Objective To evaluate the immediate and mid-term effectiveness of hybrid procedures (combined open surgery and endovascular therapy) for multilerel femoral and popliteal artery occlusive disease. Methods Between June 2009 and June 2012, 22 cases of severe femoral and popliteal artery occlusive disease were treated by hybrid surgery. There were 15 men and 7 women with an age range of 52-78 years (mean, 66.2 years) and with a disease duration of 6 months to 5 years (mean, 1.5 years). Of 22 patients, 13 had a history of smoking; 8 were classified as Fontaine III and 14 as Fontaine IV. The complications included diabetes (8 patients), hypertension (16 patients), hyperlipemia (10 patients), coronary heart disease (11 patients), and chronic kidney failure (1 patient). Patency analyses were performed using Kaplan-Meier life tables and log-rank test. Results All patients underwent successfully procedures. The time of operation was 70-160 minutes (mean, 137 minutes). Acute myocardial infarction, hematoma of incision, fracture of stent, and stent thrombosis occurred in 1 case, respectively. At 6 months after surgery, the ankle brachial index (ABI), the transcutaneous oxygen pressure (TcpO2), and the average intermittent claudication distance were significantly increased when compared with preoperative ones [0.79 ± 0.33 vs. 0.32 ± 0.18, (42.7 ± 15.7) kPa vs. (17.6 ± 11.6) kPa, and (420 ± 80) m vs. (160 ± 54) m, P lt; 0.05]. The patients were followed up 6-24 months (mean, 14.5 months). The primary patency rate, primary assisted patency rate, and second patency rate were 77.3% (17/22), 90.9% (20/22), and 95.5% (21/22) respectively, showing no significant difference among them (P gt; 0.05). No significant difference was found in various-stage patency rates between patients at Fontaine III and IV (P gt; 0.05). Conclusion Hybrid procedures provide an effective treatment of multilevel femoral artery and popliteal artery disease while there is good outflow.
Objective To compare canine decel luarized venous valve stent combining endothel ial progenitor cells (EPC) with native venous valve in terms of venous valve closure mechanism in normal physiological conditions. Methods Thirty-six male hybrid dogs weighing 15-18 kg were used. The left femoral vein with valve from 12 dogs was harvested to prepare decelluarized valved venous stent combined with EPC. The rest 24 dogs were randomly divided into the experimental group and the control group (n=12 per group). In the experimental group, EPC obtained from the bone marrowthrough in vitro ampl ification were cultured, the cells at passage 3 (5 × 106 cells/mL) were seeded on the stent, and the general and HE staining observations were performed before and after the seeding of the cells. In the experimental group, allogenic decelluarized valved venous stent combined with EPC was transplanted to the left femoral vein region, while in the control group, the autogenous vein venous valve was implanted in situ. Color Doppler Ultrasound exam was performed 4 weeks after transplantation to compare the direction and velocity of blood flow in the distal and proximal end of the valve, and the changes of vein diameter in the valve sinus before and after the closure of venous valve when the dogs changed from supine position to reverse trendelenburg position. Results General and HE staining observations before and after cell seeding: the decelluarized valved venous stent maintained its fiber and collagen structure, and the EPC were planted on the decelluarized stent successfully through bioreactor. During the period from the reverse trendelenburg position to the starting point for the closure of the valve, the reverse flow of blood occurred in the experimental group with the velocity of (1.4 ± 0.3) cm/s; while in the control group, there was no reverse flow of blood, but the peak flow rate was decreased from (21.3 ± 2.1) cm/s to (18.2 ± 3.3) cm/s. In the control group, the active period of valve, the starting point for the closure of the valve, and the time between the beginning of closure and the complete closure was (918 ± 46), (712 ± 48), and (154 ± 29) ms, respectively; while in the experimental group, it was (989 ± 53), (785 ± 43), and (223 ± 29) ms, respectively. There was significant difference between two groups (P lt; 0.05).After the complete closure of valve, no reverse flow of blood occurred in two groups. The vein diameter in the valve sinus of the experimental and the control group after the valve closure was increased by 116.8% ± 2.0% and 118.5% ± 2.2%, respectively, when compared with the value before valve closure (P gt; 0.05). Conclusion Canine decelluarized venous valve stent combined with EPC is remarkably different from natural venous valve in terms of the valve closure mechanism in physiological condition. The former rel ies on the reverse flow of blood and the latter is related to the decreased velocity of blood flow and the increased pressure of vein in the venous sinus segment.
Objective To study the cellular biocompatibility, adhesion and proliferation of endothelial outgrowth cells (EOCs) isolated and expanded from rabbit peripheral blood cultured with aligned poly-L-lactic acid (PLLA) nanofibrous scaffolds in vitro so as to provide a basis for the applications of scaffolds biomaterials in tissue repair. Methods Nanofibrous scaffolds of PLLA by electrostatic spinning were modified by hypothermal plasmas body and type Ⅰ collagen was coated onto the materials physically. In vitro, EOCs were cultured on the modified PLLA scaffold. Adhesion and proliferation were surveyed and morphological changes and biocompatibility of seeding cells on PLLA scaffold were observed by growth curves of the cells, fluorescent microscope and scanning electron microscope respectively. Results Fibers with diameters ranging from 300 nm to 400 nm were included in the nanofibrous scaffolds, whose porosities were more than 90%. Absorbance (A) of each scaffold increased gradually after EOCs grew in the absence or presence of random, aligned, or super-aligned PLLA nanofibrous scaffold. Although there was no detectable effect of the random PLLA scaffold on the growth EOCs (Pgt;0.05), both aligned and super-aligned PLLA nanofibrous scaffold had significantly enhanced their growth since the 5th day (P＜0.05). The rates of adhesion in both aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA scaffold after 12 h and 24 h incubation (P＜0.01). The rates of proliferation after 1 d, 3 d and 7 d incubation in aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA nanofibrous scaffold (P＜0.05, P＜0.01). EOCs grew well with PLLA scaffold, yet confused and disorderly in random nanofibers. EOCs could attach, extend and proliferate following fibrous orientation in aligned and super-aligned PLLA nanofibrous scaffold, in majority of the fibers were oriented along the longitudinal axis so that a unique aligned topography was formed. Especially super-aligned PLLA nanofibrous had advantageous to keep well on cell morphology. Conclusion EOCs are ideal seeding cells for tissue engineering. EOCs can be adhered well to aligned and super-aligned PLLA nanofibrous scaffold and proliferate, keep well on cell morphology. So this type of PLLA nanofibrous scallfold is proposed to be an optimal candidate material for EOCs transplantation in tissue repair.
Objective To observe the occurrence condition of endoleak after endovascular aneurysm repair (EVAR) operation for abdominal aortic aneurysm (AAA), and to analyze the factors of the endoleak. Methods Between July 2005 and June 2013, 210 cases of AAA were treated with EVAR. Of 210 patients, 175 were male and 35 were female, aging 42-89 years (mean, 65.7 years). The patients were all proved to have infrarenal AAA by computed tomography angiography (CTA). The disease duration ranged from 1 week to 2 years (median, 11.3 weeks). The maximum diameter of the aneurysms was 44-72 mm (mean, 57.3 mm). The proximal landing zone was longer than 1.5 cm. CTA was performed routinely at 2 months after operation to detect the endoleak of contrast agent. If endoleak was found, CTA was performed again at 6 months. If obvious endoleak still existed, digital subtraction angiography (DSA) would be performed to clarify the character and the degree of the endoleak, and EVAR should be done if necessary. Results Endoleak occurred in 31 cases (14.8%) during operation, including 11 cases of type I endoleak (8 cases of type IA and 3 cases of type IB), 18 cases of type II endoleak, and 2 cases of type III endoleak (type IIIB). The patients were followed up 2-8 months (mean, 3.1 months). At 2 months after operation, contrast agent endoleak was found in the remnant aneurysm cavity of 12 cases (5.7%). At 6 months after eperation, contrast agent endoleak was found in 10 cases (4.8%) by CTA. In 8 patients receiving DSA, there were 4 cases of type I endoleak (3 cases of type IA and 1 case of type IB), 3 cases of type II endoleak, and 1 case of type III (type IIIB) endoleak. In 5 patients having type I and type III endoleak, collateral movement of stent graft was observed in different degree; after increased stent graft was implanted, the endoleak disappeared after 2-4 months. The patients having type II endoleak were not given special treatment, endoleak still existed at 2 months after reexamination of CTA, but the maximum diameter of AAA had no enlargement. Conclusion The collateral movement of stent graft is a very important factor to cause type I and type III endoleak in the patients of AAA after EVAR, and endoleak can be plugged by EVAR again.
Objective To compare the early compl ications of carotid stenting (CAS) and carotid endarterectomy (CEA) in treatment of carotid artery stenosis. Methods Between January 2005 and December 2007, 63 patients with carotid artery stenosis were treated with CEA in 36 cases (CEA group) and with CAS in 27 cases (CAS group). There were 42 males and 21 females with an average age of 67.5 years (range, 52-79 years). The locations were the left side in 28 cases and the rightside in 35 cases. The carotid stenosis was 60%-95% (mean, 79%). The major cl inical symptoms were stroke and transient ischemic attack. The cranial CT showed old cerebral infarction in 24 cases, lacunar infarction in 22 cases, and no obvious abnormal change in 17 cases. The encephalon, heart, and local compl ications were compared between 2 groups within 7 days after operation. Results In CEA group, encephalon compl ications occurred in 3 cases (8.3%), heart compl ications in 2 cases (5.6%), and local compl ications in 5 cases (13.9%); while in CAS group, encephalon compl ications occurred in 8 cases (29.6%), heart compl ications in 1 case (3.7%), and local compl ications in 3 cases (11.1%). The encephalon compl ication ratio of CAS group was significantly higher than that of CEA group (χ2=4.855, P=0.028); and there was no significant difference in other compl ications ratios between 2 groups (P gt; 0.05). Conclusion CEA is the first choice to treat carotid artery stenosis.
Objective To investigate the efficacy of autologous bone marrow mononuclear cells transplantation in treating lower l imb thromboangiitis obl iterans (TAO). Methods From January 2005 to November 2008, 25 patients (27 l imbs) with lower l imb TAO were treated. There were 24 males (26 l imbs) and 1 female (1 l imb), aging 16-44 years (33 years on average). Fifteen left l imbs and 12 right l imbs were involved. The median duration of disease was 2 years (from 3 months to9 years). Intermittent claudication was observed in 5 cases (5 l imbs), 16 patients (17 l imbs) had symptom of rest pain, 4 patients (5 l imbs) suffered ulcer on the distal l imbs. The results of visual analogue scale (VAS), maximum walking distance (MWD), ankle/brachial index (ABI), and transcutaneous oxygen pressure (TcPO2) before operation were (7.16 ± 1.12) points, (0.098 ± 0.043) km, 0.20 ± 0.09, and (11.78 ± 3.46) mm Hg (1 mm Hg=0.133 kPa), respectively. A total of 300 mL bone-marrow blood was extracted from the il iac bone. And then the mononuclear cells were isolated from the bone-marrow blood. All patients received cell transplantation only one time. The amount of transplantation bone marrow mononuclear cells was (1.82-29.46) × 109 (mean 13.33 × 109). Results All patients were followed up for 1 years. After 4 weeks of implantation, the results of VAS, MWD, ABI, and TcPO2 were (2.39 ± 0.51) points, (0.783 ± 0.176) km, 0.28 ± 0.16, (21.33 ± 6.57) mm Hg, respectively, showing significant difference compared with preoperative results (P lt; 0.05). The VAS, MWD, ABI, and TcPO2 increased to (2.44 ± 0.67) points, (1.199 ± 0.304) km, 0.37 ± 0.09, (27.90 ± 5.23) mm Hg after 1 year of implantation, showing significant differences compared with preoperative results (P lt; 0.05). One ulcer healed well and the improvement was obtained in other 3 cases after 4 weeks of implantation (80%). Four ulcers healed well after 1 year of implantation (80%). After 1 year of implantation, angiography revealed 37.04% affected limbs had a satisfactory neovascularization. The angiographic levels were grade 0 in 5 cases, grade 1 in 12 cases, grade 2 in 4 cases, and grade 3 in 6 cases. Conclusion Autologous bone marrow mononuclear cells transplantation could be a simple, safe, effective method to treat TAO.
Objective To investigate the etiology, diagnosis, revascularization of upper l imb ischemia and the compl ications. Methods From March 2003 to February 2008, 72 cases of upper l imb ischemia were treated. There were 44males and 28 females, aged 19-90 years old (median 63 years old). The duration of the disease was 1 hour to 2 years. All cases had symptoms of l imb ischemia such as paleness, coldness, paralysis. According to individual condition, 72 patients accepted revascularizations including thromboembolectomy, reconstruction after traumatic injuries, pseudoaneurysm excision and angioplasty, balloon dilatation and stent implant, arterial repair, patch, vascular prosthesis or vein bypass/transplantation, and l igation or coarctation of fistula. Results Sixty patients (83.3%) recovered well after operation. Re-occlusion following thromboemboletomy was found in 6 patients (8.3%). And there were 4 patients (5.6%) with l imbs disturbance and muscles contracture and 2 patients (2.8%) with compartment syndrome in this series. The affected l imb had to be amputated in 2 patients (2.8%). And 1 patient (1.4%) died of cerebral hemorrhage because of anticoagulation 3 days after operation. All patients were followed up 1-6 years (mean 52 months) after operation. Four patients recurred and got improved after retreatments. The others got a good result with normal skin color and temperature, restoration of the radial and ulnar pulses, normal saturation of blood oxygen of finger ti p (gt; 90%) and patent blood flow of affected arteries was shown by color Doppler ultrasound. Conclusion The study indicates that identifying the etiology of upper l imb ischemia before operation and active revascularizations consistent with different causes are the key to treat the upper l imb ischemia.
Objective To observe the adhesion and prol iferation of late endothel ial progenitor cells (EPCs) planted on nanoporous PLLA scaffold in vitro and to provide a new approach that optimizes tissue engineered material. Methods Male and female New Zealand rabbits (weight 2.5-3.0 kg) were used. Isolated late EPCs from rabbit peri pheral blood were cultured. Electrostatic spinning technique was adopted to prepare misal igned nanofibers, al igned nanofibers and super-al igned nanofibers, and low temperature plasma technique was appl ied to prepare misal igned membrane, al igned membrane and super-al igned membrane. After being divided into group A (cells only), B (misal igned membrane), C (normal membrane), D (al igned membrane) and E (super-al igned membrane), the primary late EPCs (1 × 105/mL) werecultured on scaffolds and MTT method was used to detect cell prol iferation abil ity at 3, 5, 7, 9, 11, 13, 15 and 17 days afterculture. After being divided into group A (misal igned membrane), B (normal membrane), C (al igned membrane) and D (superal igned membrane), precipitation method was appl ied to detect cell adhesion rate at 4, 12 and 24 hours after compound culture, and the morphologic changes of cells were observed at 4, 24 and 72 hours after compound culture. Results Fiber diameters in nanofibrous PLLA scaffolds were 300-400 nm, with a porosity rate of above 90%. At 3, 5, 7, 9, 11, 13, 15 and 17 days after culture, A value of each group was increased with time and the cells in each group grew well, showing there was no significant difference between group A and group B at each time point (P gt; 0.05 ); during the period of 7-15 days after culture, the difference between groups C, D and E and groups A and B was significant (P lt; 0.05). At 4 hours after compound culture, the adhesion rate of group A was superior to that of groups B, C and D (P lt; 0.05); at 12 and 24 hours after compound culture, the adhesion rate of groups B, C and D was remarkably higher than that of group A (P lt; 0.05); significant difference was noted in each group between the time point of 4 hours and the time point of 12 and 24 hours after compound culture (P lt; 0.05), but no significant difference between 12 hours and 24 hours was detected (P gt; 0.05). Morphology observation demonstrated that cells grew well on the scaffolds, the cells in groups A and B grew sporadically and disorderly, while the cells in groups C and D attached and al igned along fiber and prol iferated, with an excretion of ECM. Group D was better at maintaining cell morphology. Conclusion Al igned and superal igned nanofibers of PLLA scaffold can promote the adhesion and prol iferation of seed cells on the scaffold and maintain good cell morphology, which is an appropriate candidate scaffold material for blood vessel tissue engineering. Late EPCs is an ideal cell source for blood vessel tissue engineering.
【Abstract】 Objective To design a novel small-cal iber vascular graft using a decellularized allogeneic vascularscaffold pre-loaded with bFGF. Methods The decellularized canine common carotid were obtained by a detergent-enzymatic procedure, then the scaffolds were covalently l inked with heparin and pre-loaded with bFGF, the amount of binding bFGF and releasing curve were assayed by ELISA. Canine BMSCs expanded in vitro were seed on the scaffolds to observe the effects of binding bFGF on prol iferation. Both bFGF pre-loaded and non-pre-loaded decellularized grafts were implanted in canines as carotid artery interposition for 8 weeks, the patency was examined by digital subtraction angiography and histological method. Results Histology and electron microscopic examination of the decellularized scaffolds showed that cellular components were removed completely and that the extracellular matrix structure remained intact. The amount of binding bFGF positively related to the concentration of bFGF. There was a significant difference in the amount of binding bFGF between two different scaffoldsthroughout all bFGF concentrations(P lt; 0.05), and up to 100 ng/mL, the local and sustained release of bFGF from the heparin treated scaffolds were assayed up to 20 days. Additionally, MTT test showed the bFGF-preloaded scaffolds significantly enhanced the prol iferation of seeded BMSCs in vitro compared with non-bFGF-preloaded scaffolds at 3 days after seeding and thereafter(P lt; 0.01). Furthermore, in vivo canine experiments revealed that all 8 bFGF-pre-loaded scaffolds remained patent after 8 weeks of implantation, and host cell l ined the lumen and populated the wall. Only 1 non-bFGF-pre-loaded scaffold was patent, and the other 7 grafts were occluded because of thrombsus formation. Conclusion This study provides a new strategy to develop a small diameter vascular graft with excellent biocompatibil ity and high patency rate.