Human Multipotent Mesenchymal Stromal Cells from Distinct Sources Show Different In Vivo Potential to Differentiate into Muscle Cells When Injected in Dystrophic Mice

Limb-girdle muscular dystrophies are a heterogeneous group of disorders characterized by progressive degeneration of skeletal muscle caused by the absence or deficiency of muscle proteins. The murine model of Limb-Girdle Muscular Dystrophy 2B, the SJL mice, carries a deletion in the dysferlin gene. Functionally, this mouse model shows discrete muscle weakness, starting at the age of 4-6 weeks. The possibility to restore the expression of the defective protein and improve muscular performance by cell therapy is a promising approach for the future treatment of progressive muscular dystrophies (PMD). We and others have recently shown that human adipose multipotent mesenchymal stromal cells (hASCs) can differentiate into skeletal muscle when in contact with dystrophic muscle cells in vitro and in vivo. Umbilical cord tissue and adipose tissue are known rich sources of multipotent mesenchymal stromal cells (MSCs), widely used for cell-based therapy studies. The main objective of the present study is to evaluate if MSCs from these two different sources have the same potential to reach and differentiate in muscle cells in vivo or if this capability is influenced by the niche from where they were obtained. In order to address this question we injected human derived umbilical cord tissue MSCs (hUCT MSCs) into the caudal vein of SJL mice with the same protocol previously used for hASCs; we evaluated the ability of these cells to engraft into recipient dystrophic muscle after systemic delivery, to express human muscle proteins in the dystrophic host and their effect in functional performance. These results are of great interest for future therapeutic application.

Striatum brain-derived neurotrophic factor levels are decreased in dystrophin-deficient mice

Brain dystrophin is enriched in the postsynaptic densities of pyramidal neurons specialized regions of the subsynaptic cytoskeletal network, which are critical for synaptic transmission and plasticity. Lack of dystrophin in brain structures have been involved with impaired cognitive functions. The brain-derived neurotrophic factor (BDNF) is a regulator of neuronal survival, fast synaptic transmission, and activity-dependent synaptic plasticity. The present study investigated BDNF protein levels by Elisa analysis in prefrontal cortex, cerebellum, hippocampus, striatum and cortex tissues from male dystrophic mdx (n = 5) and normal C57BL10 mouse (n = 5). We observed that the mdx mouse display diminution in BDNF levels in striatum (t = 6.073; df = 6; p = 0.001), while a tendency of decrease in BDNF levels was observed in the prefrontal cortex region (t = 1.962; df = 6; p = 0.096). The cerebellum (t = 1.258; df = 7; p = 0.249), hippocampus (t = 0.631; df = 7; p = 0.548) and cortex (t = 0.572; df = 7; p = 0.586) showed no significant alterations as compared to wt mouse. In conclusion, we demonstrate that only striatum decreased BDNF levels compared with wild-type (wt) mouse, differently to the other areas of the brain. This dystrophin deficiency may be affecting BDNF levels in striatum and contributing, in part, in memory storage and restoring. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Animal models for genetic neuromuscular diseases

The neuromuscular disorders are a heterogeneous group of genetic diseases, caused by mutations in genes coding sarcolemmal, sarcomeric, and citosolic muscle proteins. Deficiencies or loss of function of these proteins leads to variable degree of progressive loss of motor ability. Several animal models, manifesting phenotypes observed in neuromuscular diseases, have been identified in nature or generated in laboratory. These models generally present physiological alterations observed in human patients and can be used as important tools for genetic, clinic, and histopathological studies. The mdx mouse is the most widely used animal model for Duchenne muscular dystrophy (DMD). Although it is a good genetic and biochemical model, presenting total deficiency of the protein dystrophin in the muscle, this mouse is not useful for clinical trials because of its very mild phenotype. The canine golden retriever MD model represents a more clinically similar model of DMD due to its larger size and significant muscle weakness. Autosomal recessive limb-girdle MD forms models include the SJL/J mice, which develop a spontaneous myopathy resulting from a mutation in the Dysferlin gene, being a model for LGMD2B. For the human sarcoglycanopahties (SG), the BIO14.6 hamster is the spontaneous animal model for delta-SG deficiency, whereas some canine models with deficiency of SG proteins have also been identified. More recently, using the homologous recombination technique in embryonic stem cell, several mouse models have been developed with null mutations in each one of the four SG genes. All sarcoglycan-null animals display a progressive muscular dystrophy of variable severity and share the property of a significant secondary reduction in the expression of the other members of the sarcoglycan subcomplex and other components of the Dystrophin-glycoprotein complex. Mouse models for congenital MD include the dy/dy (dystrophia-muscularis) mouse and the allelic mutant dy(2J)/dy(2J) mouse, both presenting significant reduction of alpha 2-laminin in the muscle and a severe phenotype. The myodystrophy mouse (Large(myd)) harbors a mutation in the glycosyltransferase Large, which leads to altered glycosylation of alpha-DG, and also a severe phenotype. Other informative models for muscle proteins include the knockout mouse for myostatin, which demonstrated that this protein is a negative regulator of muscle growth. Additionally, the stress syndrome in pigs, caused by mutations in the porcine RYR1 gene, helped to localize the gene causing malignant hypertermia and Central Core myopathy in humans. The study of animal models for genetic diseases, in spite of the existence of differences in some phenotypes, can provide important clues to the understanding of the pathogenesis of these disorders and are also very valuable for testing strategies for therapeutic approaches.

Implant-retained thumb prosthesis with anti-rotational attachment for a geriatric patient

This report presents the use of a dental implant with an anti-rotational attachment for the retention of a thumb prosthesis. A retention system was manufactured with an attachment (UCLA) screwed into the implant with a two-bar system that was cast in metallic silver palladium. A substructure made from heat-cured acrylic resin was joined to the retention system by clips to join the thumb to the finger (bar clip) in the cast with implant rejoinder. The silicone material, Silastic-MDX 44210, was used to achieve function and aesthetics. Following osseointegration, no skin problems were observed. Whilst the implant-retained digital prosthesis presented some motor limitations, it allowed the patient to return to entertainment and achieve social conviviality.

Biocompatibility Evaluation of 3 Facial Silicone Elastomers

The failure of facial prostheses is caused by limitations in the properties of existing materials, especially the biocompatibility. This study aimed to evaluate the biocompatibility of maxillofacial silicones in subcutaneous tissue of rats. Thirty Wistar rats received subcutaneous implants of 3 maxillofacial silicone elastomers (LIM 6050, MDX 4-4210, and industrial Silastic 732 RTV). A histomorphometric evaluation was conducted to analyze the biocompatibility of the implants. Eight areas of 60.11 mm(2) from the surgical pieces were analyzed. Mesenchymal cells, eosinophils, and foreign-body giant cells were counted. Data were submitted to analysis of variance and Tukey test. Initially, all implanted materials exhibited an acceptable tissue inflammatory response, with tissue reactions varying from light to moderate. Afterward, a fibrous capsule around the silicone was observed. The silicones used in the current study presented biocompatibility and can be used for implantation in both medical and dental areas. Their prosthetic indication is conditioned to their physical properties. Solid silicone is easier to adapt and does not suffer apparent modifications inside the tissues.

Bond strength between acrylic resin and maxillofacial silicone

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

Effect of Disinfection and Accelerated Aging on Color Stability of Colorless and Pigmented Facial Silicone

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of Pigment and Opacifier on Dimensional Stability and Detail Reproduction of Maxillofacial Silicone Elastomer

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

Color Stability Comparison of Silicone Facial Prostheses Following Disinfection

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

Influence of opacifiers on dimensional stability and detail reproduction of maxillofacial silicone elastomer

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

Evaluation of dimensional change and detail reproduction in silicones for facial prostheses.

The aim of this study was to evaluate the dimensional stability and detail reproduction of two silicones used for facial prosthesis, under the influence of chemical disinfection and storage time. Twenty-eight test specimens were obtained, half made of Silastic MDX 4-4210 silicone, and the other half of Silastic 732 RTV silicone. The test specimens were divided into 4 groups: Silastic 732 RTV and Silastic MDX 4-4210, with disinfection 3 times a week with Efferdent and without disinfection. Dimensional change was analyzed using an electronic comparison microscope and detail reproduction was observed under a stereo microscope, immediately and 2 months after the test specimens were made. Once the results were obtained, an analisis of variance (ANOVA) was applied, followed by the Tukey's Test with 1% confidence. The storage time factor had a statistical influence on dimensional stability: Silastic MDX 4-4210 had less contraction than Silastic 732 RTV. Chemical disinfection did not significantly alter the dimensional stability of the materials used. Regarding detail reproduction, no alteration of values was observed in any of the materials analyzed, regardless of storage period or disinfection.

Visual evaluation of color stability after accelerated aging of pigmented and nonpigmented silicones to be used in facial prostheses

Objectives: The objective of this study was to evaluate by a visual method of comparison the color stability of nonpigmented and pigmented facial silicones after accelerated aging. Materials and Methods: Two kinds of silicones were used in this study; one specifically formulated for facial prostheses and the other an acetic silicone for industrial use. Twenty-four trial bodies were made for each silicone. These were divided into colorless and intrinsically pigmented groups: ceramic, make-up, and iron oxide. The groups were submitted to accelerated aging for nonmetallic materials. An initial reading and subsequent readings were made at 163, 351, 692, and 1000 hours using a visual method of comparison. The values were annotated in a spreadsheet by two observers, according to scores elaborated for this study. Results: All groups presented color stability in the visual method. According to the results obtained and analyzed in this study, we can conclude that both silicones, Silastic 732 RTV and Silastic MDX 4-4210, behaved similarly, they can therefore be indicated for use in maxillofacial prosthesis. The time factor of aging influenced negatively, independently of the pigmentation, or lack of it, and of silicones and no group had visually noticeable alterations in any of the accelerated aging time, independently of the addition or not of pigments.

Evaluation of hardness and surface roughness of two maxillofacial silicones following disinfection

The purpose of this study was to evaluate the Shore A hardness and surface roughness of two silicones for maxillofacial prosthetic treatment, under the influence of chemical disinfection and storage. Twenty-eight specimens were obtained, half of which were made of Silastic MDX 4-4210 silicone and, the other half were made of Silastic 732 RTV silicone. The specimens were divided into four groups: Silastic 732 RTV and MDX 4-4210 with disinfection 3 times a week with Efferdent tablets and the same materials without disinfection. The hardness of the materials was analyzed with a Shore A Durometer. The surface roughness was established by a digital portable roughness tester, initially and 2 months after the confection of the specimens. A variance test was applied (2-way ANOVA), followed by Tukey test (the level of significance was set at 1%). The storage time factor statistically influenced (p < 0.01) the materials' properties of hardness and roughness. MDX 4-4210 (28.59 Shore A, 0.789 Ra) presented higher values than Silastic 732 RTV (18.08 Shore A, 0.656 Ra) for both properties. Regarding the disinfection period, there was no significant difference in any of the materials tested. © 2009 Sociedade Brasileira de Pesquisa Odontológica.

Hardness evaluation of prosthetic silicones containing opacifiers following chemical disinfection and accelerated aging

We evaluated the effects of disinfection and aging on the hardness of silicones containing opacifiers and intended for use in facial prosthetics. A total of 90 samples were produced using a cylindrical metal mold 3 mm in height and 30 mm in diameter. The samples were fabricated from Silastic MDX 4-4210 silicone in three groups: GI contained no opacifier, GII contained barium sulfate (Ba), and GIII contained titanium dioxide (Ti). The samples were disinfected using effervescent tablets (Ef), neutral soap (Ns), or 4% chlorhexidine (Cl) 3 times a week for 60 days. After this period the samples underwent 1,008 hours of accelerated aging. The hardness was measured using a durometer immediately following the disinfection period and after 252, 504, and 1,008 hours of aging. The data were statistically analyzed using 3-way ANOVA and the Tukey test (p < .05). The GIII group exhibited the greatest variation in hardness regardless of elapsed time. All groups displayed greater hardness after 1,008 hours of accelerated aging independent of disinfectant type. All of the hardness values were within the clinically acceptable range.

Effect of disinfection and accelerated ageing on dimensional stability and detail reproduction of a facial silicone with nanoparticles

The aim of the present study was to evaluate the effect of disinfection and accelerated ageing on the dimensional stability and detail reproduction of a facial silicone with different types of nanoparticle. A total of 60 specimens were fabricated with Silastic MDX 4-4210 silicone and they were divided into three groups: colourless and pigmented with nanoparticles (make-up powder and ceramic powder). Half of the specimens of each group were disinfected with Efferdent tablets and half with neutral soap for 60 days. Afterwards, all specimens were subjected to accelerated ageing. Both dimensional stability and detail reproduction tests were performed after specimen fabrication (initial period), after chemical disinfection, and after accelerated ageing periods (252, 504 and 1008 hours). The dimensional stability test was conducted using AutoCAD software, while detail reproduction was analysed using a stereoscope magnifying glass. Dimensional stability values were statistically evaluated by analysis of variance (ANOVA) followed by Tukey's test (p < 0.01). Detail reproduction results were compared using a score. Chemical disinfection and also accelerated ageing affected the dimensional stability of the facial silicone with statistically significant results. The silicone's detail reproduction was not affected by these two factors regardless of nanoparticle type, disinfection and accelerated ageing. © 2012 Informa UK, Ltd.