Animal models of vascularized nerve grafts:a systematic review

The aim of this review is to present and compare the various animal models of vascularized nerve grafts described in the literature as well as to summarize preclinical evidence for superior functional results compared to non-vascularized free nerve grafts. We also will present the state of the art on prefabricated vascularized nerve grafts. A systematic literature review on vascularized nerve graft models was conducted via the retrieval with the Pub Med database on March 30, 2019. Data on the animal, nerve, and vascularization model, the recipient bed, the evaluation time points and methods, and the results of the study results were extracted and analyzed from selected articles. The rat sciatic nerve was the most popular model for vascularized nerve grafts, followed by the rabbit;however, rabbit models allow for longer nerve grafts, which are suitable for translational evaluation, and produced more cautious results on the superiority of vascularized nerve grafts. Compared to free nerve grafts, vascularized nerve grafts have better early but similar long-term results, especially in an avascular bed. There are few studies on avascular receiving beds and prefabricated nerve grafts. The clinical translation potential of available animal models is limited, and current experimental knowledge cannot fully support that the differences between vascularized nerve grafts and free nerve grafts yield a clinical advantage that justifies the complexity of the procedure.

3D printed grafts with gradient structures for organized vascular regeneration

Synthetic vascular grafts suitable for small-diameter arteries(<6 mm) are in great need.However,there are still no commercially available small-diameter vascular grafts(SDVGs) in clinical practice due to thrombosis and stenosis after in vivo implantation.When designing SDVGs,many studies emphasized reendothelization but ignored the importance of reconstruction of the smooth muscle layer(SML).To facilitate rapid SML regeneration,a high-resolution 3D printing method was used to create a novel bilayer SDVG with structures and mechanical properties mimicking natural arteries.Bioinspired by the collagen alignment of SML,the inner layer of the grafts had larger pore sizes and high porosity to accelerate the infiltration of cells and their circumferential alignment,which could facilitate SML reconstruction for compliance restoration and spontaneous endothelialization.The outer layer was designed to induce fibroblast recruitment by low porosity and minor pore size and provide SDVG with sufficient mechanical strength.One month after implantation,the arteries regenerated by 3D-printed grafts exhibited better pulsatility than electrospun grafts,with a compliance(8.9%) approaching that of natural arteries(11.36%) and significantly higher than that of electrospun ones(1.9%).The 3D-printed vascular demonstrated a three-layer structure more closely resembling natural arteries while electrospun grafts showed incomplete endothelium and immature SML.Our study shows the importance of SML reconstruction during vascular graft regeneration and provides an effective strategy to reconstruct blood vessels through 3D-printed structures rapidly.

Treatment of Synovial Cysts in Relation to the Tibial Tunnel of Anterior Cruciate Ligament Grafts by Filling the Tunnel with Acrylic Cement

Introduction: Synovial cyst of the tibial tunnel in connection with anterior cruciate ligament (ACL) reconstruction is a rare but particularly troublesome complication. Medical treatment is often doomed to failure, and surgical treatment usually consists of excising the cyst and filling the tunnel with bone. The aim of this study was to evaluate the results of filling the tunnel with acrylic cement. Hypothesis: Filling the tibial bone tunnel with acrylic cement should eliminate communication between the joint cavity and the pre-tibial surface and prevent cyst recurrence. Patients and Methods: This retrospective series is composed of 13 patients, 9 men and 4 women, mean age 48.5 years (31 to 64) operated on between 2011 and 2019 for an intra- and extraosseous synovial cyst consecutive to the tibial tunnel of an ACL graft. Between 1983 and 2016, 12 of the patients had had a bone graft without bone block fixation (DI-DT or Mac Intosh) and one patient, a bone-bone transplant (KJ). The cyst was of variable size, located on the anteromedial aspect of the proximal end of the tibia, and often painful, warranting consultation. At the time of the initial operation, 9 patients had undergone meniscectomies (6 medial, 2 lateral, 1 double). In 7 knees, there were 7 cartilage lesions in the femorotibial and/or patellofemoral compartments (one stage 1 lesion, 2 stage 2 lesions, 4 stage 3 lesions, and no stage 4 lesions). Only 2 knees had neither cartilage nor meniscus lesions. After curettage of the bone tunnel /− removal of the non-resorbed or PEEK interference screw, the tunnel was filled with acrylic cement /− reinforced with a ligament staple to prevent expulsion. All patients underwent regular follow-up consultations until recovery. Results: At a maximum follow-up of 8 years, only 1 cyst recurred, representing a 7.69% failure rate. It was reoperated with another technique, which involved filling the tibial bone tunnel with bone graft taken from a half-bank head. After recovery, the cyst healed definitively. All patients were able to return to their previous activity within 15 days of surgery. Conclusion: Filling the tibial tunnel with acrylic cement reinforced /− with a ligament staple is a reliable and rapid solution for the treatment of intra- and extra-articular synovial cysts in relation to the tibial tunnel of ACL grafts.

Endoplasmic Reticulum Stress-induced Endothelial Dysfunction Promotes Neointima Formation after Arteriovenous Grafts in Mice on High-fat Diet

Objective Endothelial dysfunction is one candidate for triggering neointima formation after arteriovenous grafts(AVGs),but the factors mediating this process are unclear.The purpose of this study was to investigate the role of endoplasmic reticulum stress(ERS)-induced endothelial dysfunction in neointima formation following AVGs in high-fat diet(HFD)mice.Methods CCAAT-enhancer-binding protein-homologous protein(CHOP)knockout(KO)mice were created.Mice were fed with HFD to produce HFD model.AVGs model were applied in the groups of WT ND,WT HFD,and CHOP KO HFD.Human umbilical vein endothelial cells(HUVECs)were cultured with oxidized low density lipoprotein(ox-LDL)(40 mg/L)for the indicated time lengths(0,6,12,24 h).ERS inhibitor tauroursodeoxycholic acid(TUDCA)was used to block ERS.Immunohistochemical staining was used to observe the changes of ICAM1.Changes of ERS were detected by real-time RT-PCR.Protein expression levels and ERS activation were detected by Western blotting.Endothellial cell function was determined by endothelial permeability assay and transendothelial migration assay.Results HFD increased neointima formation in AVGs associated with endothelial dysfunction.At the same time,ERS was increased in endothelial cells(ECs)after AVGs in mice consuming the HFD.In vitro,ox-LDL was found to stimulate ERS,increase the permeability of the EC monolayer,and cause endothelial dysfunction.Blocking ERS with TUDCA or CHOP siRNA reversed the EC dysfunction caused by ox-LDL.In vivo,knockout of CHOP(CHOP KO)protected the function of ECs and decreased neointima formation after AVGs in HFD mice.Conclusion Inhibiting ERS in ECs could improve the function of AVGs.

Appearance of aseptic vascular grafts after endovascular aortic repair on[(18)F]fluorodeoxyglucose positron emission tomography/computed tomography

BACKGROUND Diagnosis of prosthetic vascular graft infection with[(18)F]fluorodeoxyglucose positron emission tomography/computed tomography(18F-FDG PET/CT)allows for early detection of functional changes associated with infection,based on increased glucose utilization by activated macrophages and granulocytes.Aseptic vascular grafts,like all foreign bodies,can stimulate an inflammatory response,which can present as increased activity on 18F-FDG PET/CT.Consequently,distinguishing aseptic inflammation from graft infection,though important,can be difficult.In the case of endovascular aneurysm repair(EVAR),a minimally invasive procedure involving the transfemoral insertion of an endoprosthetic stent graft,the normal postoperative appearance of these grafts on 18F-FDG PET/CT can vary over time,potentially confounding study interpretation.AIM To investigate the visual,semiquantitative,and temporal characteristics of aseptic vascular grafts in patients status post EVAR.METHODS In this observational retrospective cohort study,patients with history of EVAR who underwent 18F-FDG PET/CT for indications other than infection were identified retrospectively.All patients were asymptomatic for graft infection-no abdominal pain,fever of unknown origin,sepsis,or leukocytosis-at the time of imaging and for≥2 mo after each PET/CT.Imaging studies such as CT for each patient were also reviewed,and any patients with suspected or confirmed vascular graft infection were excluded.One hundred two scans performed on 43 patients(34 males;9 females;age=77±8 years at the time of the final PET/CT)were retrospectively reviewed.All 43 patients had an abdominal aortic(AA)vascular graft,40 patients had a right iliac(RI)limb graft,and 41 patients had a left iliac(LI)limb graft.Twentytwo patients had 1 PET/CT and 21 patients had from 2 to 9 PET/CTs.Grafts were imaged between 2 mo to 168 mo(about 14 years)post placement.Eight grafts were imaged within 6 mo of placement,including three that were imaged within three months of placement.The mean interval between graft placement and PET/CT for all 102 scans was 51±39 mo.PET/CT data was reconstructed with region-of-interest analysis of proximal,mid and distal portions of the grafts and background ascending aorta.Maximum standardized uptake value(SUVmax)was recorded for each region.SUVmax-to-background uptake ratios(URs)were calculated.Visual assessment was performed using a 2-pattern grading scale:Diffuse(homogeneous uptake less than liver uptake)and focal(one or more areas of focal uptake in any part of the graft).Statistical analysis was performed.RESULTS In total,there were 306 AA grafts,285 LI grafts,282 RI grafts,and 306 ascending aorta background SUVmax measurements.For all 102 scans,mean SUVmax values for AA grafts were 2.8-3.0 along proximal,mid,and distal segments.Mean SUVmax values for LI grafts and RI grafts were 2.7-2.8.Mean SUVmax values for background were 2.5±0.5.Mean URs were 1.1-1.2.Visual analysis of the scans reflected results of quantitative analysis.On visual inspection,98%revealed diffuse,homogeneous 18F-FDG uptake less than liver.Graft URs and visual pattern categories were significantly associated for AA graft URs(F-ratio=21.5,P<0.001),LI graft URs(F-ratio=20.4,P<0.001),and RI graft URs(F-ratio=30.4,P<0.001).Thus,visual patterns of 18F-FDG uptake corresponded statistically significantly to semiquantitative URs.The age of grafts showing focal patterns was greater than grafts showing diffuse patterns,87±89 vs 50±37 mo,respectively(P=0.02).URs were significantly associated with graft age for AA grafts(r=0.19,P=0.001).URs were also significantly associated with graft age for LI grafts(r=0.25,P<0.0001),and RI grafts(r=0.31,P<0.001).Quartiles of similar numbers of graft(n=25-27)grouped by graft age indicated that URs were significantly higher for 4th quartile vs 2nd quartile URs(F-ratio=19.5,P<0.001).When evaluating URs,graft SUVmax values within 10%-20%of the ascending aorta SUVmax is evident in aseptic grafts,except for grafts in the oldest quartiles.In this study,grafts in the oldest quartiles(>7 years post EVAR)showed SUVmax up to 30%higher than the ascending aorta SUVmax.CONCLUSION Characteristics of an aseptic vascular stent graft in the aorta and iliac vessels on 18F-FDG PET/CT include graft SUVmax values within 10%-20%of the ascending aorta background SUVmax.The SUVmax of older aseptic grafts can be as much as 30%above background.The visual uptake pattern of diffuse,homogeneous uptake less than liver was seen in 98%of aseptic vascular grafts,making this pattern particularly reassuring for clinicians.

Current Strategies of Surface Modifications to Polyurethane Biomaterials for Vascular Grafts

As the number of patients suffering from cardiovascular diseases and peripheral vascular diseases rises,the constraints of autologous transplantation remain unavoidable.As a result,artificial vascular grafts must be developed.Adhesion of proteins,platelets and bacteria on implants can result in stenosis,thrombus formation,and postoperative infection,which can be fatal for an implantation.Polyurethane,as a commonly used biomaterial,has been modified in various ways to deal with the adhesions of proteins,platelets,and bacteria and to stimulate endothelium adhesion.In this review,we briefly summarize the mechanisms behind adhesions,overview the current strategies of surface modifications of polyurethane biomaterials used in vascular grafts,and highlight the challenges that need to be addressed in future studies,aiming to gain a more profound understanding of how to develop artificial polyurethane vascular grafts with an enhanced implantation success rate and reduced side effect.

Treatment of proximal humeral fractures accompanied by medial calcar fractures using fibular autografts:A retrospective,comparative cohort study

BACKGROUND Severe proximal humerus comminuted fractures are often accompanied by medial calcar comminuted fractures and loss of medial support,which are important factors that lead to internal fixation failure.The appropriate treatment for proximal humerus comminuted fractures has not been established.Therefore,this study assessed the outcomes of using a fibular autograft with locking plates to treat severe proximal humerus comminuted fractures.AIM To investigate the outcomes of using a fibular autograft with locking plates to treat severe proximal humerus comminuted fractures.METHODS This retrospective,comparative cohort study included two groups of patients.Group 1 comprised 22 patients and group 2 comprised 25 patients with complete follow-up data.Group 1 was treated with a fibular autograft with open reduction and locking plates to enable internal fixation.Group 2 was treated with open reduction and locking plates to enable internal fixation.The intraoperative blood loss volume from the shoulder wound,operative time,shoulder wound pain,bone fracture healing time,Constant-Murley score of the shoulder joint,preoperative Holden walking function score,Mallet score of the shoulder joint,and humeral neck-shaft angle during surgery of the two groups were compared,and the differences were analysed using an independent sample t-test.RESULTS Group 1 had a shorter mean operative time than group 2(2.25±0.30 h vs 2.76±0.44 h;P=0.000).Group 1 had a lower shoulder wound pain score on the first day after surgery than group 2(7.91±1.15 points vs 8.56±1.00 points;P=0.044).Group 1 had a shorter fracture healing time than group 2(2.68±0.48 mo vs 3.64±0.64 mo;P=0.000).Group 1 had higher Constant-Murley scores of the shoulder joint at 3,6,and 12 mo after surgery than group 2(76.64±4.02 points vs 72.72±3.02 points,86.36±3.53 points vs 82.96±3.40 points,and 87.95±2.77 points vs 84.68±2.63 points,respectively;P=0.000,0.002,and 0.000,respectively).Group 1 had higher Mallet scores of the shoulder joint at 3,6,and 12 mo after surgery than group 2(10.32±0.57 points vs 9.96±0.54 points,13.36±1.00 points vs 12.60±0.87 points,and 13.91±0.75 points vs 13.36±0.70 points,respectively;P=0.032,0.007,and 0.013,respectively).CONCLUSION Using locking plates with a fibular autograft can recreate medial support,facilitate fracture healing,and improve shoulder function;therefore,this may be an effective treatment option for severe proximal humerus comminuted fractures.

Liver grafts from hepatitis B surface antigen-positive donors: A review of the literature

The scarcity of available organs and the gap between supply and demand continue to be the main limitations of liver transplantation. To relieve the organ shortage, current transplant strategies have implemented extended criteria, which include the use of liver from patients with signs of past or present hepatitis B virus(HBV) infection. While the use of liver grafts from donors with evidence of past HBV infection is quite limited, some data have been collected regarding the feasibility of transplanting a liver graft from a hepatitis B surface antigen(HBs Ag) positive donor. The aim of the present work was to review the literature regarding liver transplants from HBs Ag-positive donors. A total of 17 studies were identified by a search in Medline. To date, HBs Ag positive grafts have preferentially been allocated to HBs Ag positive recipients. The large majority of these patients continue to be HBs Ag positive despite the use of immunoglobulin, and infection prevention can only be guaranteed by using antiviral prophylaxis. Although serological persistence is evident, no significant HBV-related disease has been observed, except in patients coinfected with delta virus. Consistently less data are available for HBs Ag negative recipients, although they are mostly promising. HBs Agpositive grafts could be an additional organ source for liver transplantation, provided that the risk of reinfection/reactivation is properly prevented.

Impact of small-for-size liver grafts on medium-term and long-term graft survival in living donor liver transplantation: A meta-analysis

BACKGROUND Small-for-size grafts (SFSGs) in living donor liver transplantation (LDLT) could optimize donor postoperative outcomes and also expand the potential donor pool. Evidence on whether SFSGs would affect medium-term and long-term recipient graft survival is lacking. AIM To evaluate the impact of small-for-size liver grafts on medium-term and longterm graft survival in adult to adult LDLT. METHODS A systematic review and meta-analysis were performed by searching eligible studies published before January 24, 2019 on PubMed, EMBASE, and Web of Science databases. The primary outcomes were 3-year and 5-year graft survival. Incidence of small-for-size syndrome and short term mortality were also extracted. RESULTS This meta-analysis is reported according to the guidelines of the PRISMA 2009 Statement. Seven retrospective observational studies with a total of 1821 LDLT recipients were included in the meta-analysis. SFSG is associated with significantly poorer medium-term graft survival. The pooled odds ratio for 3-year graft survival was 1.58 [95% confidence interval 1.10-2.29, P = 0.014]. On the other hand, pooled results of the studies showed that SFSG had no significant discriminatory effect on 5-year graft survival with an odds ratio of 1.31 (95% confidence interval 0.87-1.97, P = 0.199). Furthermore, incidence of small-for-size syndrome detected in recipients of SFSG ranged from 0-11.4% in the included studies. CONCLUSION SFSG is associated with inferior medium-term but not long-term graft survival. Comparable long-term graft survival based on liver graft size shows that smaller grafts could be accepted for LDLT with appropriate flow modulatory measures. Close follow-up for graft function is warranted within 3 years after liver transplantation.

Living donor liver transplantation using dual grafts:Ultrasonographic evaluation

AIM: To evaluate the dual-graft living donor liver transplantation (LDLT) with ultrasonography, with special emphasis on the postoperative complications. METHODS: From January 2002 to August 2007, 110 adult-to-adult LDLTs were performed in West China Hos- pital of Sichuan University. Among them, dual-graft implantations were performed in six patients. Sonographic findings of the patients were retrospectively reviewed. RESULTS: All the six recipients survived the dual-graft adult-to-adult LDLT surgery. All had pleural effusion. Four patients had episodes of postoperative abdominal complications, including fluid collection between the grafts in three patients, intrahepatic biliary dilatation in two, hepatofugal portal flow of the left lobe in two, and atrophy of the left lobe in one. CONCLUSION: Although dual-graft LDLT takes more efforts and is technically complicated, it is safely feasible. Postoperative sonographic monitoring of the recipient is important.

Feasibility of using marginal liver grafts in living donor liver transplantation

Liver transplantation(LT) is one of the most effective treatments for end-stage liver disease caused by related risk factors when liver resection is contraindicated. Additionally,despite the decrease in the prevalence of hepatitis B virus(HBV) over the past two decades,the absolute number of HBs Ag-positive people has increased,leading to an increase in HBV-related liver cirrhosis and hepatocellular carcinoma. Consequently,a large demand exists for LT. While the wait time for patients on the donor list is,to some degree,shorter due to the development of living donor liver transplantation(LDLT),there is still a shortage of liver grafts. Furthermore,recipients often suffer from emergent conditions,such as liver dysfunction or even hepatic encephalopathy,which can lead to a limited choice in grafts. To expand the pool of available liver grafts,one option is the use of organs that were previously considered "unusable" by many,which are often labeled "marginal" organs. Many previous studies have reported on the possibilities of using marginal grafts in orthotopic LT; however,there is still a lack of discussion on this topic,especially regarding the feasibility of using marginal grafts in LDLT. Therefore,the present review aimed to summarize the feasibility of using marginal liver grafts for LDLT and discuss the possibility of expanding the application of these grafts.

Structural design and mechanical performance of composite vascular grafts

This study reviews the state of the art in structural design and the corresponding mechanical behaviours of composite vascular grafts. We critically analyse surface and matrix designs composed of layered, embedded, and hybrid structures along the radial and longitudinal directions;materials and manufacturing techniques, such as tissue engineering and the use of textiles or their combinations;and the corresponding mechanical behaviours of composite vascular grafts in terms of their physical–mechanical properties, especially their stress–strain relationships and elastic recovery. The role of computational studies is discussed with respect to optimizing the geometrics designs and the corresponding mechanical behaviours to satisfy specialized applications, such as those for the aorta and its subparts. Natural and synthetic endothelial materials yield improvements in the mechanical and biological compliance of composite graft surfaces with host arteries. Moreover,the diameter, wall thickness, stiffness, compliance, tensile strength, elasticity, and burst strength of the graft matrix are determined depending on the application and the patient. For composite vascular grafts, hybrid architectures are recommended featuring multiple layers, dimensions, and materials to achieve the desired optimal flexibility and function for complying with user-specific requirements. Rapidly emerging artificial intelligence and big data techniques for diagnostics and the threedimensional(3D) manufacturing of vascular grafts will likely yield highly compliant, subject-specific, long-lasting, and economical vascular grafts in the near-future.

TrkA regulates the regenerative capacity of bone marrow stromal stem cells in nerve grafts

We previously demonstrated that overexpression of tropomyosin receptor kinase A(TrkA)promotes the survival and Schwann celllike differentiation of bone marrow stromal stem cells in nerve grafts,thereby enhancing the regeneration and functional recovery of the peripheral nerve.In the present study,we investigated the molecular mechanisms underlying the neuroprotective effects of TrkA in bone marrow stromal stem cells seeded into nerve grafts.Bone marrow stromal stem cells from Sprague-Dawley rats were infected with recombinant lentivirus vector expressing rat TrkA,TrkA-shRNA or the respective control.The cells were then seeded into allogeneic rat acellular nerve allografts for bridging a 1-cm right sciatic nerve defect.Then,8 weeks after surgery,hematoxylin and eosin staining showed that compared with the control groups,the cells and fibers in the TrkA overexpressing group were more densely and uniformly arranged,whereas they were relatively sparse and arranged in a disordered manner in the TrkA-shRNA group.Western blot assay showed that compared with the control groups,the TrkA overexpressing group had higher expression of the myelin marker,myelin basic protein and the axonal marker neurofilament 200.The TrkA overexpressing group also had higher levels of various signaling molecules,including TrkA,pTrkA(Tyr490),extracellular signal-regulated kinases 1/2(Erkl/2),pErk1/2(Thr202/Tyr204),and the anti-apoptotic proteins Bcl-2 and Bcl-xL.In contrast,these proteins were downregulated,while the pro-apoptotic factors Bax and Bad were upregulated,in the TrkA-shRNA group.The levels of the TrkA effectors Akt and pAkt(Ser473)were not different among the groups.These results suggest that TrkA enhances the survival and regenerative capacity of bone marrow stromal stem cells through upregulation of the Erk/Bcl-2 pathway.All procedures were approved by the Animal Ethical and Welfare Committee of Shenzhen University,China in December 2014(approval No.AEWC-2014-001219).

Hypothermic machine perfusion with metformin-University of Wisconsin solution for ex vivo preservation of standard and marginal liver grafts in a rat model

AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex vivo by hypothermic machine perfusion(HMP).METHODS Eighteen young(4 mo old) and 18 aged(17 mo old)healthy male SD rats were selected and randomly divided into three groups: control group, UW solution perfusion group(UWP), and UW solution with metformin perfusion group(MUWP). Aspartate aminotransferase(AST), alanine aminotransferase(ALT), lactate dehydrogenase(LDH), interleukin-18(IL-18), and tumor necrosis factor-alpha(TNF-α) in the perfused liquid were tested. The expression levels of AMPK and endothelial nitric oxide synthase(e NOS) in liver sinusoidal endothelial cells were also examined.Additionally, microscopic evaluation of the harvested perfused liver tissue samples was done. RESULTS AST, ALT, LDH, IL-18 and TNF-α levels in the young and aged liver-perfused liquid were, respectively,significantly lower in the MUWP group than in the UWP group(P < 0.05), but no significant differences were found between the young and aged MUWP groups.Metformin increased the expression of AMPK and e NOS protein levels, and promoted the extracellular release of nitric oxide through activation of the AMPK-e NOS mediated pathway. Histological examination revealed that in the MUWP group, the extent of liver cells and tissue damage was significantly reduced compared with the UWP group.CONCLUSION The addition of metformin to the UW preservative solution for ex vivo HMP can reduce rat liver injury during cold ischemia, with significant protective effects on livers, especially of aged rats.

High-risk corneal allografts: A therapeutic challenge

Corneal transplantation is the most common surgical procedure amongst solid organ transplants with a high survival rate of 86% at 1-year post-grafting. This high success rate has been attributed to the immune privilege of the eye. However, mechanisms originally thought to promote immune privilege, such as the lack of antigen presenting cells and vessels in the cornea, are challenged by recent studies. Nevertheless, the immunological and physiological features of the cornea promoting a relatively weak alloimmune response is likely responsible for the high survival rate in "low-risk" settings. Furthermore, although corneal graft survival in "lowrisk" recipients is favourable, the prognosis in "high-risk" recipients for corneal graft is poor. In "high-risk" grafts, the process of indirect allorecognition is accelerated by the enhanced innate and adaptive immune responses due to pre-existing inflammation and neovascularization of the host bed. This leads to the irreversible rejection of the allograft and ultimately graft failure. Many therapeutic measures are being tested in pre-clinical and clinical studies to counter the immunological challenge of "high-risk" recipients. Despite the prevailing dogma, recent data suggest that tissue matching together with use of systemic immunosuppression may increase the likelihood of graft acceptance in "high-risk" recipients. However, immunosuppressive drugs are accompanied with intolerance/side effects and toxicity, and therefore, novel cell-based therapies are in development which target host immune cells and restore immune homeostasis without significant side effect of treatment. In addition, developments in regenerative medicinemay be able to solve both important short comings of allotransplantation:(1) graft rejection and ultimate graft failure; and(2) the lack of suitable donor corneas. The advances in technology and research indicate that wider therapeutic choices for patients may be available to address the worldwide problem of corneal blindness in both "low-risk" and "high-risk" hosts.

Preparation of acellular nerve grafts with triton X-100

BACKGROUND: The source of nerve allograft enriches. We may choose expediently nerve allograft to repair injured nerve and the structure of choice nerve homology or similar with the injured nerve, but the immunological rejection limits the clinical application of nerve allograft. The ideal substitute of autograft never is researching. OBJECTIVE: In this experiment, Triton X-100 was used to extract the Schwann cells and myelin sheaths of allograft nerve and obtain the inartificial and eliminated antigenicity nerve-transplanter (nerve grafts). DESIGN: Controlled experiment. SETTING: Department of Hand Surgery, the Third Affiliated Hospital of Hebei Medical University; Second Department of Orthopedics, Fourth Center Hospital of Tianjin. MATERIALS: Thirty health New Zealand big ear white rabbit, of either sex (gender), weighing 2000-3000 g, were provided by the Center of Experimental Animal of Hebei Medical University. TritonX-100 was offered by SIGMA Company. METHODS: The experiment was carried out at the Central Laboratory of the Third Affiliated Hospital of Hebei Medical University from December 2003 to December 2004. Sixty pieces of sciatic nerves, 10-mm-long nerve segment, which were taken from 30 rabbits, were incised. They were randomly divided into chemical extraction group (n =50) and control group (n =10). In the chemical extraction groups, the nerves were put into 3% Triton X-100 solution. They were treated with Triton X-100 for 12 hours, 24 hours, 48 hours, 96 hours and 1 week, respectively. They were examined in every period. The control groups did not treated with anything. ① Respectively two segments of nerve by 2 mm length were taken from each nerve in the every periods. ② The laminin immunohistochemical stained sections were performed with image acquisition and analyzed with multicolor pathological image analysis system. Measured the laminin antibody reaction part of each section and computed laminin average gray degrees of the unit area. All dates were analyzed by SPSS 10.0 software. MAIN OUTCOME MEASURES: ① General observation and histological observation in two groups; ② Compared with laminin average gray degrees of the unit area in each section. RESULTS: ① General observation: In the control groups, fresh nerve was polish, rigidity and elasticity. After the nerves were chemical extracted, the floccules was seen at two ends and around of the nerves. The nerves being extraction presented ivory and lackluster. Its diameter and length compared reduced, tenderness and tenacity with the fresh nerve. Observed by light microscope, Schwann cells, myelin sheaths and basement membrane distribute uniformly in control groups. After the nerves were extracted, Schwann cells and myelin sheaths disappeared. Basement membrane presented barrier array in longitudinal sections. Between the membranes was the basement membrane tube. Observed with scanning electron microscope, the basement membrane tubes composed by collagen fibers were remained and collagen fibers maintained their former position, form and structure. Further, the structure of membrane was seen in the tubes. It was Schwann cells basement membrane. ② In chemical extraction groups, laminin average gray degrees of the unit area were 140.1±3.41 (12 hours), 142.1±3.14 (24 hours), 142.1±3.14 (48 hours), 140.4±4.03 (96 hours), 141.7±2.62 (1 week). In the control groups, laminin average gray degree of the unit area was 142.7±7.24. There were not significant differences among the groups (P > 0.05). CONCLUSION: The method of chemical extraction by using of Triton X-100 may be an ideal measure for preparing tissue-engineered nerve-transplanter and reserved the live of laminin in the basement membrane.

Retrograde percutaneous coronary intervention via critically degenerated saphenous vein grafts for chronic total occlusion in native coronary arteries

We report the case of a 78-year-old woman with saphenous vein graft (SVG) disease and chronic total occlusion (CTO) in three native coronary arteries [left anterior descending artery (LAD), left circumflex artery, and the right coronary artery], who was successfully treated by percutaneous coronary intervention (PCI) using the retrograde approach via the critically degenerated SVGs. The patient, a 78-year-old woman, presented with sudden chest pain and dyspnea. She had previously undergone coronary artery bypass surgery using SVGs for the three vessels and percutaneous coronary intervention with sirolimuseluting stent placement in the posterolateral branch 13 and 3 years ago, respectively. Electrocardiography revealed ST-segment elevation in leads V1-4, whereas emergent coronary angiography revealed total occlusion in her native coronary arteries. Primary PCI was scheduled. A channel dilator was advanced very smoothly and safely into the distal site of the CTO lesion in the LAD, which showed complete occlusion in the proximal region, via an SVG that was temporally occluded four days earlier. A reverse controlled antegrade and retrograde tracking technique was used to successfully perform percutaneous recanalization. Subsequently, the other two native CTO lesions protected by critically degenerated SVGs were treated with retrograde intervention via the SVGs. The retrograde approach via critically degenerated SVGs is safe, reliable, and fast. If an SVG bypassing the native CTO lesion is critically degenerated, percutaneous coronary intervention should be performed via the SVG.

Structural bone allografts with intramedullary vascularized fibular autografts for the treatment of massive bone defects in extremities

Objective:To report the clinical outcome of repairing massive bone defects biologically in limbs by homeochronous using structural bone allografts with intramedullary vascularized fibular autografts. Methods: From January 2001 to December 2005, large bone defects in 19 patients (11 men and 8 women, aged 6 to 35 years) were repaired by structural bone allografts with intramedullary vascularized fibular autografts in the homeochronous period. The range of the length of bone defects was 11 to 25 cm (mean 17.6 cm), length of vascularized free fibular was 15 to 29 cm (mean 19.2 cm), length of massive bone allografts was 11 to 24 cm (mean 17.1 cm). Location of massive bone defects was in humerus(n=1), in femur(n=9) and in tibia(n=9), respectively. Results: After 9 to 69 months (mean 38.2 months) follow-up, wounds of donor and recipient sites were healed inⅠstage, monitoring-flaps were alive, eject reaction of massive bone allografts were slight, no complications in donor limbs. Fifteen patients had the evidence of radiographic union 3 to 6 months after surgery, 3 cases united 8 months later, and the remained one case of malignant synovioma in distal femur recurred and amputated the leg 2.5 months, postoperatively. Five patients had been removed internal fixation, complete bone unions were found one year postoperatively. None of massive bone allografts were absorbed or collapsed at last follow-up. Conclusion: The homeochronous usage of structural bone allograft with an intramedullary vascularized fibular autograft can biologically obtain a structure with the immediate mechanical strength of the allograft, a potential result of revascularization through the vascularized fibula, and accelerate bone union not only between fibular autograft and the host but also between massive bone allograft and the host.

Gene expression profiles of human adipose-derived mesenchymal stem cells dynamically seeded on clinically available processed nerve allografts and collagen nerve guides

It was hypothesized that mesenchymal stem cells(MSCs) could provide necessary trophic factors when seeded onto the surfaces of commonly used nerve graft substitutes. We aimed to determine the gene expression of MSCs when influenced by Avance■ Nerve Grafts or Neura Gen■ Nerve Guides. Human adipose-derived MSCs were cultured and dynamically seeded onto 30 Avance■ Nerve Grafts and 30 Neura Gen■ Nerve Guides for 12 hours. At six time points after seeding, quantitative polymerase chain reaction analyses were performed for five samples per group. Neurotrophic [nerve growth factor(NGF), glial cell line-derived neurotrophic factor(GDNF), pleiotrophin(PTN), growth associated protein 43(GAP43) and brain-derived neurotrophic factor(BDNF)], myelination [peripheral myelin protein 22(PMP22) and myelin protein zero(MPZ)], angiogenic [platelet endothelial cell adhesion molecule 1(PECAM1/CD31) and vascular endothelial cell growth factor alpha(VEGFA)], extracellular matrix(ECM) [collagen type alpha I(COL1A1), collagen type alpha III(COL3A1), Fibulin 1(FBLN1) and laminin subunit beta 2(LAMB2)] and cell surface marker cluster of differentiation 96(CD96) gene expression was quantified. Unseeded Avance■ Nerve Grafts and Neura Gen■ Nerve Guides were used to evaluate the baseline gene expression, and unseeded MSCs provided the baseline gene expression of MSCs. The interaction of MSCs with the Avance■ Nerve Grafts led to a short-term upregulation of neurotrophic(NGF, GDNF and BDNF), myelination(PMP22 and MPZ) and angiogenic genes(CD31 and VEGFA) and a long-term upregulation of BDNF, VEGFA and COL1A1. The interaction between MSCs and the Neura Gen■ Nerve Guide led to short term upregulation of neurotrophic(NGF, GDNF and BDNF) myelination(PMP22 and MPZ), angiogenic(CD31 and VEGFA), ECM(COL1A1) and cell surface(CD96) genes and long-term upregulation of neurotrophic(GDNF and BDNF), angiogenic(CD31 and VEGFA), ECM genes(COL1A1, COL3A1, and FBLN1) and cell surface(CD96) genes. Analysis demonstrated MSCs seeded onto Neura Gen■ Nerve Guides expressed significantly higher levels of neurotrophic(PTN), angiogenic(VEGFA) and ECM(COL3A1, FBLN1) genes in the long term period compared to MSCs seeded onto Avance■ Nerve Grafts. Overall, the interaction between human MSCs and both nerve graft substitutes resulted in a significant upregulation of the expression of numerous genes important for nerve regeneration over time. The in vitro interaction of MSCs with the Neura Gen■ Nerve Guide was more pronounced, particularly in the long term period(> 14 days after seeding). These results suggest that MSC-seeding has potential to be applied in a clinical setting, which needs to be confirmed in future in vitro and in vivo research.

Repair of Rat Segmental Defects with Mineralized Collagen Grafts Combined with or without Mesenchymal Stem Cells and BMP-2

The aim of the present study was to investigate and compare the bone formation capacity with three different grafts. Four millimeter segmental defects were created in adult rat tibias and were either left empty （control defects） or implanted with （1） nano-hydroxyapatite/collagen/PIA （nHAC/PIA） composite, （2） nHAC/ PIA composite added with bone marrow mesenchymal tem cells （ BMSCs ）, （ 3 ） nHAC/ PIA composite added with bone morphogenetic protein 2 （ BMP- 2）. Radiographs of the defects were taken weekly post-surgery. After 1 or 2 months, the rats were eathaaized. Histologic analyses were performed on the harvested tissue. nHAC/ PIA composite could enhance the repair of rat tibia segmental defects. Addition of BMSCs or BMP- 2 to nHAC/ PIA led to an increase in osteogenesis, nHAC/ PIA composite could be an Meal alternative bone-grafi material and it could also be used as an Meal carrier of BMSCs or BMP- 2.