Experimental study on autologous injectable platelets rich fibrin combined with bone mesenchymal stem cells in treating sciatic nerve injury in rats / 中国修复重建外科杂志

Objective: To investigate the effectiveness of autologous injectable platelet rich fibrin (i-PRF) combined with bone marrow mesenchymal stem cells (BMSCs) for sciatic nerve injury in rats. Methods: BMSCs were isolated and cultured from tibial bone marrow of Sprague Dawley (SD) neonatal rats aged 10-15 days and passaged to the 4th generation. i-PRF was prepared from posterior orbital venous blood of adult SD rats by improved low-speed centrifugation. Twenty-four adult SD rats were selected and randomly divided into 4 groups with 6 rats in each group after the sciatic nerve Ⅲ degree injury model was established by modified crush injury method. Groups A, B, C, and D were injected with BMSCs suspension+autologous i-PRF, autologous i-PRF, BMSCs suspension, and normal saline, respectively. The Basso-Beattie-Bresnahan (BBB) score was used to evaluate the recovery of neurological function of the affected limb of rats every week from 1 to 8 weeks after operation. At 2 months after operation, the rats were sacrificed and the histological changes of sciatic nerve were observed by HE staining. The microstructural changes of nerve fibers, myelin sheath, and nucleus were observed by transmission electron microscope. The expressions of N-cadherin, Nestin, and glial fibrillary acidic protein (GFAP) were detected by Western blot. Results: No immune rejection or death occurred in the rats after operation. There was no significant difference in BBB scores between groups at 1 week after operation ( P>0.05); at 2-8 weeks after operation, BBB scores in group A were significantly higher than those in groups B, C, and D, and in groups B, C than in group D ( P0.05). HE staining showed that the nerve fibers in group A arranged in order, without defect or demyelination; the nerve fibers in group B were not clear and slightly swollen; some of the nerve fibers in group C were disordered and demyelinated; the nerve fibers in group D were not continuous, obviously demyelinated, and some of the nerve adventitia damaged. Transmission electron microscope showed that the structure of nerve fibers in group A was clear, myelin sheath was complete, and nucleus was dense; group B was slightly less than group A; group C had fuzzy structure, demyelination, and hollowing out; group D had disorder structure, demyelination, and hollowing out, and the middle part of nerve adventitia continuity. Western blot detection results showed that there was no significant difference in the relative expression of Nestin between groups ( P>0.05). The relative expression of N-cadherin was significantly lower in groups B, C, and D than in group A, in groups C and D than in group B, and in group D than in group C ( P0.05). Conclusion: Autologous i-PRF combined with BMSCs can effectively treat sciatic nerve tissue injury in rats.

Effect of the local application of stem cells on repairing facial nerve defects: a systematic review / 华西口腔医学杂志

OBJECTIVE@#To systematically evaluate the repairing effect of stem cells on facial nerve defects.@*METHODS@#Articles regarding the regenerating effect of stem cells on facial nerves in animals were collected from the databases of Pubmed, Cochrane Library, Web of Science, Embase, Scopus, and CBM. Two professionals independently completed the article screening, data extraction, and bias risk assessment. RevMan 5.3 and random-effects models were used for the statistical analysis, and the results were presented in the form of mean differences (MD) with a 95%CI. The results of functional evaluation (vibrissae movement, facial paralysis) and histological evaluation (density of myelinated fibers, diameter of fibers, thickness of myelin sheath, G ratio) of facial nerve were Meta-analyzed.@*RESULTS@#A total of 4 614 articles were retrieved from the 6 databases, and 15 of these articles were included in the Meta-analysis. For vibrissae movement and facial paralysis, the stem cell group scored significantly higher than the non-stem cell group (P<0.05). The density of myelinated fibers and thickness of the myelin sheath in the stem cell group were higher than those in the non-stem cell group (P<0.05). The G ratio in the stem cell group was smaller than that in the non-stem cell group (P=0.001). There was no significant difference in fiber diameter (P=0.08).@*CONCLUSIONS@#Stem cells have potential in promoting facial nerve regeneration.

Effectiveness Of Intermittent Pelvic Traction With And Without Self Neural Tissue Mobilization In Lumbar Radiculopathy

Background: Low back pain is a very common problem in adults. The clinical application of intermittent pelvic traction has become the common treatment for lumbar radiculopathy and is also used by clinicians in day to day practice. Only few authors have studied about the effectiveness of self neural tissue mobilization during intermittent pelvic traction. Hence the study was undertaken with an intention to find out the effect of intermittent pelvic traction with and without self neural tissue mobilization on pain and functional disability in patients with lumbar radiculopathy. Methodology: 74 patients with lumbar radiculopathy were included in the study. Subjects were randomly divided into two groups. Group A received only IPT and Group B received IPT with SNTM for 20 minutes with 10 second hold and rest time respectively. The treatment was given initially for 6 days continuously thereafter the treatment was given on alternate days for 1 week. Pre and post intervention pain (VAS) and functional disability (ODI) measures were noted. Results: On comparison between pre and post interventional values of pain and functional disability using paired ‘t’ test, a significant difference (p<0.001) were found in both the groups. As well as the present study showed significant decrease in the pain measurement (p=0.023) and functional disability measures (p=0.043) in group treated with IPT with SNTM. Conclusions: This study concluded that both IPT and IPT with SNTM are effective in both pain and functional disability measures. But when both groups were compared with each other, the IPT with SNTM was more effective than that of IPT alone. So it can be chosen as a treatment for lumbar radiculopathy.

Determination of the Effect of Diameter of the Sac on Prognosis in 64 Cases Operated for Meningomyelocele

OBJECTIVE: To examine the effect of meningomyelocele sac size on prognosis by retrospective review of 64 cases operated for meningomyelocele between January 2009 and December 2012. METHODS: We evaluated newborn babies operated for meningomyelocele by retrospectively reviewing their files for head circumference, location and with of the defect, accompanying anomalies, treatments administered, drugs that mother used during pregnancy. Based on the defect size, 3 patient groups were created as 0–24 cm² (group I), 25–39 cm² (group II), and 40 cm² and above (group III). RESULTS: Throughout the study, 64 babies were evaluated. Mean head circumference was 37.4 cm (range, 30.7–50 cm). Based on their location, 49 of the defects (76.5%) were lumbar, 7 (10.9%) were thoracolumbar, 4 (6.2%) were thoracic, 3 (3.1%) were sacral, 1 (1.5%) was cervical. Mean size of the meningomyelocele sac was 4.7 cm×5.8 cm (range, 1 cm×1 cm—10 cm×8 cm), 13 of the babies (20.3%) had skin defect requiring flap. According to accompanying anomalies, 47 of the babies (73.4%) had hydrocephalus, 7 (10.9%) had club foot, 1 (1.5%) had diastematomyelia, 1 (1.5%) had tethered cord. Thirty-nine of the babies (60.9%) had paraplegia, 10 (15.6%) had paraparesis, 8 (12.5%) had monoplegia; neurological examination in the remaining 7 babies was normal. CONCLUSION: In our study, increased diameter of meningomyelocele sac was associated with greater amount of neural tissue within the sac, which worsens the prognosis. Sac localization was not changing prognosis but infection rates, hospitalization duration were increased in babies with bigger diameter of sacs.

Comparative Study of Poly(L-Lactic Acid) Scaffolds Coated with Chitosan Nanoparticles Prepared via Ultrasonication and Ionic Gelation Techniques

In this study, an attempt was made to develop bi-functional constructs serving both as scaffolds and potential delivery systems for application in neural tissue engineering. The constructs were prepared in two steps. In the first step, the bulks of poly (L-lactic acid) (PLLA) in 1, 4-dioxane/water (87:13) were fabricated using liquid-liquid thermally induced phase separation technique. In the next step, the prepared bulks were coated with chitosan nanoparticles produced by two different techniques of ultrasonication and ionic gelation by grafting-coating technique. In ultrasonication technique, the chitosan solution (2 mg/mL) in acetic acid/sodium acetate buffer (90:10) was irradiated by an ultrasound generator at 20 kHz and power output of 750 W for 100 s. In ionic gelation technique, the tripolyphosphate in water solution (1 mg/mL) was added to the same chitosan solution. The physicochemical properties of the products were characterized by Scanning Electron Microscopy, Attenuated Total Reflection Fourier Transform-Infrared, liquid displacement technique, contact angle measurement, compressive and tensile tests, as well as zeta potential and particle size analysis using dynamic light scattering. Moreover, the cell proliferation and attachment on the scaffolds were evaluated through human glioblastoma cell line (U-87 MG) and human neuroblastoma cell line [BE (2)-C] culture respectively. The results showed that the samples coated with chitosan nanoparticles prepared by ultrasonication possessed enhanced hydrophilicity, biodegradation and cytocompatibility compared with pure PLLA and PLLA coated with chitosan nanoparticles prepared by ionic gelation. This study suggests successful nanoparticles-scaffold systems which can act simultaneously as potential delivery systems and tissue engineering scaffolds.

Optogenética e estimulação óptica neural: estado atual e perspectivas / Optogenetics and neural optical stimulation: current state and perspectives

Ao longo dos últimos 50 anos, o uso da luz, em especial o laser, vem promovendo grandes avanços em diversas áreas da ciência e da tecnologia. Na última década o uso de estímulos ópticos no campo da biomédica tem despertado grande interesse no meio acadêmico e na indústria. Dois ramos que se destacam pelo seu crescimento são: a estimulação óptica direta e a optogenética. A primeira utiliza diferentes parâmetros da luz para adequar o efeito desejado na interação com o tecido biológico. A segunda faz uso de engenharia genética para tornar os tecidos biológicos sensíveis à luz. A estimulação neural por infravermelho (estimulação óptica direta) não necessita de contato direto com o tecido e apresenta maior seletividade especial se comparada à estimulação elétrica, mas tem a capacidade restrita de ativar (despolarizar) os neurônios. A optogenética, entretanto, pode ser utilizada para manipular o tecido neural tornando-o sensível à luz; sendo, então, possível despolarizar ou hiperpolarizar os neurônios codificados, assim como monitorar as ativações por meio de codificação de proteínas fluorescentes sensíveis à tensão elétrica. Tanto a técnica de estimulação óptica por infravermelho ou a técnica de optogenética, vêm sendo aplicadas apenas à modelos animais. Os resultados mostram, entretanto, que há grande viabilidade de aplicação da estimulação óptica em seres humanos. Futuramente, tais técnicas poderão substituir o atual padrão ouro para a ativação neural, a estimulação elétrica, em aplicações envolvendo doenças neurológicas específicas.

Within the last 50 years the light and specially the laser has fomented great advances in several areas of science and technology. During the past decade the use of optical stimuli in the biomedical research field have been of great interest for both academy and industry. Two research branches that can be highlighted due to its growth are: direct optical stimulation and optogenetic. The first one uses different parameters of light to optimize the desired effect on the tissue interaction. The other branch works with genetic engineering technics to make cells sensitive to light. The neural stimulation by infrared (direct optical stimulation) does not require direct contact with the tissue and has higher spatial selectivity when compared to electrical stimulation, but it has restricted ability to activate (depolarize) neurons. The optogenetic, however, can be used to manipulate the neural tissue depolarizing or hyperpolarizing encoded neurons, as well as monitor activations by encoding fluorescent proteins sensitive to voltage. The stimulation by infrared optical or optogenetic, has been applied only to animal models although there is a great possibility for human applications. In the future, it may even replace existing techniques such as electrical brain stimulation to treat specific neurological diseases.

Distribution and differentiation of transplanted neural tissue committed stem cells to the brain after ischemic stroke / 解放军医学杂志

Objective To investigate the distribution and differentiation of neural tissue committed stem cells (NTCSCs) by venous transplantation to the brain after middle cerebral artery occlusion in rats. Methods Monocytes were collected from bone marrow and cultured in DMEM/F12 serum free medium plus EGF, bFGF and 2% B27 to obtain NTCSCs. Expressions of nestin, CXCR4, CD31, β -HI-tubulin and GFAP mRNA in NTCSCs were detected by RT-PCR. Modified Zea-Longa suture method was used to prepare middle cerebral artery occlusion (MCAO) model. The NTCSCs were labeled with DAPI and were injected through the caudal vein of the rats. At 1,3,5,7 days after injection, the rats were sacrificed and the brain tissues were prepared with frozen section in transplantation group. The distribution and differentiation of transplanted NTCSCs were observed under fluorescent microscope and laser confocal microscope respectively after immunofluorescent staining. Results The cultured NTCSC formed neural stem cell spheres in neural stem cell culture medium, and they expressed Nestin and SDF-1 receptor CXCR4 at protein level. RT-PCR showed that NTCSCs expressed Nestin, CXCR4, CD31, β - III-tubulin and GFAP mRNA. DAPI-labelled NTCSCs could be observed in the brain 3 days after transplantation and a few GFAP expressing cells could be found on the 7th day. Conclusion The NTCSCs by intravenous transplantation can enter rat brain, and they distribute mainly in ischemic tissues. Some NTCSCs labelled with DAPI in the brain tissue could express GFAP.

Evidencia científica de los efectos terapéuticos atribuidos al deslizamiento lateral cervical / Scientific evidence of the therapeutic effects attributed to the cervical lateral glide

El deslizamiento Lateral Cervical es una técnica de Terapia Manual Ortopédica muy usada para el manejo del dolor y la disfunción vertebral de origen cervical, aunque existen estudios experimentales acerca de los fundamentos neurofisiológicos sobre los cuales ejerce su acción; se realizará una síntesis de la evidencia relativa a los efectos clínicos de la técnica de deslizamiento lateral cervical, a través de una Revisión Sistemática de Ensayos Clínicos Aleatorizados. Objetivo: Determinar si existe evidencia científica que avale los efectos terapéuticos atribuidos a la técnica de deslizamiento lateral cervical usado por los kinesiólogos para el manejo clínico de diversas condiciones músculo esqueléticas; Estrategia de búsqueda; sólo se incluyeron en la búsqueda de Ensayos Clínicos Aleatorizados (ECAs), las bases de datos usadas fueron : MEDLINE (Pubmed/PMC), OVID, Cochrane, PEDro y EMBASE (Science-Direct); Resultados; se seleccionaron 4 ECAs que cumplían con los criterios de elegibilidad; Conclusiones; Existe Moderada Evidencia que el deslizamiento lateral cervical produce Hipoalgesia inmediata en pacientes con Dolor Cervicobraquial Neurogénico de tipo Sub agudo, Epicondilalgia unilateral crónica y dolor de hombro unilateral con mala respuesta al tratamiento tradicional.

The Cervical Lateral Glide is a technique of Therapy Manual Orthopedic very used to the pain management and vertebral dysfunction of origin cervical, althought there esperimental studies about the basics neurophysiological on which exercises its action, There will be a synthesis of the evidence concerning the effects of clinical the technique of the cervical lateral glide, side through a systematic review of randomized controlled trials; Objective; Determine if there es evidence that scientific endorse the therapeutics effects attributed to the technique of cervical lateral glide used by the Physical Therapies for clinics handling of various conditions muscle skeletal; Strategy of Search; Only were included in the search Randomized Controlled Trials (ECAs), the databases used were: MEDLINE (Pubmed/PMC), Cochrane, OVID, PEDro and EMBASE (ScienceDirect); Results; Only 4 ECAs with their eligibility criteria of our review; Conclusions; There moderate evidence that cervical lateral glide producesinmediate hypoalgesia in patients with subacute Neurogenic Cérvicobrachial Pain, chronic lateral epicondylalgia and unilateral shoulder pain with bad responses to traditional treatment.

Advance in Biocompatibility of Biological Material for Neural Tissue Engineering(review) / 中国康复理论与实践

@#Biological scaffolds imitate the structure and function of extracellular matrix,and so good biocompatibility is essential for it.The materials in neural tissue engineering mainly include natural biomaterial and artificial biodegradable materials presently.This article has reviewed the biological function of materials mostly used in neural tissue engineering.

Self-assembled IKVAV Peptide Nanofibers Promote Adherence of PC12 Cells / 华中科技大学学报（医学）（英德文版）

Lack of biocompatibility and bioactivity is a big problem for the synthetic materials that have been generated for neural tissue engineering. To get around the problem and generate better scaffold for neural tissue repair, we intended to generate nano-fibers by self-assembly of polypeptide IKVAV. Bioactive IKVAV Peptide-Amphiphile (IKVAV-PA) was first synthesized and purified, the property of which was analyzed and determined by high-performance liquid chromatography (HPLC)and mass spectrometry (MS). Then, by addition of hydrogen chloride (HCl), self-assembly of IKVAV-PA was induced in vitro and nano-fibers formed as shown by transmission electron microscopy (TEM). The effect of IKVAV nanofibers on adherence of PC12 cells was assayed in cell culture and the results showed that the rates of adherence of PC12 increased significantly when the density of IKVAV was within a certain range (0.58 μg/cm2 to 15.6 μg/cm2). However, its effect on the rates of adherence did not significantly alter with time, whether after 1 hour or 3 hours of culture. In general,we showed that IKVAV-PA can successfully self-assemble to form nanofiber, and promote rapid and stable adherence of PC12 cells, and the effect of the self-assembled IKVAV to promote PC12 cells adherence is dosage-dependent within a certain range of densities.

The Influence of Different Concentrations of Cryoprotectants on Neuronal Cell Viability

Apromising technique for the treatment of Parkinson's disease and other various neurodegenerative disorders is the transplantation of fetal neural tissue. There must, however, be a prompt and reliable source, and one solution is cryopreservation, where tissue viability can maintained for prolonged periods. Fetal neural tissue is, however, known to be susceptible to freeze-storage damage during cryopreservation. In this study, we examined the influence of different concentrations of cryoprotectants upon the survival of rat fetal neurones. Fetal rat brain tissue was frozen with 7-15% dimethyl sulfoxide(DMSO) and 10-50% fetal bovine serum(FBS) as cryoprotectants, then stored for a period of 5 months. Post-storage neuronal cell viabilty was assessed by vital staining followed by determination of cell density. Average total viability of frozen cells with 7% DMSO and 10-50% FBS was less than 50%. Cryopreserved cells with 10-50% DMSO and 10-50% FBS showed almost the same viability(around 70%). The highest viability was obtained with 15% DMSO+20% FBS combination(76%) and 10% DMSO+10% FBS combination(75%). Theoretically, the higher the concentration of cryoprotectants, the higher the viability: however, the best result was achieved stated above, when the combination of cryoprotectants was at the concentrations stated above.