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2.
Brain Behav ; 8(7): e01027, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29920989

RESUMO

BACKGROUND: The aim was to evaluate the regenerative effect of epineural injection of rat ASCs (rASCs) in three different settings of acute and chronic compression in a rat sciatic nerve model. METHODS: Acute compression (60 s) with a vessel clamp over a distance of 1 mm (group 1) or 10 mm (group 2), as well as chronic compression with a permanent remaining, nonabsorbable polymeric clip over a distance of 1 mm (group 3) was performed. Depending on the group, either 5 × 106 rASCs or the same volume (25 µl) of culture medium (CM) was injected with a 30G needle in the epineurium at the time of compression. Outcome measures were functional gait evaluations, imaging analysis, histomorphometric analyses, and muscle weight. RESULTS: The rats in group 2 had a better function than those with group 1 at one and especially at 2 weeks. After 4 weeks however, almost all rats were close to a normal function. There was a similar Muscle Weight Ratio (MWR) after 2 weeks in all groups, whereas after 4 weeks, the MWR in group 3 was lower compared with group 1 and 2. Histomorphometric analysis showed a better myelination in group 1 & 2 compared to group 3 after 4 weeks. ASCs have a beneficial effect on myelin thickness (G-Ratio). CONCLUSIONS: We successfully evaluated the regenerative effect of epineural injection of rASCs in three different settings of acute and chronic compression. However, there were no significant differences in outcomes between the ASC-treated groups and control groups.


Assuntos
Adipócitos/transplante , Células-Tronco Adultas/transplante , Regeneração Nervosa/fisiologia , Nervo Isquiático/fisiologia , Animais , Constrição , Modelos Animais de Doenças , Feminino , Marcha/fisiologia , Injeções , Músculo Esquelético/fisiologia , Síndromes de Compressão Nervosa/terapia , Tamanho do Órgão/fisiologia , Nervos Periféricos , Ratos Sprague-Dawley , Neuropatia Ciática/terapia
3.
J Plast Reconstr Aesthet Surg ; 71(6): 833-839, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29653898

RESUMO

The nervous system plays a key role in controlling the dynamic functions of multicellular complex organisms. Although peripheral nerves are supposed to play a pivotal role in tumor growth and dissemination, little experimental evidence exists to date. We assessed the effect of denervation on breast cancer growth by magnetic resonance imaging (MRI) in rats. Human breast cancer cells were implanted into adipofascial flaps with intact or surgically excised supplying nerve. Tumor volumes were measured 2 and 8 weeks after implantation by in vivo MRI. Results were validated by histology. Postoperative tumor volumes at 2 and 8 weeks were reduced by 76% (95% CI: 22-93%) in the denervated groups. Tumor area as determined histologically was reduced by 70% (95% CI: 60-78%). Thus, peripheral denervation may be an effective surgical approach for the palliative treatment of locally progressing or uncontrollable breast cancer.


Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/patologia , Denervação , Nervos Periféricos/cirurgia , Retalhos Cirúrgicos/inervação , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Imageamento por Ressonância Magnética , Transplante de Neoplasias , Ratos , Ratos Sprague-Dawley , Carga Tumoral
4.
Aesthet Surg J ; 38(8): 900-910, 2018 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-29596609

RESUMO

BACKGROUND: Autologous fat grafting (AFG) to the breast is a frequent procedure in aesthetic and reconstructive surgery. Despite pure volume gain, questions remain regarding the engraftment rate, quality, and longevity. Little is known about the role of recipient tissue or innervation of the grafted area. OBJECTIVES: The goal of this study was to determine the optimal recipient layer and muscular pretreatment of AFG. METHODS: Fat was grafted to the breast, pectoralis muscle, or adjacent subcutaneous tissue of 42 rats. Nerve treatment included excision of a nerve segment, botulinum toxin (BTX) injection, or no treatment. Magnetic resonance imaging (MRI) and histological workup were carried out after 2 and 6 weeks. RESULTS: Six weeks after AFG, the proportion of viable fat cells within the grafted fat stayed high (median, [IQR]: 81% [72% to 85%]). The signs of inflammation decreased over time. Intramuscular grafting with intact nerves had a decreasing effect on the viability of the grafted cells compared with subcutaneous treatment (-10.21%; 95% confidence interval [-21.1 to 0.68]). CONCLUSIONS: If utilized on an intact nerve, intramuscular injection may lead to inferior results. If the nerve was cut or treated with BTX; however, intramuscular injection tends to be superior. These findings may prove interesting for future studies and eventual clinical application.


Assuntos
Tecido Adiposo/transplante , Denervação/métodos , Mamoplastia/métodos , Músculos Peitorais/cirurgia , Animais , Toxinas Botulínicas/administração & dosagem , Mama/inervação , Mama/cirurgia , Feminino , Injeções Intramusculares , Modelos Animais , Neurotoxinas/administração & dosagem , Músculos Peitorais/inervação , Ratos , Ratos Sprague-Dawley , Nervos Torácicos/efeitos dos fármacos , Nervos Torácicos/cirurgia , Transplante Autólogo/efeitos adversos , Transplante Autólogo/métodos
5.
Brain Behav ; 7(7): e00723, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28729931

RESUMO

INTRODUCTION: A range of behavioral testing paradigms have been developed for the research of central and peripheral nerve injuries with the help of small animal models. Following any nerve repair strategy, improved functional outcome may be the most important evidence of axon regeneration. A novel automated gait analysis system, the CatWalk™, can measure dynamic as well as static gait patterns of small animals. Of most interest in detecting functional recovery are in particular dynamic gait parameters, coordination measures, and the intensity of the animals paw prints. This article is designed to lead to a more efficient choice of CatWalk parameters in future studies concerning the functional evaluation of nerve regeneration and simultaneously add to better interstudy comparability. METHODS: The aims of the present paper are threefold: (1) to describe the functional method of CatWalk gait analysis, (2) to characterize different parameters acquired by CatWalk gait analysis, and to find the most frequently used parameters as well as (3) to compare their reliability and validity throughout the different studies. RESULTS: In the reviewed articles, the most frequently used parameters were Swing Duration (30), Print Size (27), Stride Length (26), and Max Contact Area (24). Swing Duration was not only frequently used but was also the most reliable and valid parameter. Therefore, we hypothesize that Swing Duration constitutes an important parameter to be chosen for future studies, as it has the highest level of reliability and validity. CONCLUSION: In conclusion, CatWalk can be used as a complementary approach to other behavioral testing paradigms to assess clinically relevant behavioral benefits, with the main advantage that this system demonstrates both static and dynamic gait parameters at the same time. Due to limited reliability and validity of certain parameters, we recommend that only the most frequently assessed parameters should be used in the future.


Assuntos
Marcha/fisiologia , Regeneração Nervosa/fisiologia , Traumatismos dos Nervos Periféricos/fisiopatologia , Recuperação de Função Fisiológica/fisiologia , Animais , Modelos Animais de Doenças , Reprodutibilidade dos Testes
6.
Stem Cells Dev ; 24(18): 2127-41, 2015 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-26134465

RESUMO

Tissue engineering is a popular topic in peripheral nerve repair. Combining a nerve conduit with supporting adipose-derived cells could offer an opportunity to prevent time-consuming Schwann cell culture or the use of an autograft with its donor site morbidity and eventually improve clinical outcome. The aim of this study was to provide a broad overview over promising transplantable cells under equal experimental conditions over a long-term period. A 10-mm gap in the sciatic nerve of female Sprague-Dawley rats (7 groups of 7 animals, 8 weeks old) was bridged through a biodegradable fibrin conduit filled with rat adipose-derived stem cells (rASCs), differentiated rASCs (drASCs), human (h)ASCs from the superficial and deep abdominal layer, human stromal vascular fraction (SVF), or rat Schwann cells, respectively. As a control, we resutured a nerve segment as an autograft. Long-term evaluation was carried out after 12 weeks comprising walking track, morphometric, and MRI analyses. The sciatic functional index was calculated. Cross sections of the nerve, proximal, distal, and in between the two sutures, were analyzed for re-/myelination and axon count. Gastrocnemius muscle weights were compared. MRI proved biodegradation of the conduit. Differentiated rat ASCs performed significantly better than undifferentiated rASCs with less muscle atrophy and superior functional results. Superficial hASCs supported regeneration better than deep hASCs, in line with published in vitro data. The best regeneration potential was achieved by the drASC group when compared with other adipose tissue-derived cells. Considering the ease of procedure from harvesting to transplanting, we conclude that comparison of promising cells for nerve regeneration revealed that particularly differentiated ASCs could be a clinically translatable route toward new methods to enhance peripheral nerve repair.


Assuntos
Regeneração Nervosa/fisiologia , Traumatismos dos Nervos Periféricos/terapia , Nervo Isquiático/lesões , Transplante de Células-Tronco/métodos , Engenharia Tecidual/métodos , Implantes Absorvíveis , Adipócitos/citologia , Tecido Adiposo/citologia , Animais , Terapia Baseada em Transplante de Células e Tecidos/métodos , Células Cultivadas , Feminino , Fibrina/metabolismo , Modelos Animais , Músculo Esquelético/crescimento & desenvolvimento , Músculo Esquelético/inervação , Neurogênese/fisiologia , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/citologia , Células-Tronco/citologia
7.
Cell Transplant ; 24(2): 203-11, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-24380629

RESUMO

Traumatic nerve injuries are a major clinical challenge. Tissue engineering using a combination of nerve conduits and cell-based therapies represents a promising approach to nerve repair. The aim of this study was to examine the regeneration potential of human adipose-derived stem cells (hASCs) after transplantation in a nonautogenous setting and to compare them with autogenous rat ASCs (rASCs) for early peripheral nerve regeneration. Furthermore, the use of MRI to assess the continuous process of nerve regeneration was elaborated. The sciatic nerve injury model in female Sprague-Dawley rats was applied, and a 10-mm gap created by using a fibrin conduit seeded with the following cell types: rASCs, Schwann cell (SC)-like cells from rASC, rat SCs (rSCs), hASCs from the superficial and deep abdominal layer, as well as human stromal vascular fraction (1 × 10(6) cells). As a negative control group, culture medium only was used. After 2 weeks, nerve regeneration was assessed by immunocytochemistry. Furthermore, MRI was performed after 2 and 4 weeks to monitor nerve regeneration. Autogenous ASCs and SC-like cells led to accelerated peripheral nerve regeneration, whereas the human stem cell groups displayed inferior results. Nevertheless, positive trends could be observed for hASCs from the deep abdominal layer. By using a clinical 3T MRI scanner, we were able to visualize the graft as a small black outline and small hyperintensity indicating the regenerating axon front. Furthermore, a strong correlation was found between the length of the regenerating axon front measured by MRI and the length measured by immunocytochemistry (r = 0.74, p = 0.09). We successfully transplanted and compared human and autologous stem cells for peripheral nerve regeneration in a rat sciatic nerve injury model. Furthermore, we were able to implement the clinical 3T MRI scanner to monitor the efficacy of cellular therapy over time.


Assuntos
Imageamento por Ressonância Magnética , Traumatismos dos Nervos Periféricos/terapia , Nervo Isquiático/diagnóstico por imagem , Transplante de Células-Tronco , Células-Tronco/citologia , Tecido Adiposo/citologia , Animais , Modelos Animais de Doenças , Feminino , Fibrina/química , Humanos , Imuno-Histoquímica , Regeneração Nervosa , Traumatismos dos Nervos Periféricos/diagnóstico por imagem , Traumatismos dos Nervos Periféricos/patologia , Radiografia , Ratos , Ratos Sprague-Dawley , Células de Schwann/citologia , Células de Schwann/transplante , Nervo Isquiático/patologia , Engenharia Tecidual , Transplante Autólogo , Transplante Heterólogo
9.
Cytotechnology ; 65(3): 437-45, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23095943

RESUMO

The therapeutic potential of adult stem cells may become a relevant option in clinical care in the future. In hand and plastic surgery, cell therapy might be used to enhance nerve regeneration and help surgeons and clinicians to repair debilitating nerve injuries. Adipose-derived stem cells (ASCs) are found in abundant quantities and can be harvested with a low morbidity. In order to define the optimal fat harvest location and detect any potential differences in ASC proliferation properties, we compared biopsies from different anatomical sites (inguinal, flank, pericardiac, omentum, neck) in Sprague-Dawley rats. ASCs were expanded from each biopsy and a proliferation assay using different mitogenic factors, basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) was performed. Our results show that when compared with the pericardiac region, cells isolated from the inguinal, flank, omental and neck regions grow significantly better in growth medium alone. bFGF significantly enhanced the growth rate of ASCs isolated from all regions except the omentum. PDGF had minimal effect on ASC proliferation rate but increases the growth of ASCs from the neck region. Analysis of all the data suggests that ASCs from the neck region may be the ideal stem cell sources for tissue engineering approaches for the regeneration of nervous tissue.

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