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Microcarriers provide a high surface-area-to-volume ratio that can realize high yields of cell products, including human mesenchymal stem cells (hMSCs). Here, we report a novel poly(vinyl alcohol) (PVA)-based microcarrier for hMSC expansion in suspension culture. PVA microcarriers were prepared as collagen-coated PVA hydrogels 181 µm in size and a high surface-area-to-weight ratio of 2945 cm2/g. The PVA microcarriers supported a 2.6-fold expansion of hMSCs in a 30-mL single-use stirred bioreactor after a 7 d culture period, comparable to that of commercially available microcarriers. Interestingly, we observed that hMSCs on PVA microcarriers adhered to adjacent microcarriers, resulting in the aggregation of hMSC-PVA microcarriers. Therefore, we conducted a long-term expansion culture using a bead-to-bead cell transfer method with PVA microcarriers. Fresh microcarriers were added to the cell-populated microcarriers in the bioreactor on days 7 and 14. hMSCs on PVA microcarriers continued to grow for 21 d using the bead-to-bead cell transfer method. Furthermore, magnetic PVA (PVA-mag) microcarriers were developed by loading magnetic nanoparticles into PVA microcarriers, and we demonstrated that these PVA-mag microcarriers enabled cell recovery by magnetic separation. These results suggest that these PVA microcarriers can contribute to the large-scale culture of hMSCs for regenerative medicine and cell therapy.
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Introduction: Lower-limb osteoarthritis (OA) in the elderly can be a risk factor for frailty, which is the preliminary of disability, but it may be reversible with appropriate interventions. We aimed to use the Kihon Check List (KCL) to assess multiple domains of frailty and to identify the characteristics of frailty in patients with hip or knee OA following total joint arthroplasty. Materials and Methods: This study included 136 ≥ 65-year-old patients (mean age: 73.0 years) who underwent total arthroplasty with end-stage hip and knee OA. We assessed frailty status, instrumental activities of daily living (IADL), and health-related quality of life (HRQoL) according to the KCL, functional ambulatory index (FAI) and EuroQol-5 Dimension (EQ5D), respectively, as well as the extent of pain preoperatively and at postoperative 6 months. Results: Using KCL, seventy-eight (57.4%) patients were frail preoperatively, but the prevalence significantly decreased to 52 patients (38.2%) at postoperative 6 months. Total arthroplasty intervention provided significant improvements in the total KCL scores, including the physical domain (P < .01), pain (P < .01), FAI scores (P < .01), and EQ5D (P < .01), but not the social domain. Multivariate logistic regression analysis identified age at surgery (OR: .93, 95% CI: .86-.99) and preoperative FAI score (OR: 1.10, 95% CI: 1.03-1.19) as independent predictors of postoperative frailty. Conclusions: Total arthroplasty procedures on patients with hip and knee OA reduced their KCL score, but social aspects were less improved than physical aspects in the shortterm. Older age and preoperative lower IADL score can be useful for accurately estimating less improvement of frailty in the early postoperative phase. Our results suggest that long term follow-up of OA is needed to provide comprehensive interventions, including in social aspects, especially for patients with lower activity.
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An acidic polysaccharide (APS) was isolated from the extract of Cordyceps militaris grown on germinated soybeans. Analyses of sugar composition indicated that APS consisted of d-galactose, L-arabinose, D-xylose, L-rhamnose, and D-galacturonic acid. On the basis of the result of methylation analysis, APS was considered to be mainly composed of Araf-(1-->, -->5)-Araf-(1-->, -->4)-Galp-(1--> and -->4)-GalAp-(1--> residues. When the polysaccharide was intranasally administered, it decreased virus titers in the bronchoalveolar lavage fluid and the lung of mice infected with influenza A virus and increased survival rate. Furthermore, APS increased TNF-alpha and IFN-gamma levels in mice when compared with those of untreated mice. APS enhanced nitric oxide (NO) production and induced iNOS mRNA and protein expressions in RAW 264.7 murine macrophage cells. The induction of mRNA expression of cytokines including IL-1beta, IL-6, IL-10, and TNF-alpha was also observed. These results demonstrated that APS might have beneficial therapeutic effects on influenza A virus infection at least in part by modulation of the immune function of macrophages.
Asunto(s)
Antivirales/uso terapéutico , Cordyceps/química , Glycine max , Factores Inmunológicos/farmacología , Virus de la Influenza A/efectos de los fármacos , Polisacáridos/uso terapéutico , Animales , Línea Celular , Cordyceps/crecimiento & desarrollo , Citocinas/genética , Femenino , Expresión Génica/efectos de los fármacos , Germinación , Concentración de Iones de Hidrógeno , Factores Inmunológicos/aislamiento & purificación , Ratones , Ratones Endogámicos BALB C , Infecciones por Orthomyxoviridae/tratamiento farmacológico , Infecciones por Orthomyxoviridae/inmunología , Polisacáridos/química , Polisacáridos/aislamiento & purificaciónRESUMEN
OBJECTIVE: Heat sterilization of glucose peritoneal dialysis (PD) fluid generates reactive carbonyl compounds (RCOs), which have been implicated in the formation of advanced glycation end products (AGEs) on peritoneal proteins, with an attendant deterioration of peritoneal permeability in PD patients. To reduce their levels in PD fluid, we had previously devised beads coupled with RCO-trapping agents. The hazards linked to the diffusion of RCO-trapping compounds in the systemic circulation are avoided. Hydrazine-epoxy beads proved the most effective. Still, the amount needed to trap all RCOs remained relatively large. METHODS: We developed a novel agent linking a powerful RCO-trapping AGE inhibitor, pyrazolinone-polyethyleneimine, with cellulose beads (PPCBs). We tested its effectiveness to lower RCOs and AGE formation. RESULTS: Mixed with glucose PD fluid, PPCBs markedly lowered RCOs (alpha-dicarbonyls and aldehydes) and inhibited the generation of pentosidine, an AGE, to levels similar to those of filter-sterilized PD fluid. Their effectiveness is more than one order of magnitude above those of previously developed beads. The PPCBs markedly improved PD fluid biocompatibility. Incubation of 1 L commercial glucose PD fluid at 25 degrees C for 24 hours with 10 or 30 g of wet PPCBs reduced RCO content by 75% - 90% and 100% respectively, without altering the pH or glucose and electrolyte contents of the PD fluid. CONCLUSIONS: We developed a high-affinity adsorption bead to reduce the toxic RCO content and AGE formation potential (carbonyl stress) of PD fluid.