Effect of cyclic compressive loading on the bond strength of an adhesive system to dentin after collagen removal

Purpose: The objective of this study was to verify the effect of cyclic compressive loading on the shear bond strength of an adhesive system following collagen removal. Materials and Methods: Sixty bovine teeth were divided into 4 groups based on the adhesive procedure used: groups 1 and 2 - etching with 35% phosphoric acid and application of the Single Bond adhesive system; groups 3 and 4 - after etching, a 10% sodium hypochlorite solution was applied for 1 min before the application of the adhesive. In all the specimens, a Z100 resin cylinder was built up over the bond area. Groups 2 and 4 were submitted to 500,000 cycles with a load of 100 N. Results: The mean values for the shear bond test (MPa) were: group 1: 7.37 ± 1.15; group 2: 5.72 ± 1.66; group 3: 5.95 ± 1.21; group 4: 3.66 ± 1.12. There was no difference between groups 1 and 2 (p > 0.01). Between groups 1 and 3, 2 and 4, and 3 and 4 there was a significant difference (p < 0.01). The majority of the specimens demonstrated an adhesive failure. Conclusion: The application of sodium hypochlorite on dentin decreased the values of shear bond strength, as did the load cycling in the group treated with sodium hypochlorite.

Doxazosin reduces cell proliferation and increases collagen fibers in rat prostatic lobes

We investigated the effects of doxazosin (Dox), an alpha-adrenoceptor antagonist used clinically for the treatment of benign prostatic hyperplasia (BPH), on the rat prostatic complex by assessing structural parameters, collagen fiber content, cell proliferation, and apoptosis. Adult Wistar rats were treated with Dox (25 mg/kg per day), and the ventral (VP), dorsolateral, and anterior prostate (AP) regions of the prostate complex were excised at 3, 7, and 30 days after treatment. At 24 h before being killed, the rats were injected once with 5-bromodeoxyuridine (BrdU; thymidine analog) to label mitotically active cells. The prostates were weighed and processed for histochemistry, morphometry-stereology, immunohistochemistry for BrdU, Western blotting for proliferating cell nuclear antigen (PCNA), and the TUNEL reaction for apoptosis. Dox-treated prostate lobes at day 3 presented increased weight, an enlarged ductal lumen, low cubical epithelial cells, reduced epithelial folds, and stretched smooth muscle cells. However, at day 30, the prostates exhibited a weight reduction of ∼20% and an increased area of collagen and reticular fibers in the stromal space. Dox also reduced epithelial cell proliferation and increased apoptosis in the three prostatic lobes. Western blotting for PCNA confirmed the reduction of cell proliferation by Dox, with the AP and VP being more affected than the dorsal prostate. Thus, Dox treatment alters epithelial cell behavior and prostatic tissue mechanical demand, inducing tissue remodeling in which collagen fibers assume a major role. © 2007 Springer-Verlag.

Smear layer removal and collagen fiber exposure using tetracycline hydrochloride conditioning

Aim: Smear layer removal and collagen fiber exposure may improve periodontal treatment and regeneration. This in vitro study assessed smear layer removal and collagen fiber exposure after tetracycline hydrochloride (TTC) application on root surfaces using scanning electron microscopy (SEM). Methods and Materials: Root cementum was removed with diamond burs followed by scaling and root planning. Four hundred fifty samples were divided into ten groups: a control (saline application) and nine different TTC concentrations were applied at doses of 10, 25, 50, 75, 100, 125, 150, 200, and 250 mg/ml. The TTC application was performed in all groups in three different ways (passive, brushing, and burnishing) and at three different periods of conditioning (1, 2, and 3 minutes). A previously trained, calibrated, and blind examiner evaluated photomicrographs of the samples using Sampaio's index (2005). Statistical analysis was performed using the Kruskal-Wallis' and Dunn's tests. Results: The concentrations of 50 mg/mL and 75 mg/mL applied by burnishing were the most effective in smear layer removal and collagen fiber exposure. Both the passive mode of application (p=0.0001) and 1 minute period of application (p=0.002) were the least effective. Conclusions: The concentrations of 50 mg/mL and 75 mg/mL applied by burnishing during 2 or 3 minutes were the most effective. Clinical Significance: These parameters may be applied in periodontal procedures involving TTC root conditioning to optimize results.

Treatment of gingival recession with collagen membrane and DFDBA: A histometric study in dogs

In a previous study, we evaluated the findings related to the use of resorbable collagen membranes in humans along with DFDBA (demineralized freeze-dried bone allograft). The aim of this subsequent study was to histometrically evaluate in dogs, the healing response of gingival recessions treated with collagen membrane + DFDBA (Guided Tissue Regeneration, GTR) compared to a coronally positioned flap (CPF). Two types of treatment were randomly carried out in a split-mouth study. Group 1 was considered as test (GTR: collagen membrane + DFDBA), whereas Group 2 stood for the control (only CPF). The dogs were given chemical bacterial plaque control with 0.2% chlorhexidine digluconate during a 90-day repair period. Afterwards, the animals were killed to obtain biopsies and histometric evaluation of the process of cementum and bone formation, epithelial migration and gingival level. A statistically significant difference was found between groups with a larger extension of neoformed cementum (GTR = 32.72%; CPF = 18.82%; p = 0.0004), new bone (GTR = 23.20%; CPF = 09.90%; p = 0.0401) and with a smaller area of residual gingival recession in the test group (GTR = 50.69%; CPF = 59.73%; p = 0.0055) compared to the control group. The only item assessed that showed no statistical difference was epithelial proliferation on the root surface, with means of 15.14% for the GTR group and 20.34% for the CPF group (p = 0.0890). Within the limits of this study we concluded that the treatment of gingival recession defects with GTR, associating collagen membrane with DFDBA, showed better outcomes in terms of a larger extension of neoformed cementum and bone, as well as in terms of a smaller proportion of residual recessions.

Morphology of collagen fibers and elastic system fibers in actinic cheilitis

Background: Actinic cheilitis (AC) is a premalignant condition intimately related to exposure of the lips to sun rays. Aim: The objective of this study was to evaluate the elastic and collagen fibers in the lamina propria of AC. The degree of epithelial atypia was correlated with the quantity of elastic and collagen fibers. Materials and Methods: Fifty-one cases were investigated. One slide was stained with hematoxylin-eosin for the evaluation of atypia, the second was stained with Weigert′s resorcin-fuchsin for the assessment of elastic fibers, and the third slide was stained with Mallory′s trichrome for the analysis of collagen fibers. Results: Ordinal logistic regression analysis revealed a significant correlation between the presence of atypia and collagen fibers (P<0.05). Conclusions: It was concluded that there seems to be a reduction in the quantity of collagen fibers in cases of moderate and severe atypia. No correlation was observed between the degradation of elastic system fibers and the grade of dysplasia.

Smear layer removal for collagen fiber exposure after citric acid conditionings

Aim: The aim of the present study was to compare the removal of the smear layer and exposure of collagen fibers of the root surface following the application of five citric acid solution concentrations. Methods and Materials: Two hundred seventy (270) samples were equally divided into six groups (n=45) for treatment with saline solution (control) and five different concentrations of citric acid (0.5, 1, 2, 15, and 25 percent). Three acid application methods were used (passive, brushing, and burnishing) as well as three application periods (1, 2, and 3 minutes). A previously trained, calibrated (kappa score = 0.93), and blind examiner subsequently scored scanning electron micrographs (SEMs) of the samples. Statistical analyses were performed by using Kruskal-Wallis and Dunn's post-hoc tests. Results: According to the results obtained and within the limitations of the methodology used, the citric acid applications were more effective than the control treatment of applying saline solution (p<0.05). However, no statistically significant differences were observed among the three application methods and three application periods. Descriptive analyses showed that best results for exposure of collagen fibers were obtained with the application of citric acid at 25 percent by brushing for 1 or 3 minutes. Conclusions: The best results for exposure of collagen fibers in this study were obtained with application of citric acid at 25 percent by brushing for 1 or 3 minutes, even though there were no statistically significant differences among the groups. Clinical Significance: The best results for exposure of collagen fibers on root surfaces noted in this study were obtained with application of citric acid at 25 percent by brushing for 1 or 3 minutes. © 2010 Seer Publishing LLC.

Morphological and molecular analysis of the collagen fibers in inflammatory process

Collagen makes up one third of the total protein in humans, being formed by the connection of three polypeptide chains arranged in a triple helix. This protein has fundamental importance in the formation of extracellular matrix of connective tissue. This study aimed to analyze the structural changes of collagen, which are resulting from inflammatory processes in oral mucosa, and to make the comparative analysis between the histopathology and the Raman spectra. The samples of tissues with inflammatory fibrous hyperplasia (IFH) and normal mucosa (NM) were evaluated by Raman Spectroscopy, hematoxylin-eosin and Massons trichrome stain. The histological analysis in both stains showed differences in collagen fibers, which was presented as thin fibers and arranged in parallel direction in NM and as collagen fibers are thick, mature and not organized, showing that these types of stain show morphological changes of collagen in IFH. The Raman Spectroscopy discriminate the groups of NM and IFH based on vibrational modes of proline, hydroxiproline and CH3, CH2. The histological stains only shows information from morphological data, and can be complemented by Raman spectra. This technique could demonstrate that inflammatory process caused some changes in collagen structure which is related to aminoacids such as proline and hidroxyproline. © 2011 SPIE-OSA.

Histochemical and ultrastructural analysis of hepatic glycogen and collagen fibers in alloxan-induced diabetic rats submitted to long-term physical training

Alterations in liver functions are common among diabetic patients, and many symptoms in the liver have been reported, including changes in glycogen stores and in the amount of collagen fibers. The practice of physical training and its morphological effects in this organ, however, are scarcely studied. In order to observe the morphological effects of alloxan-induced diabetes and the alterations arising from the practice of long-term chronic physical training in the liver, samples were collected and processed, and then analyzed by means of the histochemical techniques Periodic Acid-Schiff and Picrosirius-Hematoxylin, and ultrastructural cytochemical test of Afzelius. Through evaluation of the tissue, it was observed a drastic reduction in hepatic glycogen stores of sedentary diabetics, recovered in trained diabetic rats. Furthermore, it was detected a decrease in the content of perisinusoidal collagen fibers in the diabetic liver, also recovered due to the development of a training protocol. On ultrastructural level, cytochemical analysis confirmed the loss of glycogen and the recovery obtained by training. In conclusion, the practice of a long-term chronic physical training protocol may be considered an important assistant in the treatment of diabetes, mitigating the occurrence of possible damages to liver tissue. © 2011 Elsevier Ltd.

Recent developments in chalcogenide photonic crystal fibres

Elaboration of low-losses highly non linear chalcogenide optical fibers for the generation of efficient non linear effects in the infrared remains a challenge. In recent years, much work has been devoted to the study of microstructured optical fibers (MOFs) with different designs and various elaboration processes. Their background losses were typically of several dB/m. © 2011 IEEE.

Influence of dental exposure to oral environment on smear layer removal and collagen exhibition after using different conditioning agents

Although in vitro studies have shown encouraging results for root surface conditioning with demineralizing agents, in vivo studies have failed to show its benefits in periodontal healing. This can be attributed to several factors, among which, the hypermineralization of dental surface. Therefore, this in vitro study compared, using scanning electron microscopy (SEM), the effect of root surface conditioning with different conditioners (1% and 25% citric acid, 24% EDTA and 50 mg/mL tetracycline hydrochloride) in impacted teeth and in teeth that had their roots exposed to the oral environment. One trained examiner assessed the SEM micrographs using a root surface modification index. There was a tendency of more root surface modification in the group of impacted teeth, suggesting that the degree of root mineralization influences its chemical demineralization.

The role of fibres and the hypodermis in Compositae melanin secretion

Melanins are dark, insoluble pigments that are resistant to concentrated acids and bleaching by oxidising agents. Phytomelanin (or phytomelan) is present in the seed coat of some Asparagales and in the fruits of some Compositae. In Compositae fruits, melanin is deposited in the schizogenous spaces between the hypodermis and underlying fibrous layer. Phytomelanin in Compositae is poorly understood, and there are only speculations regarding the cells that produce the pigment and the cellular processes involved in the secretion and polymerisation of phytomelanin. This report describes the cellular processes involved in the secretion of phytomelanin in the pericarp of Praxelis diffusa, a species with a structure typical of the family. The ovaries and fruits at different stages were fixed and processed according to the standard methods of studies of light microscopy and transmission electron microscopy. Hypodermal cells have abundant rough endoplasmic reticulum and mitochondria, and the nuclei have chromatin that is less dense than other cells. These characteristics are typical of cells that synthesise protein/amino acids and suggest no carbohydrate secretion. The fibres, however, have a dense cytoplasm rich in the Golgi bodies that are associated with vesicles and smooth endoplasmic reticulum, common characteristics of carbohydrate secretory cells. Our results indicate that the hypodermal cells are not responsible for the secretion of phytomelanin, as previously described in the literature; in contrast, this function is assigned to the adjacent fibres, which have an organisation typical of cells that secrete carbohydrates. © 2012 Elsevier Ltd.

Corneal Absorption of a New Riboflavin-Nanostructured System for Transepithelial Collagen Cross-Linking

Corneal collagen cross-linking (CXL) has been described as a promising therapy for keratoconus. According to standard CXL protocol, epithelium should be debrided before treatment to allow penetration of riboflavin into the corneal stroma. However, removal of the epithelium can increase procedure risks. In this study we aim to evaluate stromal penetration of a biocompatible riboflavin-based nanoemulsion system (riboflavin-5-phosphate and riboflavin-base) in rabbit corneas with intact epithelium. Two riboflavin nanoemulsions were developed. Transmittance and absorption coefficient were measured on corneas with intact epithelia after 30, 60, 120, 180, and 240 minutes following exposure to either the nanoemulsions or standard 0.1% or 1% riboflavin-dextran solutions. For the nanoemulsions, the epithelium was removed after measurements to assure that the riboflavin had passed through the hydrophobic epithelium and retained within the stroma. Results were compared to de-epithelialized corneas exposed to 0.1% riboflavin solution and to the same riboflavin nanoemulsions for 30 minutes (standard protocol). Mean transmittance and absorption measured in epithelialized corneas receiving the standard 0.1% riboflavin solution did not reach the levels found on the debrided corneas using the standard technique. Neither increasing the time of exposure nor the concentration of the riboflavin solution from 0.1% to 1% improved riboflavin penetration through the epithelium. When using riboflavin-5-phosphate nanoemulsion for 240 minutes, we found no difference between the mean absorption coefficients to the standard cross-linking protocol (p = 0.54). Riboflavin nanoemulsion was able to penetrate the corneal epithelium, achieving, after 240 minutes, greater stromal concentration when compared to debrided corneas with the standard protocol (p = 0.002). The riboflavin-5-phosphate nanoemulsion diffused better into the stroma than the riboflavin-base nanoemulsion. © 2013 Bottos et al.

Tissue engineering: Using collagen type i matrix for bone healing of bone defects

Among the many tissues in the human body, bone has been considered as a powerful marker for regeneration and its formation serves as a prototype model for tissue engineering based on morphogenesis. Therefore, collagen type I is one of the most useful biomaterials used in tissue engineering as extracellular matrix components capable to promote bone healing. The literature reveals excellent biocompatibility and safety due to its biological characteristics, such as biodegradability and weak antigenicity, making collagen type I the primary resource in medical applications. Thus, it was also used for tissue engineering including skin replacement, bone substitutes, and artificial blood vessels and valves. The authors describe the treatment of an abscessed apical periodontal cyst and show good outcomes of bone healing, using tissue engineering, as collagen type I matrix. © 2013 by Mutaz B. Habal, MD.

Genotoxicological assessment of two reactive dyes extracted from cotton fibres using artificial sweat

Human eyes have a remarkable ability to recognize hundreds of colour shades, which has stimulated the use of colorants, especially for clothing, but toxicological studies have shown that some textile dyes can be hazardous to human health. Under conditions of intense perspiration, dyes can migrate from coloured clothes and penetrate into human skin. Garments made from cotton fabrics are the most common clothing in tropical countries, due to their high temperatures. Aiming to identify safe textile dyes for dyeing cotton fabrics, the genotoxicity [in vitro Comet assay with normal human dermal fibroblasts (NHDF), Tail Intensity] and mutagenicity [Salmonella/microsome preincubation assay (30 min), tester strains TA98, TA100, YG1041 and YG1042] of Reactive Blue 2 (RB2, CAS No. 12236-82-7, C.I. 61211) and Reactive Green 19 (RG19, CAS No. 61931-49-5, C.I. 205075) were evaluated both in the formulated form and as extracted from cotton fibres using different artificial sweats. Both the dyes could migrate from cotton fibres to sweat solutions, the sweat composition and pH being important factors during this extraction. However, the dye sweat solutions showed no genotoxic/mutagenic effects, whereas a weak mutagenic potential was detected by the Ames test for both dyes in their formulated form. These findings emphasize the relevance of textile dyes assessment under conditions that more closely resemble human exposure, in order to recognize any hazard. © 2013 Elsevier Ltd. All rights reserved.

The importance of size-exclusion characteristics of type I collagen in bonding to dentin matrices

The mineral phase of dentin is located primarily within collagen fibrils. During development, bone or dentin collagen fibrils are formed first and then water within the fibril is replaced with apatite crystallites. Mineralized collagen contains very little water. During dentin bonding, acid-etching of mineralized dentin solubilizes the mineral crystallites and replaces them with water. During the infiltration phase of dentin bonding, adhesive comonomers are supposed to replace all of the collagen water with adhesive monomers that are then polymerized into copolymers. The authors of a recently published review suggested that dental monomers were too large to enter and displace water from collagen fibrils. If that were true, the endogenous proteases bound to dentin collagen could be responsible for unimpeded collagen degradation that is responsible for the poor durability of resin-dentin bonds. The current work studied the size-exclusion characteristics of dentin collagen, using a gel-filtration-like column chromatography technique, using dentin powder instead of Sephadex. The elution volumes of test molecules, including adhesive monomers, revealed that adhesive monomers smaller than ∼1000 Da can freely diffuse into collagen water, while molecules of 10,000 Da begin to be excluded, and bovine serum albumin (66,000 Da) was fully excluded. These results validate the concept that dental monomers can permeate between collagen molecules during infiltration by etch-and-rinse adhesives in water-saturated matrices. © 2013 Acta Materialia Inc.