P-gp expression inhibition mediates placental glucocorticoid barrier opening and fetal weight loss

Abstract Background Prenatal adverse environments can cause fetal intrauterine growth retardation (IUGR) and higher susceptibility to multiple diseases after birth, related to multi-organ development programming changes mediated by intrauterine overexposure to maternal glucocorticoids. As a glucocorticoid barrier, P-glycoprotein (P-gp) is highly expressed in placental syncytiotrophoblasts; however, the effect of P-gp on the occurrence of IUGR remains unclear. Methods Human placenta and fetal cord blood samples of IUGR fetuses were collected, and the related indexes were detected. Pregnant Wistar rats were administered with 30 mg/kg·d (low dose) and 120 mg/kg·d (high dose) caffeine from gestational day (GD) 9 to 20 to construct the rat IUGR model. Pregnant mice were administered with caffeine (120 mg/kg·d) separately or combined with sodium ferulate (50 mg/kg·d) from gestational day GD 9 to 18 to confirm the intervention target on fetal weight loss caused by prenatal caffeine exposure (PCE). The fetal serum/placental corticosterone level, placental P-gp expression, and related indicator changes were analyzed. In vitro, primary human trophoblasts and BeWo cells were used to confirm the effect of caffeine on P-gp and its mechanism. Results The placental P-gp expression was significantly reduced, but the umbilical cord blood cortisol level was increased in clinical samples of the IUGR neonates, which were positively and negatively correlated with the neonatal birth weight, respectively. Meanwhile, in the PCE-induced IUGR rat model, the placental P-gp expression of IUGR rats was decreased while the corticosterone levels of the placentas/fetal blood were increased, which were positively and negatively correlated with the decreased placental/fetal weights, respectively. Combined with the PCE-induced IUGR rat model, in vitro caffeine-treated placental trophoblasts, we confirmed that caffeine decreased the histone acetylation and expression of P-gp via RYR/JNK/YB-1/P300 pathway, which inhibited placental and fetal development. We further demonstrated that P-gp inducer sodium ferulate could reverse the inhibitory effect of caffeine on the fetal body/placental weight. Finally, clinical specimens and other animal models of IUGR also confirmed that the JNK/YB-1 pathway is a co-regulatory mechanism of P-gp expression inhibition, among which the expression of YB-1 is the most stable. Therefore, we proposed that YB-1 could be used as the potential early warning target for the opening of the placental glucocorticoid barrier, the occurrence of IUGR, and the susceptibility of a variety of diseases. Conclusions This study, for the first time, clarified the critical role and epigenetic regulation mechanism of P-gp in mediating the opening mechanism of the placental glucocorticoid barrier, providing a novel idea for exploring the early warning, prevention, and treatment strategies of IUGR.

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Rikkunshito Ameliorates Cancer Cachexia Partly through Elevation of Glucarate in Plasma

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/871832 ◽

2015 ◽

Vol 2015 ◽

pp. 1-11 ◽

Cited By ~ 6

Author(s):

Katsuya Ohbuchi ◽

Shin Nishiumi ◽

Naoki Fujitsuka ◽

Tomohisa Hattori ◽

Masahiro Yamamoto ◽

...

Keyword(s):

Weight Loss ◽

Herbal Medicine ◽

Rat Model ◽

Cancer Cachexia ◽

Host Cells ◽

Metabolome Analysis ◽

Traditional Herbal Medicine ◽

Delayed Onset ◽

Tumor Bearing

Cancer cachexia, which is characterized by decreased food intake, weight loss and systemic inflammation, increases patient’s morbidity and mortality. We previously showed that rikkunshito (RKT), a Japanese traditional herbal medicine (Kampo), ameliorated the symptoms of cancer cachexia through ghrelin signaling-dependent and independent pathways. To investigate other mechanisms of RKT action in cancer cachexia, we performed metabolome analysis of plasma in a rat model bearing the Yoshida AH-130 hepatoma. A total of 110 metabolites were detected in plasma and RKT treatment significantly altered levels of 23 of those metabolites in cachexia model rats. Among them, glucarate, which is known to have anticarcinogenic activity through detoxification of carcinogens via inhibition ofβ-glucuronidase, was increased in plasma following administration of RKT. In our AH-130 ascites-induced cachexia rat model, administration of glucarate delayed onset of weight loss, improved muscle atrophy, and reduced ascites content. Additionally, glucarate reduced levels of plasma interferon-γ(IFN-γ) in tumor-bearing rats and was also found to suppress LPS-induced IFN-γexpression in splenocytesin vitro. These results suggest that glucarate has anti-inflammatory activity via a direct effect on immune host cells and suggest that RKT may also ameliorate inflammation partly through the elevation of glucarate in plasma.

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Therapeutic Activity of Intramuscular Peramivir in Mice Infected with a Recombinant Influenza A/WSN/33 (H1N1) Virus Containing the H275Y Neuraminidase Mutation

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00753-12 ◽

2012 ◽

Vol 56 (8) ◽

pp. 4375-4380 ◽

Cited By ~ 14

Author(s):

Yacine Abed ◽

Andrés Pizzorno ◽

Guy Boivin

Keyword(s):

Weight Loss ◽

Body Weight ◽

Influenza A ◽

Low Dose ◽

Body Weight Loss ◽

High Dose ◽

Therapeutic Activity ◽

Enzymatic Assays ◽

Oseltamivir Resistance

ABSTRACTThe therapeutic activity of intramuscular (IM) peramivir was evaluated in mice infected with a recombinant influenza A/WSN/33 virus containing the H275Y neuraminidase (NA) mutation known to confer oseltamivir resistance. Regimens consisted of single (90 mg/kg of body weight) or multiple (45 mg/kg daily for 5 days) IM peramivir doses that were initiated 24 h or 48 h postinfection (p.i.). An oral oseltamivir regimen (1 or 10 mg/kg daily for 5 days) was used for comparison. Untreated animals had a mortality rate of 75% and showed a mean weight loss of 16.9% on day 5 p.i. When started at 24 h p.i., both peramivir regimens prevented mortality and significantly reduced weight loss (P< 0.001) and lung viral titers (LVT) (P< 0.001). A high dose (10 mg/kg) of oseltamivir initiated at 24 h p.i. also prevented mortality and significantly decreased weight loss (P< 0.05) and LVT (P< 0.001) compared to the untreated group results. In contrast, a low dose (1 mg/kg) of oseltamivir did not show any benefits. When started at 48 h p.i., both peramivir regimens prevented mortality and significantly reduced weight loss (P< 0.01) and LVT (P< 0.001) whereas low-dose or high-dose oseltamivir regimens had no effect on mortality rates, body weight loss, and LVT. Our results show that single-dose and multiple-dose IM peramivir regimens retain clinical and virological activities against the A/H1N1 H275Y variant despite some reduction in susceptibility when assessedin vitrousing enzymatic assays. IM peramivir could constitute an alternative for treatment of oseltamivir-resistant A/H1N1 infections, although additional studies are warranted to support such a recommendation.

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EGR1 Modulated LncRNA HNF1A-AS1 Drives Glioblastoma Progression Via miR-22-3p/ENO1 Axis

10.21203/rs.3.rs-477104/v1 ◽

2021 ◽

Author(s):

Chunchun Ma ◽

Hongliang Wang ◽

Gang Zong ◽

Jie He ◽

Yuyang Wang ◽

...

Keyword(s):

Cell Proliferation ◽

Molecular Mechanisms ◽

Bioinformatic Analysis ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Clinical Samples ◽

Regulation Mechanism ◽

Functional Studies

Abstract Background: Accumulating evidences revealed that long noncoding RNAs (lncRNAs) have been participated in cancer malignant progression, including glioblastoma multiforme (GBM). Despite much studies have found the precise biological role in the regulatory mechanisms of GBM，however the molecular mechanisms，particularly upstream mechanisms still need further elucidated. Methods: RT-QPCR, cell transfection, western blotting and bioinformatic analysis were executed to detect the expression of EGR1, HNF1A-AS1, miR-22-3p and ENO1 in GBM. Cell proliferation assay, colony formation assay, wound healing, migration and invasion assays were performed to detect the malignant characters of GBM cell. The molecular regulation mechanism was confirmed by luciferase reporter assay, ChIP and RIP. Finally, orthotopic mouse models were established to examine the effect of HNF1A-AS1 in vivo.Results: In the current study, we analyzed clinical samples to show that the long non-coding antisense transcript of HNF1A, HNF1A-AS1, is upregulated and associated with poor prognosis in GBM. Functional studies revealed that knockdown of HNF1A-AS1 markedly inhibits cell proliferation, migration and invasion both in vitro and in vivo, whereas overexpression of HNF1A-AS1 exerts opposite effect. Mechanistically, the transcription factor EGR1 forced the transcription of HNF1A-AS1 by directly binding the promoter region of HNF1A-AS1. Furthermore, combined bioinformatics analysis with our mechanistic work, using luciferase reporter assays and RIP, we first demonstrated that HNF1A-AS1 functions as a competing endogenous RNA (ceRNA) with miR-22-3p to regulate ENO1 expression in GBM cells. HNF1A-AS1 directly binds to miR-22-3p and significantly inhibits miR-22-3p expression, while ENO1 expression was increased. miR-22-3p inhibitor offsets the HNF1A-AS1 silencing induced suppression in proliferation, migration and invasion of GBM cells, as well as promotion effect on ENO1 expression. ENO1 was verified as a direct target of miR-22-3p and its expression levels was negatively with the prognosis in GBM patients. Conclusion: Taken together, our study illuminated the definite mechanism of HNF1A-AS1 in promoting GBM malignancy, and provided a novel therapeutic target for further clinical application.

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Toxicological profile of umbilical cord blood-derived small extracellular vesicles

10.1101/2021.06.30.450343 ◽

2021 ◽

Author(s):

Silvia C Rodrigues ◽

Renato M S Cardoso ◽

Claudia F Gomes ◽

Filipe V Duarte ◽

Patricia C Freire ◽

...

Keyword(s):

Cord Blood ◽

Umbilical Cord ◽

Extracellular Vesicles ◽

Umbilical Cord Blood ◽

Mononuclear Cells ◽

High Dose ◽

Cell Therapies ◽

Cell Counts ◽

Blood Mononuclear Cells

The development and adoption of cell therapies has been largely limited by difficulties associated with their safety, handling and storage. Extracellular vesicles (EV) have recently emerged as a likely mediator for the therapeutic effect of cells, offering several advantages over cell therapies. Due to their small size and inability to expand and metastasize, EV are generally considered safer than cell transplantation. Nevertheless, few studies have scrutinized the toxicity profile of EV, particularly after repeated high dose administration. The present study aimed to evaluate a preparation of small EV obtained from umbilical cord blood mononuclear cells (UCB-MNC-sEV) for its cytotoxicity in different cell lines, as well as its differential accumulation, distribution and toxicity following repeated intravenous (IV) administrations in a rodent model. In vitro, repeated sEV exposure in concentrations up to 1x10^11 particles/ml had no deleterious impact on the viability or metabolic activity of peripheral blood mononuclear cells, THP-1 monocytes, THP-1-derived macrophages, normal dermal human fibroblasts or human umbilical vein endothelial cells. DiR-labeled sEV, injected IV for four weeks in healthy rats, were detected in clearance organs, particularly kidneys, spleen and liver, similarly to control dye. Moreover, repeated administrations during six and twelve weeks of up to 1x10^10 total particles of sEV-dye were well tolerated, with no changes in general hematological cell counts, or kidney and liver toxicity markers. Importantly, unlabeled sEV likewise did not induce significant alterations in cellular and biochemical blood parameters, nor any morphological changes in heart, kidney, lung, spleen, or liver tissue. In sum, our data shows that UCB-MNC-sEV have no significant toxicity in vitro or in vivo, even when administered repeatedly at high concentrations, therefore confirming their safety profile and potential suitability for future clinical use.

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Toxicological Profile of Umbilical Cord Blood-Derived Small Extracellular Vesicles

Membranes ◽

10.3390/membranes11090647 ◽

2021 ◽

Vol 11 (9) ◽

pp. 647 ◽

Cited By ~ 1

Author(s):

Silvia C. Rodrigues ◽

Renato M. S. Cardoso ◽

Claudia F. Gomes ◽

Filipe V. Duarte ◽

Patricia C. Freire ◽

...

Keyword(s):

Cord Blood ◽

Umbilical Cord ◽

Extracellular Vesicles ◽

Umbilical Cord Blood ◽

Mononuclear Cells ◽

High Dose ◽

Cell Therapies ◽

Cell Counts ◽

Blood Mononuclear Cells

The development and adoption of cell therapies has been largely limited by difficulties associated with their safety, handling, and storage. Extracellular vesicles (EV) have recently emerged as a likely mediator for the therapeutic effect of cells, offering several advantages over cell therapies. Due to their small size and inability to expand and metastasize, EV are generally considered safer than cell transplantation. Nevertheless, few studies have scrutinized the toxicity profile of EV, particularly after repeated high-dose administration. The present study aimed to evaluate a preparation of small EV obtained from umbilical cord blood mononuclear cells (UCB-MNC-sEV) for its cytotoxicity in different cell lines, as well as its differential accumulation, distribution, and toxicity following repeated intravenous (IV) administrations in a rodent model. In vitro, repeated sEV exposure in concentrations up to 1 × 1011 particles/mL had no deleterious impact on the viability or metabolic activity of peripheral blood mononuclear cells, THP-1 monocytes, THP-1-derived macrophages, normal dermal human fibroblasts, or human umbilical vein endothelial cells. DiR-labelled sEV, injected intravenously for four weeks in healthy rats, were detected in clearance organs, particularly the kidneys, spleen, and liver, similarly to control dye. Moreover, repeated administrations for six and twelve weeks of up to 1 × 1010 total particles of sEV dye were well-tolerated, with no changes in general haematological cell counts, or kidney and liver toxicity markers. More importantly, unlabelled sEV likewise did not induce significant alterations in cellular and biochemical blood parameters, nor any morphological changes in the heart, kidney, lung, spleen, or liver tissue. In sum, our data show that UCB-MNC-sEV have no significant toxicity in vitro or in vivo, even when administered repeatedly at high concentrations, therefore confirming their safety profile and potential suitability for future clinical use.

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Establishment of a Spontaneous Stromal Microenvironment from Cord Blood Supports Human Dynamic Erythropoietic Temporal Maturation in Long-Term Serum- and Cytokine-Free 3D Cultures and Reveals a Distinct CD44hi Population

Blood ◽

10.1182/blood-2019-131085 ◽

2019 ◽

Vol 134 (Supplement_1) ◽

pp. 2219-2219

Author(s):

Susana Brito Dos Santos ◽

Athanasios Mantalaris ◽

Nicki Panoskaltsis

Keyword(s):

Cord Blood ◽

Mononuclear Cells ◽

Ex Vivo ◽

Hla Typing ◽

High Dose ◽

Ex Vivo Model ◽

Erythroid Progenitor ◽

Erythroid Precursors

Dynamic cultures which can represent physiologic erythropoiesis in vitro require a three-dimensional (3D) architecture with a supportive microenvironment and addition of erythropoietin (EPO). We have previously reported on a 3D bone marrow (BM) biomimicry using polyurethane scaffolds to expand cord blood mononuclear cells (CBMNCs) in a serum- and cytokine-free environment, without addition of dexamethasone, for 28 days (D). CBMNCs were seeded (4x106 cells/scaffold), supplemented with 10ng/mL stem cell factor (SCF; D0-D28) and 100mU/mL EPO (D7-D28), with medium exchange every 3D and exposed to a hypoxia (5%)/normoxia (20%) schedule to mimic BM oxygen gradients. Hypoxia induced rapid erythroid commitment and established an early erythroid progenitor population in the absence of EPO. Normoxia and EPO was required at later maturational stages and enhanced the γ-globin to β-globin switch. We identified D7-D14 as crucial for endogenous cytokine production. Herein, we extended cultures to D48 using two high-dose EPO-stimulation cycles (1U/mL; D20 and D44) to enhance erythropoiesis and further define the microenvironment. Proliferation was higher after EPO pulses (p<0.05) but did not result in enhanced erythropoiesis, suggesting the absence of erythroid precursors. An allogeneic CB unit was added to "recharge" the cultured scaffolds at D39 and a new cycle of erythropoietic differentiation was initiated. Cell proliferation was 4.5-fold higher at D68, compared with that at D28. From D53-D68, CD71+CD235a+ cells were constantly produced (25-54%), corresponding with the presence of erythroid precursors supporting CFU-E and BFU-E. Erythroblastic islands were identified and maturing and enucleated reticulocytes/RBCs were abundant (19±2%; 1±0.3x106 cells) with expression of γ- and β-globin, band 3 and 4.1R RBC membrane proteins. To further evaluate the relative contributions of each CB unit to the "recharge" culture, seeded scaffolds were subjected to irradiation (or not) 48h prior to recharge. By D65, >30% of supernatant cells were CD71+/modCD235a+ and supported BFU-E and CFU-E. Proliferation in long-term cultures was attributed to the second CB unit, regardless of irradiation, as shown by HLA-typing of D68 cells; the first CBMNCs only contributed to establishment of the microenvironment. To further characterize erythroid differentiation dynamics, expression of CD44 vs CD235a was used to identify and sort three erythroid populations. Progenitors in CD44-/modCD235a- populations evolved to CD44modCD235amod, and then to CD44-/modCD235a+ cells, which constitute the most mature erythroid phenotype; CD44modCD235amod erythroid precursors supported mainly BFU-E and CFU-E. A unique CD44hiCD71mod population increased during culture, displayed myeloid progenitor morphology and only supported CFU-GM. This population did not express CD34, CD33 or CD14 but expressed c-KIT, which suggests a hematopoietic population that provides essential culture support. Further characterization of the spontaneously created microenvironment by in situ quantitative analysis of scaffold mid-sections during the 68-day culture showed varying and high dynamic expression of Nestin, STRO-1, CD146, CD68 and CD169 in separate cell populations as well as expression of RUNX2 and Osx. Osteopontin was not detected. In summary, a BM biomimicry composed of diverse stromal populations was spontaneously created in the 3D in vitro scaffold system. This microenvironment proved to effectively induce and sustain erythropoiesis to enucleation to at least D68 when supplied with a second source of CBMNCs, without addition of stroma-specific factors. A unique CD44hi immature monocyte/macrophage population was identified, which contributes to the inductive microenvironment, and distinct stages of human erythroid maturation could be identified using CD44 and CD235a. This work presents a novel and dynamic ex vivo model that can (1) recapitulate physiologic human erythropoiesis in steady-state and stress conditions, (2) capture the fetal to adult hemoglobin transition, (3) explore the direct role of oxygen on erythropoiesis, (4) assess the microenvironment relevant to erythropoiesis in the absence of serum and exogenous factors, (5) sustain long-term erythropoiesis with terminal maturation and, (6) explore the different stromal niche environments spontaneously created for the support of erythropoiesis. Disclosures Brito Dos Santos: GE Healthcare: Employment.

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Cobblestone Area-Forming Cell (CAFC) Content of Cord Blood Does Not Predict Engraftment in Double Cord Blood Transplantation.

Blood ◽

10.1182/blood.v110.11.2243.2243 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2243-2243

Author(s):

Simon N. Robinson ◽

Marcos de Lima ◽

Hong Yang ◽

William K. Decker ◽

Dongxia Xing ◽

...

Keyword(s):

Cord Blood ◽

Ex Vivo ◽

Cord Blood Transplantation ◽

Randomized Study ◽

Clinical Samples ◽

Hematopoietic Stem ◽

Blood Transplantation ◽

Qualitative Measure

Abstract INTRODUCTION: Delayed engraftment remains a serious problem following cord blood (CB) transplantation. It may be due, at least in part, to the limited dose of CB hematopoietic stem and progenitor cells (HSPC) transplanted. Limitations associated with HSPC dose may be reduced by the transplantation of 2 CB units. In an accompanying abstract, de Lima et al. report in detail on an ongoing MDACC randomized study in which patients received double CB units as either 2 unmanipulated units (2×UN), or 1 unmanipulated and 1 ex vivo expanded CB unit (UN+EX). This study has revealed that one CB unit ultimately predominates as the source of long-term, sustained hematopoiesis. We hypothesized that the number of primitive HSPC in the CB unit might ultimately predict which CB unit would ultimately prevail. The in vitro cobblestone area-forming cell (CAFC) assay was used to provide a measure of primitive components of the CB HSPC in each unit. A photomicrograph of a typical cobblestone area (CA) derived from a single CB-derived CAFC is shown. (Figure) CA persisting in in vitro culture for ≥6 weeks (derived from CAFCwk6) represent relatively primitive HSPC and their numbers may provide a qualitative profile for CB units. We hypothesized that of the two CB units transplanted, the one with the greater number of CAFCwk6 would ultimately persist long-term in the patient. METHODS: Clinical samples of 2×UN or UN+EX CB cells were plated in the in vitro CAFC assay.1 The frequency of CAFCwk6 was estimated and total CAFCwk6 numbers transplanted for each CB unit calculated. CAFCwk6 data for each CB unit and engraftment data from patients were analyzed to determine whether the number of CAFCwk6 transplanted was predictive of which CB unit would ultimately be responsible for long-term, sustained engraftment. RESULTS: Preliminary data was accrued from 10 patients.(Table) Six patients received 2×UN and 4 patients received UN+EX (EX indicated by #). CAFCwk6 content at transplant was predictive of which CB unit would ultimately be responsible for long-term, sustained engraftment in only 6 of the 10 cases (60%). CONCLUSION: CAFCwk6 represent a relatively primitive component of the HSPC compartment, however, these data suggest that CAFCwk6 numbers, although possibly a qualitative measure, are not a predictor of long-term sustained engraftment. Figure Figure

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Broad safety impact of high-dose ascorbic acid and induction chemotherapy for high-risk pancreatic cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.e15711 ◽

2017 ◽

Vol 35 (15_suppl) ◽

pp. e15711-e15711

Author(s):

Howard Bruckner ◽

Azriel Hirschfeld ◽

Daniel Gurell ◽

King Lee

Keyword(s):

Quality Of Life ◽

Weight Loss ◽

Ascorbic Acid ◽

High Risk ◽

Stage Iv ◽

High Dose ◽

Cost Treatment

e15711 Background: High dose ascorbic acid inhibits many tumors in vitro. It may reduce chemotherapy's adverse hematologic events and improve quality of life. Moderate dose gemcitabine, fluorouracil, leucovorin, irinotecan, oxaliplatin (GFLIP) is an effective regimen for patients with many cancers (Anticancer Research 17). High dose ascorbic acid may reverse half the preexisting and largely eliminate the predicted cumulative toxicity of GFLIP (ASCO 15). Methods: High dose ascorbic acid AA 75-100 grams IV was given 1-2 times per week with GFLIP every 2 weeks until progression with serial wkly and Q2wk blood tests and 3 month imaging. Eligibility: Unresectable, III, recurrent and metastatic, mod and high grade typical pancreatic ca; ECOG 0-2; +/- prior chemo; adults of any age with consent. Results: A prescheduled 2 yr analysis found 26 patients; 16 ≥ 65, (9 ≥ 70 years of age). 25 stage IV; Nine had failed at least one prior standard chemo (35%), 3 were PS 2, 9 had severe weight loss. Safety: Five had uncomplicated 3 (19%) and none had 4 neutropenia or neutropenic infection, one (4%) had a 3 anemia and thrombocytopenia without bleeding. Prophylactic growth factors were not used and limited to 1-2 as needed low doses used infrequently. There was no limiting diarrhea, enteritis, stomatitis, weight loss or coagulopathy due to GFLIP. GFLIP is on track to reproduce/retain prior response and survival benefit (ASCO 08). Conclusions: Ascorbic Acid-GFLIP can be exceptionally safe and well tolerated. It may avoid standard 20-40% rates of severe toxicities. It can be, otherwise unavailable, safe reduced cost, treatment for many elderly and prior resistant tumor patients. Given the broad multi-disease role of the GFLIP drugs, and available personalized medicine tests (Cancer Letters), further development is very attractive and feasible. Potential outcome can spare all, and especially high risk pancreatic ca patients, 20k severe toxicities every year toxicities as much as $100 million cost and make some ineligibility criteria unnecessary. Development of safe AA regimens may benefit quality of life and improve survival for many with a broad spectrum of cancers that otherwise go untreated. Clinical trial information: NCT01905150.

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Intra-articular Injection of Cell-laden 3D Microcryogels Empower Low-dose Cell Therapy for Osteoarthritis in a Rat Model

Cell Transplantation ◽

10.1177/0963689720932142 ◽

2020 ◽

Vol 29 ◽

pp. 096368972093214

Author(s):

Dan Xing ◽

Wei Liu ◽

Bin Wang ◽

Jiao Jiao Li ◽

Yu Zhao ◽

...

Keyword(s):

Cell Therapy ◽

Rat Model ◽

Low Dose ◽

High Rate ◽

Saline Control ◽

High Dose ◽

Sprague Dawley ◽

Osteoarthritic Joint

Intra-articular injection of mesenchymal stem cells (MSCs) in an osteoarthritic joint can help slow down cartilage destruction. However, cell survival and the efficiency of repair are generally low due to mechanical damage during injection and a high rate of cell loss. We, thus, investigated an improved strategy for cell delivery to an osteoarthritic joint through the use of three-dimensional (3D) microcryogels. MSCs were seeded into 3D microcryogels. The viability and proliferation of MSCs in microcryogels were determined over 5 d, and the phenotype of MSCs was confirmed through trilineage differentiation tests and flow cytometry. In Sprague Dawley rats with induced osteoarthritis (OA) of the knee joint, a single injection was made with the following groups: saline control, low-dose free MSCs (1 × 105 cells), high-dose free MSCs (1 × 106 cells), and microcryogels + MSCs (1 × 105 cells). Cartilage degeneration was evaluated by macroscopic examination, micro-computed tomographic analysis, and histology. MSCs grown in microcryogels exhibited optimal viability and proliferation at 3 d with stable maintenance of phenotype in vitro. Microcryogels seeded with MSCs were, therefore, primed for 3 d before being used for in vivo experiments. At 4 and 8 wk, the microcryogels + MSCs and high-dose free MSC groups had significantly higher International Cartilage Repair Society macroscopic scores, histological evidence of more proteoglycan deposition and less cartilage loss accompanied by a lower Mankin score, and minimal radiographic evidence of osteoarthritic changes in the joint compared to the other two groups. In conclusion, intra-articular injection of cell-laden 3D microcryogels containing a low dose of MSCs can achieve similar effects as a high dose of free MSCs for OA in a rat model. Primed MSCs in 3D microcryogels can be considered as an improved delivery strategy for cell therapy in treating OA that minimizes cell dose while retaining therapeutic efficacy.

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Successful High-Dosage Monotherapy of Tigecycline in a Multidrug-Resistant Klebsiella pneumoniae Pneumonia–Septicemia Model in Rats

Antibiotics ◽

10.3390/antibiotics9030109 ◽

2020 ◽

Vol 9 (3) ◽

pp. 109 ◽

Cited By ~ 1

Author(s):

Hessel Van der Weide ◽

Marian T. Ten Kate ◽

Denise M. C. Vermeulen-de Jongh ◽

Aart Van der Meijden ◽

Rixt A. Wijma ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Rat Model ◽

Bacterial Infections ◽

Recent Literature ◽

Respiratory Infections ◽

Multidrug Resistant ◽

High Dose ◽

The Moment

Background: Recent scientific reports on the use of high dose tigecycline monotherapy as a “drug of last resort” warrant further research into the use of this regimen for the treatment of severe multidrug-resistant, Gram-negative bacterial infections. In the current study, the therapeutic efficacy of tigecycline monotherapy was investigated and compared to meropenem monotherapy in a newly developed rat model of fatal lobar pneumonia–septicemia. Methods: A Klebsiella pneumoniae producing extended-spectrum β-lactamase (ESBL) and an isogenic variant producing K. pneumoniae carbapenemase (KPC) were used in the study. Both strains were tested for their in vitro antibiotic susceptibility and used to induce pneumonia–septicemia in rats, which was characterized using disease progression parameters. Therapy with tigecycline or meropenem was initiated at the moment that rats suffered from progressive infection and was administered 12-hourly over 10 days. The pharmacokinetics of meropenem were determined in infected rats. Results: In rats with ESBL pneumonia–septicemia, the minimum dosage of meropenem achieving survival of all rats was 25 mg/kg/day. However, in rats with KPC pneumonia–septicemia, this meropenem dosage was unsuccessful. In contrast, all rats with KPC pneumonia–septicemia were successfully cured by administration of high-dose tigecycline monotherapy of 25 mg/kg/day (i.e., the minimum tigecycline dosage achieving 100% survival of rats with ESBL pneumonia–septicemia in a previous study). Conclusions: The current study supports recent literature recommending high-dose tigecycline as a last resort regimen for the treatment of severe multidrug-resistant bacterial infections. The use of ESBL- and KPC-producing K. pneumoniae strains in the current rat model of pneumonia–septicemia enables further investigation, helping provide supporting data for follow-up clinical trials in patients suffering from severe multidrug-resistant bacterial respiratory infections.

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