Potential for Tumorigenesis and Repair of Osteochondral Defects by iPS Cell Transplantation in Rat

Abstract Articular cartilage repair remains a challenge in the field of orthopedic medicine. Cell-based therapy for cartilage repair, such as autologous chondrocyte implantation, was established in the 1990s. However, the issue of the source from which the lesion-targeting cells are harvested remains a limitation of this approach as larger lesions require more cells for repair, and thus, more healthy tissue must be damaged to harvest the needed cells. Reprogramming of induced pluripotent stem (iPS) cells is a promising tool for cell-based regenerative therapy because of their proliferative capacity and pluripotency; however, these characteristics also create a risk of tumorigenesis. This study aimed to determine the probability of iPS cell-derived tumor occurrence as a function of injection or transplantation site, and to assess whether transplanted iPS cells can promote cartilage defect repair. Pluripotent mouse iPS cells (5x10 6 cells/ml) were subcutaneously injected or transplanted into experimentally induced lesions in the knee cartilage of immunodeficient rats. Subcutaneous teratoma formation was observed in 30% of animals (3 of 10) at 4weeks, and 41% of animals (7 of 17) at 12 weeks after iPS cell injection. Cartilage repair as indicated by modified Wakitani's score was similar in the cell-free group and in the iPS cell implantation group at 4 weeks [11.8 ± 1.8 (n = 8) vs. 10.3 ± 2.8 (n = 18)]. iPS cell implantation yielded a score of 7.8 ± 2.0 (n = 10) at 12 weeks, significantly better than the cell-free group [10.5 ± 0.6 (n = 4)]. There was no macro-or microscopic evidence of tumor formation at the cartilage repair site after iPS cell implantation. Although we could not use the iPS cells directly for cartilage repair, the results of our study indicate the potential for a new therapy for cartilage repair by developing iPS reprogramming technology

Burnout and fear of COVID-19 among medical students in Japan: impact of infection history, gender, and social support

The COVID-19 pandemic caused significant changes in medical students' lives and study methods, with online learning replacing in-person classes and limited opportunities for clinical practice. However, there are few studies about burnout and fear of COVID-19 among medical students, especially in East Asia, and a need for research investigating the impact of gender, a history of COVID-19 infection, and social support. In March 2022, we conducted a cross-sectional web-based survey of 4th/5th year medical students who completed a clinical clerkship in Japan. Our survey included the Japan Burnout Scale (JBS, range 5-85, comprising of emotional exhaustion, depersonalization and reduced personal accomplishment), fear of COVID-19 scale (range, 1-4), gender, school year, COVID-19 history, household composition, online education use, and financial burden. There were 343 respondents and 42.4% were women. Multivariable adjusted linear regression analyses showed that students with a COVID-19 infection history had significantly higher overall burnout, depersonalization, reduced personal accomplishment, and lower fear of COVID�19. Students with low social support (living alone and greater financial burden) had higher overall burnout, emotional exhaustion, and depersonalization. Gender had no significant effect on burnout (mean JBS among women was 38.6 versus 39.3 among men). Gender significantly predicted fear of COVID-19, with women scoring higher (1.60 versus 1.50). The findings of the present study have implications that medical schools should provide pastoral care for their students according to students’ circumstances, especially those who live alone, have a high financial burden, and/or were infected with COVID-19

Identification of the Neogenin-Binding Site on the Repulsive Guidance Molecule A

Repulsive guidance molecule (RGM) is a membrane-bound protein that was originally identified as an axon guidance molecule in the chick retinotectal system. RGMa, one of the 3 isoforms found in mammals, is involved in laminar patterning, cephalic neural tube closure, axon guidance, and inhibition of axonal regeneration. In addition to its roles in the nervous system, RGMa plays a role in enhancing helper T-cell activation. Binding of RGM to its receptor, neogenin, is considered necessary to transduce these signals; however, information on the binding of RGM to neogenin is limited. Using co-immunoprecipitation studies, we have identified that the RGMa region required for binding to neogenin contains amino acids (aa) 259–295. Synthesized peptide consisting of aa 284–293 directly binds to the extracellular domain (ECD) of recombinant neogenin, and addition of this peptide inhibits RGMa-induced growth cone collapse in mouse cortical neurons. Thus, we propose that this peptide is a promising lead in finding reagents capable of inhibiting RGMa signaling

Nanocrystalline hydroxyapatite and zinc-doped hydroxyapatite as carrier material for controlled delivery of ciprofloxacin

In bone disorders infections are common. The concentration of majority of antibiotics is very low in the bone tissue. A high local dose can be obtained from the ciprofloxacin-loaded hydroxyapatite nanoparticles. The present study is aimed at developing the use of hydroxyapatite and zinc-doped hydroxyapatite nanoparticles as a carrier for ciprofloxacin drug delivery system. The ciprofloxacin-loaded hydroxyapatite and zinc-doped hydroxyapatite have a good antibacterial activity against Pseudomonas aeruginosa and Staphylococcus aureus. Hydroxyapatite and zinc-doped hydroxyapatite were prepared and characterized using X-ray diffraction, Transmission electron microscopy and inductively coupled plasma optical emission spectrometry. They were loaded with ciprofloxacin using optimized drug loading parameters. Drug loading, in vitro drug release and antimicrobial activity were analyzed. The influence of zinc on the controlled release of ciprofloxacin was analyzed. The results show that the presence of zinc increases the drug release percentage and that the drug was released in a controlled manner

Development of the lateral ventricular choroid plexus in a marsupial, Monodelphis domestica

<p>Abstract</p> <p>Background</p> <p>Choroid plexus epithelial cells are the site of blood/cerebrospinal fluid (CSF) barrier and regulate molecular transfer between the two compartments. Their mitotic activity in the adult is low. During development, the pattern of growth and timing of acquisition of functional properties of plexus epithelium are not known.</p> <p>Methods</p> <p>Numbers and size of choroid plexus epithelial cells and their nuclei were counted and measured in the lateral ventricular plexus from the first day of its appearance until adulthood. Newborn <it>Monodelphis </it>pups were injected with 5-bromo-2-deoxyuridine (BrdU) at postnatal day 3 (P3), P4 and P5. Additional animals were injected at P63, P64 and P65. BrdU-immunopositive nuclei were counted and their position mapped in the plexus structure at different ages after injections. Double-labelling immunocytochemistry with antibodies to plasma protein identified post-mitotic cells involved in protein transfer.</p> <p>Results</p> <p>Numbers of choroid plexus epithelial cells increased 10-fold between the time of birth and adulthood. In newborn pups each consecutive injection of BrdU labelled 20-40 of epithelial cells counted. After 3 injections, numbers of BrdU positive cells remained constant for at least 2 months. BrdU injections at an older age (P63, P64, P65) resulted in a smaller number of labelled plexus cells. Numbers of plexus cells immunopositive for both BrdU and plasma protein increased with age indicating that protein transferring properties are acquired post mitotically. Labelled nuclei were only detected on the dorsal arm of the plexus as it grows from the neuroependyma, moving along the structure in a 'conveyor belt' like fashion.</p> <p>Conclusions</p> <p>The present study established that lateral ventricular choroid plexus epithelial cells are born on the dorsal side of the structure only. Cells born in the first few days after choroid plexus differentiation from the neuroependyma remain present even two months later. Protein-transferring properties are acquired post-mitotically and relatively early in plexus development.</p

Procollagen II C propeptide level in the synovial fluid as a predictor of radiographic progression in early knee osteoarthritis

Objective: To investigate the prognostic value of procollagen type II carboxy-terminal propeptide (PIICP) level in synovial fluid in relation to early tibiofemoral joint osteoarthritis (OA). Methods: Data were collected on 172 women (age 40 to 59 years) who had knee pain and tibiofemoral joint OA in the early stage. Standing semiflexed knee radiographs were obtained by fluoroscopy at baseline and at four year follow up and a computerised, magnification corrected measurement system was applied to measurement of minimal joint space width in the tibiofemoral compartment. Synovial fluid sampling was performed at baseline and at the four year follow up. Levels of PIICP in the synovial fluid were measured by enzyme immunoassay. The outcome measures were assessed by radiographic joint space narrowing (JSN) in the tibiofemoral joints over four years. Multiple linear regression analyses were used to examine the relation between radiographic JSN and synovial fluid level of PIICP. Results: The number of women available at both baseline and at four year follow up was 110. The average of radiographic JSN over four years was 0.53 mm (range 0.00-2.01). Body mass index showed a slightly positive association with baseline PIICP level. In multiple linear regression analyses adjusted for age and body mass index, radiographic JSN over four years had a direct positive correlation with baseline PIICP level (r=0.395; 95% confidence interval (95% CI) 0.231 to 0.529; p<0.001). Conclusion: In a four year prospective study of women, quantification of synovial fluid PIICP was able to predict subsequent radiographic progression in early tibiofemoral joint OA

Gill tissue lipids of salmon (Salmo salar L.) presmolts and smolts from anadromous and landlocked populations

Composition of membrane lipids from the gills of juvenile Atlantic salmon (Salmo salar) in presmolt and smolt phases of development was compared among anadromous and non-anadromous populations. Three stocks migrating from spawning rivers to either lake (landlocked stock), brackish water or full strength sea water were grown under common garden conditions, and gill lipids and their acyl and alkenyl chains were examined in February (presmolts) and at the end of May (smolts) by mass spectrometry and gas-liquid chromatography. The most remarkable changes upon transition from the presmolt phase to the smolt phase were: (i) increase in the cholesterol/phospholipid ratio, (ii) decrease in the abundance of phosphatidylinositol (PI) content, (iii) increase in the amount of sulfatides, (iv) increase in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) species with two highly unsaturated acyl chains, and finally (v) convergence of interstock differences in PC and PE species composition towards a similar lipid composition. Increases in the gill membrane content of cholesterol and sulfatides are discussed as pre-adaptation of salmon gills for salt-secretion, which may occur by increases in membrane microdomains (rafts) harboring ion channels and pumps. The decreases of PI were likely related to adjusting the gill membrane permeability to ions by diminishing prostanoid production. The similarity of those changes among three salmon stocks and the convergence of initially (presmolt phase) different PC and PE species profiles between the stocks towards similar lipid composition suggests that smoltification process of the gill epithelium is largely similar in anadromous and landlocked populations