In vivo and in vitro evaluation of an Acetobacter xylinum synthesized microbial cellulose membrane intended for guided tissue repair

Background: Barrier materials as cellulose membranes are used for guided tissue repair. However, it is essential that the surrounding tissues accept the device. The present study histologically evaluated tissue reaction to a microbial cellulose membrane after subcutaneous implantation in mice. Furthermore, the interaction between mesenchymal stem cells and the biomaterial was studied in vitro to evaluate its ability to act as cellular scaffold for tissue engineering.Methods: Twenty-five Swiss Albino mice were used. A 10 x 10 mm cellulose membrane obtained through biosynthesis using Acetobacter xylinum bacteria was implanted into the lumbar subcutaneous tissue of each mouse. The mice were euthanatized at seven, 15, 30, 60, and 90 days, and the membrane and surrounding tissues were collected and examined by histology.Results: A mild inflammatory response without foreign body reaction was observed until 30 days post-surgery around the implanted membrane. Polarized microscopy revealed that the membrane remained intact at all evaluation points. Scanning electron microscopy of the cellulose membrane surface showed absence of pores. The in vitro evaluation of the interaction between cells and biomaterial was performed through viability staining analysis of the cells over the biomaterial, which showed that 95% of the mesenchymal stem cells aggregating to the cellulose membrane were alive and that 5% were necrotic. Scanning electron microscopy showed mesenchymal stem cells with normal morphology and attached to the cellulose membrane surface.Conclusion: The microbial cellulose membrane evaluated was found to be nonresorbable, induced a mild inflammatory response and may prove useful as a scaffold for mesenchymal stem cells.

Anterior Tooth Rehabilitation With Frozen Homogenous Bone Graft and Immediately Loaded Titanium Implant Using Computer-Guided Surgery

Computed tomographic scanning is a precise, noinvasive surveying technique that enables the professionals to improve the precision of implant placement by building a prototype that allows the confection of surgical guides. The authors present a clinical case of anterior tooth rehabilitation with frozen homogenous bone graft and immediately loaded titanium implant using computer-guided surgery. A multislice computed tomography was realized, and a prototype was built. All the procedures were previously realized in the prototype before started in the patient. This technique allows a better surgical planning, makes the procedures more accurate, and reduces surgery time.

Large Unicystic Ameloblastoma of the Mandible: Management Guided by Biological Behavior

Ameloblastoma is a true neoplasm of odontogenic epithelial origin. It is a slow-growing benign tumor of the jaw, and patients usually present late after the tumor achieves considerable size to cause facial disfigurement. Diagnosis mainly from tissue biopsy and radiograph findings does assist in differentiating between types of ameloblastoma. Unicystic ameloblastoma is a tumor with a strong propensity for recurrence. There is a difference in biological behavior between mural unicystic ameloblastoma and those which are simply cystic or show intraluminal proliferation. The challenges in the management of this tumor are to provide complete excision in addition to reconstructing the bony defect, to provide the patient with reasonable cosmetic and functional outcome. The authors report a case of a mural unicystic ameloblastoma in a 23-year-old man who was treated by partial resection of the mandible. Biomedical prototypes were used because they provide acceptable precision and are useful for treatment planning.

Computer-Guided Surgery in Implantology: Review of Basic Concepts

The aim of the present study was to conduct a critical literature review about the technique of computer-guided surgery in implantology to highlight the indications, purposes, immediate loading of implants and complications, protocol of fabrication, and functioning of virtual planning software. This literature review was based on OLDMEDLINE and MEDLINE databases from 2002 to 2010 using the key words "computer-guided surgery" and "implant-supported prosthesis." Thirty-four studies regarding this topic were found. According to the literature review, it was concluded that the computer-assisted surgery is an excellent treatment alternative for patients with appropriate bone quantity for implant insertion in complete and partially edentulous arches. The Procera Nobel Guide software (Nobel Biocare) was the most common software used by the authors. In addition, the flapless surgery is advantageous for positioning of implants but with accurate indication. Although the computer-guided surgery may be helpful for virtual planning of cases with severe bone resorption, the conventional surgical technique is more appropriate. The surgical guide is important for insertion of the implants regardless of the surgical technique, and the success of immediate loading after computer-guided surgery depends on the accuracy of clinical and/or laboratorial steps.

Lethal photosensitization and guided bone regeneration in treatment of peri-implantitis: an experimental study in dogs

The purpose of this study was to evaluate the effect of lethal photosensitization and guided bone regeneration (GBR) on the treatment of ligature-induced peri-implantitis in different implant surfaces. The treatment outcome was evaluated by clinical and histometric methods. A total of 40 dental implants with four different surface coatings (10 commercially pure titanium surface (cpTi); 10 titanium plasma-sprayed (TPS); 10 acid-etched surface; 10 surface-oxide sandblasted) were inserted into five mongrel dogs. After 3 months, the animals with ligature-induced peri-implantitis were subjected to surgical treatment using a split-mouth design. The controls were treated by debridment and GBR, while the test side received an additional therapy with photosensitization, using a GaAlAs diode laser, with a wavelength of 830 nm and a power output of 50 mW for 80 s (4 J/cm(2)), and sensitized toluidine blue O (100 mu g/ml). The animals were sacrificed 5 months after therapy. The control sites presented an earlier exposition of the membranes on all coating surfaces, while the test group presented a higher bone height gain. Re-osseointegration ranged between 41.9% for the cpTi surface and 31.19% for the TPS surface in the test sites; however differences were not achieved between the surfaces. The lethal photosensitization associated with GBR allowed for better re-osseointegration at the area adjacent to the peri-implant defect regardless of the implant surface.

Evaluation of collagen-based membranes for guided bone regeneration, by three-dimensional computerized microtomography

Objective. The aim of this study was to evaluate the use of a collagen-based membrane compared with no treatment on guided bone regeneration by 3-dimensional computerized microtomography (mu CT).Study Design. Defects were created between the mesial and distal premolar roots of the second and third premolars (beagle dogs; n = 8). A collagen-based membrane (Vitala; Osteogenics Biomedical Inc., TX, USA) was placed in one of the defects (membrane group; n = 16), and the other was left untreated (no-membrane group; n = 16). Left and right sides provided healing samples for 2 and 16 weeks. Three-dimensional bone architecture was acquired by mu CT and categorized as fully regenerated (F, bone height and width) or nonregenerated (N).Results. Chi-square tests (95% level of significance) showed that tooth did not have an effect on outcome (P = .5). Significantly higher F outcomes were observed at 16 weeks than 2 weeks (P = .008) and in membrane group than in no-membrane group (P = .008).Conclusions. The collagen-based membrane influenced bone regeneration at the furcation. (Oral Surg Oral Med Oral Pathol Oral Radiol 2012;114:437-443)

Polyurethane and PTFE membranes for guided bone regeneration: Histopathological and ultrastructural evaluation

Objective: The purpose of this study was to research a membrane material for use in guided bone regeneration. Study design: In this study, 25 male Wistar rats were used to analyze the biocompatibility and degradation process of biomembranes. The morphological changes in subcutaneous implantations were assessed after 7, 14, 21, 28 and 70 days. The materials were made of polyurethane polymer (AUG) obtained from vegetal oil (Ricinus communis) and polytetrafluoroethylene membrane (PTFE). The surface characteristics of the physical barriers in scanning electronic microscopic (SEM) were also evaluated. Results: In both groups, the initial histological analysis showed moderate inflammatory infiltrate, which was predominantly polymorphonuclear. There was also a presence of edema, which was gradually replaced by granulation tissue, culminating in a fibrous capsule. In the AUG group, some multinucleated giant cells were present in the contact interface, with the space previously occupied by the material. However, membrane degradation was not observed during the period studied. According to the present SEM findings, porosity was not detected in the AUG or PTFE membranes. Conclusion: The researched material is biocompatible and the degradation process is extremely slow or may not even occur at all.

Calcitonin in bone-guided regeneration of mandibles in ovariectomized rats: densitometric, histologic and histomorphometric analysis

The aim of the present study was to investigate bone promotion in surgical defects created in the mandible of normal and ovariectomized female rats using calcitonin associated with a polytetrafluoroethylene barrier. The 100 female rats were divided into four groups: control (C), control treated with calcitonin (CM), ovariectomized control (OV) and ovariectomized treated with calcitonin (OVM). A circumscribed bone defect 4 mm in diameter was created in the region of the mandibular angle, and covered with the barrier. Groups CM and OVM received 2 IU/kg of synthetic salmon calcitonin intramuscularly three times a week. The animals were killed 3, 7, 14, 21 and 28 days after surgery. The bone defects were submitted to densitometric, histologic and histomorphometric analysis. Groups C and CM showed higher levels of bone formation after 7 days compared to the OV and OVM groups. A significant difference was observed between groups C and OV at 3-14 days. The OV group presented slower bone regeneration of the surgical bone defect created in the mandibular angle than group C. Synthetic salmon calcitonin accelerated regeneration of the bone defect in the mandibles of OVM animals similarly to group C, and also increased the formation of new bone during the regeneration process in CM.

Immunolocalization of bone morphogenetic protein 2 during the early healing events after guided bone regeneration

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

Synthesis of chitosan/hydroxyapatite membranes coated with hydroxycarbonate apatite for guided tissue regeneration purposes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Guided bone regeneration with subperiosteal implants of PTFE and hydroxyapatite physical barriers in rats

A regeneração periodontal e do rebordo ósseo utilizando barreiras físicas são procedimentos bem estabelecidos em cirurgias reconstrutivas. As características do biomaterial e o desenho da membrana empregados na regeneração tecidual guiada desempenham um papel importante na obtenção de bons resultados. O objetivo deste estudo experimental histológico foi comparar o uso de dois tipos de barreiras físicas na regeneração óssea guiada em defeitos criados na tíbia de ratos. Quinze animais foram divididos em três grupos: grupo I (barreira não-porosa de politetrafluoretileno), grupo II (blocos de hidroxiapatita de coral) e grupo III (controle que não recebeu nenhuma barreira). A análise histológica demonstrou várias quantidades de osso neoformado com ambos os tipos de barreiras. A barreira de politetrafluoretileno mostrou melhores resultados do que a hidroxiapatita. Os resultados deste estudo sugerem que a regeneração óssea pode ser conseguida com a técnica de submersão da barreira física.

Isolation and sequence determination of peptides in the venom of the spider wasp (Cyphononyx dorsalis) guided by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry

Micro-scale (sub-pmol) isolation and sequence determination of three peptides from the venom of the solitary spider wasp Cyphononyx dorsalis is described. We isolated two novel peptides Cd-125 and Cd-146 and a known peptide Thr(6)-bradykinin from only two venom sacs of solitary spider wasp Cyphononyx dorsalis without bioassay-guided fractionation. but instead guided by MALDI-TOF MS. The MALDI-TOF MS analysis of each fraction showed the purity and molecular weight of the components, which led to the isolation of the peptides virtually without loss of sample amount. The sequences of the novel peptides Cd-125 (Asp-Thr-Ala-Arg-Leu-Lys-Trp-His) and Cd-146 (Ser-Glu-Thr-Gly-Asn-Thr-Val-Thr-Val-Lys-Gly-Phe-Ser-Pro-Leu-Arg) were determined by Edman degradation together with mass spectrometry. and finally corroborated by solid-phase synthesis. The known peptide Thr(6)-bradykinin (Arg-Pro-Pro-Gly-Phe-Thr-Pro-Phe-Arg) was identified by comparison with the synthetic authentic specimen. This is the first example for any kinins to be found in Pompilidae wasp venoms. The procedure reported here can be applicable to studies on many other components of solitary wasp venoms with limited sample availability. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

Ovulation synchronization following commercial application of ultrasound-guided follicle ablation during the estrous cycle in mares

A regimen of progesterone plus estradiol (P&E) was used as a standard for ovarian synchronization to test the efficacy and evaluate the commercial application of ultrasound-guided follicle ablation as a non-steroidal alternative for ovulation synchronization in mares. Recipient mares at a private embryo transfer facility were at unknown stages of the estrous cycle at the start of the experiment on Day 1 when they were randomly assigned to an ablation group (n = 18-21 mares) or to a ME group (n = 20-21 mares). In the ablation group, mares were lightly sedated and all follicles > 10 mm were removed by transvaginal ultrasound-guided follicle aspiration. In the ME group, a combination of progesterone (150 mg) plus estradiol (10 mg) prepared in safflower oil was given daily (im) for 10 d. Two doses of prostaglandin FZ, (PGF, 10 mg/dose, im) were given 12 h apart on Day 5 in the ablation group, or a single dose on Day 10 in the ME group. Human chorionic gonadotropin (hCG, 2500 IU/mare, im) was given at a fixed time, 6 and 10 d after PGF treatment in the ablation and ME groups, respectively, with the expectation of a follicle > 30 mm at the time of treatment. In both the ablation and P&E groups, transrectal ultrasonography was done at the start of the study (Day 1) and again on the day of hCG treatment and daily thereafter to determine the presence of a CL, measure diameter of the largest follicle and detect ovulation. The mean interval from the start of the study and from PGF treatment to ovulation was shorter (P < 0.0001) in the ablation group (13.7 and 9.7 d, respectively) compared to the P&E group (22.3 and 13.2 d, respectively). Following fixed-day treatment with hCG after PGF treatment, the degree of ovulation synchronization was not different (P > 0.05) between the ablation and P&E groups within a 2-d (56 and 70%) or 4-d (83% and 90%) period. Although ultrasound-guided follicle ablation may not be practical in all circumstances, it excluded the conventional 10-d regimen of progesterone and estradiol and was considered an efficacious and feasible, non-steroidal alternative for ovulation synchronization in mares during the estrous cycle. (C 2007 Elsevier B.V. All rights reserved.