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Objective To investigate the cflecl of microencapsulated olfactory sheathing cells transplantation on pathological pain induced by peripheral nerve injury and the expression levels of P2X7 receptor in L4., spinal cord. Methods Nash differential adherence method was used to cultivate and expand cells from olfactory bulb tissue of a SD rat. Forty healthy SD rats were randomly divided into control group, model group, OFCs group and microencapsulated OECs group (MC-OECs groups). At the 7th and 14th days after surgery, the mechanical stimulation constraining thresholds of the rats in each group were measured by behavioral method. The experssion levels of P2X7 receptor positive cells percentage and average absorbance by in situ hybridization were observed in L4-J spinal cord. Results At the 7th and 14th days after surgery, compared with the control group, the reflex threshold of mechanical withdrawl of rats in the CCI group significantly decreased(P<0. 05). The percentage and average absorbance of P2X7 receptor positive cells in L4., spinal cord significantly increased(PcO. 05) compared with the CCI group, the reflex threshold of mcchaniral withdrawl of rats in the OF.Cs group was significantly increased (/'cO. 05). The percentage and average absorbance of P2X7 receptor positive cells in L«., spinal cord significantly decreased compared with the OECs group(P<0. 05). The reflex threshold of mechanical withdrawl of rats in the MC-OECs group was higher(/,<0. 05). The percentage and average absorbance of P2X7 receptor positive cells in L4., spinal cord were lower(H<0. 05). Conclusion Microencapsulated olfactory ensheathing cells transplantation can relieve the neuropathic pain belter and reduce the expression levels of P2X7 receptor and repair peripheral nerve injury than OECs transplantate.
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Olfactory groove schwannomas (OGSs) are rare benign tumors of the anterior skull base region. Considering the lack of Schwann cells in the optic and olfactory nerves, their origin remains enigmatic. Despite the precursor cell, total resection of the lesion is curative, as long as the histopathological features of the tumor are compatible with schwannoma. We report the case of a 32-year-old woman, addicted to crack, who was brought to the hospital presenting with cognitive dysfunction after being physically assaulted, whose neuroimaging revealed a large extra-axial mass in the subfrontal sagittal region. The presentation, immunohistochemical markers and histogenesis are discussed in the present study, along with a literature review.
Subject(s)
Humans , Female , Adult , Skull Base Neoplasms/surgery , Cranial Fossa, Anterior/surgery , Neurilemmoma/surgery , Magnetic Resonance Imaging , Tomography, X-Ray Computed/methods , Skull Base Neoplasms/pathology , Skull Base Neoplasms/diagnostic imaging , Cranial Fossa, Anterior/pathology , Cranial Fossa, Anterior/diagnostic imaging , Anosmia , Neurilemmoma/pathology , Neurilemmoma/diagnostic imagingABSTRACT
Objective To study the mRNA and protein expressions of the neurofilament in the region of acute spinal cord injury (SCI) of rats after olfactory ensheating cells (OECs) transplantation combined with methyprednisolone administration and investigate the molecular mechanisms of OECs transplantation combined with methyprednisolone administration in promoting the recovery of the spinal cord.Methods Acute spinal cord injury was established in SD rats ( T10 ) by using NYU instrument. The rats were randomly divided into control group, SCI group, DF12 group, OECs transplantation group,methyprednisolone administration group and OECs + methyprcdnisolone group. The mRNA and protein expressions of the neurofilament in the SCI regions of rats in different groups at different time points were detected by using immunohistochemistry, RT-PCR and Western blotting. Results A significant increase of mRNA and protein expressions of the neurofilament could be found in the other five groups compared with the control group at days 7, 14 and 28 after SCI. The mRNA and protein expressions of the neurofilament in the injury region of the OECs group, the methyprednisolone group and the OECs + MP group were more significantly increased than that of the SCI group and the DF12 group. The expression of the neurofilament in the injury region of the OECs + MP group was more significantly increased than that of the OECs group and the MP group ( P ＜ 0. 05). Conclusions OECs transplantation or methyprednisolone administration can induce the mRNA and protein expressions of the neurofilament. Meanwhile, OECs transplantation combined with methyprednisolone administration can significantly increase the mRNA and protein expressions of the neurofilament, as may be one of mechanisms promoting spinal cord repair.
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@#ObjectiveTo explore the feasibility and potential benefit of olfactory ensheathing cell (OEC) intraspinal transplantation in the treatment of intractable chronic neuropathic pain after spinal cord injury (SCI).Methods17 patients, 15 male and 2 female, with intractable chronic neuropathic pain after spinal cord injury was treated by OEC implant from November, 2004 to November, 2007. The age ranged from 18 to 68 (mean 40.4) years. The etiology of cord impairment included car accidents, falls, radiation damage, machine extrusion, gun-shot, and diving. The patients suffered severe persistent pain for 6 to 309 (mean 102.2) months, and the time points when cell therapy were administrated in the patients ranged from 6 to 312 (mean 105.9 months) after their injuries. Olfactory bulbs were harvested and trypsinized down to single fetal OECs. They were cultured for 12～14 days before implant. The fetal OECs were transplanted by injection into spinal cord at opposing ends of the injury site. The degree of pain was assessed and compared before operation and long-term follow-up according to the International Association of Neurorestoratology Spinal Cord Injury Functional Rating Scale (IANR-SCIFRS), i.e., 0 point means extreme pain, uncontrolled; 1 point, severe pain, narcotics required; 2 points, mild pain, ordinary pain killer effective; 3 points, no pain.ResultsThe follow-up and pain reevaluation were performed at 0.5 to 88 months with an average of 17.5 months after cell transplantation. The mean score of pain amelioration is 1.2 points.ConclusionThe OEC intraspinal transplantation appears to have a promising role in treatment of intractable chronic neuropathic pain after SCI.
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Objective To explore the influence factors for the functional improvement after the fetal olfactory ensheathing cell (OEC) transplantation for chronic spinal cord injury(SCI). Methods The olfactory bulbs were harvested and trypsinized down to single fetal OEC. They were cultured for 12-17 days prepared for use. From November 2001 to December 2003, a total of 300 patients volunteered for the fetal OEC transplantation, among whom 222 suffered from complete chronic SCI and 78 suffered from incomplete chronic SCI. The procedures were performed on the patients with a disease course ranging from 6 months to 31 years (average 3.1 years) after their injuries. The fetal OEC was transplanted by the form of injections into the spinal cord at the upper and lower ends of the injury site. All the patients were assessed by the ASIA standard before the transplantation and 2-8 weeks after the transplantation. The influence factors including age, sex, duration after the injury, and injury degrees and levels were compared with those in the functional improvement after fetal OEC transplantation. Results The partially-improved neurological functions assessed by the ASIA standard were indicated by the motor scores increasing from 39.1±20.6 to 45.9±20.3 (P<0.001), the light touch scores from 51.7±24.9 to 63.4±23.0 (P<0.001), and the pin prick scores from 53.0±24.2 to 65.3±22.7(P<0.001). There was no significant difference in the functional improvement of the motor, light touch, and pin brick when compared with the age, sex, duration after the injury, and the injury degrees and levels. The motor scores and light touch scores at the cervical level were higher than the scores at the thoracic level. Conclusion The fetal OEC transplantation can partially improve the neurological functions quickly in treatment of the chronic spinal cord injury. All the influence factors except the motor scores and light touch scores, which were higher at the cervical level than at thoracic level, have no impact on the functional improvement after the fetal OEC transplantation.
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Objective To determine the safety of the fetal olfactory ensheathing cell(OEC) transplantation in patients with chronic spinal cord injury (SCI) by examination of the magnetic resonance imaging (MRI). Methods A prospective clinical study involving 16 patients with chronic SCI was designed to investigate the feasibility and biological safety of the fetal OEC transplantation in treatment of SCI. The olfactory bulbs from the 3-4-month-old aborted human fetuses following the strict ethical guidelines were harvested and trypsinized down to single fetal OEC. These cells were then cultured for 12-17 days and were prepared for a clinical use. From November 2001 to December 2002, 16 patients with chronic SCI were randomly enrolled. The patients suffered from SCI for 1.5-8 years (average 4.3 years) after the injury. The suspension (50 μl) containing about 1×106 fetal OECs was transplanted by an injection into the patients' spinal cords above and below the injury site. All the patients were assessed before the transplantation and were followed up with MRI for 29-42 months (average 38 mon) after the transplantation. Results No cell-related adverse effects were observed in any patient during the follow-up period. The follow-up with MRI did not reveal any development of optic glial tumor, tumor-like mass, new hemorrhage, edema, expanding cyst, new cyst formation, infection or disruption of the neural structure in the transplant site of all the patients. Conclusion This is the first clinical study demonstrating the long-term safety of the OEC therapy for SCI. The results indicate that our protocol is feasible and safe in treatment of patients with chronic SCI within 38 months after the injury. Although the size of the samples for our study was not big enough, the positive results of the study have encouraged us to make a further research in this field.
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Objective On the basis that olfactory ensheathing cells (OECs) transplanted into injured spinal cord may facilitate axonal regeneration, the OECs were cultured from olfactory bulb and nasal olfactory mucosa in the present study, in order to explore if the olfactory mucosa could be a new donation for transplanting the olfactory ensheathing cells. Methods OECs were harvested from olfactory bulb and mucosa based on the differing rates of attachment of the various cell types, following GFAP and NGFRp75 immunocytochemistry. Results Three morphological and immunohistochmically distinct types of cell which appeared bipolar,tripolar and flat morphology were present in primary cultures of adult rat olfactory bulb and olfactory mucosa.Conclusion The method of purification for OECs based on the different rates of attachment among the various cell types is simple, inexpensive and practical. The OECs from nasal olfactory mucosa like ones from the olfactory bulb is an accessible source of tissue for autologous grafting in human spinal paralegia in the future.
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Objective In order to explore the possibility of autologous transplantation of olfactory mucosa for spinal cord repair,the changes of autologous olfactory mucosa being transplanted into spinal cord and the effect of promoting axon regeneration in adult rats were investigated. Methods Forty male adult rats were divided into two groups randomly:olfactory mucosa transplantation and control groups.Olfactory mucosa (1*!mm+2) was taken from the posterior region of nasal septum,and this graft was transplanted into the posterior funiculus of cervical 1 segment of spinal cord and destructed the corticospinal tract(2*!mm?1
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The treatment of spinal cord injury is always a stubborn problem for neurosurgeons because nerve cell cannot regenerate by itself and the glia scar can prevent the axonal regeneration. Therefore, to decrease the neuron death and formation of scar and to increase axonal regeneration are the keys for clinical treatment. Many studies showed that the olfactory ensheathing cells(OEC) can promote axonal regeneration and prover axonal growth,bringing hope for treament of spine injury, but the result is not satisfactory in clinic. In order to improve the clinical effect of OES transplantation many studies combine it with genetic engineering and tissue engineering and some progress is made.
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Objective To explore the effects of expressing human ciliary neurotrophic factor (hCNTF) mediated by retroviral vector in olfactory ensheathing cells(OECs) on the survival and neurite outgrowth of cultured neurons. Methods S\|hCNTF fragment was digested with endonucleases(Kpn I and Xba I) from pcDNA\-3\|S\|hCNTF plasmid and cloned into pRev\|TRE vector.The harvested pRev\|TRE\|hCNTF was identified and transfected with pRev\|Tet\|On into ecotropic Ecopack\|293 cells,resulting in 2 retroviral supernatants(pRev\|TRE\|hCNTF and pRev\|Tet\|On).Primarily cultured rat olfactory ensheathing cells(OECs) were co\|infected with the 2 retroviruses,and induced to secrete hCNTF with different concentrations of doxycline.The secreted hCNTF in OEC culture supernatant was detected with Western\|blot.Dorsal root ganglion (DRG) from a postnatal rat of 2 days was co\|cultured with CNTF\|modified OECs,and the supernatant was used to culture retinal ganglion cells(RGCs).Following ?\|tubulin immunocytochemical staining,the length of DRG neurites were measured,while the numbers of surviving RGCs were counted. Results 1.Individual 630bp and 400bp fragments were digested from pRev\|TRE\|S\|hCNTF expression vector with endonucleases(Hind Ⅲ and BamH Ⅰ),and respected direction and integration of hCNTF cDNA which inserted pRev\|TRE vector were identified; 2.The expression of 24kD CNTF proteins in CNTF\|modified OEC culture supernatant was positively\|correlated with the concentration of doxycline,while no such protein expression was detected in the control groups; 3.The number of surviving RGCs in CNTF\|modified OECs group(41^34?5^4) was significantly higher than those in unmodified OEC(23^15?4^7),OECs(24^55?5^8) and blank(16^8?6^5) groups;and 4^The neurites of DRG were longer (660?67?m) and denser in CNTF\|modified OECs group,as compared with unmodified OECs(418?45?m),Mock+OECs(400?65?m) and blank (0?m) control groups.No process migrated and grew from the tissue mass in blank group.Conclusion\ hCNTF can be expressed in OECs with a doxycline concentration\|dependent manner after transfected via pRev\|TRE\|S\|hCNTF vector,and possesses a marked enhancing effect on the survival and neurite outgrowth of cultured neurons.[
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Objective To investigate the expression of GDNF in the olfactory ensheathing glia cells of the adult rats and golden hamster for studying the mechanisms of the OECs in the CNS regeneration. Methods The immunohistochemical staining was performed on the slides of adult olfactory bulb from 3 rats and 3 golden hamsters.According to the primary antibody,tissue slides were divided into 3 groups:rabbit polyclonal GDNF,the rabbit polyclonal GFAP and rabbit polyclonal NGFRp75(Santa Cruz Biotechnology)respectively,and followed by the rabbit ABC immunoreactive staining system. Results It revealed that GDNF was expressed by cells in the first two lines of olfactory bulb from rat and golden hamster.The GFAP and NGFRp75 were also expressed in the same position from other two groups of slides respectively.Conclusion The olfactory ensheathing cell could secret GDNF in the progress of mediating neuronal survival and axonal elongation.
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Objective The purpose of the present study is to illustrate the effect of olfactory ensheathing cells(OECs) on the survival and neurite outgrowth of GABAergic neurons in vitro. Methods OECs were dissociated from olfactory bulb and neurons from spinal cord of E12 mouse. On the sixth day in vitro,the Millipore cultue blank with OECs was transferred to the neuron culture mediam and continue the co-culture for another 6 days.The cultured neurons were stained with anti-GABA antibody.The neurite of neurons was observed with an image system.The number of GABAergic positive neurons was counted under the microscope. Result The number of GABAergic neurons was 39