Autogenous bone combined with anorganic bovine bone for maxillary sinus augmentation: Analysis of the osteogenic potential of cells derived from the donor and the grafted sites

Objectives: This study aimed to comparatively evaluate the in vitro osteogenic potential of cells obtained from the mandibular ramus (MR, autogenous bone donor site) and from the maxillary sinus (MS) bone grafted with a mixture of anorganic bovine bone (ABB) and MR prior to titanium implant placement (MS, grafted implant site). Material and methods: Cells were obtained from three patients subjected to MS floor augmentation with a 1: 1 mixture of ABB (GenOx Inorg®) and MR. At the time of the sinus lift procedure and after 8 months, prior to implant placement, bone fragments were taken from MR and MS, respectively, and subjected to trypsin-collagenase digestion for primary cell culturing. Subcultured cells were grown under osteogenic condition for up to 21 days and assayed for proliferation/viability, osteoblast marker mRNA levels, alkaline phosphatase (ALP) activity and calcium content/Alizarin red staining. ALP activity was also determined in primary explant cultures exposed to GenOx Inorg® (1: 1 with MR) for 7 days. Data were compared using either the Mann-Whitney U-test or the Kruskal-Wallis test. Results: MS cultures exhibited a significantly lower osteogenic potential compared with MR cultures, with a progressive increase in cell proliferation together with a decrease in osteoblast markers, reduced ALP activity and calcium content. Exposure of MR-derived primary cultures to GenOx Inorg® inhibited ALP activity. Conclusion: These results suggest that the use of GenOx Inorg® in combination with MR fragments for MS floor augmentation inhibits the osteoblast cell differentiation at the implant site in the long term. © 2013 John Wiley & Sons A/S.

Clinical and molecular study of a new form of hereditary myotonia in Murrah water buffalo

Hereditary myotonia caused by mutations in CLCN1 has been previously described in humans, goats, dogs, mice and horses. The goal of this study was to characterize the clinical, morphological and genetic features of hereditary myotonia in Murrah buffalo. Clinical and laboratory evaluations were performed on affected and normal animals. CLCN1 cDNA and the relevant genomic region from normal and affected animals were sequenced. The affected animals exhibited muscle hypertrophy and stiffness. Myotonic discharges were observed during EMG, and dystrophic changes were not present in skeletal muscle biopsies; the last 43 nucleotides of exon-3 of the CLCN1 mRNA were deleted. Cloning of the genomic fragment revealed that the exclusion of this exonic sequence was caused by aberrant splicing, which was associated with the presence of a synonymous SNP in exon-3 (c.396C>T). The mutant allele triggered the efficient use of an ectopic 5' splice donor site located at nucleotides 90-91 of exon-3. The predicted impact of this aberrant splicing event is the alteration of the CLCN1 translational reading frame, which results in the incorporation of 24 unrelated amino acids followed by a premature stop codon. Copyright © 2012 Elsevier B.V. All rights reserved.

An alternative technique to the harvesting of a connective tissue graft from a thin palate: Enhanced wound healing

This article describes an alternative surgical approach to the harvesting of subepithelial connective tissue grafts from thin palates. A partial-thickness flap is raised, and a graft composed of epithelium and connective tissue is removed from the palate. The superficial layer ( epithelium and a thin zone of connective tissue) is then dissected from the graft and replaced at the donor site to facilitate faster healing. The subjacent layer of connective tissue is placed as needed to obtain root coverage. The clinical application of this technique is described in two patients with multiple gingival recessions.

Clinical and histological evaluation of subepithelial connective tissue after collagen sponge implantation in the human palate

Rocha AL, Shirasu BK, Hayacibara RM, Magro-Filho O, Zanoni JN, Araujo MG. Clinical and histological evaluation of subepithelial connective tissue after collagen sponge implantation in the human palate. J Periodont Res 2012; 47: 758765. (c) 2012 John Wiley & Sons A/S Background and Objective: Successful root-coverage treatment depends on the thickness of the donor tissue. This study aimed to evaluate the thickness of donor tissue after augmentation of the connective tissue in the palatal area by implantation of lyophilized collagen sponge (Hemospon (R)). Material and Methods: Ten patients with an indication for root coverage, whose palate was deficient in adequate connective tissue, were recruited. The procedure was carried out in two stages. In the first stage, the palatal thickness in the donor site was measured at three standardized points (points 1, 2 and 3), from the distal of the canine to the distal of the first molar, and the lyophilized collagen sponge was inserted. In the second stage, the palatal thickness over the implant was measured (at points 1, 2 and 3), two biopsies of the palatal mucosa were collected one over the implant (experimental sample) and the other on the contralateral side (control sample) and then root-coverage treatment was performed. Analyses consisted of clinical assessment of the palatal measurements before and after sponge implantation, and histological assessment of the experimental and control biopsy samples. Data were analyzed using the Wilcoxon test. Results: Both analyses showed a significant increase in mean thickness, of 1.08 mm of neoformed tissue in the clinical analysis (the tissue at point 2 was the thickest of the three points) and of 0.53 mm in the histological analysis. Conclusion: The insertion of lyophilized collagen sponge induced a significant increase in the thickness of palatal connective tissue.

Histomorphometric analysis of the repair process of autogenous bone grafts fixed at rat calvaria with cyanoacrylate

Objective: The purpose of this study was to perform histological and histometric analyses of the repair process of autogenous bone grafts fixed at rat calvaria with ethyl-cyanoacrylate adhesive. Material and Methods: Thirty-two rats were divided into two groups (n=16), Group I - Control and Group II - Adhesive. Osteotomies were made at the right parietal bone for graft obtainment using a 4-mm-diameter trephine drill. Then, the bone segments were fixed with the adhesive in the parietal region of the opposite side to the donor site. After 10 and 30 days, 8 animals of each group were euthanized and the calvarias were laboratorially processed for obtaining hematoxylin and eosin-stained slides for histological and histometric analyses. Results: An intense inflammatory reaction was observed at the 10-day period. At 30 days, this reaction was less intense, despite the presence of adhesive at the recipient-site/graft interface. Graft incorporation to the recipient site was observed only at the control group, which maintained the highest graft size at 10 and 30 days. Conclusions: Although the fragment was stable, the presence of adhesive in Group II did not allow graft incorporation to the recipient site, determining a localized, discrete and persistent inflammatory reaction.

Low-level laser therapy (670 nm) on viability of random skin flap in rats

This study investigated the effects of 670 nm laser, at different fluences, on the viability of skin flap in rats. One hundred male animals were used. The animals were divided into control group; group treated with 3 J/cm(2); group treated with 6 J/cm(2); group treated with 12 J/cm(2) and group treated with 24 J/cm(2). The skin flap was made on the backs of all animals studied, with a plastic sheet interposed between the flap and the donor site. Laser irradiation was done immediately after the surgery and on days 1, 2, 3 and 4 after surgery. The percentage of necrosis of the flap was calculated at the 7th postoperative day. Additionally, a sample of each flap was collected to enable us to count the blood vessels. Treated animals showed a statistically significant smaller area of necrosis than did the control group. The necrosis in the treated groups was 41.82% (group 2), 36.51% (group 3), 29.45% (group 4) and 20.37% (group 5). We also demonstrated that laser irradiation at 670 nm, at all doses used, had a stimulatory effect on angiogenesis. Our study showed that the 670 nm laser was efficient to increase the viability of the skin flap, at all fluences used, with a tendency of reaching better results at higher doses.

Aesthetic correction of a hypertrophic sinus frontalis: A long-term follow-up evaluation and refinement of the method

Two cases treated for hypertrophy of the frontal sinus are presented. In both patients, the excised frontal bone was inverted, and the resultant cavity was filled with either bone dust from the parietal region or Medpor. The first patient is shown with an excellent result after a follow-up period of 16 years. The second patient, who also achieved an excellent result, is presented after a follow-up period of 5 years. Using Medpor instead of bone dust resulted in the advantage that the operation could be performed with the patient under local anesthesia with sedation and prevention of an additional donor site. The use of Medpor has certain advantages over the use of hydroxyapatite, not least of which is its significant lower cost.

UMP kinase from Mycobacterium tuberculosis: Mode of action and allosteric interactions, and their likely role in pyrimidine metabolism regulation

The pyrH-encoded uridine 5′-monophosphate kinase (UMPK) is involved in both de novo and salvage synthesis of DNA and RNA precursors. Here we describe Mycobacterium tuberculosis UMPK (MtUMPK) cloning and expression in Escherichia coli. N-terminal amino acid sequencing and electrospray ionization mass spectrometry analyses confirmed the identity of homogeneous MtUMPK. MtUMPK catalyzed the phosphorylation of UMP to UDP, using ATP-Mg 2+ as phosphate donor. Size exclusion chromatography showed that the protein is a homotetramer. Kinetic studies revealed that MtUMPK exhibits cooperative kinetics towards ATP and undergoes allosteric regulation. GTP and UTP are, respectively, positive and negative effectors, maintaining the balance of purine versus pyrimidine synthesis. Initial velocity studies and substrate(s) binding measured by isothermal titration calorimetry suggested that catalysis proceeds by a sequential ordered mechanism, in which ATP binds first followed by UMP binding, and release of products is random. As MtUMPK does not resemble its eukaryotic counterparts, specific inhibitors could be designed to be tested as antitubercular agents. © 2010 Elsevier Inc. All rights reserved.

Vascular pericranial graft: A viable resource for frontal sinus obliteration

Inappropriate treatments of frontal sinus fractures may lead to serious complications, such as mucopyocele, meningitis, and brain abscess. Assessment of nasofrontal duct injury is crucial, and nasofrontal duct injury requires sinus obliteration, which is often accomplished by autologous grafts such as fat, muscle, or bone. These avascular grafts have an increased risk of resorption and infection, as well as donor site morbidity. For these reasons, pericranial flap, which is vascular, should be used for frontal sinus obliteration. The pericranial flap presented with less morbidity procedure and has decreased infection rates, which justifies its use in frontal sinus obliteration. This paper aims to report a case of a comminuted frontal sinus fracture in a 29-year-old man who was successfully treated by frontal sinus obliteration, using pericranial local flap. The patient was followed up postoperatively for 16 months without infection. Copyright © 2013 by Mutaz B. Habal, MD.

Anterior pericranial flap for frontal sinus duct obliteration: Is it a valuable resource?

Inappropriate treatments of frontal sinus fractures may lead to serious complications, such as mucopyocele, meningitis, and brain abscess. Assessment of nasofrontal duct injury is crucial, and nasofrontal duct injury requires sinus obliteration, which is often accomplished by autogenous grafts such as fat, muscle, or bone. These avascular grafts have an increased risk of resorption and infection and donor site morbidity. For these reasons, pericranial flap, which is vascular, should be used for frontal sinus obliteration. The pericranial flap presented with less morbidity procedure and has decreased infection rates, which justifies its use in frontal sinus obliteration. This study aimed to report a case of a comminuted frontal sinus fracture with a brief literature review, regarding the use of pericranial flap. The authors report a case of a 23-year-old male subject with a severely comminuted fracture of the anterior and posterior walls of the frontal sinus. The patient was successfully treated by cranialization with frontal sinus duct obliteration, using anterior pericranial flap. The patient was followed up for 16 months with no postoperative complication, such as infection. Pericranial flap is a good resource for frontal sinus duct obliteration because it is a durable and well-vascularized flap, which determines low rates of postoperative complications. Copyright © 2013 by Mutaz B. Habal, MD.

Low level laser therapy increases angiogenesis in a model of ischemic skin flap in rats mediated by VEGF, HIF-1α and MMP-2

It is known that low level laser therapy is able to improve skin flap viability by increasing angiogenesis. However, the mechanism for new blood vessel formation is not completely understood. Here, we investigated the effects of 660 nm and 780 nm lasers at fluences of 30 and 40 J/cm2 on three important mediators activated during angiogenesis. Sixty male Wistar rats were used and randomly divided into five groups with twelve animals each. Groups were distributed as follows: skin flap surgery non-irradiated group as a control; skin flap surgery irradiated with 660 nm laser at a fluence of 30 or 40 J/cm2 and skin flap surgery irradiated with 780 nm laser at a fluence of 30 or 40 J/cm2. The random skin flap was performed measuring 10 × 4 cm, with a plastic sheet interposed between the flap and the donor site. Laser irradiation was performed on 24 points covering the flap and surrounding skin immediately after the surgery and for 7 consecutive days thereafter. Tissues were collected, and the number of vessels, angiogenesis markers (vascular endothelial growth factor, VEGF and hypoxia inducible factor, HIF-1α) and a tissue remodeling marker (matrix metalloproteinase, MMP-2) were analyzed. LLLT increased an angiogenesis, HIF-1α and VEGF expression and decrease MMP-2 activity. These phenomena were dependent on the fluences, and wavelengths used. In this study we showed that LLLT may improve the healing of skin flaps by enhancing the amount of new vessels formed in the tissue. Both 660 nm and 780 nm lasers were able to modulate VEGF secretion, MMP-2 activity and HIF-1α expression in a dose dependent manner. © 2013 Published by Elsevier B.V.

Avaliação volumétrica por meio da tomografia computadorizada de feixe cônico, dos leitos ósseos potencialmente doadores na região de ramo e mento

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Utilization of Ethyl Cyanoacrylate and 2-Octyl Cyanoacrylate Adhesives for Autogenous Bone Graft Fixation: Histomorphometric Study in Rats

The present study analyzes the repair process of autogenous bone graft in a block fixed with ethyl cyanoacrylate and 2-octyl cyanoacrylate adhesives in rat calvaria. Forty-eight rats, divided into 3 groups, received round osteotomies at the right parietal bone for the attainment of autogenous bone graft fragment, which was fixed at the opposite side to the donor site with ethyl cyanoacrylate (ethyl group) and 2-octyl cyanoacrylate (octyl group) adhesives. In the control group, bone fragment was only juxtaposed at the parietal bone surface without any fixation material. The animals were euthanized after 10 and 60 postoperative days. The calvariae were processed in a laboratory for the attainment of slides stained through the hematoxylin and eosin technique for histological and histometric analysis. The qualitative analysis showed a discrete inflammatory infiltrate in the control group and moderate inflammatory infiltrate in the ethyl and octyl groups at the 10-day period, which remained at the 60-day period, mainly in the octyl group. The bone fragment remained bonded to the recipient site through the adhesive, but graft incorporation was not observed in any of the specimens. Resorption was higher in the octyl group followed by the ethyl and control groups, both at the 10-and 60-day periods, but with no statistical significance (P < .05). Although promoting graft fixation and its maintenance at the recipient site, both studied adhesives did not allow the graft incorporation, producing a localized and discrete inflammatory reaction, which persisted at 60 days, being more intense in the octyl cyanoacrylate group.

Radical management of solid ameloblastoma of the mandible: report of a case with 5-year follow-up

Ameloblastoma is a true neoplasm of odontogenic epithelial origin. This pathology can be classified into 4 groups: unicystic, solid or multicystic, peripheral, and malignant. Solid ameloblastomas of the mandible are the most common of them, and represent a challenging group of tumours to treat; in addition the follicular histopathological subtype has a high likelihood of recurrence. Thus, the challenges in the management of this tumour are to provide complete excision in addition to reconstruct the bony defect, in order to provide the patient with reasonable cosmetic and functional outcome. With this in mind, this paper aimed to describe the management of a solid multilocular ameloblastoma of follicular subtype in a 39-year-old female. Case report The authors report a case of a solid multilocular ameloblastoma of follicular subtype in a 39-year-old female who was successfully treated by partial resection of the mandible with immediate reconstruction using an iliac crest, as a donor site. After 15 months, the patient was rehabilitated using titanium implant dentistry, and has been followed up for 5 years without signs or symptoms of recurrence. Conclusion Correct surgical planning is the key for successful management of solid ameloblastoma with multilocular features, which is best treated using radical resection with immediate reconstruction, which ensures complete tumour excision, prevents recurrence, and enables fast and safe dental rehabilitation. Biomedical prototypes should be used since they provide acceptable precision and are useful for surgical planning.

Avaliação da espessura do osso parietal como região doadora de enxertos ósseos.

Introduction: The study of graft donor sites, whether from the anatomical, physiological or morphological point of view, has become a topic of current interest, due to the increasing number of patients needing facial bone reconstruction for various reasons. Purpose: In view of the need to constantly improve surgical techniques for autogenous bone graft harvesting, still considered the best choice for facial bone reconstruction, this paper describes an anatomical study on dry skulls in order to evaluate the average thickness of the parietal bone. Material and Methods: Measurements of this bone were taken with a goniometer, at four previously defined points, in the region that is often used as a donor site, in 49 dry skulls (98 parietal bones). The results were evaluated using the T test. Results: Thickness was measured at four predetermined points. The mean values (Point A = 4898mm, B = 4517mm, C = 6185mm, D = 4280mm) show that the bone can be even thinner than previously reported in the literature in other studies of the same nature. The largest bone thickness is in the medial and posterior region. Conclusion: A knowledge of these anatomical characteristics is helpful in preventing possible surgical complications, as well as making it safer for the surgeon to remove this graft and providing more information on whether or not to indicate this region as a bone graft donor site.