Construction of brain-like spheroids containing endothelial tubular networks by an adhesive culture system.

Spheroids which are composed of several types of cells have been widely studied in the pharmaceutical field as their structure and functions are similar to human organs. Three-dimensional brain-like tissues are one of the most important tissues for the development of medicines to treat brain diseases and for in vitro brain models. In this study, spheroids mainly containing neurons, astrocytes, and endothelial cells were fabricated using a novel 3D culture plate, "MicoCell™" to construct a brain mimicking tissue. Due to the multicavity structures of MicoCell, ∼102 of attached spheroids were fabricated in a single plate. Spheroids in MicoCell were attached onto a mild cell adhesive surface, allowing for easy immunostaining and microscopic observation. Spheroid formation was improved by adding a Rho-Kinase inhibitor during cultivation. Endothelial cells formed vascular network structures in spheroids and some parts of the vascular structures attached onto the bottom of a culture plate. Co-culture of multiple cell types required optimization of the culture medium during spheroid formation. The mixture of neural stem cell medium and endothelial growth medium showed good spheroid formation and a vascular network. These results indicated that our culture plates and brain mimicking spheroids would be a suitable candidate for pharmaceutical applications such as drug screening and for in vitro brain models.

3D-cultured small size adipose-derived stem cell spheroids promote bone regeneration in the critical-sized bone defect rat model.

Adipose-derived stem cells (ADSCs), due to their regenerative ability, have beneficial effects on bone and cartilage defects. In addition, spheroid formation of ADSCs obtained using three-dimensional (3D) culture accelerates the regenerative ability of ADSCs. The study investigated the regenerative effect of 3D-cultured small size ADSC spheroids without a scaffold in rats with defects in the critical-sized calvarial bone. ADSC-single cells, ADSC-spheroids, or PBS (as control) were implanted in rats, and radiological and histological assessment of bone regeneration was performed. Bone defects were significantly regenerated in the ADSC-spheroid group compared to that in the control group. ADSC-spheroids also showed the most significant bone regeneration in histological assessment. Immunohistochemistry assessment showed that ADSC-spheroids could survive 12 weeks after cell implantation. In vitro, cell apoptosis in ADSC-spheroids was significantly suppressed compared to that in ADSC-single cells. In addition, gene expression related to bone morphogenesis, angiogenesis, and stemness in ADSC-spheroids was elevated. The scaffold-free 3D-cultured small ADSC-spheroids survived in in vitro and in vivo conditions and promoted bone regeneration. Therefore, injectable small size ADSC-spheroids are a novel and less-invasive therapeutic option for treating bone defects.

Re-evaluation of Pre-pump Arterial Pressure to Avoid Inadequate Dialysis and Hemolysis: Importance of Prepump Arterial Pressure Monitoring in Hemodialysis Patients.

Prepump arterial pressure (PreAP) is monitored to avoid generating excessive negative pressure. The National Kidney Foundation K/DOQI clinical practice guidelines for vascular access recommend that PreAP should not fall below -250 mm Hg because excessive negative PreAP can lead to a decrease in the delivery of blood flow, inadequate dialysis, and hemolysis. Nonetheless, these recommendations are consistently disregarded in clinical practice and pressure sensors are often removed from the dialysis circuit. Thus far, delivered blood flow has been reported to decrease at values more negative than -150 mm Hg of PreAP. These values have been analyzed by an ultrasonic flowmeter and not directly measured. Furthermore, no known group has evaluated whether PreAP-induced hemolysis occurs at a particular threshold. Therefore, the aim of this study was to clarify the importance of PreAP in the prediction of inadequate dialysis and hemolysis. By using different diameter needles, human blood samples from healthy volunteers were circulated in a closed dialysis circuit. The relationship between PreAP and delivered blood flow or PreAP and hemolysis was investigated. We also investigated the optimal value for PreAP using several empirical monitoring methods, such as a pressure pillow. Our investigation indicated that PreAP is a critical factor in the determination of delivered blood flow and hemolysis, both of which occured at pressure values more negative than -150 mm Hg. With the exception of direct pressure monitoring, commonly used monitoring methods for PreAP were determined to be ineffective. We propose that the use of a vacuum monitor would permit regular measurement of PreAP.

The Effect of Preoperative Nutritional Intervention for Adult Spinal Deformity Patients.

STUDY DESIGN: A prospective nutritional intervention study for adult spinal deformity (ASD) patients. OBJECTIVE: To investigate how a nutritional intervention affects the incidence of postoperative medical complications and the nutritional status. SUMMARY OF BACKGROUND DATA: The medical complication rate in ASD surgery is very high, and one risk factor is malnutrition. Nutritional intervention may improve the patient's nutritional status and reduce risk, but this is unexplored regarding ASD surgery. METHODS: Malnourished patients (i.e., a prognostic nutritional index [PNI] score of <50) scheduled for surgery after November 2018 (Group I) received nutritional intervention consisting of nutritional guidance and supplements on the surgery day. The medical complication rates between Group I and Group NI (malnourished patients who underwent surgery between January 2014 and October 2018; historical controls) were evaluated. The nutritional status courses of Group I and Group NI2 (patients who did not participate in nutritional intervention after November 2018) were assessed. RESULTS: Group I had 24 patients in (mean age, 70 yr), and Group NI had 69 patients (mean age, 68 yr). The mean intervention duration was 41 days. The preoperative PNI score did not differ between the groups, but there was a significant difference in medical complications incidences (Group I: 25%; Group NI: 53.6%; P = 0.015). The nutritional status significantly deteriorated in Group I (PNI: 47-45; P = 0.011) and Group NI2 (61 patients; mean age, 68 yr; PNI: 52-48; P = 0.000), but the PNI changes were significantly smaller in Group I (ΔPNI: Group I: -1.9, Group NI 2: -3.5; P = 0.027). CONCLUSION: Nutritional intervention with guidance and supplements reduced postoperative medical complications in malnourished patients. The nutritional status of ASD patients requiring surgery also naturally worsened, suggesting that ASD may contribute to malnutrition. Nutritional intervention may reduce the nutritional status deterioration.Level of Evidence: 3.

Fabrication of Spheroids with Dome-Shaped Endothelial Tube Networks by an Adhesive Culture System.

3D functional tissues, such as spheroids fabricated by mesenchymal stem cells (MSCs), which can mimic parts of tissues and organs, have recently been extensively studied in the fields of regenerative medicine and drug discovery. In this study, spheroids containing endothelial tubular structures are fabricated by use of a novel 3D culture plate, "MicoCell." As MicoCell has a mild cell adhesive surface and multicavity structures, it can provide multiple attached spheroids at the same time (about ≈102 to ≈104 spheroids). Spheroids can be fabricated without using serum, and are easily collected by simple pipetting and no use of enzyme. For the fabrication of spheroids containing endothelial tubular structures, MSCs and endothelial cells are co-cultured with MicoCell. Surprisingly, endothelial tubular structures are found to extend upward from the bottom where the spheroids attach onto, forming a dome-shaped morphology. Notably, some tubular structures in the spheroids have a basement membrane and markedly improved oxygen level of the inner part of spheroids. Moreover, as spheroids attach onto the bottom, they do not require any pre-treatment such as embedding into gel before microscopic observation using an optical clearing reagent. These results indicate that the culture plates will be suitable for clinical and pharmaceutical applications.