Objective To study the microstructural change of detrusor muscle and neuromuscular junction (NMJ) after bladder functional reconstruction for atonic bladder caused by medullary cone injury and to discuss the feasibility of bladder functional reconstruction for improving the detrusor muscle degeneration. Methods A total of 104 adult female Sprague-Dawley rats (weighing, 200-250 g) were randomized divided into 3 groups: normal group (n=8), control group (n=48), and experimental group (n=48). No treatment was given in normal group; the medullary cone injury was established by sharp transection of spinal cord at L4, 5 levels in control group; and the anastomosis of bilateral L5 ventral root (VR)-S2 VR and L5 dorsal root (DR)-S2 DR was performed for bladder functional reconstruction after modeling of medullary cone injury in experimental group. After operation, the survival condition of rats was observed. At 3 days and 3 consecutive days before 1, 2, 3, 4, 5, and 6 months after operation, the residual urine volume was measured; at 1, 2, 3, 4, 5, and 6 months after operation, the detrusor muscle was harvested to measure the muscle fiber cross-sectional area by HE staining, to calculate the percentage of connective tissue by Masson trichrome staining, and to observe the ultrastructure of the detrusor muscle and the NMJ by transmission electron microscope (TEM). Results Eleven rats were supplemented because of death after operation. In control group, a significant increase of the residual urine volume was observed with the extension of time (P lt; 0.05); in experimental group, an increase was observed at the first 3 months after operation, and then gradually decreased, showing significant differences between the other time point (P lt; 0.05) except between at 3 days and at 5 months after operation (P gt; 0.05); there was significant difference between control and experimental groups at other each time point (P lt; 0.05) except at 3 days, 1 month, and 2 months (P gt; 0.05). HE staining and Masson trichrome staining indicated that the muscle fibers arranged in disorder with gradually aggravated atrophy and gradually increased connective tissue in control group, while the shape of the detrusor muscle recovered with no increased connective tissue at 4, 5, and 6 months after operation in experimental group; there was significant difference in cross-sectional area of detrusor muscle and percentage of connective tissue between normal group and experimental group, and between normal group and control group at each time point (P lt; 0.05). In control group, the cross-sectional area of detrusor muscle decreased and the percentage of connective tissue increased with the extension of time (P lt; 0.05). In experimental group, the cross-sectional area of detrusor muscle decreased at the first 3 months and then increased, and the percentage of connective tissue increased slowly with the extension of time. There was no significant difference of cross-sectional area of detrusor muscle at the first 3 months between control and experimental groups (P gt; 0.05), but the values in experimental group were significantly higher than those in control group at 4, 5, and 6 months after operation (P lt; 0.05). There were significant differences of the percentage of connective tissue between control and experimental groups at each time point (P lt; 0.05). In control group, the amount of synaptic vesicles decreased in the NMJ with time passing; vacuole like structure was observed in NMJ at 3 months; there was almost no nerve ending at 6 months. In experimental group, the amount of synaptic vesicles decreased at 1 and 3 months after operation, but obviously increased at 6 months. Conclusion The reconstruction of bladder function with L5 nerve roots above the paraplegic plane can effectively inhibit the degeneration of detrusor muscle and improve its microstructural changes after medullary cone injury.
Objective To investigate the effects of chitosan/polyvinyl alcohol (PVA) nerve conduits for repairing radial nerve defect in Macaques. Methods Twelve adult Macaques weighing 3.26-5.35 kg were made the models of radial nerve defect (2 cm in length) and were randomly divided into 3 groups according to nerve grafting, with 4 Macaques in each group. Chitosan/PVA nerve conduit, non-graft, and autografts were implanted in the defects in groups A, B, and C, respectively. And the right radial nerves were used as normal control. At 8 months postoperatively, the general observation,electrophysiological methods, and histological examination were performed. Results At 8 months postoperatively, theregenerated nerve bridged the radial nerve defect in group A, but no obvious adhesion was observed between the tube and the peripheral tissue. The regenerated nerve had not bridged the sciatic nerve defect in group B. The adhesions between the implanted nerve and the peri pheral tissue were significant in group C. Compound muscle action potentials (CMAP) were detected in group A and group C, and no CMAP in group B. Peak ampl itude showed a significantly higher value in normal control than in groups A and C (P lt; 0.05), but there was no significant difference between groups A and C (P gt; 0.05). Nerve conduction velocity and latency were better in normal control than in groups A and C, and in group C than in group A, all showing significant differences (Plt; 0.05). The density of myl inated fibers in groups A and C was significantly lower than that in normal control (P lt; 0.05), but there was no significant difference between groups A and C (P gt; 0.05). The diameter and the myel in sheath thickness of the myl inated fibers in normal control were significantly higher than those in groups A and C, and in group C than in group A, all showing significant differences (P lt; 0.05). Conclusion The chitosan/PVA nerve conduits can promote the peripheral nerve regeneration, and may promise alternative to nerve autograft for repairing peripheral nerve defects.
Objective To investigate the effects of carboxymethylchitosan- carboxymethylcellulose (CMCH-CMC) film on the adhesion and heal ing of colonic anastomosis. Methods Sixty-four healthy adult male SD rats was randomly divided into control group and experimental group (n=32). The model of colonic anastomosis was made according to Buckenmaier’ smethod in all rats. The experimental group was treated by wrapping anastomosis with CMCH-CMC film (3 cm × 2 cm) and the control group was not treated. At 7 days and 14 days after operation, the adhesion formation of colonic anastomosis was observed, the tensile strength of the anstomosis was assessed and compared with 6 normal rats, and the hydroxyprol ine (HP) content of the anastomotsis was detected. Results There were 3 deaths in the experimental group and 2 deaths in the control group. The adhesive scores of the experimental group on the 7th and 14th postoperative day [(0.50 ± 0.16) points and (0.45 ± 0.14) points, (Plt; 0.05)] were significantly lower than those of the control group [(1.67 ± 0.15) points and (2.29 ± 0.18) points, (P lt; 0.05)], (Plt; 0.01). Tensile strength were more marked on the 14th postoperative day than on the 7th postoperative day in the control group (Plt; 0.05), but there was no significant difference between the 7th day and the 14th day in the experimental group. The tensile strength of thecontrol group and the experimental group on the 14th postoperative day [(178.36 ± 20.10) and (172.74 ± 22.18) mmHg] were respectively higher than those on the 7th postoperative day [(138.67 ± 16.65) and (130.81 ± 18.38) mmHg] (Plt; 0.01). The tensile strength of the control group and the experimental group on the 7th postoperative day were respectively significantly lower than that of the normal rats (P lt; 0.01). The level of HP in the anastomosis was significantly higher on the 7th postoperative day in the experimental group [(84.47 ± 11.87) μg/mg dried weight] than that of the control group [(55.47 ± 12.89) μg/mg dried weight), (Plt; 0.05)], but there was no significant difference between the experimental group and the control group on the 14th postoperative day [(146.07 ± 14.81) μg/mg dried weight, (137.14 ± 16.81) μg/mg dried weight, (P gt; 0.05)]. Conclusion The CMCH-CMC film can decrease adhesion the formation of colonic anastomosis, but does not interfere with the heal ing of colonic anastomosis.
Objective To investigate the effects of chitosan on the cell cycle of the human fibroblasts and on the Ki-67 antigen expression in vitro and to investigate the mechanism of chitosan preventing the postoperative tissue adhesion. Methods The cultured fibroblasts were treated for 48 hours with 0，0.01，0.1，1.0，10.0 mg/ml of chitosan, respectively;then, the cell cycle of the fibroblasts was measured by the flow cytometry. The cultured fibroblasts were treated for 24 hours with the chitiosan at the above concentrations; then, the Ki-67 antigen in the cell nucleus was detected with the immunohistochemical staining toobserve its expression. Results The growth of the fibroblastswas obviously suppressed by chitosan, especially in the cell morphology. When the concentrations of chitosan were 1.0 mg/ml and 10.0 mg/ml, the percentages of the fibroblasts in the proliferation stage were 32.3%±5.2% and 14.7%±2.9%, respectively,which were significantly smaller than the percentage of the fibroblasts when the concentration of chitosan was 0 mg/ml (the control group) (41.9%±5.8%， P＜0.05). When the concentrations were 0.01 mg/ml and 0.1 mg/ml, the percentages of the fibroblasts in the proliferation stage were 39.0%±6.0% and 35.5%±3.4%, respectively, which were smaller than that of the control, but not significantly different from that of the control (P＞0.05). When the concentrations of chitosan were 0.1 mg/ml,1.0 mg/ml and 10.0 mg/ml, the percentages of the fibroblasts that had the positiveKi-67 antigen were 37.3%±3.4%, 30.5%±6.2% and 17.8%±3.0%，respectively, which were significantly smaller than that of the control (57.6%±8.9%， P＜0.05). When the concentration was 0.01 mg/ml, the percentage of the fibroblasts that had the positive Ki-67 antigen was 54.1%±8.0%, which was smaller than that of the control, but not significantly different from that of the control (P＞0.05). ConclusionChitosan can inhibit the proliferation of the fibroblasts and increase the percentage of the fibroblasts in the quiescent stage, which can be considered as one of the mechanisms that chitosan can prevent the postoperative tissueadhesion.
Objective To study the prevention of postoperative adhesion using carboxymethylcellulose(CMC). Methods The literature was reviewed extensively, concerning the physical and chemical characters of CMC, the mechanism of preventing adhesion, the effect on healing of anastomoses, the experimental researches and the clinical application. Results CMC was a polysaccharide with favorable biocompatibility, biodegradation, stable physical and chemical characters, and it can prevent postoperative adhesion. Conclusion CMC is with a wide outlook in prevention of postoperative adhesion.
Objective To introduce the clinical experience of localdistally based turnover adipofascial flap for small to medium size wound of the extremities. Methods From 1994 to 2003, 33 cases of distally based longitudinal neuro-veno-adipofascial turnover adipofascial flap (axial perforator pattern in26, random pattern in 7) were transferred in the forearm (19) and lower leg (14).These flaps were all raised in the forearm or lower leg as local flaps. The length (pedicle plus flap) was 9 to 18 cm and the width was 3 to 4 cm, with L/W ratio of 3 to 5∶1. After transferring by 180° turnover, a splitthickness skin graft was used tocover the fascia surface. Results All the 33 flaps survived. The donor sites were closed directly, and the recipient sites were covered with full-or splitthickness skin grafts. Both donor and recipient sites healed primarily. Conclusion Turnover adipofascial flap is a simple and reliable method for small to medium size wound of the extremities.
Objective To design a combined flap of subscapular axis including vascularized lateral scapular,rib and latissimus dorsi to repair the large defect of tibia. Methods The patient was a 39-year-old man who got a posttraumatic 12 cm defect of tibiaafter primary debridement and external fixation because of open fracture 5 months ago. There was a 12 cm×6 cm scar involved the proximal medial segment of tibia.After resection of scar and fibular tissue over the bone defect floor, alatissimus dorsi myocutaneous flap 14 cm×5 cm pedicled with subscapular artery-thoracodorsal artery,a flap 12.5 cm on the outside of the scapular pedicled with thoracodorsal artery, and 6th rib flap 13 cm by serratus were prepared.The tibialis posterior and saphenous vein were used for astomosis. A proximalanatomic plate was applied to the fixation of tibia. Results Thecompound flap survived the operation. The follow-up period was 2 years. Bone union occurred 6 months after operation. Conclusion This combined flap is successful and can provide alternative to the resolution of large defect of tibia.
Objective To compare the effect and coverage of bacteriostasis of chitosan and sodium hyaluronate. Methods Each of the five bacteria, Proteus mirabilis, Escherichia coli, Candida albicans, Pseudomonas aeruginosa, Staphylococcus aureus, was cultivated for 33 tubes of broth culture. Leaving three tubes each group as control group, ploidy diluted concentration of high relative molecular weight chitosan, low relative molecular weight chitosan and sodium hyaluronate were added respectively in the broth culture. All the tubes were cultivated for 18 hours at 37 ℃ with homeothermia. Then the growth of bacteria was observed. ResultsThe minimal inhibitory concentrations (MIC) of high relative molecular weight chitosan were : Proteus mirabilis 0.031%, Escherichia coli 0.063%, Candida albicans 0.063%, Pseudomonas aerugionosa 0.063%, Staphylococcus aureus 0.063%; and the MIC of low relative molecular weight chitosan were: Proteus mirabilis 0.125%, Escherichia coli 0.025%, Candida albicans 0.25%, Pseudomonas aeruginosa 0.25%, Staphylococcus aureus 0.125%; bacteria grew well in each tube of sodium hyaluronate group and control group. Conclusion The above results show that sodium hyaluronate has no bacteriostasis, while chitosan has bacteriostasison broad spectrum and high relative molecular weight chitosan has ber effect.
OBJECTIVE: To investigate an alternative procedure for complete denervation of bladder in the supra-cone cord injury to restore the bladder function. METHODS: Sixteen dogs were included in this study after their spinal cords were transected above the cone. They were divided into 6 groups and performed the rhizotomy of L7 to S3 root in different combination respectively. The bladder and urethra pressure change by electrostimulation during operation and cystometrogram change after operation were tested. RESULTS: 1. Electrostimulation study: for bladder innervation, S2was the most important and S1 was secondary. While for urethra innervation, S1 was more important than S2. When the anterior and posterior roots of S1 and S2 were intact with rhizotomy of posterior roots of L7 and S3, stimulated the common or posterior root of S1 and S2, the change of pressure in bladder and urethra was the same. When the anterior roots of S1 and S2 were resected with rhizotomy of posterior roots of L7 and S3, the pressure in bladder and urethra was significant decreased compared to stimulating the corresponding posterior roots. 2. Cystometrogram (CMG) study: in the complete deafferented group, resecting the posterior roots of L7 to S3, the bladder became flaccid. While resecting the posterior root of S2 and anterior root of S1 or, resecting the posterior root of S1 and anterior root of S2, combining with rhizotomy of posterior roots of L7 and S3, the CMG curve was similar to the complete deafferented group. In the S1 and S2 intact group, the bladder became spastic. CONCLUSION: Combining rhizotomy of anterior and posterior sacral root in different level has the same effects on bladder as complete deafferentation.
Objective To establish the finite element model of Y-shaped patellar fracture fixed with titanium-alloy petal-shaped poly-axial locking plate and to implement the finite element mechanical analysis. Methods The three-dimensional model was created by software Mimics 19.0, Rhino 5.0, and 3-Matic 11.0. The finite element analysis was implemented by ANSYS Workbench 16.0 to calculate the Von-Mises stress and displacement. Before calculated, the upper and lower poles of the patella were constrained. The 2.0, 3.5, and 4.4 MPa compressive stresses were applied to the 1/3 patellofemoral joint surface of the lower, middle, and upper part of the patella respectively, and to simulated the force upon patella when knee flexion of 20, 45, and 90°. Results The number of nodes and elements of the finite element model obtained was 456 839 and 245 449, respectively. The max value of Von-Mises stress of all the three conditions simulated was 151.48 MPa under condition simulating the knee flexion of 90°, which was lower than the yield strength value of the titanium-alloy and patella. The max total displacement value was 0.092 8 mm under condition simulating knee flexion of 45°, which was acceptable according to clinical criterion. The stress concentrated around the non-vertical fracture line and near the area where the screws were sparse. Conclusion The titanium-alloy petal-shaped poly-axial locking plate have enough biomechanical stiffness to fix the Y-shaped patellar fracture, but the result need to be proved in future.