Objective To provide theoretical evidence for clinical application of the epidermal stem cells after an investigation on changes of the epidermal stem cells during the survival process after the fullthickness skin autograft. Methods On the backs of 42 Wistar rats, orthotopic transplantation models (1.5 cm×1.5 cm) of the fullthickness skin autograft were made. According to the time of the specimen taking, at 1, 3, 5, 7, 14, 21 and 30 days after operation, the rats were randomly divided in 7 groups (Groups 1-7). Specimens taken in each group before operation were used as controls. At each time point, the gross observation was made on the transplanted skin flaps, from which the skin tissues were harvested at each time point before and after operation. The routine pathological and the immunohistochemical examinations were performed on the specimens, which were stained by HE and were observed for immunohistochemical changes and the changes in the cells positive for integrinβ-1 and p63. Results All the fullthickness skin autografts survived 3 days after operation except the skin autograft in 1 rat in both Group 5 and Group 6, which was infected around the transplanted skin flap. In Groups 1-4, cell edema, inflammatory cell infiltration, and increased fibrocytes were observed. In Groups 5-7, the maturity degree of the epithelial cells became higher and higher, and the fibrocyte proportion was lowered. In each group the cell positivity rate for integrin β1 was lower than the cell positivity rate for p63. The positive cells were arranged in disorder, distributed into the layers of the epidermis and gradually concentrated in the basal layer of the epidermis and the bulge of the folliculus pili. The positive cells were also found in the other layers of the epidermis.The positive cells were gradually decreased in number, and reached the lowest level in Group 2. There was a significant difference in the above variables in Groups 1,2,3,5,6 and 7 between before and after operations (P＜0.05). Conclusion During the survival process of the fullthickness skin autograft, the proportion of theepidermal stem cells is gradually decreased at first; Then, the proportion isgradually increased, even beyond the normal level; finally, the proportion is decreased again. The distribution of the epidermal stem cells appear in disorder, almost distributed in the layers of the epidermis; finally, the almost normal distribution can be found.
Objective To investigate the survival effect and reaction mechanismsof motor neurons after reimplantation of the avulsed root into the spinal cord,and to observe the survival and differentiation in the spinal cord after brachial plexus roots avulsion. Methods Thirty adult Wistar rats were randomly devided into the control group and the experimental group (n=15). Laminectomy of C4-6 was performed via a posterior approach. The ventral and dorsal roots of C5，6 were both avulsed from the spinal cord outside the dura mater and within the vertebral canal.For the experimental group, the ventral root of C6 wasreimplanted into the ventralhorn under microscope. The dorsal root was left. The ventral and dorsal roots of C5 were placed inside the nearby muscles. For the control group, the ventral and dorsal roots of both C5 and C6 were placed inside the nearby muscles. At 2, 4, 6, 8, 12 weeks postoperatively, the C6 spinal cord was stained with HE. The changes of the number and morphology of motor neurons were observed onHEstained sections. The C6 spinal nerve root was stained with silver nitrate, andthe regeneration of nerve fiber was observed. Results All rats were recovered well and their wounds were healed at primary stage. The gross observation showed that the avulsed nerve roots in control group adhered to adjacent muscles, however the one in experimental groups which had been implanted into spinal cord adhered to scar tissues and were not separated from spinal cord. At each time point postoperatively, the HEstained transverse sections showed that the number of motor neurons decreased significantly with soma swollen and atrophied, Nissle bodies decreased or disappeared. The survival rates of motor neurons in the control group were 60.9％±5.8%,42.3％±3.5％,30.6％±6.1％27.5％±7.9％ and 20.4％±6.8％ respectively；in the experimental group,the survival rates were 67.1％±7.4％,56.3％±4.6％,48.7％±8.8％,44.2％±5.5％ and 42.5％±8.3％ respectively. The survival rates of motor neurons in the experimental group was higher than those in the control group at all time points,showing statistically significant difference（Plt;0.01）.At 12 weeks postoperatively, thesilver nitrate stained specimen from the C6 nerve root showed regeneration of the motor neurons in the ventral horn into the reimplanted nerve root through axon in the experimental group,but the degeneration of the nerve fiber appeared and the number of the myelinated nerve fiber decreased in the control group. Conclusion Through reimplantationof the avulsed ventral nerve root into the ventral horn, degeneration of the motor neurons in the ventral horn can be reduced. After reimplantation of avulsed nerve root, there is axonal regrowth of motor neurons into the spinal nerve root and regeneration of the myelinated nerve fiber also appears.
Objective To study the allograft effect of two kinds of tissue engineered oral mucosa lamina proprias on skin fullthickness wounds. Methods The cultured Wistar rat oral mucosa fibroblasts (OMF) were incorporated into collag en or chitosancollagen to construct the tissue engineered oral mucosa laminaproprias, and then the OMFs were labeled with BrdU. The fullthickness round skin defects were made with a round knife (diameter, 0.8 cm) on the backs of 36 Wistar rats (2125 weeks old), which were divided into 2 experimental groups: the fibroblastpopulated collagen lattices (FPCL) group (grafted by FPCLs) and the fibroblastpopulated chitosan collagen lattices (FPCCL) group (grafted by FPCCLs), and the control group (only covered with gauges). All the wounds were observed by the naked eyes or the light microscope, and were measured 4, 7, 14, and 21 days postoperatively. Results There were no infection during the wound healing period. At 7 days after the grafting, the wounds in the 3 groups were covered by scab and/or gauze; at 14 days, the gauze and scab on the wounds in the three groups were all replaced by the new epidermis naturally except one scab each in the FPCCL group and the control groups,which was replaced at 17 days.All the centers of the new epidermis were measurable as the pink red points. At 21 days, all the new skins were smooth without hairs, and their color was similar to the normal one. At 4, 7, and 14 days,there was an indication that the wound diameters became significantly smaller in the three groups; but after the 14th day, there was no significant indication of this kind. At 7 days, the wound diameter in the FPCL group was significantly smaller than that in the FPCCL group and the control group (Plt;0.01). Under the lightmicroscope, at 4 days postoperatively, the decayed tissue on the surfaces of the recipient wounds in the FPCL group and the FPCCL group was separated from the lower granular tissue in which there were many inflammatory cells, fibroblasts, and new vessels. There was a similar-phenomenon in the control group. Each skin wound in the three groups was only partly keratinocyted at 7 days postoperativel y. The recipient wounds were wholly keratinocyted with when rete ridges observed at 14 and 21 days, but in the control group the wounds were keratinocyted with no rete ridges. Fibers in the new dermis were thin. The OMFs with Brdu appeared in the granular tissue and new dermis at 4, 7, 14, and 21 days postoperatively, which could be illustr ated by the immunohistochemical staining. The positive OMFs and the granular tissue joined in the repair of the skin defe cts without any allergic reaction during the period of the wound healing. Conclusion The oral mucosa fibroblasts as the new seed cells can join i n the repair of the skin defects effectively and feasibly. The fibroblastpopul ated collagen lattices and the fibroblastpopulated chitosan collagen lat tices can repair skin defects effectively and feasibly, too. And the quality of the new skins was better in the two experimental groups than in the control group.
Objective To investigate the quantity and distribution of motor fiber of rat’s C7 nerve root. Methods Motor fiber quantity and section area in the main nerves of the upper extremity and the fascicles of C7 in 30 SD rats were analyzed.Results Fascicles and certain amount (207) of motor fibers from the anterior division of C7 were distributed to musculocutaneous nerve and median nerve, the orientation of these fibers were not clear. The ones (323) from posterior division were to the axillary, radial, and dorsal thoracic nerves, thus the orientation of these fascicles was relatively definite. Conclusion Thedistribution of the motor fibers and fascicles in the divisions of C7 in rat is similar to human beings, so rat is a relatively good model for the study of selective C7 nerve root transfer.