Healing process after surgical treatment with scalpel, electrocautery and laser radiation: histomorphologic and histomorphometric analysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A study of histological responses from Ti-6Al-7Nb alloy dental implants with and without plasma-sprayed hydroxyapatite coating in dogs

Titanium alloys are hoped to be used much more for applications as implant materials in the medical and dental fields because of their basic properties, such as biocompatibility, corrosion resistance and specific strength compared with other metallic implant materials. Thus, the Ti-6Al-7Nb alloy that has recently been developed for biomedical use, that is, primarily developed for orthopaedic use, is to be studied in this paper, for application in dental implants. The biocompatibility test in vivo was carried out in dogs and the osseointegration was verified through histological analysis of the samples of the Ti-6Al-7Nb alloy with and without hydroxyapatite coating that were inserted in the alveoli. Within the controlled conditions the samples did not show any toxic effects on the cells. (C) 2001 Kluwer Academic Publishers.

Densitometry, radiography, and histological assessment of collagen as methods to evaluate femoral bones in an experimental model of osteoporosis

All methods to detect experimental loss of bone present technique limitations. The sensitivities of image and histological analyses to detect the effects of teriparatide in rats with bone loss after ovariectomy were evaluated. All methods were qualitatively valid.The standardization of methods to assess bone loss after ovariectomy is crucial to establish the degree of experimental osteoporosis. In general, methods per image or histological techniques are used. To validate these two ways to determine the degree of bone loss in ovariectomized rats, we evaluated the sensitivities of bone densitometry, conventional radiography, and histological analysis of the area occupied by collagen, detecting the effects of teriparatide treatment in the femur of ovariectomized rats with bone loss.Wistar rats were divided into three groups: a control group, in which the animals were only subjected to laparotomy; an ovariectomized group, in which bilateral removal of the ovaries was performed; and an ovariectomized + teriparatide group, in which bilateral removal of the ovaries was performed, and the animals were treated with 3 mu g/100 g/day of teriparatide. Three months following the ovariectomy, bone densitometry, radiographic densitometry, and histological analysis of the area occupied by collagen fibers were carried out in the femur diaphysis.The bone densitometry revealed 11.2% reduction in femur density; in the conventional radiography, the loss of bone mass was 14.5%, and with the histological analysis, a 40.9% reduction in the area occupied by collagen was detected in the femur diaphysis.In conclusion, histological analysis could not be quantitatively compared with the methods of bone densitometry and conventional radiography; however, all of these methods were qualitatively valid for assessing the loss of bone stemming from ovariectomy and the therapeutic effect of teriparatide in the prevention of osteoporosis.

Self-fertility and polyembryony in South American yellow trumpet trees (Handroanthus chrysotrichus and H. ochraceus, Bignoniaceae): a histological study of postpollination events

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hydroxylapatite implants with or without collagen in the zygomatic arch of rats. Histological study.

The authors studied the behavior of calcium phosphate materials used as inlay implants into bone cavities prepared in the zygomatic arch of rats. Fifty male albino rats were divided into four groups as follows: group I-preparation of bone cavities which did not receive any implant material as controls; group II-implants of Interpore 200; group III-implants of experimental hydroxylapatite; group IV-implants of experimental hydroxylapatite combined with collagen. The animals were sacrificed after 5, 15, 30, 60 and 120 days and the specimens were submitted to histological analysis. Results showed that the experimental hydroxylapatite used in group III presented better osteogenic properties compared to the other materials. All tested materials were biocompatible, although group IV presented a more intense inflammatory response.

Histological study of hamster buccal mucosa following topical application of DMBA and exposition to low power 337 nm laser light

The aim of the present work is to analyze the histological changes on hamster buccal mucosa caused by the topical use of 7,12-dimethylbenzanthracene (DMBA) and exposition to a 220 μJ/pulse nitrogen laser light (@ 337 nm) at an average power of 2,3 mW. Twenty-one hamsters divided into two experimental groups were treated six times with DMBA. One hamster was kept as control. Group I was composed by ten hamsters and was submitted only to DMBA. Group II, also with ten hamsters, received the same treatment as group I and was exposed to the laser radiation. The time duration of each irradiation section was 10 seconds. All the treatment happened in alternated days. The histological analysis took place twice, after the end of the treatment and after sixty days. Both experimental groups presented dilatation of vessels, thickening of the epithelial tissue and the presence of inflammatory infiltrates. The preliminary results indicates that in group II the number of dilated vessels and its new area are much more significant than in group I.

Analysis of the nucleolar cycle in the seminiferous epithelium of rodents

The nucleolus is a subcompartment of the nucleus and the site of ribosome biogenesis. During the mitotic and meiotic cell cycles, a disorganization and later reorganization of the nucleolar material occur, an event called nucleologenesis. In the spermatogenesis of mammals and other vertebrates, there is evidence of the disorganization of the nucleolus at the end of meiosis I, which supplies material for the cytoplasmic formation of an organelle called the chromatoid body (CB). The CB is a structure characteristic of spermatogenic cells and seems to be responsible for RNA metabolism in these cells and for some events of spermiogenesis, such as the formation of the acrosome, cellular communication between spermatids, and the formation of the spermatozoon middle piece and tail. The aim of this paper was to obtain information about the cytochemical and ultrastructural nature of the nucleolar cycle and the distribution of cytoplasmic RNAs in the seminiferous tubule cells of Rattus novergiucus, Mus musculus and Meriones unguiculatus. The testis was fixed in Bouin and Karnovsky solutions for conventional histological analysis and for cytochemical study that included: periodic acid-Schiff, hematoxylin-eosin, Feulgen reaction, silver-ion impregnation, Gomori's reticulin stain, toluidine blue, modified method of critical electrolyte concentration, and basic and acid fast green. The blocks of testis fixed in glutaraldehyde were used for ultrastructural analysis by transmission electron microscopy. Ultrathin sections were double-stained with uranyl acetate and lead citrate. All the techniques used provided information on the origin and function of the CB in the spermatogenic cells. Therefore, considering the persistence of the RNA and nucleolar ribonucleoproteins during spermatogenesis of Rattus novergicus, Mus musculus and Meriones unguiculatus, our findings corroborate the statement that these molecular complexes are very important in the spermiogenesis phases. It can be suggested that these ribonucleoprotein corpuscles (chromatoid bodies) are of nuclear origin and have a role in the successive series of events that occur in the formation of the spermatozoon. Furthermore, these results reinforce the conservation of the mechanisms involved in preserving necessary levels of protein stocks in different stages of cell differentiation, from spermatid to spermatozoon, in these rodent species. ©FUNPEC-RP.

Regional odontodysplasia in early childhood: A clinical and histological study

Regional odontodysplasia (RO) is a rare disorder of dental development. The affected teeth are clinically hypoplastic and hypocalcified, presenting a ghost-like appearance radiographically. The aim of this work was to report a clinical case of a child with both primary and permanent dentition affected by RO. The conducted therapy was based on a conservative approach, which consisted of follow-up clinical evaluations of the anomalous teeth. However, the endodontic treatment of the primary incisors failed. Then, the chosen option for patient rehabilitation became extraction followed by removable of prosthesis confection. The extracted teeth were processed for histological analysis. In spite of the uncertain prognosis, but taking into account the psychological aspects of the patient, a conservative approach in an attempt to maintain those viable teeth in the oral cavity should be established.

Evaluation of bone ingrowth into porous titanium implant: Histomorphometric analysis in rabbits

A porous material for bone ingrowth with adequate pore structure and appropriate mechanical properties has long been sought as the ideal bone-implant interface. This study aimed to assess in vivo the influence of three types of porous titanium implant on the new bone ingrowth. The implants were produced by means of a powder metallurgy technique with different porosities and pore sizes: Group 1 = 30% and 180 μm; Group 2 = 30% and 300 μm; and Group 3 = 40% and 180 μm. Six rabbits received one implant of each type in the right and left tibiae and were sacrificed 8 weeks after surgery for histological and histomor-phometric analyses. Histological analysis confirmed new bone in contact with the implant, formed in direction of pores. Histomorphometric evaluation demonstrated that the new bone formation was statistically significantly lower in the group G1 than in group G3, (P = 0.023). Based on these results, increased porosity and pore size were concluded to have a positive effect on the amount of bone ingrowth.

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.

Tomographic, Histological, and Immunohistochemical Evidences on the Use of N-Butyl-2-Cyanoacrilate for Onlay Graft Fixation in Rabbits

Background: The bone tissue responses to Cyanoacrylate have been described in the literature, but none used N-butyl-2-cyanoacrilate (NB-Cn) for bone graft fixation. Purpose: The aims of the study were: (a) to analyze the bone grafts volume maintenance fixed either with NB-Cn or titanium screw; (b) to assess the incorporation of onlay grafts on perforated recipient bed; and (c) the differences of expression level of tartrate-resistant acid phosphatase (TRAP) protein involved in bone resorption. Materials and Methods: Eighteen New Zealand White rabbits were submitted to calvaria onlay grafting on both sides of the mandible. On one side, the graft was fixed with NB-Cn, while on the other hand the bone graft was secured with an osteosynthesis screw. The computed tomography (CT) was performed just after surgery and at animals sacrifice, after 1 (n=9) and 6weeks (n=9), in order to estimate the bone grafts volume along the experiments. Histological sections of the grafted areas were prepared to evaluate the healing of bone grafts and to assess the expression of TRAP protein. Results: The CT scan showed better volume maintenance of bone grafts fixed with NB-Cn (p≤0.05) compared with those fixed with screws, in both experimental times (analysis of variance). The immunohistochemical evaluation showed that the TRAP expression in a 6-week period was significantly higher compared with the 1-week period, without showing significant difference between the groups (Wilcoxon and Mann-Whitney). Histological analysis revealed that the NB-Cn caused periosteum damage, but provided bone graft stabilization and incorporation similar to the control group. Conclusion: The perforation provided by screw insertion into the graft during fixation may have triggered early revascularization and remodeling to render increased volume loss compared with the experimental group. These results indicate that the NB-Cn possesses equivalent properties to titanium screw to be used as bone fixation material in osteosynthesis. © 2010 Wiley Periodicals, Inc.

Genome-Wide Analysis Reveals Selection for Important Traits in Domestic Horse Breeds

Intense selective pressures applied over short evolutionary time have resulted in homogeneity within, but substantial variation among, horse breeds. Utilizing this population structure, 744 individuals from 33 breeds, and a 54,000 SNP genotyping array, breed-specific targets of selection were identified using an FST-based statistic calculated in 500-kb windows across the genome. A 5.5-Mb region of ECA18, in which the myostatin (MSTN) gene was centered, contained the highest signature of selection in both the Paint and Quarter Horse. Gene sequencing and histological analysis of gluteal muscle biopsies showed a promoter variant and intronic SNP of MSTN were each significantly associated with higher Type 2B and lower Type 1 muscle fiber proportions in the Quarter Horse, demonstrating a functional consequence of selection at this locus. Signatures of selection on ECA23 in all gaited breeds in the sample led to the identification of a shared, 186-kb haplotype including two doublesex related mab transcription factor genes (DMRT2 and 3). The recent identification of a DMRT3 mutation within this haplotype, which appears necessary for the ability to perform alternative gaits, provides further evidence for selection at this locus. Finally, putative loci for the determination of size were identified in the draft breeds and the Miniature horse on ECA11, as well as when signatures of selection surrounding candidate genes at other loci were examined. This work provides further evidence of the importance of MSTN in racing breeds, provides strong evidence for selection upon gait and size, and illustrates the potential for population-based techniques to find genomic regions driving important phenotypes in the modern horse. © 2013 Petersen et al.

Histological and immunohistochemical analyses of the chronology of healing process after immediate tooth replantation in incisor rat teeth

Dental tissues have special characteristics, and its regenerative capacity is noteworthy. However, understanding the circumstances that lead to regeneration is challenging. In this study, the chronology of the healing process after immediate replantation of rat incisor teeth was examined by histological and immunohistochemical analyses within a 60-day period. Thirty-six male Wistar rats had their maxillary right incisors extracted and replanted after 15min in saline storage. The rats were sacrificed immediately 3, 7, 15, 28, and 60days after replantation. The histological analysis showed rupture of the periodontal ligament and formation of a blood clot, which started being replaced by a connective tissue after 3days. At 7days, the gingival mucosa epithelium was reinserted and areas of root resorption could be seen. At 15days, the periodontal ligament was repaired. At 3days, the pulp presented an absence of the odontoblast layer, which started being replaced by a connective tissue. This tissue suffered gradual calcification, filling the root canal at 28 and 60days. The root ends were closed. The immunohistochemical analysis revealed greater expression of OP, OPG, and RANK proteins in the initial periods (0 and 3days), while TRAP expression predominated at 28 and 60days (P<0.05). In conclusion, in delayed tooth replantation, there is great new bone formation activity in the earlier periods of the repair process, while a predominance of bone resorption and remodeling is observed in the more advanced periods. © 2012 John Wiley & Sons A/S.

Clinical similarities and histological diversity comparing fresh frozen onlay bone blocks allografts and autografts in human maxillary reconstruction

Background: In the absence of autologous bone for harvesting, fresh-frozen bone allografts turned into an alternative for bone reconstruction procedures. Purpose: The purpose of this study was to make a histological analysis of fresh-frozen onlay bone allografts (ALs), compared with autografts, in patients who needed maxillary reconstruction prior to dental implants placement. Materials and Methods: Twelve patients with bone deficiencies (width inferior to 4mm) in the sites where the implants were planned were enrolled in the study. From these, six were elected to be treated with autogenous (AT) bone grafts and six with fresh-frozen bone AL. This last group included the patients who had absence of a convenient amount of bone in donor sites. Each patient received from one to six graft blocks, totalling to 12 ATs and 17 ALs. Seven months after grafting procedures, biopsies of the grafts were made using 2-mm internal diameter trephine burs, and processed for histological analysis. One biopsy was retrieved from each patient. Results: Clinically, all grafts were found to be firm in consistency and well-incorporated to the receptor bed. Histological analysis showed a large amount of necrotic bone surrounded by few spots of new-formed bone in the AL group, suggesting low rate of graft remodeling. In the AT group, an advanced stage of bone remodeling was seen. Conclusions: Human fresh-frozen bone block AL showed clinical compatibility for grafting procedures, although associated to slow remodeling process. Further studies are needed to define, at long term, the remodeling process chronology the clinical longitudinal results for fresh-frozen bone AL. Copyright © 2013 Wiley Periodicals, Inc.

Performance of race instrumentation system in curved root canals: a comprehensive analysis by three study methods

In this study, curved maxillary molar root canals were instrumented with RaCe rotary system to evaluate: 1. the occurrence of canal transportation using a radiographic platform; 2. the action of the instruments on the dentin walls, centering ability and canal enlargement by analysis of digital images; and the percentage of regular dentin surfaces and debris within the canal by histological analysis. Ten mesiobuccal roots of extracted human maxillary molars were embedded in acrylic resin and sectioned at the middle and apical thirds. Root canal shaping was performed using the RaCe rotary system at 250 rpm and 1 Ncm torque. Each instrument set was used five times according to a crowndown technique in the following sequence: 40/0.10, 35/0.08, 25/0.06, 25/0.04, 25/0.02 (working length - WL), 30/0.02 (WL) and 35/0.02 (WL). Each instrument was inserted until resistance was felt and then pulled back, followed by brushing movements towards all canal walls. Each specimen was assessed by three study methods: radiographic platform, digitized image assessment and histological analysis. The radiographic platform showed lack of apical transportation. No statistically significant difference (Wilcoxon test, p>0.05) was found between the middle and apical thirds regarding instrument action on dentin walls, centering ability, area of root canal enlargement, percentage of regular dentin surfaces and debris within the root canal. It may be concluded that RaCe system is a suitable method for the preparation of curved root canals, regarding the maintenance of root canal original path, action on dentin walls, canal enlargement and removal of debris from the root canal lumen.