Objective To compare their competence of olfactory epithel ial gl iacytes, olfactory globular nerve layer (OGNL) gl iacytes and SC in repair nerve defect of sciatic nerve, and select the best gl iacytes for repair of peri pheral nerve defect. Methods Olfactory epithel ial gl iacytes, OGNL gl iacytes and SC were extracted from 20 female Wistar rats aged 2-3 months and cultured in vitro for 2 weeks, then purified and condensed for transplantation. Eighty adult female Wistar rats were randomized into groups A, B, C and D (n=20). The left sciatic nerves were excised 25 mm axons and retained epineuriumlumen anastomosed to proximal ends. The culture mediums, SC, OGNL gl iacytes, and olfactory epithel ial gl iacytes weretransplanted into the epineurium lumen of groups A, B, C and D, respectively. Three months postoperatively, the injured sciatic nerve regeneration was evaluated by methods of macroscopic observation, photomicroscope, transmission electron microscope, retro-marked fluorescence transportation distance, the gl ial fibrillary acidic protein (GFAP) and nerve growth factor (NGF) were assayed by immunofluorescence, and the myel in basic protein (MBP) and neurofilament (NF) protein were assayed by ELISA. Results The scores of ankle joint were (3.325 ± 0.963), (4.200 ± 1.005), (5.143 ± 0.635) and (5.950 ± 0.154) in groups A, B, C and D, respectively; showing statistically significant difference between groups (P lt; 0.05). The obse vations of gross, sections under microscope and transmission electron microscope showed the regeneration of defect nerve was best in group D, followed by group C, and group B was superior to group A. The transportation distance of retro-marked fluorescence was longest in group D, followed by group C, and group B was superior to group A. The concentrations of GFAP and NGF were largest in group D, followed by group C, and group B was superior to group A. The MBP concentrations were (9.817 ± 3.267), (12.347 ± 3.091), (14.937 ± 2.075) and (22.757 ± 0.871) ng/mL in groups A, B, C and D, respectively; showing statistically significant difference between other groups (P﹤0.05) except between group A and group B (P gt; 0.05). And the NF concentrations were (13.869 ± 5.677), (18.498 ± 3.889), (23.443 ± 2.260) and (27.610 ± 1.125) ng/mL in groups A, B, C and D, respectively; showing statistically significant difference between groups (P﹤0.05). Conclusion Olfactory epithel ial gl iacytes, OGNL gl iacytes and SC transplantation could repair injured nerve. The competence of olfactory epithel iums is superior to the OGNL gl iacytes andSC, and the OGNL gl iacytes is better than SC.
Objective To investigate the synergetic effect and possibil ity of repairing spinal cord injury (SCI) by transplantation of olfactory ensheathing cells (OECs) and chondroitinase ABC (ChABC) in adult rats. Methods Three adult male SD rats were used to isolated olfactory bulb and primarily cultured OECs. In the 8th or 9th day, OECs were transplanted, the concentration of cells was modulated to 1 × 105/μL. Fifty-four SD rats were made the models of T8 spinal cord crush injury and divided into 4 groups. In group A (control, n=36), injured site was not treated; in groups B, C and D (n=6), OECs, ChABC and OECs+ChABC were injected into injured site, respectively. At 1, 2, 3, 7 and 14 days after injury, the BBB score system was used to evaluate the motion function. At 0, 1, 2, 3, 7, 14 days in group A and at 14 days in groups B, C, D after injury, the maximal transverse diameter and gross area of necrosis were evaluated on HE stained sections. The immunofluorescence double label ing staining for gl ial-fibrillary acidic protein (GFAP)/CS56, GFAP/growth associated protein 43(GAP-43) and GFAP/neurofilament 160(NF160) was carried out to evaluate the regeneration of nerve fiber. Results At 14 days after injury, there were significant difference in the BBB scores between group A and groups B, C, D (P lt; 0.05), and between groups B, C and group D (P lt; 0.05), HE staining showed that the formation of cavity was observed in each group at 14 days after injury. There were significant difference in the maximal transverse diameter and gross area of necrosis between groups B, C, D and group A (P lt; 0.01), and between groups B, C and group D (P lt; 0.01). The immunofluorescence staining indicated that expression of GFAP were more intense in group A than in other groups, and the cavity of the lesion site was apparent, but it was moderate in groups B and C. The expression of GAP-43 was more intense in group D than in groups B and C. The expression of NF160 was more intense in group D. Conclusion Transplantation strategy of OECs combined with ChABC was effective in the repair of SCI in some extent.