Expression of platelet endothelial cell adhesion molecule-1 between pancreatic microcirculation and peripheral circulation in rats with acute edematous pancreatitis

OBJECTIVE: To study the ehanges of platelet endothelial cell adhesion molecule-1 (PECAM-1) expression on polymorphonuclear leukocytes (PMNs) in peripheral circulation anti pancreatic microcirculation in rats with acute edematous pancreatitis (AEP). METHODS: The model of AEP was established with 50 Wistar rats, and the changes of PECAM-1 expression on PMNs from the splenic vein and inferior vena cava were determined by flow cytometry. RESULTS: PECAM-I expression on PMNs showed no significant difference between pancreatic microcirculation and peripheral circulation at AEP2h and AEP4h time points. From the AEP4h to the AEP8h time point, PECAM-1 expression in peripheral circulation was up-regulated, but PECAM-1 expression in pancreatic microcirculation was down-regulated. PECAM-1 expression had a significant difference between pancreatic microcirculation and peripheral circulation at the AEP8h time point (P<0.05). CONCLUSION: PECAM-1 expression on PMNs is in a converse way between pancreatic microcirculation and peripheral circulation in AEP.

Effects of TCMP-1 on the changes of platelet endothelial cell adhesion molecule-1 expression in acute edematous pancreatitis

BACKGROUND: Traditional Chinese medicine is a potent agent in the management of clinical and experimental acute pancreatitis (AP), but the molecular mechanism of its the- rapeutic action is unclear. Numerous experimental and clinical studies have shown that platelet endothelial cell ad- hesion molecule-1 (PECAM-1) is pivotal to leukocyte re- cruitment, which results in microcirculatory injury during inflammation, but its role in acute pancreatitis is poorly un- derstood. We investigated the effects of a compound of tra- ditional Chinese medicine pancreatitis-1 (TCMP-1) on the changes of platelet endothelial cell adhesion molecule-1 (PECAM-1) expression on polymorphonuclear leukocytes (PMNs) in acute edematous pancreatitis (AEP). METHODS: The model of acute pancreatitis was estab- lished by subcutaneous injection of caerulein, and TCMP-1 treated groups were given TCMP-1 by catheterization from mouth to stomach (20 ml/kg) immediately after first time subcutaneous injection of caerulein. The changes of expres- sion of PECAM-1 on leukocytes from the blood of the splenic vein and inferior vena cava were determined by flow cytometry. RESULTS: In the AEP group, expression of PECAM-1 on PMNs was not significantly different between pancreatic microcirculation and systemic circulation at AEP2h and AEP4h time point. Then from AEP4h time point to AEP8h time point, expression of PECAM-1 was up-regulated in systemic circulation while it was down-regulated in pancre- atic microcirculation and was significantly different be- tween pancreatic microcirculation and systemic circulation at AEP8h time point (P<0.05). In the TCMP-1 treated group, compared with the AEP group, expression of PE-CAM-1 on PMNs decreased in different levels between pan- creatic microcirculation and systemic circulation and was of significant difference at AEP8h time point (P <0.05). CONCLUSION: Inhibition of PECAM-1 expression on PMNs may prevent PMNs from transmigration through the endo- thelium and may be one of the treatment mechanisms of TCMP-1 decoction on AEP.

Platelet Adhesion onto Nitinol Surfaces Modified by Microwave Cold-plasma

The macromolecular structures on nitinol surfaces were prepared by ECR microwave cold-plasma of tetraglyme conditions. The surface chemistry was characterized by high resolution ESCA. The results showed that the modified nitinol surfaces were built up mainly of -CH2-CH2-O- linkages and were particularly effective in preventing platelet adhesion.

Differences in platelet endothelial cell adhesion molecule-1 expression between peripheral circulation and pancreatic microcirculation in cerulein-induced acute edematous pancreatitis

AIM: To investigate the changes of platelet endothelial cell adhesion molecule-1 (PECAM-1) expression on polymorphonuclear leukocytes (PMNs) in peripheral circulation and pancreatic microcirculation in cerulein-induced acute edematous pancreatitis (AEP).METHODS: Fifty Wistar rats were randomly divided into control group (n=10) and AEP group (n=40). A model of AEP was established by subcutaneous injection of cerulein 5.5 and 7.5 μg/kg at 0 and 1 h after the beginning of experiment respectively. PECAM-1 expression on PMNs from splenic vein and inferior vena cava was determined by RT-PCR at mRNA level and determined by flow cytometry at protein level.RESULTS: In experimental rats, an increased PECAM-1mRNA expression was seen from 4 to 8 h of AEP in peripheral circulation (0.77±0.25%, 0.76±0.28%, 0.89±0.30%,1.00±0.21% ), while in pancreatic microcirculation,expression decreased from 2 h and reached the lowest level at 6 h of AEP (0.78±0.29%, 0.75±0.26%, 0.62±0.28%,0.66±0.20%). There were significant differences at 8-h time point of AEP between peripheral circulation and pancreatic microcirculation (1.00±0.21% vs0.66±0.20%, P＜0.05).Meanwhile,the difference at protein level was also found.CONCLUSION: A reverse expression of PECAM-1 on PMNs was found between peripheral circulation and pancreatic microcirculation, suggesting that inhibition of PECAM-1expression may improve the pathological change of AEP.

The relationship between platelet endothelial cell adhesion molecule-1 and paraquat-induced lung injury in rabbits

BACKGROUND:Platelet endothelial cell adhesion molecule-1(PECAM-1),also known as CD31,is mainly distributed in vascular endothelial cells.Studies have shown that PECAM-1 is a very significant indicator of angiogenesis,and has been used as an indicator for vascular endothelial cells.The present study aimed to explore the relationship between the expression of PECAM-1 and the degree of acute lung injury(ALI) and fibrosis in paraquat(PQ) induced lung injury in rabbits.METHODS:Thirty-six adult New Zealand rabbits were randomly divided into three groups(12rabbits in each group) according to PQ dosage:8 mg/kg(group A),16 mg/kg(group B),and 32 mg/kg(group C).After PQ infusion,the rabbits were monitored for 7 days and then euthanized.The lungs were removed for histological evaluation.Masson staining was used to determine the degree of lung fibrosis(LF),and semi-quantitative immune-histochemistry analysis to determine the expression of PECAM-1.Pearson's product-moment correlation analysis was performed to evaluate the relationship between the expression of PECAM-1 and the extent of lung injuries expressed by ALI score and degree of LF.RESULTS:Rabbits in the three groups showed apparent poisoning.The rabbits survived longer in group A than in groups B and C(6.47±0.99 days vs.6.09±1.04 days vs.4.77±2.04 days)(P<0.05).ALI score was lower in group A than in groups B and C(8.33±1.03 vs.9.83±1.17 vs.11.50±1.38)(P<0.05),and there was statistically significant difference between group B and group C(P=0.03).LF was slighter in group A than in groups B and C(31.09%±2.05%vs.34.37%±1.62%vs.36.54%±0.44%)(P<0.05),and there was statistically significant difference between group B and group C(P=0.026).The PEACAM-1 expression was higher in group A than in groups B and C(20.31%±0.70%vs.19.34%±0.68%vs.18.37%±0.46%)(P<0.05),and there was statistically significant difference between group B and group C(P=0.017).Pearson's correlation analysis showed that the expression of PECAM-1 was negatively correlated to both ALI score(Coe=-0.732,P=0.001)and degree of LF(Coe=-0.779,P<0.001).CONCLUSIONS:The PECAM-1 expression significantly decreases in New Zealand rabbits after PQ poisoning,and the decrease is dose-dependent.The PECAM-1 expression is negatively correlated with ALI score and LF,showing a significant role in the development of lung injuries induced by PQ.

Association of G+1688A Polymorphism of Platelet Endothelial Cell Adhesion Molecule-1 Gene with Myocardial Infarction in the Chinese Han Population

In order to investigate the association of G＋1688A （Ser563Asn） polymorphism of platelet endothelial cell adhesion molecule-1 （PECAM-1） gene with myocardial infarction （MI） in the Chinese Han population, the G＋1688A polymorphism in PECAM-1 gene was detected by polymerase chain reaction-restriction fragment-length polymorphism （PCR-RFLP） method among 502 subjects, including 218 patients with MI and 284 controls. The results showed that there was significant difference in AA frequencies of genotype G＋1688A polymorphism between case and control groups （39% vs 24%, P〈0.001）. A similar trend was observed on the allele frequencies （A/G： 62% vs 49%, P〈0.001）. Among the subjects with high serum total cholesterol level or high systolic blood pressure level, the variant AA genotype was associated with high risk of MI （adjusted OR, 2.13; 95% CI, 1.08 -4.41 and adjusted OR, 2.53; 95%CI, 1.63-3.63）. The single nucleotide polymorphism （SNP） at position ＋1688 in the exon 8 of PECAM-1 gene was associated with MI and the allele A might be a risk factor for MI in the Chinese Han population.

Platelet adhesion to the surface of a sudden tubular expansion tube under swirling flow condition

The size mismatch in an end-to-end vascular anastomosis between the host vessel and the graft may cause flow disturbance and predispose to thrombosis [1].Although a number of techniques have been employed to reduce the risk of anastomotic thrombosis due to the size mismatch。

Characterization of Ti-Cu Films Deposited by HPPMS and Effect on NO Catalytic Release and Platelet Adhesion Behavior

Ti-Cu films with different Cu concentrations were fabricated by high-power pulsed magnetron sputtering（HPPMS） to release copper ions and catalyze NO to improve the blood compatibility. The Cu concentrations of films were 25.7 at% and 68.8 at%. Pure Ti films were also fabricated. Copper release, catalytic release of nitric oxide（NO）, and blood platelet adhesion of Ti-Cu films were studied. Ti-Cu films released copper ions in PBS solution and more Cu ions were released from films with 68.8 at% Cu. Ti-Cu films had excellent ability of catalytical decomposition of exogenous donor S-nitroso-N-acetyl-DL-penicillamine（SNAP） and as a result, nitric oxide（NO） was generated. The NO generation catalyzed by Ti-Cu films was significantly higher than that by pure Ti films. This was more eminent in the Ti-Cu films with 68.8 at% Cu. The platelet adhesion and activation of Ti-Cu films were significantly inhibited compared to that of pure Ti films in the presence of SNAP. The Ti-Cu film fabricated by HPPMS showed the ability of releasing Cu ions to catalyze SNAP to generate NO to inhibit platelet adhesion and activation.

Integrin binding peptides facilitate growth and interconnected vascular-like network formation of rat primary cortical vascular endothelial cells in vitro

Neovascularization and angiogenesis in the brain are important physiological processes for normal brain development and repair/regeneration following insults. Integrins are cell surface adhesion receptors mediating important function of cells such as survival, growth and development during tissue organization, differentiation and organogenesis. In this study, we used an integrin-binding array platform to identify the important types of integrins and their binding peptides that facilitate adhesion, growth, development, and vascular-like network formation of rat primary brain microvascular endothelial cells. Brain microvascular endothelial cells were isolated from rat brain on post-natal day 7. Cells were cultured in a custom-designed integrin array system containing short synthetic peptides binding to 16 types of integrins commonly expressed on cells in vertebrates. After 7 days of culture, the brain microvascular endothelial cells were processed for immunostaining with markers for endothelial cells including von Willibrand factor and platelet endothelial cell adhesion molecule. 5-Bromo-2′-dexoyuridine was added to the culture at 48 hours prior to fixation to assess cell proliferation. Among 16 integrins tested, we found that α5β1, αvβ5 and αvβ8 greatly promoted proliferation of endothelial cells in culture. To investigate the effect of integrin-binding peptides in promoting neovascularization and angiogenesis, the binding peptides to the above three types of integrins were immobilized to our custom-designed hydrogel in three-dimensional(3 D) culture of brain microvascular endothelial cells with the addition of vascular endothelial growth factor. Following a 7-day 3 D culture, the culture was fixed and processed for double labeling of phalloidin with von Willibrand factor or platelet endothelial cell adhesion molecule and assessed under confocal microscopy. In the 3 D culture in hydrogels conjugated with the integrin-binding peptide, brain microvascular endothelial cells formed interconnected vascular-like network with clearly discernable lumens, which is reminiscent of brain microvascular network in vivo. With the novel integrin-binding array system, we identified the specific types of integrins on brain microvascular endothelial cells that mediate cell adhesion and growth followed by functionalizing a 3 D hydrogel culture system using the binding peptides that specifically bind to the identified integrins, leading to robust growth and lumenized microvascular-like network formation of brain microvascular endothelial cells in 3 D culture. This technology can be used for in vitro and in vivo vascularization of transplants or brain lesions to promote brain tissue regeneration following neurological insults.

A Novel Approach for the Surface Modification of Polymeric Membrane with Phospholipid Polymer

A new economic and convenient method to modify the surface of microporous polypropylene (PP) membranes with phospholipid polymer was given. The process included the photo-irradiated graft polymerization of N,N-dimethylaminoethyl methacrylate (DMAEMA) and the ring-opening reaction of the grafted polyDMAEMA with 2-alkyloxy-2-oxide-1,3,2-dioxo-phospholanes (AOP). Four AOPs, whose alkyloxy groups consisted of dodecyl, tetradecyl, hexadecyl and octadecyl moieties, were used to convert the grafted polyDMAEMA to phospholipid polymers. FT-IR spectra confirmed the chemical change of membrane surface. Platelets adhesion experiment indicated that PP membrane with excellent blood compatible surface could be fabricated by this method.

Temporal alterations in pericytes at the acute phase of ischemia/reperfusion in the mouse brain

Pericytes,as the mural cells surrounding the microvasculature,play a critical role in the regulation of microcirculation;however,how these cells respond to ischemic stroke remains unclear.To determine the temporal alterations in pericytes after ischemia/reperfusion,we used the 1-hour middle cerebral artery occlusion model,which was examined at 2,12,and 24 hours after reperfusion.Our results showed that in the reperfused regions,the cerebral blood flow decreased and the infarct volume increased with time.Furthermore,the pericytes in the infarct regions contracted and acted on the vascular endothelial cells within 24 hours after reperfusion.These effects may result in incomplete microcirculation reperfusion and a gradual worsening trend with time in the acute phase.These findings provide strong evidence for explaining the“no-reflow”phenomenon that occurs after recanalization in clinical practice.

Silk Fibroin-modified Poly(butadiene) urethane Films and Their Effects on Fibroblast Viability

Surface-modified poly(butadiene)urethane (PBTU) films with silk fibroin (SF) were prepared by simple chemical method under the normal temperature. The physical properties and biological behaviour of the SF-modified PBTU film were evaluated. The results showed that the SF-modified PBTU films kept the tenacity and pliability very well, and could overcome rigid and brittle weaks of silk fibroin films. The morphology of SF in the PBTU film was dendritic aggregations, and the water-contact angle measurement indicated that the surface hydrophilicity of modified films was apparently enhanced. The biocompatibility of PBTU films was improved due to the change of surface components. The degree of platelet adhesion and the cell viability of rat embryo dermal fibroblasts seeded on PBTU films, SF films, and SF-modified PBTU films were measured by counting platelets before and after they contacted the films and MTT assay, respectively. The results indicated that platelet adhesion resistance and cell viability on the modified film were greatly superior to those on the PBTU film and the compound interface had good stability in the air.

Effect of Yiqitongyanghuatan Recipe on Experimental Atherosclerosis in Rabbits

In order to investigate the roles of Yiqitongyanghuatan (YQTYHT) recipe in reducing the levels of serum cholesterol and plasma lipid peroxidation(LPO), platelet aggregation function (PAgF) and platelet adhesion function (PAdF), the area of atherosclerotic plague coverage in aorta and the thickness of plague, 32 male Japanese white rabbits were divided into 4 groups. The results showed that the YQTYHT recipe could significantly lower the levels of serum cholesterol and tryglyceride, plasma LPO, and PAgF and PadF. The area of atherosclerotic plague coverage in aorta and the thickness of plague in the YQTYHT fed rabbits were decreased as compared with that in the high cholesterol fed rabbits. The above roles might contribute to the main mechanism of YQTYHT against atherosclerosis.

SYNTHETIC STUDIES ON BLOOD COMPATIBLE BIOMATERIALS 13:A NOVEL SEGMENTED POLYURETHANE CONTAINING PHOSPHORYLCHOLINE STRUCTURE:SYNTHESIS,CHARACTERIZATION AND BLOOD COMPATIBILITY EVALUATION

A new phosphorylcholine, (6-hydroxy) hexyl-2-(trimethylammonio) ethyl phosphate (HTEP), was synthesized andcharasterized. Segmented polyurethane (SPU) containing phosphorylcholine structure was synthesized based ondiphenylmethane diisocyanate (MDI), soft segment polytetramethylene glycol (PTMG) and HTEP, with 1,4-butanediol (BD)as a chain extender. The existence of phosphorylcholine structure on the surface of SPU was revealed by attenuated totalreflectance Fourier transform infrard spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS) and water contactangle measurements. The blood compatibilities of the polymers were evaluated by hemolytic testing and a platelet-richplasma (PRP) adhesion experiment, which was viewed by scanning electron microscopy (SEM) with polyurethane as areference. The novel segmented polyurethane containing phosphorylcholine structure showed improved blood compatibility.

IMPROVING NONTHROMBOGENICITY OF CHITIN WITH ZWITTERIONIC STRUCTURE OF SULFOBETAINE

In order to improve the nonthrombogenicity of chitin,a new monomer,N,N-dimethyl(β-hydroxyethyloxyethyl) ammonium propanesulfonate(DHAPS)was designed,synthesized and grafted onto the chitin membrane by using hexamethylene diisocyanate(HDI)as a coupling agent.Surface analysis of the grafted membranes by ATR-FTIR and XPS confirms that DHAPS has been successfully grafted onto the membrane surface.The platelet resistant property of the grafted membranes was evaluated by a platelet-rich plasma adhesion method.The results showed that platelet-adhesive resistance of the modified membrane has been greatly improved.

Syntheses and Surface Properties of Polyacrylonitrile-based Copolymer Membranes Containing Sugar Moieties

To improve the hydrophilicity of polyacrylonitrile-based membranes, sugar moieties were incorporated into acrylonitrile-based copolymers via the radical copolymerization of α-allyl glucoside(AG) with acrylonitrile(AN) with 2,2′-azobis-iso-butyronitrile(AIBN) as the initiator in dimethyl sulphoxide(DMSO). It was found that the yield increased with the increase of the initiator concentration and reaction time, while it decreased with the increase of the monomer molar ratio of AG to AN. Raising the AG proportion in the monomer mixture resulted in the increase of the AG content in the copolymer. M_v of the copolymers decreased with increasing the AG monomer fraction in feed. The copolymers were fabricated into dense membranes and their surface properties were studied by means of the water contact angle measurement and platelet adhesion tests. It was found that the static water contact angle on the membrane decreased significantly from 70° to 33° with the increase of the AG content. The adhesive number of platelets on the membrane surface also decreased significantly with increasing AG content in the copolymers. These results demonstrate that the hydrophilicity and biocompatibility of the acrylonitrile-based copolymer membranes could be improved efficiently by the copolymerization of acrylonitrile with vinyl carbohydrates.

GRAFTING OF SULFOBETAINE ONTO A POLYURETHANE SURFACE TO IMPROVE BLOOD COMPATIBILITY

On the molecular level, it is believed that polymers containing zwitterionic structures should be compatible withblood. In this work polyurethane films were grafted with sulfobetaine by a three-step procedure. In the first step, the films'surfaces were treated with hexamethylene diisocyanate (HDI) in toluene at 50℃ in the presence of di-n-butyl tin dilaurate(DBTDL) as a catalyst. The extent of the reaction was monitored by ATR-IR spectra; a maximum number of free NCOgroups was obtained after a reaction time of 90 min. In the second step, the hydroxyl groups of N,N-dimethylethylethanolamine (DMEA) were allowed to react in toluene with NCO groups bound on the surface. In the thirdstep, sulfobetaines were formed on the surface through the ring-opening reaction between tertiary amine of DMEA and 1,3-propanesultone (PS). The surfaces of the films were characterized by ATR-IR and XPS showing that the grafted surfaceswere composed of sulfobetaine. The results of the contact angle measurement show that the surface was strongly hydrophilic.The platelet adhesion test demonstrated that the films grafted with sulfobetaine have excellent blood compatibility.

Surface Modification of Silk Fibroin Films with Zwitterionic Phosphobetaine to Improve the Hemocompatibility

Zwitterionic phosphobetaine bearing a hydroxyl and a zwitterionic group,8-hydroxy-2-octyl phosphorylcholine（HOPC）,was synthesized and constructed to the surface of silk fibroin（SF） films in order to improve the hemocompatibility of fibroin films by a an isocyanate head group.The surface characteristics of the modified films were measured by attenuated total reflection Fourier transform infrared spectroscopy（ATR-FTIR） and electron spectroscopy for chemical analysis（ESCA）,displaying the successful immobilization of Zwitterionic phosphobetaine on the surface of these fibroin films.Moreover,the further platelet adhesion test in platelets rich plasma（PRP） of human beings showed the zwitterionic phosphobetaine led mainly to good nonthrombogenicity.The experimental results indicated a reasonable approach to improve the blood compatibility of fibroin films.

Construction of tissue-engineered heart valves by using decellularized scaffolds and endothelial progenitor cells

Background Tissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. This study was designed to develop a tissue engineering heart valve by using human umbilical cord blood-derived endothelial progenitor cells （EPCs） and decellularized valve scaffolds. Methods Decellularized valve scaffolds were prepared from fresh porcine heart valves. EPCs were isolated from fresh human umbilical cord blood by density gradient centrifugation, cultured for 3 weeks in EGM-2-MV medium, by which time the resultant cell population became endothelial in nature, as assessed by immunofluorescent staining. EPC-derived endothelial cells were seeded onto the decellularized scaffold at 3 × 10^6 cells/cm^2 and cultured under static conditions for 7 days. Proliferation of the seeded cells on the scaffolds was detected using the MTT assay. Tissue-engineered heart valves were analyzed by HE staining, immunofluorescent staining and scanning electron microscopy. The anti-thrombogenic function of the endothelium on the engineered heart valves was evaluated by platelet adhesion experiments and reverse transcription-polymerase chain reaction （RT-PCR） analysis for the expression of endothelial nitric oxide synthase （eNOS） and tissue-type plasminogen activator （t-PA）.Results EPC-derived endothelial cells showed a histolytic cobblestone morphology, expressed specific markers of the endothelial cell lineage including von Willebrand factor （vWF） and CD31, bound a human endothelial cell-specific lectin, Ulex Europaeus agglutinin-1 （UEA-1）, and took up Dil-labeled low density lipoprotein （Dil-Ac-LDL）. After seeding on the decellularized scaffold, the cells showed excellent metabolic activity and proliferation. The cells formed confluent endothelial monolayers atop the decellularized matrix, as assessed by HE staining and immunostaining for vWF and CD31. Scanning electron microscopy demonstrated the occurrence of tight junctions between cells forming the confluent monolayer. Platelets adhesion experiments suggested that the neo-endothelium was non-thrombogenic. The expression levels of eNOS and t-PA genes in the neo-endothelium were quite similar to those in human umbilical vein endothelial cells. Conclusions EPCs isolated from the human umbilical cord blood can differentiate into endothelial cells in vitro and form a functional endothelium atop decellularized heart valve scaffolds. Thus, EPCs may be a promising cell source for constructing tissue-engineered heart valves.

Biocompatibility, biodegradation, and neovascularization of human single-unit platelet-rich fibrin glue： an in vivo analysis

Background The clinical applications of fibrin glue span over several surgical modalities.The aim of this study was to evaluate the biocompatibility and biodegradation of different formulations of platelet-rich fibrin glue in vivo and examine its effects on the neovascularization of wound sites.Methods Human-derived single-unit fibrin glue was prepared.Incisions were made on the backs of rats,and these were coated with homemade glues containing different concentrations of aminomethylbenzoic acid （Groups A-F） or commercial adhesives （Group G）.A sham control group was included （Group H）.The wounds were examined by histological analysis and immunohistochemistry at several time points.Results Successful wound closure was achieved in all groups by day 12.Acute inflammation occurred during the first six days,but gradually disappeared.The longest sealant duration was achieved using the lowest concentration of antifibrinolytic agent in a 1：10 volume ratio with cryoprecipitate.Expression levels of the platelet endothelial cell adhesion molecule-1 were significantly higher in Groups A and C compared to the control groups （Groups G and H） on day 3 （P ＜0.05）.Conclusions Single-unit platelet-rich fibrin glue has excellent biocompatibility and is associated with the upregulation of neovascularization.The addition of aminomethylbenzoic acid could prevent the degradation of fibrin glue.