Objective Vascular bundle and sensory nerve bundle implantation can promote the osteogenesis of tissue engineered bone. To investigate whether vascular bundle and sensory nerve bundle implantation will affect the expressions of neurokinin 1 receptor (NK1R) and vasoactive intestinal peptide type 1 receptor (VIPR1). Methods Fifty-four 5-montholdNew Zealand rabbits were selected. Autologous bone marrow was aspirated from the posterior il iac spine of rabbits, and the bone marrow mesenchymal stem cells (BMSCs) were prol iferated in vitro. At the 3rd passage, the BMSCs were cultured in the osteogenic culture medium for 7 days. The tissue engineered bone was prepared by the combined culture of these osteoblastic induced BMSCs and β tricalcium phosphate scaffold material. A 1.5 cm segmental bone defect was created at the right femur of rabbits. After the plate fixation, defects were repaired with sensory nerve bundle plus tissue engineered bone (group A, n=18), with vascular bundle plus tissue engineered bone (group B, n=18), and tissue engineered bone only (group C, n=18). X-ray examination was used to evaluate the degree of the ossification. The expression levels of NK1R and VIPR1 were measured by the immuohistochemistry analysis and the mRNA expression of NK1R and VIPR1 by real-time PCR at 4, 8, and 12 weeks after operation. Results The better osteogenesis could be observed in group A and group B than in group C at all time points. X-ray scores were significantly higher in group B than in groups A and C (P lt; 0.05) at 4 weeks, and in groups A and B than in groupC (P lt; 0.05) at 8 and 12 weeks. The mRNA expressions of NK1R and VIPR1 were highest at 8 weeks in groups A and B and gradually decreased at 12 weeks (P lt; 0.05); the expressions were higher in groups A and B than that in group C (P lt; 0.05), and in group B than group A (P lt; 0.05). Immunohistochemistry analysis showed that the expressions of NK1R and VIPR1 were highest at 8 weeks in 3 groups, and the expressions were higher in groups A and B than in group C. Conclusion Implanting vascular bundles into the tissue engineered bone can significantly improve the expression levels of NK1R and VIPR1. It is an ideal method to reconstruct composite tissue engineered bone.
To establ ish the animal model of the artificial physiological reflex arc with the reconstruction of the sensory and the motorial functions of atonic bladder simultaneously in the rats, and to provide the foundation to furtherinvestigate the repairing effectiveness of this technique. Methods There were 20 adult male SD rats (weighing 280-300 g)which were randomly divided into 2 groups (n=10): group A and group B. Group A was anastomosis of the ventral roots(VR) and the dorsal roots (DR) between L6 and S2 simultaneously to establ ish the model of the artificial physiological reflex arc. Group B was anastomosis of the main trunk between L6 and S2 to establ ish the model. The contents of the observation included: ① To measure the external diameter of the VR, DR and the main trunk of L6 and S2 with the sl iding cal iper; and to measure not only the distance between L6 and S2 but also the separable length of L6 with the ruler. ② Fast Blue dyeing of the VR, DR and the main trunk of L6 and S2 was performed to count their nerve fibers assisted by the Leica FW4000 system 2 weeks after opertation. ③ The observation of the urination of the rat and BBB scoring to evaluate the motorial function of the lower l imbs was performed postoperatively. Results ① L6 located in the lateral side of the S1-4 in the vertebral body of L6. The external diameters of the VR, DR and the main trunk of L6 were (0.68 ± 0.13), (0.88 ± 0.10) and (1.54 ± 0.33) mm, respectively, while those of S2 were (0.62 ± 0.08), (0.79 ± 0.14) and (1.39 ± 0.42) mm, respectively. The distance between L6 and S2 was (14.21 ± 1.95) mm, and the separable length of L6 was (10.76 ± 2.11) mm. Furthermore, the microdissection indicated the VR and the DR between L6 and S2 could be anastomosed respectively with no-tension at the level of the vertebral body of L6; and the main trunk of L6 and S2 could be anastomosed with no-tension at the level of the confluens of L5, 6. ② With Fast Blue dyeing, there were 892 ± 32, 354 ± 26 and 532 ± 17 nerve fibers of the VR, DR and the main trunk of L6, respectively. And there were 788 ± 29, 325 ± 19, and 478 ± 22 nerve fibers of the VR, DR and the main trunk of S2, respectively. There were no volar ulcer,trichomadesis and self-eating of the affected l imbs in the both groups postoperatively. The urinations of the rats after operationwere not different from those before operation. The mean BBB scores of pre- and postoperation in group A were 20.20 ± 0.35 and 19.80 ± 0.23, respectively; the mean BBB scores of pre- and postoperation in group B were 20.20 ± 0.35 and 19.20 ± 0.31, respectively. There was no significant difference of the above indexes between group A and group B (P gt; 0.05). Conclusion Anastomosis of the VR and the DR between L6 and S2 simultaneously in rats is an ideal animal model to establ ish the artificial physiological reflex arc owing to its simple and reproducible procedures.
Repairing degloving injury of fingers by transplantation of ateriolized venous network flap with sensory nerve for six cases (7 fingers). The flaps were all gotsurvived. The procedure of the operation was performed as following: 3～5 supperficial veins and the medial or lateral cutaneous nerve were separated on the palmar side of the forearm as pedicle. According to the defect, the corresponding flaps was designed and was transferred to the injuried finger. Anastomosed the veins with the two digital arteries and veins. Anastomosed the cutaneous nerve with the digital nerves. The patients were followed up for two years. The flaps were soft and wearresisting. The joint movements of the fingers were normal. The twopoints discrimination was 5 to 10mm. The contour of the fingers was satisfactory. The procedure has the following advantages: 1. carried out one operation; 2. good sensation, 3. good appearance and satisfactory function. The indications and factors affecting the survival of the flap were discussed.
ObjectiveTo investigate the mechanism of Semaphorin 3A (Sema3A) in fracture healing after nerve injury by observing the expression of Sema3A in the tibia fracture healing after traumatic brain injury (TBI). MethodsA total of 192 Wistar female rats, 8-10 weeks old and weighing 220-250 g, were randomly divided into tibia fracture group (group A, n=48), TBI group (group B, n=48), TBI with tibia fracture group (group C, n=48), and control group (group D, n=48). The tibia fracture model was established at the right side of group A; TBI model was made in group B by the improved Feeney method; the TBI and tibia fracture model was made in group C; no treatment was given in group D. The tissue samples were respectively collected at 3, 5, 7, 14, 21, and 28 days after operation; HE staining, immunohistochemistry staining, and Western blot method were used for the location and quantitative detection of Sema3A in callus tissue. ResultsHE staining showed that no obvious changes were observed at each time point in groups B and D. At 3 and 5 days, there was no obvious callus growth at fracture site with inflammatory cells and fibrous tissue filling in groups A and C. At 7 and 14 days, fibrous tissue grew from periosteum to fracture site in groups A and C; the proliferation of chondrocytes in exterior periosteum gradually formed osteoid callus at fracture site in groups A and C. The chondrocyte had bigger size, looser arrangement, and more osteoid in group C than group A. Group B had disorder periosteum, slight subperiosteal bone hyperplasia, and no obvious change of bone trabecula in group B when compared with group D. At 21 and 28 days, cartilage callus was gradually replaced by new bone trabecula in groups A and C. Group C had loose arrange, disorder structure, and low density of bone trabecula, big callus area and few chondrocyte and osteoid when compared with group A; group B was similar to Group D. Immunohistochemistry staining showed that Sema3A expression in chondrocytes in group C was higher than that in group A, particularly at 7, 14, and 21 day. Sema3A was significantly higher in osteoblasts of new bone trabecula in group A than group C, especially at 14 and 21 days (P<0.05). Western blot results showed that the Sema3A had the same expression trend during fracture healing in groups A and C. However, the expression of Sema3A protein was significantly higher in group C than group A (P<0.05) and in group B than group D (P<0.05) at 7, 14, 21, and 28 days. ConclusionAbnormal expression of Sema3A may play a role in fracture healing after nerve injury by promoting the chondrocytes proliferation and reducing the distribution of sensory nerve fibers and osteoblast differentiation.