ABSTRACT
Background & Aim: Liposomes are spherical-shaped vesicles composed of one or more lipid bilayers. The ability of liposomes to encapsulate hydro- or lipophilic drugs allowed these vesicles to become a useful drug delivery system. Natural cell membranes, such as Bioxome, have newly emerged as new source of materials for molecular delivery systems. Bioxome are biocompatible and GMP-compliant liposome-like membrane that can be produced from more than 200 cell types. Bioxome self-assemble, with in-process self-loading capacity and can be loaded with a variety of therapeutic compounds. Once close to the target tissue, Bioxome naturally fuse with the cell membrane and release the inner compound. Orgenesis is interested in evaluating the potential of Bioxome as new drug delivery system for treatment of several diseases, including skin repair, local tumour or COVID19. Methods, Results & Conclusion(s): Bioxome were obtained from adipose- derived Mesenchymal Stem Cells, with a process of organic- solvent lipid extraction, followed by lyophilization and sonication assemblage. During the sonication process, Bioxome were charged or not with several cargos. Size distribution of empty Bioxome was detected by Particle Size Analyzer (NanoSight). Electron Microscopy (EM) was performed to assess Bioxome morphology. Lipid content was evaluated by electrospray ionization system. Dose response in vitro test on human lung fibroblasts treated or not with Bioxome encapsulating a specific cargo (API) against COVID19 were performed. NanoSight analysis showed that nanoparticle size in Bioxome samples ranged between 170+/-50 nm, with a concentration ranging between 109-1010+/-106 particles/mL. EM clearly showed the double phospholipid layers that composes the Bioxome. Stability study demonstrated that Bioxome are stable in size and concentration up to 90 days at +4Cdegree or even at RT. No change in size between encapsulated Bioxome with small size (~340 Da) cargo vs empty Bioxome was observed up to 30 days storage. Lipidomic analysis approach revealed that the yield of lipids and their composition are satisfactory for a therapeutic product using Bioxome. Lastly, in the in vitro model of COVID19, Bioxome encapsulating API effectively saved cells from death (20x vs untreated cells) and at lower doses of API than these of non-encapsulated cargo (0.005 microM vs 0.1 microM). Bioxome seems to be an excellent candidate for liposome mimetic tool as drug delivery system for targeting specific organs and diseases treatment.Copyright © 2023 International Society for Cell & Gene Therapy
ABSTRACT
Background & Aim: From SARS-CoV-1 outbreak in 2002 to the most recent SARS-CoV-2 pandemic (COVID-19), emergence of viral diseases has repeatedly threatened humanity over the recent decades. These viral diseases mainly cause respiratory symptoms, which can even lead to death when appropriate measures are not taken. In this study, we investigated whether adipose tissue-derived mesenchymal stem cell EVs (ASC-EVs) can attenuate acute lung injury (ALI) induced by H1N1 influenza A virus and SARS-CoV-2 and by what mechanism the ant-viral effect may occurs. Methods, Results & Conclusion: EVs were isolated from ASC or HEK293T conditioned media by tangential flow filtration, and were characterized according to MISEV recommendation. Influenza A/ Puerto Rico/08/1934 (H1N1) and SARS-CoV-2 (NCCP43326) were used to model highly pathogenic human influenza A and SARS-CoV-2 virus infection, respectively, in mice and Syrian hamsters respectively. Treatment of ASC-EVs, from 0.15 x 109 to 5.0 x 109 particles/mL, showed inhibitory activities on cytopathic effects and replication of H1N1 and SARS-CoV-2 in MDCK cells and Vero E6 cells, respectively. In the mouse H1N1 influenza A virus induced acute lung injury (ALI) model, total of 4 daily injections of 1 x 1010 particles of ASC-EVs administration resulted in significantly increased survival rate by 30 – 40%, recovery of body weight, and improved clinical disease score from 9 dpi. In addition, ASC-EV treatment downregulated various inflammatory cytokines such as IL-1β, IL-6 and TNFα in lung tissue by up to 77%. In the Syrian hamster SARS-CoV-2 induced ALI model, total of 4 daily injections of ASC-EVs at a dose of 3 x 1010 or 1 x 1010 particles resulted recovery of body weights from 5 dpi, in a dose-dependent manner, by 9.7% - 12.75%. Further, ASC-EV treatment resulted in significant downregulation of viral genes and IL-1 beat in lung tissue. To elucidate the molecular mechanisms of the observed anti-viral effects of ASC-EVs, the role of multiple miRNAs and proteins present in the ASC-EVs were assessed in vitro. We identified one specific protein that conveyed anti-viral efficacy against the two studied viruses including SARS-CoV-2. Loss and gain of function studies revealed that this protein may be involved in the anti-viral efficacy of the ASC-EVs. Our findings support the concept that that ASC-EVs have anti-viral effects against virus induced ALI, which may have implications for the treatment of not only treatment COVID-19, but also future ALI-inducing virus diseases.
ABSTRACT
Background: Despite spontaneous recovery, 60% of stroke patients have residual impairment. Thawed allogenic adipose-derived mesenchymal stem cells (ADSCs) were used in cell therapy studies with promising results. However, fresh allogenic ADSCs, obtained after a short (oneweek) culture from a frozen cell stock, have a better viability and functionality. We assessed feasibility and safety of intravenous (IV) injection of fresh ADSCs in subacute ischemic stroke. Methods: RESSTORE 1a is a first-in-human, open, dose-escalation toxicity 3+3 trial. Four doses of ADSCs were assessed: 1, 1.5, 2 or 3 million/ kg injected <10 days after stroke onset. Dose-limiting toxicity (DLT) events were predefined: recurrent stroke, myocardial infarction, pulmonary embolism, life-threatening event within one week after treatment. Results: 17 patients were included (mean age +/-SD = 57 +/-18 years;inclusion NIHSS = 14.5 +/-3.5). As 2 patients were not treated (non conformity of ADSC culture and technical problem during infusion), 2 additional patients were included. A DLT event (recurrent minor stroke) occurred in the 2-million/kg group. Among the 15 treated patients, 3 patients received 1, 1.5 or 3 million ADSCs /kg, and 6 patients 2 million ADSCs /kg. A lung carcinoma was diagnosed in an active smoker (1.5 year after treatment) and a patient died after Covid-19 pneumopathy (1 year after treatment). No dose effect on safety was noted. Conclusions: IV injection of fresh allogenic ADSC seems safe in subacute stroke. A multicenter randomized trial is planned.