Análise citogenética de células-tronco derivadas do subendotélio da veia umbilical humana

Mesenchymal stem cells (MSCs) are known as a population of multi-potential cells able to proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat and stroma. Several applications of the study of EC can be emphasized the therapeutic techniques such as guided bone regeneration by implantation of EC in the affected site, without the need for bone grafts, using titanium as a vehicle. The process of cryopreservation is essential for the maintenance of cell cultures, since the cell line is frozen, it can be maintained in liquid nitrogen for an indefinite period and then thawed for therapeutic or experimental purposes. The aim of this study was to isolate a population of MSCs derived from the subendothelium of the umbilical vein human (MSCs-SUVH) to assess cytogenetic analysis by the possibility of appearance of chromosomal changes in two different situations: MSCs-SUVH regarding the process of cryopreservation and MSCs-SUVH grown on the surface of titanium. Flow cytometry analysis revealed that, this cell population was positive for the markers CD29, CD73 and CD90, but there was no expression of hematopoietic lineage markers, such as CD14, CD34 and CD45 and demonstrated capacity for osteogenic differentiation. The chromosomes obtained from the primary culture of MSCs-SUVH were analyzed by GTW banding technique, and results are described as guidelines to ISCN 2005. There was not the emergence of clonal chromosomal changes in the MSCs-SUVH in different situations analyzed. However one of the strings presented a balanced paracentric inversion, probably a cytogenetic constitutional alterations, which was present before and after the experimental situations that the MSCs-SUVH was submitted

Avaliação microscópica dos fragmentos ósseos obtidos por diferentes métodos de osteotomia e de irrigação em aloenxertos irradiados e congelados de coelho

Oral and facial bone defects can undertake appearance, psychosocial well-being and stomathognatic function of its patients. Over the yerars several strategies for bone defect regeneration have arised to treat these pathologies, among them the use of frozen and irradiated bone allograft. Manipulation of bone grafts it s not determined yet, and several osteotomy alternatives can be observed. The present work evaluated with a microscope the bone fragments obtained from different osteotomy methods and irrigation on rings and blocks allografts irradiated and frozen at 80° negative in a rabbit model. The study is experimental in vitro and it sample was an adult male New Zealand rabbit. The animal was sacrificed to obtain long bones, that were submitted to freezing at 80º negative and irradiated with Cobalt- 60. Then the long bones were sectioned into 24 bone pieces, divided into 4 groups: G1 (n=06) osteotomy was performed with bur No. 6 forming rings with 5 mm thickness with high-speed handpiece with manual irrigation; G2 (n=06) osteotomy was performed with bur No. 6 forming rings with 5 mm thick with surgical motor with a manual irrigation rotation 1500 rpm; GA (n=06), osteotomy with trephine using manual irrigation with saline; and GB (n=06), osteotomy with trephine using saline from peristaltic pumps of surgical motor. Five bone pieces of each group were prepared for analysis on light microscopy (LM) and one on electronic scan electronic microscopy (SEM). On the SEM analysis edges surface, presence of microcracks and Smear Layer were evaluated. Analyzing osteotomy technics on SEM was observed: increased presence of microcracks cutting with high speed; increased presence of areas covered by Smear Layer when cutting with motor implant. The irrigation analysis with SEM was observed: that the presence of microcracks does not depend on the type of irrigation; on manual irrigation, there was greater discrepancy between the cutting lines. The descriptive analysis of the osteotomy and irrigation process on LM showed: histological analysis showing the bony margins with clear tissue changed layer, composed of blackened tissue of charred appearance near to the cortical bone; on the edges of the bony part, bone fragments that were displaced during the bone cut and bone irregularities were observed. After analysis of results we can conclude: that there was greater regularity of the bone cut using high-speed handpiece than using motor implant; the cut with trephine using saline irrigated from peristaltic pumps of surgical motor showed greater homogeneity when compared with manual irrigation; charred tissue was found in all obtained bone samples, whit no significant statistically difference on the proportion of carbonization of the two analysed technics

Estudo da descelularização tecidual na produção de arcabouços biológicos para enxertos

The regeneration of bone defects with loss of substance remains as a therapeutic challenge in the medical field. There are basically four types of grafts: autologous, allogenic, xenogenic and isogenic. It is a consensus that autologous bone is the most suitable material for this purpose, but there are limitations to its use, especially the insufficient amount in the donor. Surveys show that the components of the extracellular matrix (ECM) are generally conserved between different species and are well tolerated even in xenogenic recipient. Thus, several studies have been conducted in the search for a replacement for autogenous bone scaffold using the technique of decellularization. To obtain these scaffolds, tissue must undergo a process of cell removal that causes minimal adverse effects on the composition, biological activity and mechanical integrity of the remaining extracellular matrix. There is not, however, a conformity among researchers about the best protocol for decellularization, since each of these treatments interfere differently in biochemical composition, ultrastructure and mechanical properties of the extracellular matrix, affecting the type of immune response to the material. Further down the arsenal of research involving decellularization bone tissue represents another obstacle to the arrival of a consensus protocol. The present study aimed to evaluate the influence of decellularization methods in the production of biological scaffolds from skeletal organs of mice, for their use for grafting. This was a laboratory study, sequenced in two distinct stages. In the first phase 12 mice hemi-calvariae were evaluated, divided into three groups (n = 4) and submitted to three different decellularization protocols (SDS [group I], trypsin [Group II], Triton X-100 [Group III]). We tried to identify the one that promotes most efficient cell removal, simultaneously to the best structural preservation of the bone extracellular matrix. Therefore, we performed quantitative analysis of the number of remaining cells and descriptive analysis of the scaffolds, made possible by microscopy. In the second stage, a study was conducted to evaluate the in vitro adhesion of mice bone marrow mesenchymal cells, cultured on these scaffolds, previously decellularized. Through manual counting of cells on scaffolds there was a complete cell removal in Group II, Group I showed a practically complete cell removal, and Group III displayed cell remains. The findings allowed us to observe a significant difference only between Groups II and III (p = 0.042). Better maintenance of the collagen structure was obtained with Triton X-100, whereas the decellularization with Trypsin was responsible for the major structural changes in the scaffolds. After culture, the adhesion of mesenchymal cells was only observed in specimens deccelularized with Trypsin. Due to the potential for total removal of cells and the ability to allow adherence of these, the protocol based on the use of Trypsin (Group II) was considered the most suitable for use in future experiments involving bone grafting decellularized scaffolds

Uso de câmara úmida em enxertia convencional de maracujazeiro-amarelo sobre três porta-enxertos

A propagação do maracujazeiro através da enxertia poderá ser a solução para o controle de doenças do sistema radicular. Assim, com o objetivo de avaliar o desenvolvimento e a sobrevivência de plantas enxertadas de maracujazeiro-amarelo (Passiflora edulis Sims) em três porta-enxertos, utilizando-se de câmara úmida, instalou-se este experimento em um viveiro de mudas no município de Adamantina-SP, no período de dezembro de 2005 a abril de 2006. O delineamento experimental utilizado foi em blocos ao acaso, em esquema fatorial 3 x 2, com quatro repetições e 10 plantas/parcela. Os fatores avaliados foram três porta-enxertos (Passiflora edulis, P. alata e P. gibertii)em dois ambientes (com e sem câmara úmida). Foi utilizada a enxertia convencional pelo método de garfagem tipo fenda cheia. Avaliaram-se a sobrevivência e a altura dos enxertos, o diâmetro do caule do porta-enxerto e do enxerto, o número de folhas e a fitomassa seca da parte aérea e das raízes. Verificaram-se melhores resultados com os porta-enxertos P. edulis e P. gibertii, superiores ao P. alata em todas as variáveis estudadas. Os maiores índices de sobrevivência foram observados em P. edulis e P. gibertii, com 100% e 98,8%, respectivamente. Apesar de o P. alata ter-se mostrado inferior, é viável o uso deste material como porta-enxerto para o maracujazeiro-amarelo. O uso da câmara úmida favoreceu o pegamento dos enxertos em P. alata.

Uso de câmara úmida em enxertia hipocotiledonar de maracujazeiro-amarelo sobre três porta-enxertos

Com o objetivo de avaliar o desenvolvimento e a sobrevivência de plantas de maracujazeiro-amarelo (Passiflora edulis Sims) enxertadas sobre três porta-enxertos com uso de câmara úmida, instalou-se este experimento em um viveiro de mudas no município de Adamantina - SP, no período de dezembro de 2005 a abril de 2006. O delineamento foi em blocos ao acaso, em esquema fatorial 3 x 2, com quatro repetições. Os fatores avaliados foram três porta-enxertos (Passiflora edulis, P. alata e P. gibertii) em dois ambientes (com e sem câmara úmida). Foi utilizada a enxertia hipocotiledonar pelo método de garfagem tipo fenda cheia. Avaliaram-se a altura das plantas, o diâmetro do caule do porta-enxerto e do enxerto, o número de folhas, o comprimento do entrenó, a porcentagem de sobrevivência das plantas e a fitomassa seca da parte aérea e das raízes. Observou-se que o método de enxertia utilizado foi bem-sucedido para as três espécies de porta-enxertos estudadas. P. gibertii e P. edulis proporcionaram melhores resultados como porta-enxertos. O porta-enxerto P. alata proporcionou menor altura de planta, menor diâmetro do enxerto e do porta-enxerto e menor índice de sobrevivência. O uso da câmara úmida não favoreceu nenhuma das variáveis estudadas.

Desenvolvimento e produtividade do maracujazeiro-amarelo enxertado em três porta-enxertos

O objetivo deste trabalho foi avaliar o desenvolvimento vegetativo e a produtividade do maracujazeiro-amarelo (Passiflora edulis Sims) enxertado sobre três porta-enxertos em área sem histórico de morte prematura de plantas. O experimento foi conduzido no município de Adamantina-SP, no período de abril de 2006 a junho de 2007, adotando-se o delineamento em blocos ao acaso, em esquema fatorial 3 x 2 + 1 tratamento adicional, com quatro repetições. Os fatores avaliados foram três porta-enxertos: Passiflora edulis, P. alata e P. gibertii, em dois sistemas de enxertia, a hipocotiledonar e a convencional por garfagem tipo fenda cheia, e um tratamento adicional, sem enxertia. Utilizou-se como copa o maracujazeiro-amarelo (Passiflora edulis Sims). Avaliaram-se o diâmetro do caule do porta-enxerto, o diâmetro do caule do enxerto, o comprimento de entrenó, o comprimento dos ramos secundários, o número de ramos terciários, o número de frutos, a massa média de frutos, a produtividade e as fitomassas verde e seca das plantas. Observou-se que as três espécies estudadas podem ser utilizadas como porta-enxertos para o maracujazeiro-amarelo, com diferentes níveis de compatibilidade. P. edulis demonstrou maior compatibilidade como porta-enxerto por apresentar-se superior aos demais na maioria dos parâmetros analisados. Plantas enxertadas sobre P. gibertii apresentaram menor vigor, menor crescimento vegetativo, frutos com menor massa e menor produtividade. A enxertia hipocotiledonar resultou em maior massa média de frutos e maior produtividade no maracujazeiro-amarelo.

Muscle graft as a substitute for peripheral nerve graft in rats

OBJETIVO: Avaliar a aplicabilidade do uso de músculo autógeno, tratado de diversas maneiras, em substituição aos enxertos de nervo. MÉTODOS: Os ratos foram separados em sete grupos que receberam, como tratamento a uma lesão nervosa padronizada, os seguintes tipos de enxertos: músculo fresco, músculo fixado com formol 10%, músculo congelado em freezer, músculo congelado em refrigerador, músculo denervado, nervo periférico e um grupo ficou sem qualquer tratamento. Foi avaliado o aspecto histológico das fibras nervosas no segmento reparado. RESULTADOS: A avaliação do segmento nervoso reparado mostrou que existiam axônios em quase todos os grupos, mas a metodologia empregada não possibilitou caracterizar adequadamente as diferenças entre os grupos. CONCLUSÃO: Este estudo mostrou a migração de axônios por meio de todos os enxertos utilizados.

Total Endovascular Debranching of the Aortic Arch

Background: Significant morbidity and mortality are related to conventional aortic replacement surgery. Endovascular debranching techniques, fenestrated or branched endografts are time consuming and costly.Objective: We alternatively propose to use endovascular approach with parallel grafts for debranching of aortic arch.Methods: Under general anesthesia, 12 F sheaths were inserted in the femoral, axillary and common carotid arteries for vascular accesses. ViaBahn grafts 10 - 15 cm in length were placed into the aortic arch from right common carotid, left common carotid and left axillary arteries, until the tip of each graft reached into the ascending aorta. Through one femoral artery, the aortic stent-graft was positioned and delivered. Soon after, the parallel grafts were sequentially delivered. Self-spanding Wallstents(R) were used for parallel grafts reinforcement. Ballooning was routinely used for parallel grafts and rarely for aortic graft.Results: This technique was used in 2 cases. The first one was a lady with 72 years old, with an aortic retrograde dissection from left subclavian artery and involving remaining arch branches. Through right common carotid artery a stent-graft was placed in the ascending aorta and through the left common carotid artery a ViaBahn was inserted parallel to the former. A thoracic endograft then covered all the aortic arch dissection extending into the ascending aorta close to the sinu-tubular junction. The second case was a 82 year old male patient with a 7 cm aortic arch aneurysm. Through both common carotid arteries ViaBahn grafts were introduced and positioned into the ascending aorta. Soon after, the deployment of the thoracic stent graft covered all parallel grafts of the aortic arch, excluding the aneurysm. Both cases did not have neurologic or cardiac complications and were discharged 10 days after the procedure.Conclusions: This technique may be a good minimal invasive off-the-shelf technical option for aortic arch "debranching". More data and further improvements are required before this promising technique can be widely advocated. (C) 2011 European Society for Vascular Surgery. Published by Elsevier Ltd. All rights reserved.

Glutaraldehyde-treated homologous vein graft as a vein substitute: experimental study in rabbits

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modified 'Stent-Graft Sandwich' Technique for Treatment of Isolated Common Iliac Artery Aneurysm in Patient With Marfan Syndrome

Isolated iliac artery aneurysms are rare in the general population (0.03%) and represent 2% of all abdominal aneurysms, and the association with Marfan syndrome is even rarer. We report a Marfan syndrome case with an isolated common iliac artery aneurysm treated by using a modified 'stent-graft sandwich' technique, with preservation of the internal iliac artery perfusion. The modified 'stent-graft sandwich' technique involves building an appropriate proximal neck just in the common iliac artery for fittingly housing two new stent-grafts inside, both deployed simultaneously and each one going to both distal iliac arteries (internal and external).

Trombectomia na angioplastia primária em ponte de safena

O emprego da trombectomia na angioplastia primária demonstra bons resultados clínicos e angiográficos imediatos, por meio de redução da embolização distal e do fenômeno do no-reflow e otimização da perfusão miocárdica. Essa técnica, principalmente usada em artérias coronárias, em raras oportunidades é descrita quando empregada em pontes de safena aortocoronarianas. Relatamos o caso de um paciente de 67 anos com infarto agudo do miocárdio relacionado a oclusão de ponte de safena e submetido a trombectomia com bons resultados.

Fixação de enxerto cutâneo em malha de espessura total com sutura ou cola de fibrina

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

Clinical evaluation and induced corneal vascularization study by native and anionic collagen membranes in rabbits corneas

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

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.

The effect of BMP-2 on the osteoconductive properties of beta-tricalcium phosphate in rat calvaria defects

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