• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.30 no.1
• /
• pp.1-16
• /
• 2004

The use of artificial nerve conduit containing viable Schwann cells is one of the most promising strategies to repair the peripheral nerve injury. To fabricate an effective nerve conduit whose microstructure and internal environment are more favorable in the nerve regeneration than existing ones, a new three-dimensional Schwann cell culture technique using $Matrigel^{(R)}$. and dorsal root ganglion (DRG) was developed. Nerve conduit of three-dimensionally arranged Schwann cells was fabricated using direct seeding of freshly harvested DRG into a $Matrigel^{(R)}$ filled silicone tube (I.D. 1.98 mm, 14 mm length) and in vitro rafting culture for 2 weeks. The nerve regeneration efficacy of three-dimensionally cultured Schwann cell conduit (3D conduit group, n=6) was assessed using SD rat sciatic nerve defect of 10 mm, and compared with that of silicone conduit filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method (2D conduit group, n=6). After 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were examined using image analyzer and electromicroscopic methods. The SFI and ankle stance angle (ASA) in the functional evaluation were $-60.1{\pm}13.9$, $37.9^{\circ}{\pm}5.4^{\circ}$ in 3D conduit group (n=5) and $-87.0{\pm}12.9$, $32.2^{\circ}{\pm}4.8^{\circ}$ in 2D conduit group (n=4), respectively. And the myelinated axon was $44.91%{\pm}0.13%$ in 3D conduit group and $13.05%{\pm}1.95%$ in 2D conduit group to the sham group. In the TEM study, 3D conduit group showed more abundant myelinated nerve fibers with well organized and thickened extracellular collagen than 2D conduit group, and gastrocnemius muscle and biceps femoris tendon in 3D conduit group were less atrophied and showed decreased fibrosis with less fatty infiltration than 2D conduit group. In conclusion, new three-dimensional Schwann cell culture technique was established, and nerve conduit fabricated using this technique showed much improved nerve regeneration capacity than the silicone tube filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method.

• Journal of Korean Medicine Rehabilitation
• /
• v.19 no.3
• /
• pp.15-32
• /
• 2009

Objectives : The purpose of this Study was to identify the effect of Dokhwalgisaeng-tang(Duhuoqisheng-tang) and Jungsongouhyul pharmacopuncture on pain control and nerve regeneration after crush injury in rat sciatic nerve. Methods : Neuropathic pain was induced by crush-induced model of right sciatic nerve. Animal groups were divided as follows; Group I: no treatment control group, Group II : experimental group treated with Dokhwalgisaeng-tang(Duhuoqisheng-tang), Group III : experimental group treated with Jungsongouhyul pharmacopuncture, and Group IV : experimental group treated with Dokhwalgisaeng-tang(Duhuoqisheng-tang) and Jungsongouhyul pharmacopuncture. To evaluate pain intensity, each group was observed paw withdrawal threshold and immunoreactivity on the c-fos before and after respective treatments in five hours, first, third, and fifth day. To evaluate nerve regeneration, those were observed SFI(Sciatic Functional Index) and GAP-43(Growth Associated Protein 43) after each treatment in seventh and thirteenth day. Results : 1. Paw withdrawal threshold to the mechanical stimuli made the significant difference between group IV and the control group after five days of the experiment. 2. Paw withdrawal threshold to the thermal stimuli made the significant difference between group I and III, I and IV, II and IV. 3. In immunohistochemical response of c-fos, as time passes, the immunoreactivity of all groups was decreased gradually. Especially, group IV was observed the lowest after three days. 4. The differences of sciatic function indexes in each group were significantly between group I and III, I and IV, II and IV after 14 days, and between group I and III, I and IV, II and IV, III and IV after 21 days. 5. In immunohistochemical response of GAP-43, all groups had higher GAP-43 immunoreactivity at the 14 days from post-injury and group IV showed highest immunoreactivity. Conclusions : Based on above the results, it is proposed that Dokhwalgisaeng-tang(Duhuoqisheng-tang) and Jungsongouhyul pharmacopuncture may be helpful as a treatment in neuropathic pain and nerve regeneration in rat model.

• Archives of Plastic Surgery
• /
• v.34 no.3
• /
• pp.279-284
• /
• 2007

Purpose: The vein graft was considered as a useful conduit for nerve defect. But the problem is that it might be collapsed in long vein graft state. A new experimental model using vein graft filled with hyaluronic acid was considered. Methods: Thirty rats were used for the experimental animal. In group I, one side of the femoral nerve was exposed and a segment was removed about 15mm. The neural gap was connected with nerve graft. In group II, the nerve gap was connected with vein graft only. In group III, the nerve gap was connected with vein graft filled with hyaluronic acid. A walking track analysis was made periodically for 2 months and NCV(nerve conduction velocity) was executed at the end of the experiment. And morphologic studies were also done for all groups Results: In a walking track analysis, the toe-spread was widen and the foot-length was lengthened. The recovery of the toe-spread and foot length was checked 2 weeks interval, periodically for two months. The SFI (sciatic function index) was $-52.5{\pm}8.2$ in group I, $-68.1{\pm}4$ in group II, $-55.3{\pm}7.9$ in group III. In electrophysiological study, NCV(nerve conduction velocity) was $26.71{\pm}3.11m/s$ in group I, $17.94{\pm}4.35m/s$ in group II, $25.69{\pm}2.81m/s$ in group III. The functional recovery in group I and III was superior to that the group II statistically(p < 0.05) Under electromicroscopic study, the number of the myelinated axons were $1419.1{\pm}240$ in group I, $921.7{\pm}176.8$ in group II, $1322.2{\pm}318$ in group III. The number of the myelinated axons were much more in group I and III than group II statistically (p<0.05). Conclusion: This study suggested that the vein graft filled with hyaluronic acid is more effective than vein graft only for the conduit of the nerve gap. It was thought that the technique could be used in clinical cases with nerve defects as an alternative method to classical nerve grafts.