


Vol 6, No 3 (2011)
- Year: 2011
- Articles: 13
- URL: https://genescells.ru/2313-1829/issue/view/6165
Articles
MNENIYa SPETsIALISTOV
Abstract
В связи с тем, что настоящий выпуск жур-
нала посвящен перспективному направлению
фундаментальной и клинической ангиологии -
терапевтическому ангиогенезу, редакция обра-
тилась к практикующим хирургам со следующи-
ми вопросами: «Удовлетворены ли Вы как врач-
клиницист имеющимся арсеналом хирургических и фармацевтических средств лечения поражений
сосудов нижних конечностей? Какие пути до-
стижения прогресса в этой области - продления
жизни пациентов, уменьшения инвалидизации,
повышения качества жизни Вы видите и како-
во среди них место методов «терапевтического
ангиогенеза»?
Genes & Cells. 2011;6(3):7-12



Interv'yu s Prezidentom Rossiyskogo obshchestvaangiologov i sosudistykh khirurgovakademikom RAMN professoromAnatoliem Vladimirovichem Pokrovskim
Abstract
Глубокоуважаемый Анатолий Владимиро-
вич! Основную аудиторию нашего журнала со-
ставляют клеточные биологи, гистологи, био-
технологи и др. Для них многие клинические
дисциплины, в том числе и сосудистая хирургия,
остаются terra incognita. Но наш спецвыпуск по-
священ проблемам терапевтического ангиогене-
за, биотехнологическим способам воздействия
на восстановление сосудистой
сети. Скажите, пожалуйста,
как развивалась ангиология
и сосудистая хирургия, какие
проблемы здравоохране-
ния решаются Вами, есть ли
в практике ангиолога место
генным и клеточным техно-
логиям?
Genes & Cells. 2011;6(3):13-14



Cell-based therapy for peripheral arterial diseases
Abstract
The number of peripheral artery disease remains in the
world. Search for new methods of treatment is to improve
the condition of the distal blood stream, reduce peripheral
resistance and stimulating capillary network. One possible
solution to this problem is to use cellular technologies stimulate
angiogenesis. A new promising treatments is autologous cell
therapy using BMMNC or PBMNC. This overview focuses on
the exploration of autologous cell therapy in treating diseases
of the peripheral arteries.
Genes & Cells. 2011;6(3):15-23



Some possible molecular mechanisms of VEGF encoding plasmids functioning
Abstract
Gene therapeutic approaches to the restoration of the
ischemic tissue perfusion are considered very promising,
but to this time the molecular mechanisms which allow the
therapeutic gene encoding plasmid to transfect the target
cell and underlie the positive clinical effects remain unknown.
In this review the possible molecular mechanisms of the
angiogenic factor VEGF encoding plasmid penetration into the
cytoplasm and the nucleus of the target cell are discussed,
and also the methods for better transfection and the gene of
interest expression are proposed.
Genes & Cells. 2011;6(3):24-28



Transfer of recombinant nucleic acids into cells (transfection) by means of histones and other nuclear proteins
Abstract
Currently a protein/peptide-mediated gene delivery has
been considered a promising approach in non-viral gene
transfer. The previous investigations have shown that histones
and other nuclear proteins might be effective vectors for gene
transfer into cells. Transfection of eukaryotic cells by nucleic
acid and histone complexes (histonefection) effectively occurs
with various histone proteins. The presence of DNA-binding
domains and specific signal sequences of nuclear location
allows to use histones (Н1/Н5, Н2А, Н2В, Н3, Н4) and other
nuclear proteins (such as HMG family proteins and histonelike
prokaryotic proteins) for recombinant genes transfer.
The positive charge of histone protein molecules enables
electrostatic interaction with negatively charged molecules
of nucleic acids and charge neutralization that facilitates
the complexes penetration through a negatively charged
cell membrane. Thus, histonefection is a promising method
for non-viral transfer of recombinant nucleic acids in gene
therapy.
Genes & Cells. 2011;6(3):29-40



Search of an optimum variant of cell therapy of a critical limbs ischemia in experiment
Abstract
The search of new way of cell injection that can provide
maximal angiogenesis in ischemic limbs is urgent aim of
investigation for the right way of cell delivery other things
being equal is the main request for cell therapy efficiency.
On experimental model of critical ischemia (by angiographic
method and evaluation of capillary density on histological
sections) it was shown that cell therapy independently from
the way of cell injection (intramuscularly, intra-arterialy and
combined) induces angiogenesis, the rate and intensity of
which depend on the way of cell delivery. The optimum variant
of cell therapy of critical ischemia is combined (intramusculararterial)
method of cell injection, uniting the advantages of
intramuscular and intra-arterial ways.
Genes & Cells. 2011;6(3):41-45



Age effects on angiogenic properties of adipose tissue mesenchymal stem cells
Abstract
Мезенхимальные стволовые клетки жировой ткани
(МСК-ЖТ) способны стимулировать ангиогенез посред-
ством продукции факторов роста и стабилизации растущих
сосудов. МСК-ЖТ являются перспективным материалом
для аутологичной клеточной терапии. Однако с возрас-
том пациентов активность их МСК-ЖТ, а, следовательно, и
эффективность клеточной терапии могут снижаться. Целью
нашей работы было оценить ангиогенные свойства МСК-ЖТ
при старении.
Материал и методы. МСК-ЖТ были выделены из под-
кожной жировой ткани мышей линии BalB/с возраста 1-2,
12, 18 и 24 мес., а также 12 доноров трех возрастных
групп (12 (5; 12) лет, 45 (41; 45) лет и 65 (59; 76) лет).
МСК-ЖТ 2-го пассажа помещали на 48 ч в условия 1%
или 20% содержания кислорода. Оценивали способность
клеток стимулировать ангиогенез in vitro и in vivo.
Результаты. Способность МСК-ЖТ мышей стимулиро-
вать формирование капилляроподобных структур in vitro и
васкуляризацию подкожных имплантатов Матригеля in vivo
с возрастом снижалась в среднем на 60%. Содержание
мРНК сосудистого эндотелиального фактора роста (VEGF)
и плацентарного фактора роста (PlGF) в этих клетках сни-
жалось, а фактора роста гепатоцитов (HGF) и компонентов
системы внеклеточного протеолиза (рецептор к урокиназе,
металлопротеиназы 2 и 9 типов, ингибитор активатора
плазминогена-1) - возрастало с возрастом животных. Сти-
муляция экспрессии VEGF, PlGF и HGF в условиях гипоксии
была выражена слабее в случае МСК-ЖТ старых живот-
ных. Экспрессия PlGF и ангиопоэтина-1, а также секреция
VEGF и HGF МСК-ЖТ человека отрицательно коррелирова-
ли с возрастом донора.
Таким образом, при старении способность МСК-ЖТ сти-
мулировать ангиогенез снижается в результате подавления
экспрессии ключевых ангиогенных факторов.
Genes & Cells. 2011;6(3):48-57



Using autologous bone marrow cells to improve myocardial perfusion in surgical treatmentof patients with valvular defect heart
Abstract
The paper presents the results of a pilot clinical study of
the use of injection intramiokardial mononuclear fraction of
cells autologous bone marrow as an additional procedure to
the standard surgical treatment in 10 patients of heart valve
defects in order to assess opportunities to improve blood flow
in areas of severe hypoperfusion and myocardial scarring.
In terms of monitoring of patients to 72 months after cell
transplantation demonstrated the safety of this treatment,
the improvement of regional perfusion and myocardial
contractility in the areas of injection autologous mononuclear
fraction of bone marrow cells.
Genes & Cells. 2011;6(3):60-66



Stimulation of rats sciatic nerve post-traumatic regeneration using plasmids expressing vascular endothelialgrowth factor and basic fibroblast growth factor
Abstract
The development of effective treatments for patients
with peripheral nerve injury is an urgent task of biomedicine.
«Gold» standard in restoring the integrity of nerve conduits is
auto-nerve transplantation in which a peripheral nerve defect
is corrected with autologous nerve graft. Here we propose
a method for stimulating revascularization and regeneration
of auto-nerve graft by a local injection of plasmid pBud-VEGFFGF2,
expressing vascular endothelial growth factor (VEGF)
and basic fibroblast growth factor (FGF2). It is shown that
direct injection of plasmid pBud-VEGF-FGF2 in the proximal
and distal segments of nerve, as well as in the auto-nerve
graft, stimulates the regeneration of the rats sciatic nerve
and restores motor activity.
Genes & Cells. 2011;6(3):67-70



Effect of G-CSF on proangiogenic properties mobilized peripheral blood cells in patients with chronic heart failure
Abstract
The aim is to study the phenotypic characteristics and
cytokine-producing properties mobilized drug administration
G-CSF from bone marrow mononuclear cells of peripheral
blood of patients with heart failure, which developed after
acute myocardial infarction, in connection with the efficiency
of intramyocardial stem cell therapy. The study included 67
patients with CHD and III-IV functional class congestive
heart failure (NYHA), receiving current standard therapy.
Shown that the introduction of the drug G-CSF results in
mobilization of endothelial progenitor cells (EPC) from bone
marrow into peripheral blood (CD34+/CD133+and CD34+/
KDR+populations). Intramyocardial cell injection resulted in
improved perfusion at the injection site in 76% of patients.
Patients who responded to improved perfusion, the number of
CD34+CD133+PC in 3,2 times higher than in patients without
effect or impairment. Mononuclear cells after administration
of G-CSF in a 48 hour culture secrete cytokines, Epo,
GM-CSF, TNF-, contribute to the improvement of myocardial
perfusion. Peripheral blood is a readily available source of EPС,
and mononuclear cells after mobilization are able to exert
reparative effects on the ischemic myocardium.
Genes & Cells. 2011;6(3):71-75



Efficacy and safety of application «Neovasculgen» in the complex treatment patients with chronic lower limbischemia (IIb-III phase of clinical trials)
Abstract
The article deals with the results of a 6 month
comparative follow-up of non-resectable patients with
lower limb atherosclerosis having clinical symptoms as
claudication (IIa-III stages according to the Fontaine-
Pokrovsky classification) who received Neovasculgen
as a part of conservative therapy. Neovasculgen, a gene
therapy drug, is a plasmid construction with a vascular
endothelial growth factor gene. 100 patients were enrolled
into the study: 75 patients comprised the clinical group,
while 25 ones being the controls. The patients underwent
treatment in three clinical centers. The drug was injected
intramuscular, maximally close to ischemia zones, twice in
doses of 1.2 mg with the interval of 14 days. The following
factors were controlled: pain free walking distance (PlWD),
ankle-brachial index (ABI), transcutaneous partial pressure
of oxygen (TcPO2), linear blood flow rate (LBFR) in the
involved segment, angiography; also the integral treatment
result on the scale «success/failure», quality of life SF-36
questionnaire were assessed. The study duration was 6
months. PlWD was determined to increase by 110% that
shows statistically significant difference versus the control
(P=0.000).
Genes & Cells. 2011;6(3):76-83



Complex therapy of patients with chronic lower limb ischemia by angiogenesis gene inductors:immediate and long-term results
Abstract
The controlled study results of using therapeutic
angiogenesis in multimodality therapy of patients with chronic
lower limb ischemia (CLLI) are presented. 134 patients (74
ones comprised the target group and 60 patients were the
controls) with CLLI IIb-III stages according to the Fontaine-
Pokrovsky classification were included into the study.
Angiogenesis stimulation was used both as a separate
treatment modality in combination with routine conservative
therapy and as an adjuvant to reconstructive vascular
surgery. It is proved that genetically engineered angiogenesis
stimulators can be used in a multimodality therapy of patients
with CLLI, demonstrating satisfactory tolerance and safety.
Therapeutic angiogenesis technologies have been shown to
be applied as an independent therapy and as in combination
with reconstructive vascular surgery to improve a long-term
outcome of the latter.
Genes & Cells. 2011;6(3):84-88



Efficacy of therapeutic angiogenesis in patients with chronic lower limb ischemia
Abstract
Therapeutic angiogenesis is a promising treatment
modality in patients with chronic lower limb ischemia, for
whom revascularization is not indicated. A number of studies
demonstrated satisfactory therapeutic effects of gene and
cell-based therapy targeted to stimulate angiogenesis. At the
same time the factors affecting the efficacy of angiogenesis
stimulation in clinical settings are under-investigated.
In the given study 30 patients, aged 49-78 years with
chronic lower limb ischemia of atherosclerotic origin (Fontaine
IIb-IV) were randomized into groups of gene therapy (GT) or
the control group (C). Plasmid DNA containing an encoding
part of the vascular endothelial growth factor (VEGF-165)
gene was inserted into ischemic limb muscles of the GT
patients. All the patients had sessions of treadmill training
walking. Besides clinical findings, angiography, scintigraphy,
transcutaneous oxymetry, laser Doppler examination, duplex
ultrasonography, treadmill test were carried out to evaluate
the invention efficacy. Also circulating progenitor cell count
was assessed by flow cytometry, ELISA was used to evaluate
angiogenic growth factors. In 3 months limb perfusion
rate, painless walking distance, collateral circulation indices
increased. In 50% of patients the therapeutic effect was
assessed as moderate (Rutherford +2), in 30% as minimal
(Rutherford +1), 20% of patients showed no dynamics.
Clinical laboratory tests had no significant dynamics in the
group of standard therapy. No serious complications were
noted. In a correlation test the efficacy of angiogenic therapy
was associated with a patent ileac segment, shorter smoking
history and higher ankle-brachial index.
Genes & Cells. 2011;6(3):89-98


