Meniscal allograft transplantation in rabbit.

This study evaluated the technique for meniscal allograft transplantation using allografts preserved in glycerin 98% in rabbits. Euthanasia was performed at 70 days to compare the transplanted (TM1 to TM16) versus the contralateral meniscus (OM1 to OM16). Sixteen menisci, 8 transplanted and 8 contralateral, were submitted to gross examination, histomorphometric analysis for identification and quantification of cellular type, and for quantification and distribution of collagen fibers. A revascularization study was conducted in all of the other samples. Lengths of the OM varied from 0.9 to 1.0 cm and two TM were smaller. All TM were completely attached to the synovial membrane, except for one case that presented partial fixation. Both, TM and OM had similar amounts of chondrocytes, fibroblasts and fibrocytes, and at the horns, chondrocytes were predominant. The collagen fibers in TM were well organized throughout the body, and disorganized at the horns. These fibers in OM were organized. The amounts of collagen type I and III, and the vascularization of the perimeniscal tissue and of the edge were similar in OM and TM. These results demonstrated graft integration and thus this transplantation technique and preservation method may be recommended.

Treatment of critical defects produced in calvaria of mice with mesenchymal stem cells.

Mesenchymal stem cells (MSC) are present in specialized niches in perivascular regions of adult tissues and are able to differentiate into various cell types, such as those committed to repairing. Bone marrow derived MSC from eight young mice C57BL/ 6 gfp(+) were expanded in culture for repairing critical defects in calvarial bone produced in twenty-four young isogenic adult C57BL/6 mice. The animals were subjected to a cranial defect of 6.0mm diameter and divided into two equal experimental groups. Control group did not receive any treatment and the treated group received a MSC pellet containing 1.0 x 10(7) cells/mL into the defects. The group treated with MSC showed increased angiogenesis and amount of new bone deposited on the defect limits than that observed in the control group. The results demonstrated that transplantation of bone marrow-derived MSC of C57BL/6 gfp(+) mice to bone critical defects produced in mice calvarial contributes positively to the bone repair process. MSC presets ability to influence the correct functioning of osteoblasts, increases the amount of mobilized cells for the repairing process, speeds up growth, and increases deposition of bone matrix.

Effects of exercise training and stem cell therapy on the left ventricle of infarcted rats.

INTRODUCTION AND OBJECTIVES: Stem cell therapy and aerobic exercise are non-pharmacological therapies following myocardial infarction. The aim of this study was to test whether aerobic exercise training enhances the benefits of mesenchymal stem cell (MSC) therapy on remodeling of the extracellular matrix and fetal gene expression in the left ventricle of infarcted rats. METHODS: Myocardial infarction was surgically induced in six-week old male Wistar rats. Animals were divided into four groups: sedentary control (SC) and sedentary and stem cell treated (SCMSC); exercised (EX) and exercised and stem cell treated (EXMSC). Bone marrow-derived MSCs were immediately transplanted via the tail vein (concentration: 1×106 cells). Exercise training (five days/week, 60 min/day; 60% of maximal running speed) started 24 hours after myocardial infarction and lasted for 12 weeks. RESULTS: Exercise capacity was higher in exercised than in sedentary groups. Animals in the SCMSC, EX and EXMSC groups exhibited better cardiac function than those in SC. Collagen content was lower in the SCMSC, EX and EXMSC groups than in SC and skeletal α-actin expression was lower in EX and EXMSC than in SC. The α/ß-MHC ratio was higher in EX and EXMSC than in SC. The combination of therapies further reduced collagen content in the remote region of the infarct (∼24%) and skeletal α-actin expression (∼30%). CONCLUSION: Aerobic exercise training appears to enhance the beneficial effects of stem cell therapy on remodeling of the extracellular matrix and fetal gene expression in the left ventricle of rats with moderate infarction.

Acute and chronic observations of complete atrioventricular block in rats.

The mechanisms of production, and gross, microscopic and electrocardiograhic findings of surgically-induced complete heart block (CHB) in the adult rat are presented. This is an effective in vivo model for establishing alternative methods to electronic pacemakers and for providing detailed information aimed at replacement, reduction and refinement of the technique. Sternal thoracotomy was employed to identify the epicardial fat pad by the aortic root, used as a landmark for cauterization of the atrioventricular (AV) node. Stable CHB was produced in 60 rats with a 70% survival rate. The best survival rate was observed in 8-week-old animals weighing 221 ± 27.6 g. Heart rate before cauterization was 387 ± 55 bpm, reduced after cauterization to 126 ± 40 bpm in the survival and to 65 ± 19 bpm in the non-survival groups. At 30 days findings were: elevated left ventricular end-diastolic pressure (21 ± 5.4 mmHg, P < 0.05); maximal rate of rise of left ventricular pressure (LVP) during isovolumetric contraction (2192 ± 235 mmHg/s, P < 0.05); maximal rate of decrease of LVP (-1658 ± 191 mmHg/s, P < 0.05); isovolumetric relaxation constant (5.7 ± 0.8 ms, P < 0.05) with wet-to-dry lung-weight ratio (78.1 ± 0.4, P < 0.05); heart weight/body weight (0.6 ± 0.1, P < 0.05); heart volume (1.8 ± 0.3 mL, P < 0.05); longitudinal diameter (20.2 ± 1.91 mm, P < 0.05); and transversal diameter (17.0 ± 1.4 mm, P < 0.05) with supported dilated cardiomyopathy which culminated in chronic heart failure. CHB hearts had increased preload and replacement of myofibrils by collagen. CHB was achieved reproducibly by cauterization of the rat AV node and/or His bundle. This led to electrophysiological, hemodynamic, and structural remodeling, and could be useful in long-term cardiac remodeling assessments and potential therapy development.

Role of the autologous mesenchymal stem cells compared with platelet rich plasma on cicatrization of cutaneous wounds in diabetic mice / Papel das células-tronco mesenquimais autólogas em comparação com plasma rico em plaquetas na cicatrização de feridas cutâneas em camundongos diabéticos

Chronic cutaneous lesions affect 15% of diabetic human patients and represent a risk 15 to 46 times larger of limb amputations compared to people with normal glycemia. It is assumed that half of these amputations could be prevented by early treatment of wounds, for example, with proper cell therapy. Objectives: In this study, the action of the autologous transplant of mesenchymal stem-cells (MSC) was evaluated compared to the treatment with autologous platelet rich plasma (PRP) in the cicatrization of cutaneous lesions induced in diabetic mice. These animals were previously treated with streptozootocin to induce diabetes mellitus and round wounds of 1.5cm in diameter were created in the posterior region. Diameters of the wounds and healing time were evaluated during 30 days and the results were submitted to variance analysis and Tukey's test average. It was noticed that the animals treated with MSC presented a more accelerated cicatrization of the cutaneous lesion than the animals treated with PRP. However, the treatment with PRP presented better results than just the daily asepsis of the lesions with saline or covering them with semi-permeable bandage. Besides, the use of semi-permeable bandage kept the cutaneous lesions of diabetic mice did not interfere negatively with cicatrization, proved to be harmless to use, but kept the cutaneous lesions more hydrated than the ones exposed to the environment.(AU)

Lesões cutâneas crônicas afetam 15% dos pacientes diabéticos e humanos representam um risco 15 a 46 vezes maior de amputações de membros em comparação com as pessoas com a glicemia normal. Supõe-se que a metade destas amputações poderia ser evitada por meio do tratamento precoce das feridas cutâneas com, por exemplo, uma adequada terapia celular. Objetivos: Neste estudo, a ação do transplante autólogo de células estaminais mesenquimais (MSC) foi avaliada em comparação com o tratamento com plasma rico em plaquetas autólogo (PRP) na cicatrização de lesões cutâneas induzidas em camundongos diabéticos. Estes animais foram previamente tratados com estreptozotocina para induzir diabetes mellitus e feridas redondas de 1,5 cm de diâmetro foram criadas na região posterior. Os diâmetros dos ferimentos e tempo de cicatrização foram avaliados durante 30 dias e os resultados foram submetidos à análise de variância e média pelo teste de Tukey. Verificou-se que os animais tratados com MSC apresentam uma cicatrização mais acelerada da lesão cutânea que do que os animais tratados com PRP. No entanto, o tratamento com PRP apresentou melhores resultados do que apenas a assepsia das lesões diariamente com solução salina ou cobrindo-os com atadura semi-permeável. Além disso, a utilização de atadura semi-permeável mantidas as lesões cutâneas de camundongos diabéticos não interfere negativamente com a cicatrização, provou ser inofensiva para usar, mas manteve as lesões cutâneas hidratadas mais do que os expostos ao meio ambiente.(AU)

Avaliação das fibras colágenas de meniscos frescos e preservados em glicerina: estudo experimental em coelhos (Oryctolagus cuniculus) / Evaluation of the collagen fibers of the fresh meniscus and the meniscuses preserved in glycerine: experimental study in rabbits (Oryctolagus cuniculus)

No presente estudo foi avaliado o efeito da glicerina 98 por cento sobre as fibras colágenas, arquitetura tecidual e o tamanho de meniscos mediais de coelhos da raça Nova Zelândia. Os meniscos foram separados em três grupos: (1) Grupo MF com meniscos frescos (grupo controle), (2) Grupo MG com meniscos preservados em glicerina 98 por cento por 30 dias, e (3) Grupo MR com meniscos preservados em glicerina 98 por cento por 30 dias e reidratados em NaCl 0,9 por cento, por 12 horas. Os cortes histológicos foram corados com sirius red para identificação dos tipos de colágenos e observados em microscópio de luz polarizada, avaliando-se a concentração total de colágeno Tipo I e III e a disposição das fibras. Os meniscos frescos apresentaram significativamente maior concentração de fibras colágenas Tipo I e menor concentração de fibras colágenas Tipo III que os meniscos preservados (MG e MR); isto ocorreu devido à perda de água e conseqüente redução do tamanho dos meniscos e retração das fibras colágenas dos meniscos dos Grupos MG e MR; isto pode ter feito com que as fibras Tipo I, mais espessas e em maior quantidade, se tornassem mais evidentes do que as fibras colágenas Tipo III, que são mais delgadas e frágeis (fibrilas). Nos três grupos estudados, as fibras colágenas apresentaram-se de forma circunferencial, interpostas por fibras orientadas radialmente. Entretanto, nos grupos tratados (MG e MR) foi observado, em pequenas áreas, leve desorganização das fibras colágenas, o que correspondeu a 42,8 por cento e 14,3 por cento dos meniscos, respectivamente. O grupo de meniscos em glicerina apresentou redução significativa (p<0,05) no tamanho em relação ao Grupo MF. No Grupo MR, 85,7 por cento dos meniscos retornaram ao seu tamanho inicial após a reidratação. A glicerina 98 por cento é eficaz na preservação de meniscos, mantendo o tamanho, a arquitetura estrutural, a integridade e percentagem do colágeno dos meniscos preservados semelhante à de...

In the present study was evaluated the effect of 98 percent glycerin on the collagen fibers, tissue architecture and size of medial meniscuses of New Zealand rabbits. The animals were separated into three groups: (1) Group MF of fresh meniscus, (2) Group MG of meniscus preserved in glycerin for 30 days, and (3) Group MR of meniscus preserved in glycerin for 30 days and rehydrated in NaCl 0.9 percent for 12 hours. Histological sections were stained with sirius red for identification of collagen types, which were examined with a polarized light microscope. The total collagen concentration and the fiber arrangement were evaluated. Group MF presented higher Type I collagen concentration and lower Type III collagen concentration when compared with Group MG and MR. This fact is due to the water loss and consequent reduction in size and subsequent retraction of the collagen fibers of the meniscus from these groups, caused by dehydration. This may have occurred because those Type I fibers, thicker and in larger quantities, become more evident than Type III collagen fibers, which are more slender and fragile (fibrils). In the three groups studied, the collagen fibers presented themselves in a circumference form, interposed by radially oriented fibers. All of the meniscuses from Group MF presented fibers arranged obliquely, while in the treated groups, a slight disorganization of collagen fibers could be observed in some areas, what corresponds to 42.8 percent and 14.3 percent of the meniscuses, respectively. The MG group presented a significant decrease (p<0.05) in size if compared with the MF group. In the MR group, 85.7 percent of the meniscuses went back to the original size after rehydratation. The 98 percent glycerin is effective in preserving the meniscus, following rehydratation, maintaining size, structural architecture, integrity and percentage of collagen of the meniscus preserved similar to the fresh one.

Estrutura e celularidade de meniscos frescos de coelhos (Oryctolagus cuniculus) preservados em glicerina / Structure and cellularity of the fresh menisci (Oryctolagus cuniculus) of rabbits and the menisci preserved in glycerin

No presente estudo foi avaliada a arquitetura tecidual, a população celular, assim como a integridade e a distribuição dos tipos celulares em meniscos frescos de coelhos e preservados em glicerina 98 por cento. Foram analisados meniscos mediais de coelhos recém abatidos, que foram distribuídos em três grupos: o grupo MF (n=7), composto por meniscos frescos, correspondeu ao grupo controle; o grupo MG (n=7), composto por meniscos preservados em glicerina 98 por cento, por 30 dias, e o grupo MR (n=7), por meniscos preservados em glicerina 98 por cento e reidratados em NaCl 0,9 por cento, por 12 horas. Em todos os meniscos foram identificados e quantificados os diferentes tipos celulares: fibroblastos/fibrócitos e condrócitos. A população celular foi estatisticamente semelhante nos três grupos de meniscos, sendo que os meniscos preservados, grupos MG e MR, apresentaram menor intensidade de coloração e retração das fibras colágenas, diminuição de volume e maior intensidade de coloração dos núcleos (condensação da cromatina), em relação aos meniscos frescos (MF), caracterizando o fenômeno de lise celular. A matriz fibrocartilaginosa dos meniscos preservados revelou- se bem preservada mantendo a arquitetura tecidual dos meniscos. Conclui-se que a glicerina 98 por cento é uma opção de meio de preservação para meniscos objetivando aloenxerto, com matriz colágena desvitalizada.

In the present study was evaluated the tissue architecture, the percentage of cellular population, as well as viability and distribution of cells in fresh menisci of rabbits and preserved in 98 percent glycerin. Were analyzed medial menisci of rabbits freshly slaughtered, which were distributed into three groups: the MF group (n=7), composed of fresh menisci, corresponded to the control group; the MG group (n=7), composed by menisci preserved in 98 percent glycerin, for 30 days, and the MR group (n=7) by menisci preserved in 98 percent glycerin and rehydrated in NaCl 0.9 percent for 12 hours. In all menisci were identified and quantified the different cell types: fibroblasts/fibrocytes and condrocytes. The cell population percentage was statistically similar in all groups. All menisci preserved in the MG and MR groups showed a lower intensity of color and shrinkage of collagen fibers, reduced volume and higher intensity of staining of nucleus (chromatin condensation), as compared to fresh menisci (MF), featuring the phenomenon of cell lysis. The cartilaginous matrix of preserved menisci proved to be well preserved because the tissue architecture was maintained. It was concluded that 98 percent glycerin is an optional preservation mean for meniscal allografts with a devitalized collagenous matrix.