From In Vitro to In Situ Tissue Engineering

In vitro tissue engineering enables the fabrication of functional tissues for tissue replacement. In addition, it allows us to build useful physiological and pathological models for mechanistic studies. However, the translation of in vitro tissue engineering into clinical therapies presents a number of technical and regulatory challenges. It is possible to circumvent the complexity of developing functional tissues in vitro by taking an in situ tissue engineering approach that uses the body as a native bioreactor to regenerate tissues. This approach harnesses the innate regenerative potential of the body and directs the appropriate cells to the site of injury. This review surveys the biomaterial-, cell-, and chemical factor-based strategies to engineer tissue in vitro and in situ.     

CTLA4Ig抑制T细胞增殖的体外研究

目的 观察CTLA4Ig在人外周血T细胞增殖反应中的作用。方法 植物血凝素（PHA）、脂多糖（LPS）、金葡菌肠毒素B（SEB）刺激T细胞增殖,在加入或未加入CTLA4Ig情况下,观察细胞形态并检测^3H-TdR掺入量。结果ＣＴＬＡ４Ｉg对ＰＨＡ、ＬＰＳ的抑制作用呈剂量依赖关系。而CTLA4Ig浓度只高于5ug/mL时,对SEB才有抑制作用。且可完全抑帛。结论CTLA4Ig对PHA、LPS、SEB刺激下的T细胞增殖反应有明显的抑制作用。但反应模式不一致。     

In Memoriam

In the emergency department, continuity of care is constant. ED high utilizers have social needs unmet and limited places to turn for help.     

Bordetella avium Virulence Measured In Vivo and In Vitro

Bordetella avium causes an upper-respiratory-tract disease called bordetellosis in birds. Bordetellosis shares many of the clinical and histopathological features of disease caused in mammals by Bordetella pertussis and Bordetella bronchiseptica. In this study we determined several parameters of infection in the domestic turkey, Meleagris galapavo, and compared these in vivo findings with an in vitro measure of adherence using turkey tracheal rings. In the in vivo experiments, we determined the effects of age, group size, infection duration, and interindividual spread of B. avium. Also, the effect of host genetic background on susceptibility was tested in the five major commercial turkey lines by infecting each with the parental B. avium strain and three B. avium insertion mutants. The mutant strains lacked either motility, the ability to agglutinate guinea pig erythrocytes, or the ability to produce dermonecrotic toxin. The susceptibilities of 1-day-old and 1-week-old turkeys to B. avium were the same, and challenge group size (5, 8, or 10 birds) had no effect upon the 50% infectious dose. Two weeks between inoculation and tracheal culture was optimal, since an avirulent mutant (unable to produce dermonecrotic toxin) persisted for a shorter time. Communicability of the B. avium parental strain between confined birds was modest, but a nonmotile mutant was less able to spread between birds. There were no host-associated differences in susceptibility to the parental strain and the three B. avium mutant strains just mentioned: in all turkey lines tested, the dermonecrotic toxin- and hemagglutination-negative mutants were avirulent whereas the nonmotile mutants showed no loss of virulence. Interestingly, the ability of a strain to cause disease in vivo correlated completely with its ability to adhere to ciliated tracheal cells in vitro.     

Anti-septic Effects of Fisetin In Vitro and In Vivo

Sepsis is a state of disrupted inflammatory homeostasis that is initiated by infection. High mobility group box 1 (HMGB1) protein acting as a late mediator of severe vascular inflammatory conditions, such as sepsis and endothelial cell protein C receptor (EPCR), is involved in vascular inflammation. Fisetin, an active compound from the family Fabaceae, was reported to have antiviral, neuroprotective, and anti-inflammatory activities. Here, we determined the anti-septic effects of fisetin on HMGB1-mediated inflammatory responses and on the shedding of EPCR in vitro and in vivo, for the first time. First, we monitored the effects of post-treatment fisetin on lipopolysaccharide (LPS) and cecal ligation and puncture (CLP)-mediated release of HMGB1 and HMGB1-mediated regulation of pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs) and septic mice. Post-treatment fisetin was found to suppress LPS-mediated release of HMGB1 and HMGB1-mediated cytoskeletal rearrangements. Fisetin also inhibited HMGB1-mediated hyperpermeability and leukocyte migration in septic mice. Fisetin induced potent inhibition of phorbol-12-myristate 13-acetate (PMA) and CLP-induced EPCR. Fisetin also inhibited the expression and activity of tumor necrosis factor-α converting enzyme, induced by PMA in endothelial cells. In addition, fisetin inhibited the production of tumor necrosis factor-α and the activation of AKT, nuclear factor-κB, and extracellular regulated kinases 1/2 by HMGB1 in HUVECs. Fisetin also down-regulated CLP-induced release of HMGB1, production of interleukin 1β, and reduced septic mortality. Collectively, these results suggest that fisetin may be a candidate therapeutic agent for the treatment of vascular inflammatory diseases via inhibition of the HMGB1 signaling pathway.     

In Vivo Responses of Alloreactive Lymphocytes Stimulated In Vitro

Populations of mouse lymphocytes enriched in specific alloreactive cells by priming in a mixed lymphocyte response (MLR) include cells which, when injected into congenic nude mice, enable them to make alloantibody after immunization. Helper cells for the priming H-2 alloantigens (H-2b or H-2k) were enriched relative to helper cells for the other H-2 type. Furthermore, the alloantibody responses of nude mice reconstituted with lymphocytes primed twice in vitro were virtually monospecific for the priming alloantigens. These studies suggest that lymphocytes that proliferate in MLR include lymphocytes capable of giving specific help for H-2 antigens in vivo. Nude mice reconstituted with MLR-primed lymphocytes made less antibody to bacteriophage T4 and phix than mice reconstituted with unprimed cells, and fewer mice responded. Priming of cells a second time in MLR further depleted the population of phage helper cells. Similar results were sometimes, but not always, obtained when testing reconstituted nude mice for their ability to make anti-sheep erythrocyte (SRBC) responses. These results suggest that lymphocytes primed against H-2b or H-2k alloantigens do not have specificity for antigens of T4 or phix. These alloreactive cells may also lack specificity for SRBC. However, the results do not allow a definitive conclusion to be drawn.     

In Vitro and In Vivo Characterization of MEMS Microneedles

Transdermal drug delivery TDD systems have many advantages but are conventionally limited by the low permeability of skin. The idea of using microneedles to painlessly penetrate the topmost impermeable stratum corneum has previously been put forward. In this paper, the fabrication of solid and hollow silicon microneedles with straight side-walls and with the following dimensions: 20–100 m in diameter and 100–150 m in length is described. In vitro tests demonstrate that with prior solid microneedle application, transdermal drug transport is significantly increased by 10–20 times, with the degree of enhancement being related to needle diameter. In vivo tests in diabetic animals, however, were unable to demonstrate any delivery of insulin through the hollow microneedles. It is proposed that two factors, microneedle length and tip sharpness, have to be improved for systemic drug delivery to be seen in vivo.