Ultra-short laser processing of 3D bioceramic, porous scaffolds designed by freeze foaming method for orthopedic applications.

Bone substitutes are widely employed for applications in orthopedic surgery for the replacement of injured bone. Among the diverse methods that are used to design 3D bioceramic matrices, Freeze Foaming has gained attention, since it provides the ability to tune the shape of the created structures. One of the major problems related to these constructs is the lack of porosity at the outwards sides (holder) of the scaffold, thus reducing the cellular affinity and creating a rejection of the implant. In this research, we aimed to develop a bone scaffold with enhanced surface properties and improved cellular affinity. The main aim was to alter the biocompatibility characteristics of the 3D bioceramic constructs. We have produced three-dimensional, complex-shaped hollow shell structures, manufactured by Additive Manufacturing processes and as a second step, filled with a ceramic suspension by the Freeze-Foaming process. 3D constructs from HAP-derived TCP and TCP/ZrO2 were synthesized by freeze-foaming method and subsequently irradiated with a fs-laser (λ = 800 nm) spanning a range of parameters for achievement of optimal surface processing conditions. The designed scaffolds demonstrated enhanced topographical properties with improved porosity examined by SEM, EDX, and 3D profilometry after laser treatment. Wettability and computer tomography (CT) evaluation was also performed. The results from X-ray diffraction (XRD) and micro-Raman analysis did not show photochemical and surface or volume defects and changes after laser processing of the ceramic samples. Preliminary results from MG-63 osteoblast-like cell tests showed good cell affinity on the processed surfaces and no cytotoxic effect on the cells.

Toward Smart Biomimetic Apatite-Based Bone Scaffolds with Spatially Controlled Ion Substitutions.

Biomimetic apatites exhibit a high reactivity allowing ion substitutions to modulate their in vivo response. We developed a novel approach combining several bioactive ions in a spatially controlled way in view of subsequent releases to address the sequence of events occurring after implantation, including potential microorganisms' colonization. Innovative micron-sized core-shell particles were designed with an external shell enriched with an antibacterial ion and an internal core substituted with a pro-angiogenic or osteogenic ion. After developing the proof of concept, two ions were particularly considered, Ag+ in the outer shell and Cu2+ in the inner core. In vitro evaluations confirmed the cytocompatibility through Ag-/Cu-substituting and the antibacterial properties provided by Ag+. Then, these multifunctional "smart" particles were embedded in a polymeric matrix by freeze-casting to prepare 3D porous scaffolds for bone engineering. This approach envisions the development of a new generation of scaffolds with tailored sequential properties for optimal bone regeneration.

Investigating Potential Effects of Ultra-Short Laser-Textured Porous Poly-ε-Caprolactone Scaffolds on Bacterial Adhesion and Bone Cell Metabolism.

Developing antimicrobial surfaces that combat implant-associated infections while promoting host cell response is a key strategy for improving current therapies for orthopaedic injuries. In this paper, we present the application of ultra-short laser irradiation for patterning the surface of a 3D biodegradable synthetic polymer in order to affect the adhesion and proliferation of bone cells and reject bacterial cells. The surfaces of 3D-printed polycaprolactone (PCL) scaffolds were processed with a femtosecond laser (λ = 800 nm; τ = 130 fs) for the production of patterns resembling microchannels or microprotrusions. MG63 osteoblastic cells, as well as S. aureus and E. coli, were cultured on fs-laser-treated samples. Their attachment, proliferation, and metabolic activity were monitored via colorimetric assays and scanning electron microscopy. The microchannels improved the wettability, stimulating the attachment, spreading, and proliferation of osteoblastic cells. The same topography induced cell-pattern orientation and promoted the expression of alkaline phosphatase in cells growing in an osteogenic medium. The microchannels exerted an inhibitory effect on S. aureus as after 48 h cells appeared shrunk and disrupted. In comparison, E. coli formed an abundant biofilm over both the laser-treated and control samples; however, the film was dense and adhesive on the control PCL but unattached over the microchannels.

Bone Cell Exosomes and Emerging Strategies in Bone Engineering.

Bone tissue remodeling is a highly regulated process balancing bone formation and resorption through complex cellular crosstalk between resident bone and microenvironment cells. This cellular communication is mediated by direct cell and cell-matrix contact, autocrine, endocrine, and paracrine receptor mediated mechanisms such as local soluble signaling molecules and extracellular vesicles including nanometer sized exosomes. An impairment in this balanced process leads to development of pathological conditions. Bone tissue engineering is an emerging interdisciplinary field with potential to address bone defects and disorders by synthesizing three-dimensional bone substitutes embedded with cells for clinical implantation. However, current cell-based therapeutic approaches have faced hurdles due to safety and ethical concerns, challenging their clinical translation. Recent studies on exosome-regulated bone homeostasis and regeneration have gained interest as prospective cell free therapy in conjugation with tissue engineered bone grafts. However, exosome research is still in its nascent stages of bone tissue engineering. In this review, we specifically describe the role of exosomes secreted by cells within bone microenvironment such as osteoblasts, osteocytes, osteoclasts, mesenchymal stem cell cells, immune cells, endothelial cells, and even tumor cells during bone homeostasis and crosstalk. We also review exosome-based osteoinductive functionalization strategies for various bone-based biomaterials such as ceramics, polymers, and metals in bone tissue engineering. We further highlight biomaterials as carrier agents for exosome delivery to bone defect sites and, finally, the influence of various biomaterials in modulation of cell exosome secretome.

Preoperative risk factors for massive transfusion, prolonged ventilation requirements, and mortality in patients undergoing liver transplantation.

BACKGROUND: Despite improvements in techniques and management of liver transplant patients, numerous perioperative complications that contribute to perioperative mortality remain. Models to predict intraoperative massive blood transfusion, prolonged mechanical ventilation, or in-hospital mortality in liver transplant recipients have not been identified. In this study we aim to identify preoperative factors associated with the above mentioned complications. METHODS: A retrospective observational analysis was conducted on data collected from 124 orthotopic liver transplants performed at a single institution between 2014 and 2017. A multivariable logistic regression using backwards elimination was performed for three defined outcomes (massive transfusion ≥ 10 units packed red blood cells (PRBC), prolonged mechanical ventilation > 24 h, and in-hospital mortality) to identify associations with preoperative characteristics. RESULTS: Statistically significant (P < 0.05) associations with massive transfusion ≥ 10 units PRBC were hepatocellular carcinoma and preoperative transfusion of PRBC. Significant associations with prolonged mechanical ventilation > 24 h were hepatitis C, alcoholic hepatitis, elevated preoperative ALT, and hepatorenal syndrome. Male gender was protective for requiring prolonged mechanical ventilation. End-stage renal disease and hepatitis B were significantly associated with increased in-hospital mortality. CONCLUSIONS: This study identified risk factors associated with common perioperative complications of liver transplantation. These factors may assist practitioners in risk stratification and may form the basis for further investigations of potential interventions to mitigate these risks.

Synergistic role of 5-azacytidine and ascorbic acid in directing cardiosphere derived cells to cardiomyocytes in vitro by downregulating Wnt signaling pathway via phosphorylation of ß-catenin.

BACKGROUND: Cardiosphere derived cells (CDCs) represent a valuable source in stem cell based therapy for cardiovascular diseases, yet poor differentiation rate hinders the transplantation efficiency. The aim of this study is to check the ability of 5-Azacytidine (Aza) alone and in combination with ascorbic acid (Aza+AA) in delineating CDCs to cardiomyogenesis and the underlying Wnt signaling mechanism in induced differentiation. METHODS: CDCs were treated with Aza and Aza+AA for a period of 14 days to examine the expression of cardiac specific markers and Wnt downstream regulators by immunofluorescence, real time PCR and western blot. RESULTS: Results revealed that Aza+AA induced efficient commitment of CDCs to cardiomyogenic lineage. Immunofluorescence analysis showed significant augment for Nkx 2.5, GATA 4 and α-Sarcomeric actinin markers in Aza+AA group than control group (p = 0.0118, p = 0.009 and p = 0.0091, respectively). Relative upregulation of cardiac markers, Nkx 2.5 (p = 0.0156), GATA 4 (p = 0.0087) and down regulation of Wnt markers, ß-catenin (p = 0.0107) and Cyclin D1 (p = 0. 0116) in Aza+AA group was revealed by RNA expression analysis. Moreover, the Aza+AA induced prominent expression of GATA 4, α-Sarcomeric actinin and phospho ß-catenin while non phospho ß-catenin and Cyclin D1 expression was significantly suppressed as displayed in protein expression analysis. Generation of spontaneous beating in Aza+AA treated CDCs further reinforced that Aza+AA accelerates the cardiomyogenic potential of CDCs. CONCLUSION: Combined treatment of Aza along with AA implicit in inducing cardiomyogenic potential of CDCs and is associated with down regulating Wnt signaling pathway. Altogether, CDCs represent a valuable tool for the treatment of cardiovascular disorders.

Creating a Standardized Operating Room Management Curriculum for Anesthesia Trainees.

As the healthcare environment evolves, physicians are taking on new roles and responsibilities. In addition to clinical care, physicians must now be knowledgeable about administrative concepts and understand healthcare finances. However, these nonclinical subjects usually are not taught during residency training. Introducing these topics into all residencies in the form of standardized lectures, reading materials, and management rotations is a way to bridge the knowledge gap and better prepare future physicians as they enter the workforce. Currently, very few programs have a curriculum that addresses the required elements. Implementing a specific operating room management curriculum will, ideally, allow residents to gain practica and theoretical experience in this area and develop a better understanding of the scope of administrative medicine.

Taking disclosure seriously: disclosing financial conflicts of interest at the American College of Surgeons.

BACKGROUND: The accurate disclosure of financial conflicts of interest has come to light as a sound component of managing relationships between surgeons and industry. STUDY DESIGN: In this study, we summarize and categorize 4 years of disclosures (2006-2009) given by presenters at the Annual Clinical Congress of the American College of Surgeons. RESULTS: We report 3,122 disclosures by 480 individuals. "Colorectal surgeon" was the most common profession among disclosers. The most common type of disclosure was "consulting." The company with the highest number of disclosures was Covidien. Disclosers used 195 different terms to describe their relationships. CONCLUSIONS: We propose a standard nomenclature for use by surgeons when disclosing future conflicts of interest. As the attention to disclosures increases, sound policy decisions would be facilitated by such a standardized nomenclature system.