Biomedical subjects

Leo Tjäderhane

Publications and source records attributed to Leo Tjäderhane.

18 recordsLinked to original sources

PubMed2007

Collagen XVIII modulation is altered during progression of oral dysplasia and carcinoma.

BACKGROUND: Collagen XVIII is a ubiquitous basement membrane (BM) component and a precursor of endostatin. METHODS: Using immunohistochemistry and in situ hybridization, we studied the expression and localization of collagen XVIII in different stages of normal oral wound healing, epithelial dysplasia and squamous cell carcinoma (SCC). RESULTS: In mild epithelial dysplasias collagen XVIII appeared as a continuous signal in the BM, whereas in severe epithelial dysplasias and in the invasive areas of oral SCCs collagen XVIII was absent. In situ hybridization showed that collagen XVIII mRNA expression did not decrease in severe dysplasia or oral carcinoma samples when compared with the mild dysplasias. CONCLUSIONS: The results indicate that the absence of collagen XVIII protein in severe oral dysplasias is related to the processing of the protein rather than to changes in mRNA expression.

Angiogenesis Inhibitors↗

PubMed2006

Matrix metalloproteinase-8 (MMP-8) is the major collagenase in human dentin.

OBJECTIVE: Previously an unidentified collagenolytic metalloprotease together with gelatinase (matrix metalloproteinase-2, MMP-2), and enamelysin (MMP-20) have been detected in human dentin. The aim of the study was to characterize dentinal collagenolytic enzymes. Furthermore, we hypothesized that the dentinal MMPs are protected by the mineral phase, and studied the stability of dentinal MMPs. DESIGN: To characterize dentinal collagenolytic enzymes, we used Western blotting with specific antibodies against MMP collagenases (MMP-1, -8, and -13) and cathepsin K. MMP-8 immunofluorometric assay (IFMA) was also used for MMP-8 detection, and functional collagenase activity was examined with type I collagen degradation assay. The stability of dentinal MMPs was examined by autoclaving dentin blocks before protein extraction and subsequent examination of protein levels and the activities of dentin collagenase and gelatinases. RESULTS: MMP-8 (collagenase-2) was detected in dentin both with Western blot and IFMA, and dentinal samples also cleaved the intact type I collagen into characteristic 3/4(alphaA)-cleavage products in vitro. No other collagenases or cathepsin K were detected. In autoclaved samples no MMP-8 was found, but gelatinase activity was observed in protein fractions of mineralized dentin. CONCLUSIONS: MMP-8 represents the major collagenase in human dentin. Unlike MMP-8, dentinal gelatinases can be detected after autoclave treatment of dentin, indicating their high resistance to external sample treatment procedures.

Adult↗

PubMed2006

Reactivation of inactivated endogenous proteolytic activities in phosphoric acid-etched dentine by etch-and-rinse adhesives.

Auto-degradation of collagen matrices occurs in resin-infiltrated dentine by the slow action of host-derived matrix metalloproteinases. As phosphoric acid-etching inactivates these endogenous enzymes, it is puzzling how hybrid layers created by simplified etch-and-rinse adhesives can degrade in vivo. This study tested the null hypothesis that there are no differences in the relative proteolytic activities of mineralised dentine, acid-etched dentine, and etch-and-rinse adhesive-treated acid-etched dentine. Powdered dentine prepared from extracted human teeth was treated with 17% EDTA, 10% phosphoric acid, or with five simplified etch-and-rinse adhesives that were applied to 10% phosphoric acid-etched dentine. The gelatinolytic activity of the dentine powder was assayed using fluorescein-labelled gelatine. TEM examination of the air-dried, treated dentine powder was performed to confirm the presence of remnant mineralised dentine after acid-etching. 17% EDTA significantly reduced the relative proteolytic activity (73.2%) of the untreated mineralised dentine powder (control), while 10% phosphoric acid-etched dentine exhibited the highest reduction (98.1%). Treating the acid-etched dentine powder with any of the five simplified etch-and-rinse adhesives resulted in the reactivation of the proteolytic activity, with a significant negative linear correlation (P<0.05) between the increases in fluorescence and the corresponding pH values of the adhesives. It is concluded that simplified etch-and-rinse adhesives can reactivate endogenous enzymatic activities in dentine that are previously inactivated by phosphoric acid-etching. The amount of enzyme reactivated may even exceed the original quantity present in untreated mineralised dentine. This provides an explanation for the degradation of hybrid layers after acid-etched dentine matrices are infiltrated with these adhesives.

Dentin↗

PubMed2006

Matrix metalloproteinases: contribution to pathogenesis, diagnosis and treatment of periodontal inflammation.

Matrix metalloproteinases (MMPs) form a family of enzymes that mediate multiple functions both in the tissue destruction and immune responses related to periodontal inflammation. The expression and activity of MMPs in non-inflamed periodontium is low but is drastically enhanced to pathologically elevated levels due to the dental plaque and infection-induced periodontal inflammation. Soft and hard tissue destruction during periodontitis and peri-implantitis are thought to reflect a cascade of events involving bacterial virulence factors/enzymes, pro-inflammatory cytokines, reactive oxygen species and MMPs. However, recent studies suggest that MMPs can also exert anti-inflammatory effects in defence of the host by processing anti-inflammatory cytokines and chemokines, as well as by regulating apoptotic and immune responses. MMP-inhibitor (MMPI)-drugs, such as doxycycline, can be used as adjunctive medication to augment both the scaling and root planing-treatment of periodontitis locally and to reduce inflammation systematically. Furthermore, MMPs present in oral fluids (gingival crevicular fluid (GCF), peri-implant sulcular fluid (PISF), mouth-rinses and saliva) can be utilized to develop new non-invasive, chair/bed-side, point-of-care diagnostics for periodontitis and dental peri-implantitis.

Cytokines↗

PubMed2006

Activation of gelatinolytic/collagenolytic activity in dentin by self-etching adhesives.

Mild acids are known to activate dentin matrix metalloproteinase (MMPs). All self-etching dental adhesives are acidic (pH 1.5-2.7) and may activate dentin MMPs. The purpose of this study was to compare the ability of several all-in-one adhesives to activate gelatinolytic and collagenolytic activities in powdered mineralized dentin. Powdered dentin made from human teeth was mixed with all-in-one adhesives (Clearfil Tri-S Bond, G-Bond, Adper Prompt L-Pop) or a self-etching primer (Clearfil SE Bond primer) for varying times and then the reaction was stopped by extracting the adhesives using acetone. Fresh untreated mineralized dentin powder had a gelatinolytic activity of 3.31 +/- 0.39 relative fluorescent units (RFU) per mg dry weight (24 h) that increased, over storage time, to 87.5 RFU mg(-1) (24 h) after 6-8 wk. When fresh powder was treated with acidic Tri-S Bond, the gelatinolytic activity increased from 3.24 +/- 0.70 RFU mg(-1) to > 112.5 RFU mg(-1) (24 h) after 20 min and then remained unchanged. Monomers with lower pH values produced less activity. There was a significant, direct correlation between gelatinolytic activity and pH, with Tri-S giving the highest activity. Coating dentin powder with Tri-S resin prevented fluorescent substrates from gaining access to the enzyme, even though it activated the enzyme. In conclusion, self-etch adhesives may activate latent MMP and increase the activity to near-maximum levels and contribute to the degradation of resin-dentin bonds over time.

Adhesives↗

PubMed2005

Mechanical stability of resin-dentin bond components.

OBJECTIVES: To evaluate the effects of long-term storage on the mechanical properties of the components of resin-dentin bonds, that is, resin composite, adhesive system, demineralized and mineralized dentin. METHODS: Specimens of resin composite (Z250) and adhesive systems (Single Bond-SB; One-Step-OS and Clearfil Liner Bond 2V-CL) were cast in molds. Dentin specimens were prepared from dentin discs obtained from the crowns of extracted human molars. Specimens of demineralized dentin were obtained by immersion of dentin discs for 6 days in 0.5 mol/l EDTA (pH 7.0). Both dentin and resin-based substrates were shaped to hourglass or I-beam specimens that were used to determine the true stress (TS) or apparent modulus of elasticity (E), respectively. Control specimens were subjected to tensile testing at 0.6 mm/min after 24 h of immersion in distilled water. Experimental specimens were stored at 37 degrees C in either distilled water or mineral oil and tested after 12 months. The data of each group were individually analyzed by ANOVA and Tukey's test. RESULTS: Both TS and E of the resin-based materials decreased significantly after 12 months of storage in water (p < 0.05) except the TS of SB (p > 0.05). No changes were observed for specimens of mineralized dentin, regardless of storage condition (p > 0.05). Storage of demineralized dentin in water did not cause any significant effect in either TS or E (p > 0.05), however, significant reductions of TS and E of demineralized dentin occurred after storage in oil for 1 year (p < 0.05). SIGNIFICANCE: Storage time and medium may be deleterious to the mechanical properties of the resin-dentin bond components, which ultimately could compromise the durability of resin-dentin bonds.

Analysis of Variance↗

PubMed2005

Analysis of gene and protein expression in healthy and carious tooth pulp with cDNA microarray and two-dimensional gel electrophoresis.

Complementary DNA (cDNA) microarray and two-dimensional (2-D) gel electrophoresis, combined with mass spectrometry, enable simultaneous analysis of expression patterns of thousands of genes, but their use in pulp biology has been limited. Here we compared gene and protein expression of pulp tissues from sound and carious human teeth using cDNA microarray and 2-D gel electrophoresis to evaluate their usefulness in pulp biology research and to identify the genes with changes in carious teeth. The cDNA microarray revealed several differentially expressed genes and genes with a high expression in both tissues. These genes have various functions, e.g. effects on vascular and nerve structures, inflammation, and cell differentiation. Variability between cDNA hybridizations indicates that the overall gene expression pattern may vary significantly between individual teeth. The 2-D gel electrophoresis revealed no change between healthy and diseased tissue. The identification of 96 proteins in the pulp tissue revealed none of the gene products with corresponding high/different mRNA expression in cDNA microarray. Interestingly, we detected also a hypothetical protein (putative nucleoside diphosphate kinase), and present therefore the first evidence for the existence of this protein. Even though the methods reveal potentially important gene expression, they may currently have only limited value in in vivo pulp biology research.

Adolescent↗

PubMed2004

Expression of collagen XVIII and MMP-20 in developing teeth and odontogenic tumors.

Collagen XVIII is a basement membrane (BM) component, whereas MMP-20 (enamelysin) is a matrix metalloproteinase predominantly expressed in teeth. Since MMP-20 was found to degrade collagen XVIII, we studied the co-expression of these proteins in dental tissues. Collagen XVIII surrounded the developing tooth during early and late bell stages and was also present in developing enamel. Western blotting indicated that developing enamel contains collagen XVIII N-terminal fragments of the frizzled variant. Enamelysin was co-localized with collagen XVIII in the developing enamel matrix and stratum intermedium. Electron microscope analysis showed that total mineral, calcium and phosphorus contents of enamel were slightly increased in collagen XVIII null mice but the analysis revealed no visible defects in the enamel or dentin structures. In odontogenic tumors MMP-20 and collagen XVIII were co-localized in the enamel-like tumor matrix. Our results show that collagen XVIII is present in developing teeth, but its absence seems not to be critical for the development of the teeth.

Animals↗

PubMed2004

Matrix metalloproteinase-13 (MMP-13, collagenase-3) is highly expressed in human tooth pulp.

Matrix metalloproteinases (MMPs) and their specific tissue inhibitors (TIMPs) participate into extracellular matrix degradation in physiological and pathological conditions. We hypothesized that MMP expression in pulp tissue changes in response to caries attack and investigated the gene expression profiles of MMPs and TIMPs in pulp tissue of sound and carious teeth with cDNA microarray. cDNA microarray demonstrated an extremely high MMP-13 (collagenase-3) mRNA expression in pooled pulp samples of sound and carious teeth, with less pronounced expression of MMP-16 (MT3-MMP) and TIMP-1. Real-time quantitative polymerase chain reaction of individual pulp samples revealed a wide range of the MMP-13 expression level between pulp samples with possible downregulation of MMP-13 expression during caries progression. Western blot and immunohistochemical staining confirmed the presence of MMP-13 with no observable differences between sound and carious teeth pulp tissues. The results reveal that MMP-13 is expressed and synthesized in pulp tissue, an interesting feature considering the very limited expression of MMP-13 in normal adult tissues. Further studies with a larger sample size are needed to clarify the changes in MMP-13 expression during caries progression.

Adult↗

PubMed2004

Physical and chemical properties of chlorhexidine and calcium hydroxide-containing medications.

This study was performed to evaluate the physicochemical properties (pH, contact angle, working time, radio-opacity, and viscosity) of chlorhexidine (CHX) and calcium hydroxide-containing medications in gel form in different concentrations. The pH value was assessed with a pH meter. The contact angle was measured with a goniometer. The radio-opacity and working time measurements were taken in accordance with the standards of the International Organization for Standardization. The viscosity was evaluated using a Brookfield RVDV viscometer. The results showed that CHX did not affect the pH, radio-opacity, and working time of the calcium hydroxide-containing medications (p < 0.05). However, adding CHX lowered the contact angle and increased the viscosity of calcium hydroxide significantly. This research showed that CHX in different concentrations and in combination with calcium hydroxide has satisfactory physicochemical properties to be used as an intracanal medication.

Calcium Hydroxide↗

PubMed2004

The benefit of the operating microscope for access cavity preparation by undergraduate students.

Although studies show the operating microscope (OM) provides benefits for endodontists, the benefit to undergraduates has not been evaluated as completely as for specialists. The purpose of this study was to determine whether using the OM would improve students' performance in endodontic access cavity preparation and canal identification. Thirty-six dental students prepared access cavities and located canals in extracted maxillary molars, before and after training, which varied according to group. The standard group received a lecture and practice in preparation of access cavities. The microscope group received identical instruction using the OM. The control group received lectures only. All groups received equal content and instruction time (2 hr 20 min). Faculty graded preparations according to a multidimensional 5-point rating scale. Using the OM, the microscope group improved significantly in access cavity preparation (p <0.05) and significantly outperformed both standard and control groups in accuracy of identifying canals (p <0.05).

Analysis of Variance↗

PubMed2003

Efficacy of chlorhexidine- and calcium hydroxide-containing medicaments against Enterococcus faecalis in vitro.

OBJECTIVE: We sought to assess the efficacy of chlorhexidine (CHX) and calcium hydroxide, Ca(OH)(2), against Enterococcus faecalis in vitro. STUDY DESIGN: The effect of CHX (0.2% and 2% in gel or solution) and Ca(OH)(2) (alone or with 0.2% CHX gel) was evaluated by using the agar diffusion test and an in vitro human root inoculation method, to measure zone of inhibition or bacterial growth with optical density analysis, respectively. For optical density analysis, samples from infected root canals were collected after 7 days of medication and were cultured for 24 hours in brain-heart infusion to detect viable bacteria. RESULTS: In the agar diffusion test, CHX was effective against E faecalis in a concentration-dependent fashion, but Ca(OH)(2) alone had no effect. In the root canal inoculation test, CHX was significantly more effective against E faecalis than Ca(OH)(2) was (P < .05), but there were no significant differences between the modes of medication or concentrations of CHX. CONCLUSIONS: CHX is effective against E faecalis in vitro. Further in vivo studies are needed to confirm the value of CHX in clinical treatment.

Anti-Infective Agents, Local↗

PubMed2003

Laminin-5 gamma 2 chain is colocalized with gelatinase-A (MMP-2) and collagenase-3 (MMP-13) in odontogenic keratocysts.

BACKGROUND: Odontogenic keratocyst (KC) differs from other epithelial odontogenic cysts in regard to increased epithelial proliferation and a strong tendency to recur. Laminin-5 (Ln-5) is an epithelial anchoring filament component, which after modulation by certain matrix metalloproteinases (MMPs), like MMP-2 and MMP-13, induces epithelial cell migration. METHODS: Using in situ hybridization and immunohistochemistry, we studied the Ln-5 gamma-2 chain expression related to the expression of MMP-2, -8, and -13 in different odontogenic cysts, including radicular cysts (RC; n = 11), follicular cysts (FC; n = 11), and odontogenic keratocysts (KC; n = 16). RESULTS: Ln-5 mRNA was present in all cysts examined, while less than half of KCs and RCs (33 and 40%, respectively) demonstrated MMP-2 mRNA. MMP-13 mRNA was present in all KC samples. Ln-5 protein was located as a continuous ribbon in BM zone of all KCs, and MMP-2 and MMP-13 immunoreactivities colocated significantly with Ln-5 in that area. MMP-8 was expressed by stromal macrophages and epithelial goblet cells, but never located in BM zone. CONCLUSIONS: Our results indicate that the colocalization of Ln-5 with MMP-2 or MMP-13, but not with MMP-8, in BM zone of KCs, may be related to special characteristics of KC.

Basement Membrane↗

PubMed2003

Expression profile of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs in mature human odontoblasts and pulp tissue.

Previous studies have demonstrated that (at least) matrix metalloproteinase (MMP)-2, -8, -9, -14 and -20 are expressed by human odontoblasts. Here, we analysed the expression of 19 MMPs and their specific tissue inhibitors (TIMP)-1, -2 and -3) -1, -2 and -3 in mature human odontoblasts and pulp tissue. Since MMP-20 is almost exclusively expressed by the dentin-pulp complex cells, we further analysed the effect of transforming growth factor (TGF)-beta1 and bone morphogenetic protein (BMPs)-2 on its expression. Matrix metalloproteinase-9 served as a positive control for growth factor responsiveness. It was found that MMP-1, -2, -9, -10, -11, -13, -14, -15, -16, -17, -19, -20 and -23, in addition to TIMP-1, -2 and -3 were expressed by both odontoblasts and pulp tissue. Neither MMP-3 nor MMP-12 were expressed in odontoblasts or pulp tissue, and MMP-7, -8, -24 and -25 were expressed only in the odontoblasts; MMP-2, -10, -11, -14 and -20 were expressed more abundantly by odontoblasts, whereas pulp tissue expressed more MMP-13 and MMP-17. Transforming growth factor-beta1 (1 ng ml(-1)) and BMP-2 (100 ng ml(-1)) did not markedly affect MMP-20 mRNA expression. In contrast, TGF-beta1 alone and with BMP-2 significantly upregulated MMP-9 mRNA by 2.4-fold and by 2.6-fold, respectively, in odontoblasts, while in pulp tissue no effects could be detected. The wide-scale expression of MMPs and TIMPs by mature human odontoblasts and pulp tissue suggests that they may participate in dentin matrix organization prior to mineralization, and that growth factors may further control dentin matrix modeling by differentially regulating individual MMPs.

Adolescent↗

PubMed2003

Periapical inflammation affecting coronally-inoculated dog teeth with root fillings augmented by white MTA orifice plugs.

Placement of orifice plugs has been suggested to augment the seal of conventional root canal fillings. This study assessed in vivo the efficacy of white mineral trioxide aggregate (MTA) plugs in preventing periapical inflammation subsequent to coronal inoculation of root-filled teeth. The two-rooted mandibular premolars of six beagle dogs were conventionally prepared and filled with gutta-percha and sealer. A white MTA orifice plug was placed into one canal in each tooth. Pulp chambers were inoculated with plaque except for 12 teeth (negative control), and restored. Radiographs were taken at regular intervals. At 10 months, dogs were killed and jaw blocks processed for histology. None of the roots revealed radiographic or histologic evidence of severe inflammation. Mild inflammation was observed in 17% and 39% of the roots with and without an orifice plug, respectively (McNemar, p > 0.05). Without development of severe inflammation, the seal augmentation efficacy of MTA orifice plugs could not be determined.

Aluminum Compounds↗

PubMed2002

Effect of dietary carbamyl phosphate on dentine apposition in rat molars.

In rats, sucrose increases dental caries and impairs odontoblastic function by reducing dentine apposition during primary dentinogenesis. A high-sucrose diet also affects negatively a pulp or dentine function that appears to regulate solute or fluid movement within rat dentinal tissue. In earlier work it was found that carbamyl phosphate could significantly reverse sucrose-induced cariogenesis and also stimulate sucrose-depressed movement of dentinal fluid through a mechanism involving parotid function(s). In the current study, the possibility that carbamyl phosphate could overcome the sucrose-induced reduction in dentine apposition was examined. Weanling rats were fed a high-sucrose diet supplemented or not with carbamyl phosphate for 5 weeks. Dentine apposition was measured planimetrically in sagittal sections of the molars. The effect of carbamyl phosphate was similarly tested in parotidectomized animals. Carbamyl phosphate significantly reduced the deleterious effect of sucrose on dentine apposition by 58% in the first molars. However, the reduction in dentine apposition that followed parotidectomy was not altered by carbamyl phosphate supplementation. The possibility that the beneficial effect of carbamyl phosphate on dentinogenesis involves a parotid function is entertained.

Analysis of Variance↗

PubMed2002

Substantive antimicrobial activity in chlorhexidine-treated human root dentin.

OBJECTIVE: The aim of this in vitro study was to assess the substantive antimicrobial activity of different medicaments in human root dentin. STUDY DESIGN: Canals of 98 roots were enlarged to standard size and medicated for 7 days with the following: (1) 2% chlorhexidine (CHX) gel, (2) 0.2% CHX gel, (3) 2% CHX solution, (4) Ca(OH)(2), (5) Ca(OH)(2)+ 0.2% CHX gel, (6) 2% CHX solution + a 25% CHX-containing controlled-release device, (7) saline, and (8) gel vehicle. After medication, canals were inoculated with Enterococcus faecalis for 21 days. Dentin samples were collected with Gates-Glidden burs into brain heart infusion broth, and bacterial growth was assessed with spectrophotometric analysis of optical density after 72 hours of incubation. RESULTS: Mean optical densities were significantly lower for groups with 2% CHX (1, 3, and 6) when compared with those of the controls (P < .05, analysis of variance with the Tukey test). Other groups did not differ significantly from the controls. CONCLUSIONS: Canal dressing for 1 week with 2% CHX may provide residual antimicrobial activity against E faecalis.

Analysis of Variance↗

PubMed2002

The mechanism of pulpal wound healing.

The favourable response of exposed pulp tissue against a variety of materials used for pulp capping in experimental conditions, as observed by hard tissue (reparative dentine) formation, demonstrates an intrinsic capacity of pulp tissue for healing. However, in the clinical situation, in which a pulpal exposure is usually accompanied by a long-term external irritation with the subsequent long-term inflammatory response to that irritation, the outcome of pulp capping procedures is not as predictable. While some of the factors related to the defensive reactions and healing after pulp exposure and capping procedures are well understood, the mechanisms and importance of others remain less well-known. Understanding the mechanisms regulating the spread of inflammation and necrosis in pulp tissue, and the factors regulating healing after closure of the wound, would facilitate the development of new and better treatment procedures with more predictable outcomes. In this review, some of the aspects considered to be important in pulpal wound healing are discussed.

Animals↗