Influência da laserterapia na proliferação de células-tronco do ligamento periodontal humano

Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm² - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm² promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm²) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm² exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm². No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration

Uso de membrana biossintética a base de celulose na regeneração tecidual guiada

Guided Tissue Regeneration (GTR) is a regenerative treatment modality that requires the placement of a physical barrier over a bone defect in such a way that the proliferation of the surrounding soft tissues into the barrier-protected area is prevented. Thereby in the guided osseous regeneration allowing bone-forming cells from the existent bone edges to invade the space and produce bone. The physical barriers should be biocompatible, allow cellular occlusion, maintain adequate space, tissue integration and facility in the application. They can be occlusive or permeable, absorbable or non-absorbable. Among various types of physical barriers in the market, the cellulose biosynthetic membrane is emphasizing by the necessary characteristics for GTR and to be a national product developed in low cost.

Análises histológica e morfométrica do uso de membrana biossintética de celulose em trocleoplastia experimental de cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biomechanical and histologic analysis in aortic endoprosthesis using fibrin glue

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

Bone healing in critical-size defects treated with either bone graft, membrane, or a combination of both materials: a histological and histometric study in rat tibiae

Objectives: The aim of the present investigation was to histologically analyze the effect of using lyophilized bovine bone (GenOx (R) organic matrix) with (or without) guided tissue regeneration (using a decalcified cortical osseous membrane [GenDerm (R)]) on bone healing in surgically created critical-size defects created in rat tibia.Material and methods: Surgical critical-size bone defects were created in 64 animals that were randomly divided into four groups: group I (control); group II (defect filled with GenOx (R)); group III (defect covered by GenDerm (R)); group IV (defect filled with GenOx (R) and covered by GenDerm (R)). Animals were killed at 30 or 90 days post-surgery. The specimens were embedded in paraffin, serially cut, and stained with hematoxylin and eosin for analysis under light microscopy. The formation of new bone in the cortical area of the defect was histomorphometrically evaluated.Results: All experimental groups demonstrated superior bone healing compared with the control group. However, group IV samples showed evidence of more advanced healing at both 30 and 90 days post-surgery as compared with the other experimental groups.Conclusions: The bovine organic bone graft GenOx (R) associated with GenDerm (R) this produced the best treatment results in the case of critical-size defects in rat tibia.

Hypoxia Enhances the Angiogenic Potential of Human Dental Pulp Cells

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Intrusion of teeth with class III furcation: a clinical, histologic and histometric study in dogs

Aim: To assess orthodontic intrusion effects on periodontal tissues in dogs' pre-molars with class III furcations treated with open flap debridement (OFD) or with guided tissue regeneration (GTR) associated to bone autograft (BA).Material and Methods: Class III furcations were created in the pre-molars of seven mongrel dogs. After 75 days, teeth were randomly treated with OFD or GTR/BA. After 1 month, metallic crowns were assembled on pre-molars and connected apically to mini-implants by nickel-titanium springs. Teeth were randomly assigned to orthodontic intrusion (OFD+I and GTR/BA+I) groups or no movement (OFD and GTR/BA) groups. Dogs were sacrificed after 3 months of movement and 1 month retention.Results: All class III furcations were closed or reduced to class II or I in the intrusion groups, while 50% of the lesions in non-moved teeth remained unchanged. Intruded teeth presented higher probing depth and lower gingival marginal level than non-moved teeth (p < 0.01). Clinical attachment gain was reduced in the intrusion groups by the end of retention (p < 0.05). OFD+I presented smaller soft tissue area and larger bone tissue area than other groups (p < 0.05).Conclusion: Orthodontic intrusion with anchorage via mini-implants improved the healing of class III furcation defects after OFD in dogs. GTR/BA impaired those results.

PDGF-B gene therapy accelerates bone engineering and oral implant osseointegration

Platelet-derived growth factor-BB (PDGF-BB) stimulates repair of healing-impaired chronic wounds such as diabetic ulcers and periodontal lesions. However, limitations in predictability of tissue regeneration occur due, in part, to transient growth factor bioavailability in vivo. Here, we report that gene delivery of PDGF-B stimulates repair of oral implant extraction socket defects. Alveolar ridge defects were created in rats and were treated at the time of titanium implant installation with a collagen matrix containing an adenoviral (Ad) vector encoding PDGF-B (5.5 x 10(8) or 5.5 x 10(9) pfu ml (1)), Ad encoding luciferase (Ad-Luc; 5.5 x 10(9) pfu ml (1); control) or recombinant human PDGF-BB protein (rhPDGF-BB, 0.3 mg ml (1)). Bone repair and osseointegration were measured through backscattered scanning electron microscopy, histomorphometry, microcomputed tomography and biomechanical assessments. Furthermore, a panel of local and systemic safety assessments was performed. Results indicated that bone repair was accelerated by Ad-PDGF-B and rhPDGF-BB delivery compared with Ad-Luc, with the high dose of Ad-PDGF-B more effective than the low dose. No significant dissemination of the vector construct or alteration of systemic parameters was noted. In summary, gene delivery of Ad-PDGF-B shows regenerative and safety capabilities for bone tissue engineering and osseointegration in alveolar bone defects comparable with rhPDGF-BB protein delivery in vivo. Gene Therapy (2010) 17, 95-104; doi: 10.1038/gt.2009.117; published online 10 September 2009

Fresh-frozen human bone graft for repair of defect after adenomatoid odontogenic tumour removal

The aim of this paper was report the clinical, radiographic, and histological case of adenomatoid odontogenic tumour (AOT) in adolescent woman as well as present the reconstructive treatment of AOT using fresh-frozen human bone graft with guided bone regeneration. AOT is a benign, noninvasive lesion with slow but progressive growth. Biopsy and microscopic examination confirmed the presence of an AOT. Treatment was conservative and the prognosis was excellent. The patient has been followed-up for without recurrence. The use of fresh-frozen human bone graft can be a safe choice for reconstruction of the bone defects to treat AOT.

Vertical Bone Augmentation Using a Polytetrafluoroethylene Nonporous Barrier for Osseointegrated Implants Partially Inserted in Tibiae of Rabbits

Purpose: The aim of this study was to evaluate the possibility of obtaining guided bone regeneration using a poly-tetrafluoroethylene (PTFE) nonporous barrier for 2 endosseous implants, partially inserted in tibiae of rabbits.Materials and Methods: Histologic characteristics of the interface between titanium implants (one group with titanium plasma-coated implants and the other group with acid-treated surfaces) and of the regenerated bone were also studied. Twenty screw-vent implants were placed in tibiae of 5 male New Zealand rabbits, 2 at the right side and 2 at the left side, protruding 3 mm from the bone level, to create a horizontal bone defect. At the experimental group the implants were with a PTFE nonporous barrier, whereas no barriers were used in contralateral implants. Animals were sacrificed 3 months after surgery and biopsy specimens were evaluated histologically and histomorphometrically under light microscopy. Student's t test was used for statistical analysis.Results: The histologic measurements showed a mean gain in bone height of 2.15 and 2.42 mm for the barrier group and 1.95 and 0.43 mm for the control group, for the titanium plasma-spray and acid-treated implant surfaces, respectively.Conclusion: The results of the investigation revealed that the placement of implants protruding 3 nun from crestal bone defects may result in vertical bone augmentation using a nonporous PTFE barrier. (Implant Dent 2009;18:182-191)

Assessment of bovine biomaterials containing bone morphogenetic proteins bound to absorbable hydroxyapatite in rabbit segmental bone defects

OBJETIVO: Avaliar a capacidade osteo-regenerativa de dois biomateriais utilizando um modelo de defeito segmentar efetuado nas diáfises do rádio de coelhos. MÉTODOS: O defeito direito foi preenchido com pool de proteínas morfogenéticas ósseas (pBMPs) e hidroxiapatita em pó ultrafina absorvível (HA) combinada com matriz óssea inorgânica desmineralizada e colágeno, derivados do osso bovino (Grupo A). O defeito esquerdo foi preenchido com matriz óssea desmineralizada bovina com pBMPs e hidroxiapatita em pó ultrafina absorvível (Grupo B). em ambos os defeitos utilizou-se membrana reabsorvível de cortical bovina desmineralizada para reter os biomateriais no defeito ósseo e guiar a regeneração tecidual. Os coelhos foram submetidos à eutanásia aos 30, 90 e 150 dias após a cirurgia. Foram efetuados exames radiográficos, tomográficos e histológicos em todos os espécimes. RESULTADOS: Aos 30 dias de pós-cirúrgico, o osso cortical desmineralizado foi totalmente reabsorvido em ambos os grupos. A HA tinha reabsorvido nos defeitos do Grupo A, mas persistiu nos do Grupo B. Uma reação de corpo estranho foi evidente com ambos os produtos, porém mais pronunciada no Grupo B. Aos 90 dias os defeitos do grupo B tinham mais formação óssea que os do Grupo A. Entretanto, aos 150 dias após a cirurgia, nenhum tratamento havia promovido o completo reparo do defeito. CONCLUSÃO: Os biomateriais testados contribuíram pouco ou quase nada para a reconstituição do defeito segmentar.

Polímeros biorreabsorvíveis como substrato para cultura de células e engenharia tecidual

Biomateriais poliméricos são desenvolvidos para uso como substitutos de tecidos danificados e/ou estimular sua regeneração. Uma classe de biomateriais poliméricos são os biorreabsorvíveis, compostos que se decompõem tanto in vitro quanto in vivo. São empregados em tecidos que necessitam de um suporte temporário para sua recomposição tecidual. Dentre os vários polímeros biorreabsorvíveis, destacam-se os alfa-hidróxi ácidos, entre eles, diferentes composições do poli(ácido lático) (PLA), como o poli(L-ácido lático) (PLLA), poli(D-ácido lático) (PDLA), poli(DL-ácido lático) (PDLLA), além do poli(ácido glicólico) (PGA) e da policaprolactona (PCL). Estes polímeros são considerados biorreabsorvíveis por apresentarem boa biocompatibilidade e os produtos de sua decomposição serem eliminados do corpo por vias metabólicas. Diversas linhas de pesquisa mostram que os diferentes substratos à base de PLA estudados não apresentam toxicidade, uma vez que as células são capazes de crescer e proliferar sobre eles. Além disso, diversos tipos de células cultivadas sobre diferentes formas de PLA são capazes de se diferenciarem sobre os diferentes polímeros e passar a produzir componentes de matriz extracelular. Neste trabalho, é revisada a utilização de substratos à base de alfa-hidróxi ácidos, com destaque para diferentes formas de PLA, utilizados como substratos para cultura de células, bem como suas aplicações.

Alguns eventos morfológicos do processo de reparo de falha óssea diafisária no rádio esquerdo de ratos tratados com fenobarbital

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biocompatibility studies of anionic collagen membranes with different degree of glutaraldehyde cross-linking

The work describes the biocompatibility and biodegradation studies of anionic collagen membranes casted form collagen gels collagen, that were selective hydrolyzed at the carboxyamide groups, as a function of the degree of cross-links induced by glutaraldehyde. Independently from the degree of cross-links, all membranes studied were characterized by a similar inflammatory response, inversely dependent on glutaraldehyde reaction time, that decreased from the time of the implant. Cell alterations, mineralization or contact necrosis were not observed in any of the membranes studied. Rates for membrane tissue biodegradation were directly related to glutaraldehyde reaction time, and ranged from 30 to periods longer than 60 days, associated with good biocompatibility. Although other properties must be considered, their use in the treatment of periodontal diseases, the biological behavior observed with the 8 h GA cross-linked membrane suggests that, anionic collagen membrane described in this work may be of potential use, not only in association with guided tissue regeneration technique for periodontal tissue reconstruction, but also in other collagen biomaterial applications where controlled biodegradability is required. (C) 1998 Published by Elsevier B.V. Ltd. All rights reserved.

Evaluation of anionic collagen membranes in the treatment of class II furcation lesions: an histometric analysis in dogs

This study aims to evaluate the effect of using anionic collagen membranes in guided tissue regeneration treatment of Class II furcation lesions in dogs. The defects were created in the buccal furcation of 16 mandibular premolars of four dogs. After 56 days without plaque control, the sites were scaled and divided into two groups according to the treatment applied: control sites, open flap debridement; and test sites, guided tissue regeneration treatment. The animals were killed after 3 months. Histological and histometrical analyses showed that the collagen membrane was better than open flap debridement in terms of newly formed cementum and epithelial migration prevention. It provided effective blockade of epithelial tissue and promoted regeneration of lost periodontal tissues, suggesting that the membrane warrants further study. (C) 1997 Elsevier B.V. Limited. All rights reserved.