Inhibition of the JAK/STAT Pathway With Baricitinib Reduces the Multiple Organ Dysfunction Caused by Hemorrhagic Shock in Rats.

OBJECTIVE: The aim of this study was to investigate (a) the effects of the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway inhibitor (baricitinib) on the multiple organ dysfunction syndrome (MODS) in a rat model of hemorrhagic shock (HS) and (b) whether treatment with baricitinib attenuates the activation of JAK/STAT, NF-κB, and NLRP3 caused by HS. BACKGROUND: Posttraumatic MODS, which is in part due to excessive systemic inflammation, is associated with high morbidity and mortality. The JAK/STAT pathway is a regulator of numerous growth factor and cytokine receptors and, hence, is considered a potential master regulator of many inflammatory signaling processes. However, its role in trauma-hemorrhage is unknown. METHODS: An acute HS rat model was performed to determine the effect of baricitinib on MODS. The activation of JAK/STAT, NF-κB, and NLRP3 pathways were analyzed by western blotting in the kidney and liver. RESULTS: We demonstrate here for the first time that treatment with baricitinib (during resuscitation following severe hemorrhage) attenuates the organ injury and dysfunction and the activation of JAK/STAT, NF-κB, and NLRP3 pathways caused by HS in the rat. CONCLUSIONS: Our results point to a role of the JAK/STAT pathway in the pathophysiology of the organ injury and dysfunction caused by trauma/hemorrhage and indicate that JAK inhibitors, such as baricitinib, may be repurposed for the treatment of the MODS after trauma and/or hemorrhage.

Inhibition of Bruton's Tyrosine Kinase Activity Attenuates Hemorrhagic Shock-Induced Multiple Organ Dysfunction in Rats.

OBJECTIVE: The aim of this study was to investigate (a) the potential of the Bruton's tyrosine kinase (BTK) inhibitors acalabrutinib and fenebrutinib to reduce multiple organ dysfunction syndrome (MODS) in acute (short-term and long-term follow-up) hemorrhagic shock (HS) rat models and (b) whether treatment with either acalabrutinib or fenebrutinib attenuates BTK, NF-κB and NLRP3 activation in HS. BACKGROUND: The MODS caused by an excessive systemic inflammatory response following trauma is associated with a high morbidity and mortality. The protein BTK is known to play a role in the activation of the NLRP3 inflammasome, which is a key component of the innate inflammatory response. However, its role in trauma-hemorrhage is unknown. METHODS: Acute HS rat models were performed to determine the influence of acalabrutinib or fenebrutinib on MODS. The activation of BTK, NF-κB and NLRP3 pathways were analyzed by western blot in the kidney. RESULTS: We demonstrated that (a) HS caused organ injury and/or dysfunction and hypotension (post-resuscitation) in rats, while (b) treatment of HS-rats with either acalabrutinib or fenebrutinib attenuated the organ injury and dysfunction in acute HS models and (c) reduced the activation of BTK, NF- kB and NLRP3 pathways in the kidney. CONCLUSION: Our results point to a role of BTK in the pathophysiology of organ injury and dysfunction caused by trauma/hemorrhage and indicate that BTK inhibitors may be repurposed as a potential therapeutic approach for MODS after trauma and/or hemorrhage.

Optimizing cell seeding and retention in a three-dimensional bioengineered cardiac ventricle: The two-stage cellularization model.

Current cell seeding techniques focus on passively directing cells to a scaffold surface with the addition of dynamic culture to encourage cell permeation. In 3D tissue engineered constructs, cell retention efficiency is dependent on the cell delivery method, and biomaterial properties. Passive cell delivery relies on cell migration to the scaffold surface; biomaterial surface properties and porosity determine cell infiltration capacity. As a result, cell retention efficiencies remain low. The development of an effective two-stage cell seeding technique, coupled with perfusion culture, provides the potential to improve cellularization efficiency, and retention. This study, uses a chitosan bioengineered open ventricle (BEOV) scaffold to produce a two-stage perfusion cultured ventricle (TPCV). TPCV were fabricated by direct injection of 10 million primary rat neonatal cardiac cells, followed by wrapping of the outer scaffold surface with a 3D fibrin gel artificial heart muscle patch; TPCV were perfusion cultured for 3 days. The average biopotential output was 1.731 mV. TPCV cell retention following culture was approximately 5%. Cardiac cells were deposited on the scaffold surface and formed intercellular connections. Histological assessment displayed localized cell clusters, with some dissemination, and validated the observed presence of intercellular and gap-junction interactions. The study demonstrates initial effectiveness of our two-stage cell delivery concept, based on function and biological metrics. Biotechnol. Bioeng. 2016;113: 2275-2285. © 2016 Wiley Periodicals, Inc.

Inhibition of Macrophage Migration Inhibitory Factor Activity Attenuates Haemorrhagic Shock-Induced Multiple Organ Dysfunction in Rats.

Objective: The aim of this study was to investigate (a) macrophage migration inhibitory factor (MIF) levels in polytrauma patients and rats after haemorrhagic shock (HS), (b) the potential of the MIF inhibitor ISO-1 to reduce multiple organ dysfunction syndrome (MODS) in acute (short-term and long-term follow-up) HS rat models and (c) whether treatment with ISO-1 attenuates NF-κB and NLRP3 activation in HS. Background: The MODS caused by an excessive systemic inflammatory response following trauma is associated with a high morbidity and mortality. MIF is a pleiotropic cytokine which can modulate the inflammatory response, however, its role in trauma is unknown. Methods: The MIF levels in plasma of polytrauma patients and serum of rats with HS were measured by ELISA. Acute HS rat models were performed to determine the influence of ISO-1 on MODS. The activation of NF-κB and NLRP3 pathways were analysed by western blot in the kidney and liver. Results: We demonstrated that (a) MIF levels are increased in polytrauma patients on arrival to the emergency room and in rats after HS, (b) HS caused organ injury and/or dysfunction and hypotension (post-resuscitation) in rats, while (c) treatment of HS-rats with ISO-1 attenuated the organ injury and dysfunction in acute HS models and (d) reduced the activation of NF-κB and NLRP3 pathways in the kidney and liver. Conclusion: Our results point to a role of MIF in the pathophysiology of trauma-induced organ injury and dysfunction and indicate that MIF inhibitors may be used as a potential therapeutic approach for MODS after trauma and/or haemorrhage.

Potential Role of Diabetes Mellitus-Associated T Cell Senescence in Epithelial Ovarian Cancer Omental Metastasis.

Epithelial ovarian cancer (EOC) is one of the most common causes of cancer-related deaths among women and is associated with age and age-related diseases. With increasing evidence of risks associated with metabolic inflammatory conditions, such as obesity and type 2 diabetes mellitus (T2DM), it is important to understand the complex pathophysiological mechanisms underlying cancer progression and metastasis. Age-related conditions can lead to both genotypic and phenotypic immune function alterations, such as induction of senescence, which can contribute to disease progression. Immune senescence is a common phenomenon in the ageing population, which is now known to play a role in multiple diseases, often detrimentally. EOC progression and metastasis, with the highest rates in the 75-79 age group in women, have been shown to be influenced by immune cells within the "milky spots" or immune clusters of the omentum. As T2DM has been reported to cause T cell senescence in both prediabetic and diabetic patients, there is a possibility that poor prognosis in EOC patients with T2DM is partly due to the accumulation of senescent T cells in the omentum. In this review, we explore this hypothesis with recent findings, potential therapeutic approaches, and future directions.

The Bioengineered Cardiac Left Ventricle.

Left ventricle and aortic valve underdevelopment are presentations in the congenital cardiac condition hypoplastic left heart syndrome (HLHS); current clinical treatments involve right ventricle refunctionalization. Cardiac organoid models provide simplified open chambers engineered into a flow loop, to ameliorate ventricle-type function. Complete bioengineered ventricle development presents a significant advancement in cardiac organoids. This study provides the foundation for bioengineered complete ventricle (BECV) fabrication. Bioengineered trileaflet valve (BETV) molds and chitosan scaffolds were developed to emulate human neonate aortic valve geometry. Bioengineered complete ventricle were fabricated by fitting BETV into a bioengineered open ventricle (BEOV); the chamber was cellularized using a two-stage cellularization strategy, and BETV were passively seeded with rat neonatal cardiac fibroblasts and perfusion cultured for 3 days. Average pressure generated ranged from 0.06 to 0.12 mm Hg; average biopotential output was 1.02 mV. Histologic assessment displayed syncytial-type cardiomyocyte aggregates at the BECV chamber surface; BETV displayed randomly oriented, diffusely distributed cardiac fibroblasts. The fabrication of this novel BECV may aid in developing a functional engineered left ventricle for clinical application in HLHS.

The design and fabrication of a three-dimensional bioengineered open ventricle.

Current treatments in hypoplastic left heart syndrome (HLHS) include multiple surgeries to refunctionalize the right ventricle and/or transplant. The development of a tissue-engineered left ventricle (LV) would provide a therapeutic option to overcome the inefficiencies and limitations associated with current treatment options. This study provides a foundation for the development and fabrication of the bioengineered open ventricle (BEOV) model. BEOV molds were developed to emulate the human LV geometry; molds were used to produce chitosan scaffolds. BEOV were fabricated by culturing 30 million rat neonatal cardiac cells on the chitosan scaffold. The model demonstrated 57% cell retention following 4days culture. The average biopotential output for the model was 1615 µV. Histological assessment displayed the presence of localized cell clusters, with intercellular and cell-scaffold interactions. The BEOV provides a novel foundation for the development of a 3D bioengineered LV for application in HLHS. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2206-2217, 2017.

Pulsatile flow conditioning of three-dimensional bioengineered cardiac ventricle.

Current physical stimuli mechanical stretch bioreactor studies focus on conditioning planar and/or tubular engineered cardiac constructs. The current 3D bioreactor models in cardiac tissue engineering use differential pressure loading for structural support as opposed to conditioning. The development of the pulsatile flow conditioned ventricle (PFCV) provides a 3D mechanical stretch conditioning method to generate pump function in the engineered cardiac left ventricle. The study utilizes a chitosan bioengineered open ventricle scaffold, to produce the in vitro PFCV model. PFCV were fabricated by wrapping the outer scaffold surface with a 3D fibrin gel artificial heart muscle patch, followed by pulsatile flow conditioning for 20 h. The average contractile frequency was 57 bpm. The average pressure generated, under maintained flow, post-conditioning, was 3.1633 mmHg. The average biopotential output was 0.4881 mV. Histologically, the PFCV displayed a more disseminated presence of intercellular interactions and sarcomeric organization. The results of this study clearly demonstrate the effectiveness of pulsatile flow conditioning to improve the function of our engineered left ventricle.

Improving quality by encouraging providers to use pediatric combination vaccines.

In the last 4 decades, the number of diseases that vaccines can prevent has quadrupled, and, correspondingly, so has the number of immunizations that is to be administered before a child's second birthday. Based on current recommendations, a child may receive as many as six vaccine injections in a single visit. Vaccine technology has advanced significantly since the introduction of the first combination vaccine, diphtheria-tetanus (DT), in the 1950s. In the United States today, as many as five antigens can be administered in a single injection, and additional combination vaccines are in the pipeline. Increasing the number of antigens delivered with a single injection minimizes physical discomfort for the child, reduces associated stress for the parent, saves time for the provider, and is likely to improve vaccine coverage and timeliness of administration rates. While national immunization guidelines from the Centers for Disease Control and Prevention call for use of combination vaccines where available, provider and parent perceptions can act as barriers to their optimal use. Managed care organizations (MCOs) have the opportunity to improve quality of care and immunization rates by educating providers on the use of combination vaccines in accordance with the national guidelines. This article examines the evidence for pediatric combination vaccines, discusses barriers to their use among parents and providers, presents quality and cost implications of a managed care policy to broaden their use, and suggests ways in which MCOs can more actively promote appropriate use of combination vaccines by providers.

Establishing the Framework for Fabrication of a Bioartificial Heart.

There is a chronic shortage of donor hearts. The ability to fabricate complete bioartificial hearts (BAHs) may be an alternative solution. The current study describes a method to support the fabrication and culture of BAHs. Rat hearts were isolated and subjected to a detergent based decellularization protocol to remove all cellular components, leaving behind an intact extracellular matrix. Primary cardiac cells were isolated from neonatal rat hearts, and direct cell transplantation was used to populate the acellular scaffolds. Bioartificial hearts were maintained in a custom fabrication gravity fed perfusion culture system to support media delivery. The functional performance of BAHs was assessed based on left ventricle pressure and on electrocardiogram. Furthermore, BAHs were sectioned and stained for the whole heart cardiac tissue distribution and for cardiac molecules, such as α-actinin, cardiac troponin I, collagen type I, connexin 43, von Willebrand factor, and ki67. Bioartificial hearts replicated a partial subset of properties of natural rat hearts. The current study provided a method for fabrication of a BAH and revealed challenges toward BAH fabrication with functional performance metrics of natural mammalian hearts.

Engineering 3D bio-artificial heart muscle: the acellular ventricular extracellular matrix model.

Current therapies in left ventricular systolic dysfunction and end-stage heart failure include mechanical assist devices or transplant. The development of a tissue-engineered integrative platform would present a therapeutic option that overcomes the limitations associated with current treatment modalities. This study provides a foundation for the fabrication and preliminary viability of the acellular ventricular extracellular matrix (AVEM) model. Acellular ventricular extracellular matrix was fabricated by culturing 4 million rat neonatal cardiac cells around an excised acellular ventricular segment. Acellular ventricular extracellular matrix generated a maximum spontaneous contractile force of 388.3 µN and demonstrated a Frank-Starling relationship at varying pretensions. Histologic assessment displayed cell cohesion and adhesion within the AVEM as a result of passive cell seeding.

Establishing the Framework for Tissue Engineered Heart Pumps.

Development of a natural alternative to cardiac assist devices (CADs) will pave the way to a heart failure therapy which overcomes the disadvantages of current mechanical devices. This work provides the framework for fabrication of a tissue engineered heart pump (TEHP). Artificial heart muscle (AHM) was first fabricated by culturing 4 million rat neonatal cardiac cells on the surface of a fibrin gel. To form a TEHP, AHM was wrapped around an acellular goat carotid artery (GCA) and a chitosan hollow cylinder (CHC) scaffold with either the cardiac cells directly contacting the construct periphery or separated by the fibrin gel. Histology revealed the presence of cardiac cell layer cohesion and adhesion to the fibrin gel scaffold, acellular GCA, and synthesized CHC. Expression of myocytes markers, connexin43 and α-actinin, was also noted. Biopotential measurements revealed the presence of ~2.5 Hz rhythmic propagation of action potential throughout the TEHP. Degradation of the fibrin gel scaffold of the AHM via endogenous proteases may be used as a means of delivering the cardiac cells to cylindrical scaffolds. Further development of the TEHP model by use of multi-stimulus bioreactors may lead to the application of bioengineered CADs.