Objectives. The effects of local stem cell implantation on clinical and functional characteristics of peripheral vascular disease were studied in two SSc patients with non-healing ischaemic ulcers. Methods. The local injections of CD34⁺ cells from peripheral blood (PB) after mobilization by G-CSF (Case 1) and bone marrow (BM) (Case 2) were used for ischaemic skin ulcers in hands, while mononuclear cells (MNCs) were implanted in lower extremities of the same patients. Ischaemic status was evaluated by measuring ulcer healing, Raynaud's condition score (RCS), visual analogue pain, RP and ulcer scales. To evaluate vasculoprotective action of the implanted cells, we studied weekly the changes in endothelial function, using measurement of flow-mediated brachial artery reactivity by high-resolution ultrasonography, circulating endothelial precursors (CD34⁺VEGFR2⁺, CD133⁺VEGFR2⁺ CEP) by FACS analysis, cutaneous blood flow (laser Doppler flowmetry), skin surface temperature (thermograph), peripheral arterial diameter and blood flow characteristics by Duplex ultrasonography. Results. CD34⁺ cells and MNCs both from BM and PB showed rapid and evident beneficial effect on vascular symptoms resulting in ulcer healing, remarkably decreased daily frequency and duration of RP attacks, RCS, visual analogue scale for RP, ulcers and pain. Physical function and disability measured with HAQ and SHAQ improved. Therapeutic efficacy of stem cell therapy was associated with restoration of endothelial function, augmentation of microcirculatory blood flow and significant increase in circulating CD133⁺VEGFR2⁺ progenitors, known as cell effectors of angiogenesis. Conclusion. This first open-label pilot study demonstrates the feasibility and short-term safety of local CD34⁺ cell therapy for SSc ischaemic complications.

// <![CDATA[ $('.header-date').hide();$('#titleAuthors').hide();$('#abstractHeader').hide(); // ]]> Andreyan N. Osipov1, 2, 3, 4, Margarita Pustovalova1, 2, Anna Grekhova1, 5, Petr Eremin1, Natalia Vorobyova1, 3, Andrey Pulin1, Alex Zhavoronkov4, 6, 7, Sergey Roumiantsev3, 4, 8, Dmitry Y. Klokov9, Ilya Eremin1 1State Research Center - Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency (SRC-FMBC), Moscow, Russia 2Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, Russia 3Dmitry Rogachev Federal Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia 4Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region, Russia 5Emanuel Institute for Biochemical Physics, Russian Academy of Sciences, Moscow, Russia 6Insilico Medicine, Inc, ETC, Johns Hopkins University, Baltimore, Maryland, USA 7The Biogerontology Research Foundation, BGRF, London, UK 8N.I. Pirogov Russian National Research Medical University, Moscow, Russia 9Canadian Nuclear Laboratories, Chalk River, Ontario, Canada Correspondence to: Andreyan N. Osipov, e-mail: andreyan.osipov@gmail.com Keywords: mesenchymal stem cells, DNA double-strand breaks, DNA repair, X-rays, low doses Received: April 08, 2015 Accepted: July 10, 2015 Published: July 23, 2015

Biocompatible ceramic fillers are capable of sustaining bone formation in the proper environment. The major drawback of these scaffolding materials is the absence of osteoinductivity. To overcome this limitation, bioengineered scaffolds combine osteoconductive components (biomaterials) with osteogenic features such as cells and growth factors. The bone marrow mesenchymal stromal cells (BMMSCs) and the β-tricalcium phosphate (β-TCP) are well-known and characterized in this regard. The present study was conducted to compare the properties of novel octacalcium phosphate ceramic (OCP) granules with β-TCP (Cerasorb(®)), gingiva-derived mesenchymal stromal cells (GMSCs) properties with the BMMSCs and osteogenic and angiogenic properties of a bioengineered composite based on OCP granules and the GMSCs. This study demonstrates that GMSCs and BMMSСs have a similar osteogenic capacity. The usage of OCP ceramic granules in combination with BMMSCs/GMSCs significantly affects the osteo- and angiogenesis in bone grafts of ectopic models.

// <![CDATA[ $('.header-date').hide();$('#titleAuthors').hide();$('#abstractHeader').hide(); // ]]> Andreyan N. Osipov1, 2, 3, 4, Anna Grekhova1, 5, Margarita Pustovalova1, 2, Ivan V. Ozerov1, Petr Eremin1, Natalia Vorobyeva1, 3, Natalia Lazareva1, Andrey Pulin1, Alex Zhavoronkov4, 6, 7, Sergey Roumiantsev3, 4, 8, Dmitry Klokov9, Ilya Eremin1 1State Research Center - Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency (SRC-FMBC), Moscow 123098, Russia 2Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow 119991, Russia 3Dmitry Rogachev Federal Research Center of Pediatric Hematology, Oncology and Immunology, Moscow 117997, Russia 4Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region 141700, Russia 5Emanuel Institute for Biochemical Physics, Russian Academy of Sciences, Moscow 119991, Russia 6Insilico Medicine, Inc, ETC, Johns Hopkins University, Baltimore, MD 21218, USA 7The Biogerontology Research Foundation, BGRF, London W1J 5NE, UK 8N.I. Pirogov Russian National Research Medical University, Moscow 117997, Russia 9Canadian Nuclear Laboratories, Chalk River, ON K0J1P0, Canada Correspondence to: Andreyan N. Osipov, e-mail: andreyan.osipov@gmail.com Keywords: DNA DSB repair, homologous recombination, human fibroblasts, X-rays, continuous irradiation Received: April 08, 2015 Accepted: July 31, 2015 Published: August 13, 2015

Treatment of patients with large bone defects is a complex clinical problem. We have initiated the first clinical study of a gene-activated bone substitute composed of the collagen-hydroxyapatite scaffold and plasmid DNA encoding vascular endothelial growth factor. The first patient with two nonunions of previously reconstructed mandible was enrolled into the study. Scar tissues were excised; bone defects (5–14 mm) between the mandibular fragments and nonvascularized rib-bone autograft were filled in with the gene-activated bone substitute. No adverse events were observed during 12 months of follow-up. In 3 months, the average density of newly formed tissues within the implantation zone was 402.21 ± 84.40 and 447.68 ± 106.75 HU in the frontal and distal regions, respectively, which correlated with the density of spongy bone. Complete distal bone defect repair with vestibular and lingual cortical plates formation was observed in 6 and 12 months after surgery; thereby the posterior nonunion was successfully eliminated. However, there was partial resorption of the proximal edge of the autograft entailed to relapse of the anterior nonunion. Thus, the first clinical data on the safety and efficacy of the gene-activated bone substitute were obtained. Given a high complexity of the clinical situation the treatment, results might be considered as promising. NCT02293031.