effect in aging rats, similar to the effect we and others have reported in young, growing rats. The PGE(2)-stimulated bone formation, which augments bone mass, most likely results from recruitment of osteoblasts from their bone marrow stromal precursors.

Aging↗

PubMed1999

In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media.

The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objectives of this study were to evaluate the effectiveness of six different media: culture medium, alpha minimal essential medium (alpha-MEM), milk, Hank's balanced salt solution (HBSS), ViaSpan and conditioned medium (CM) to preserve cultured periodontal ligament fibroblasts (PDLF). Periodontal ligament fibroblasts were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at 4 degrees C. A control group was incubated with culture medium at 37 degrees C. After incubation, cell viability was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Storage of PDLF up to 24 h decreased their vitality by only 2%-14%. Vitality of the PDLF after 2, 8 and 24 h was highest when stored in milk or HBSS (91%-97%) and lowest when stored in ViaSpan or CM (82%-93%). PDLF stored for 2-8 h in various media had a mitogenic capacity comparable to the control. However, increasing the storage period to 24 h decreased the mitogenicity of the cells by 3%-39%. The highest mitogenicity was found in PDLF stored in milk or HBSS and the lowest in CM or ViaSpan. The clonogenic capacity of the cells dropped by 38%-71% after 24 h and was the best indicator of the deteriorating effect of long storage. Milk and HBSS were the most effective in preserving the clonogenic capacity. Nevertheless, reduction in the viability, mitogenicity or clonogenic capacity was statistically significant in nearly all the tested media only after 24 h of incubation. In conclusion, HBSS and milk were the most effective media for preserving the viability, mitogenicity and clonogenic capacity after storage for up to 24 h at 4 degrees C.

Adenosine↗

PubMed1998

Apical extent of rotary canal instrumentation with an apex-locating handpiece in vitro.

PROBLEM: The Tri Auto ZX (J. Morita Co., Kyoto, Japan) is a cordless endodontic handpiece with a built-in apex locator that is programmed to reverse the direction of rotation when the file reaches a predetermined apical level or when torque becomes excessive. OBJECTIVE: The purpose of this investigation was to examine the apical extent of rotary canal instrumentation and the ability to maintain apical constriction with the Tri Auto ZX at different automated settings. STUDY DESIGN: In 60 extracted teeth, canals were measured to the apical constriction, first visually and then electronically with the Tri Auto ZX; then they were instrumented with nickel titanium rotary files. For the instrumentation, the automatic apical reverse mechanism of the handpiece was set to 1, 1.5, or 2; these settings correspond to different distances from the apical foramen. Instrumentation was carried out apically until rotation was reversed by the automatic apical reverse function; the instrumented length was then measured, and the canal was filled with gutta-percha and sealer. The integrity of the apical constriction was assessed by exposing the apical 4 mm of the canal and observing the dentin-cementum junction. Paired t-tests were used to compare the visually measured length, the electronically measured length, and the instrumented length for each tooth. RESULTS: On average, the electronically measured length was 0.54 mm shorter than the visually measured length (p < 0.05). When the automatic apical reverse mechanism's setting was 1, the instrumented length was 0.1 mm shorter than the electronically measured length; when the setting was 1.5, the instrumented length was 0.36 mm shorter than the electronically measured length (p < 0.01). Results were inconsistent when the setting was 2. CONCLUSION: Instrumentation with the automatic apical reverse feature set at 1 consistently approximated the apical constriction; however, the constriction was frequently enlarged.

Dental Alloys↗

PubMed1997

Systemic administration of an anabolic dose of PGE2 in young rats increases the osteogenic capacity of bone marrow.

Prostaglandin E2 (PGE2) possesses significant anabolic properties when administered systemically (i.e., it increases bone formation and, consequently, bone mass). We recently characterized the effects of a 3 week administration of 6 mg/kg PGE2 into young rats and showed it increases cortical and cancellous bone mass and mechanical strength in long bones and bone density in the calvaria. We also found that a single dose of PGE2 induces the expression of early-response genes (c-fos, c-jun, and egr-1) in bone marrow cells within these two types of bone. These observations, together with findings by others of new cancellous bone formation in PGE2-treated animals, suggested that recruitment of osteoblasts from their precursors is a major mechanism of the anabolic effect of PGE2. To test this hypothesis directly, we injected PGE2 (6 mg/kg) or vehicle into 4-week-old rats for 2 weeks and then assessed the osteogenic potential of bone marrow in an ex vivo culture system. Primary and first-passage bone marrow cultures were established in the presence of beta-glycerophosphate, ascorbate, and dexamethasone, and osteogenic differentiation was measured by bone nodule formation and alkaline phosphatase activity. This regimen increased bone mass expressed as femoral ash weight by 4.7% and tibial cancellous bone area by 38.3%. Nodule formation at 21 days was increased in both primary and first-passage cultures from PGE2-treated rats despite seeding of the same number of marrow cells. Alkaline phosphatase activity was elevated in both primary and first-passage cultures from PGE2-treated rats beginning 6-10 days after culture initiation. Cell proliferation was only slightly elevated in cultures from PGE2-treated rats. These data strongly suggest that in vivo administration of PGE2 induces the proliferation or differentiation of osteoprogenitor cells in bone marrow, and this effect takes a major part in its anabolic effect in vivo.

Alkaline Phosphatase↗

PubMed1997

Reliability of different electronic apex locators to detect root perforations in vitro.

The aim of this study was to evaluate the accuracy of the electronic apex locator Root ZX compared to the Sono Explorer Mark II Junior and Apit III in detecting root perforations. Extracted human teeth (n = 30) were perforated artificially in the middle third of the root and embedded in a box containing alginate. Detection of all perforations were carried out with K-files attached to the apex locators tested. The teeth were radiographed after each electronic measurement. The actual location of the tip of the file in relation to the external root surface and the radiographic determination of the perforations were carried out using a stereomicroscope. The mean distance of the tip of the file from the external outline of the root surface was short for all instruments and clinically acceptable (0.06-0.25 mm). There was no clinically significant difference between the morphometric measurements and the radiographic measurements. No statistical significance was found between large perforations (0.55-0.60 mm) and small perforations (0.25-0.40 mm). Under the conditions of this study, all electronic apex locators tested were acceptable clinical tools in the detection of root perforations.

Analysis of Variance↗

PubMed1994

Bone marrow from mechanically unloaded rat bones expresses reduced osteogenic capacity in vitro.

Bone formation during mechanical unloading is reduced, mainly as a result of osteoblastic hypofunction. At the same time, the total number of osteoblasts per long bone is also markedly reduced. We tested the hypothesis that the number of osteogenic precursors present in the bone marrow stroma was concomitantly diminished by using an in vitro cell culture system in which femoral adherent bone marrow cells differentiate into active osteoblasts and produce bone-like nodules. Hindlimbs of 32-day-old male rats were either immobilized (unloaded) by sciatic neurectomy (immo) or sham operated (sham) and animals were killed after 11 days. Femora were either ashed to determine bone mass or used to generate bone marrow cultures. Adherent marrow cells were cultured in the presence of ascorbic acid, beta-glycerophosphate, and dexamethasone. Bone mass was significantly reduced in unloaded femora (by 16%) and tibiae (by 18%). The number of adherent cells (determined on day 6) was reduced by 50% in the immo group. Reduced cell number did not result from slower proliferation in culture since [3H]thymidine incorporation on days 4 and 6 was similar in the two groups. The osteogenic potential in vitro of marrow from unloaded bones was diminished compared with that from loaded ones as evidenced by (1) lower alkaline phosphatase (ALP) activity per mg protein (by 25-40%, examined on days 6 and 12), and (2) reduced nodule formation (by 70%, expressed as percentage of the dish area stained with Alizarin Red S on day 21). None of these changes occurred in the contralateral limb of operated (immobilized) animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Uncontrolled bleeding during endodontic treatment as the first symptoms for diagnosing von Willebrand's disease. A case report.

Von Willebrand's disease has been established as the most common hereditary hemorrhagic disease. Endodontic treatment, when possible, rather than extraction is the treatment of choice in patients suffering from such a disease. A case is presented in which, during regular endodontic treatment, uncontrollable bleeding occurred, suggestive of a generalized bleeding disorder. A thorough hematologic examination revealed von Willebrand's disease, which the patient was not aware of. The dental surgeon can play an important role in the diagnosis and treatment of such patients.

Adult↗

PubMed1990

Developmental anomaly of permanent teeth related to traumatic injury.

A case is presented where a 4-year-old girl was injured in an automobile accident. The dental injury, intrusive luxation of the maxillary deciduous incisors, seemed unimportant at the time, since the child was in critical condition. The situation was first evaluated 5 years later, when the child was referred for treatment of an apical periodontitis originating from the right central incisor. The present case reflects several important aspects that were overlooked at the time of injury: the potential of intruded luxated deciduous teeth to cause developmental anomalies to the permanent dentition; the necessity of a longer follow-up period to detect such anomalies; and the need to develop effective treatment methods.

Child↗

PubMed1989

Conservative treatment of root perforations using apex locator and thermatic compactor--case study of a new method.

Endodontic perforation, a serious complication in dental practice, is among the most common problems associated with endodontic procedures. Recommended methods of treatment are surgical or conservative using calcium hydroxide paste. A method in which perforations can be obturated immediately after their detection is described. The method utilizes an apex locator for measurement and detection of the perforation and a two-step treatment of thermatic compaction for filling the root canal as well as the perforation. A number of cases with a long follow-up are presented.

Adult↗

PubMed1989

Hypersensitivity to sodium hypochlorite.

Sodium hypochlorite (NaOCl), the most commonly used irrigant in endodontics, is known to produce allergic reactions. This fact, however, is rarely mentioned in endodontic texts. Until now, only two cases of adverse reaction describing accidental injection of NaOCl have been reported. In the current study a case is presented in which hypersensitivity to household bleach was proven with skin patch tests. The clinician was warned of the possible health hazard from using NaOCl in this patient by the past medical history. Endodontic therapy was carried out with an irrigant not containing NaOCl after the allergy to NaOCl was verified. Treatment was uneventful. It is suggested that before any endodontic treatment in which sodium hypochlorite is to be used, the patient should be asked about hypersensitivity to household bleaching materials.

Adolescent↗