FIZIOLOGIA (PHYSIOLOGY) 2006; vol. 16, nr. 1(49)
CONTENTS
1. EDITORIAL: Regenerative medicine – a challenging domain of research in XXIst century
Carmen Bunu
2. Romanian scientific progress in international stress research and stress-aging medicine
Petru Derevenco
3. Changes of oxidative stress markers and antioxidant status after administration of vitamin E andrespectively soy isoflavone in female rats with experimentally induced menopause
Lavinia Sabau, Soimita Suciu, Adriana Muresan, Doina Daicoviciu, Simona Clichici, David Ion,
Doina Colcear
4. Microfractures in full thickness cartilage of the knee
RF Tatu, Carmen Tatu, R Brediceanu
5. Abstracts presented at the workshop: New structures and biomaterials for life and health. Trends
and priorities in European research area – Timisoara, April 14, 2006
1. EDITORIAL: REGENERATIVE MEDICINE – A CHALLENGING DOMAIN OF
RESEARCH IN XXIST CENTURY
Carmen Bunu
Department of Physiology, University of Medicine and Pharmacy Victor Babes Timisoara
Address for correspondence: Dr. Carmen Bunu, Department of physiology, “Victor Babes” University of Medicine andPharmacy Timisoara, 2, Eftimie Murgu Sqare, 300041, Timisoara, e-mail: cbunu@umft.roRegenerative medicine stands at the intersection of a variety of rapidly developing scientificdisciplines: stem cell biology, immunology, tissue engineering, molecular biology, biomaterials,
transplantation biology, and clinical research. Patients suffering from diseased and injured organs
may be treated with transplanted organs. When organs or tissues are irreversibly injured, these could
be replaced by artificial systems or by a similar organ from a donor. There is a severe shortage ofdonor organs which is worsening yearly due to the aging population. In addition to providing hopefor the ageing and diseased, tissue engineering is already helping the large population of young
people who participate in sports, many of whom suffer injuries. The development of some methodsof tissue engineering which allow the replacement of organs through ex vivo culture of autologus
cells, could represent a valid substitute for organ transplant, thus peventing the risks of transmittingsome diseases from donors, a common problem in organ transplant.
Tissue engineering using cells, especialy stem cells, allows exploiting two important qualitiesof these cells: high proliferation capacity and multipotentiality of these imature cells. The new fieldof regenerative medicine is one of the most exciting fields in medicine and brings together for the
first time the scientific disciplines of biology, materials science and biomedical engineering directed
towards long-term repair and replacement of failing human tissues and organs. Living cells are
harvested, grown in the laboratory, often on an appropriate scaffold, and stimulated to form specifictissues that mimic the complex structures and physiological behaviour of natural tissues. Ultimately,"spare parts" developed in the laboratory can then be put into a patient, either by injection of cells orby implantation of intact tissue or an entire organ. The stem cell field is advancing rapidly, opening
new options for therapy in tissue engineering.
The goal of regenerative medicine is to repair organ pathologies such as those acquiredcongenitally or by cancer, trauma, infection, or inflammation. It is based upon the foundations of celltransplantation and materials science. The interest in modern biological technologies has
dramatically increased since it is feasible to isolate living, healthy cells from the body, expand themunder cell culture conditions, combine them with biocompatible carrier materials and re-transplantthem into patients. Therefore, tissue engineering gives the opportunity to generate living substitutes
for tissues and organs, which may overcome the drawbacks of classical tissue reconstruction: lacking
quality and quantity of autologous grafts, immunogenicity of allogenic grafts and loosening ofalloplastic implants. Due to the prerequisite for tissue engineering to ensure a sufficient number oftissue specific cells without donor site morbidity, much attention has been drawn to multipotential
progenitor cells such as embryonic stem cells, periosteal cells and mesenchymal stem cells [19].
According to Hipp J. and Atala A. [6], tissue can be engineered 1) in vivo- by stimulating the body's
own regeneration response with the appropriate biomaterial, or 2) ex vivo- cells can be expanded inculture, attached to a scaffold and then re-implanted into the host. Cells may be heterologous(different species), allogeneic (same species, different individual), or autologous (same individual).
Autologous cells are preferred because they will not evoke an immunologic response and thus thedeleterious side effects of immunosuppressive agents can be avoided. While autologous cells arerecognized as the ideal transplantation resource, many patients with end-stage organ disease areunable to yield sufficient cells for expansion and transplantation. Furthermore, some primary
autologous human cells cannot be expanded from particular organs (i.e. pancreas, liver).
Stem cells are envisioned as being an alternate source of cells from which the desired tissue
can be derived. Human embryonic stem cells (HESC) can be derived from discarded non transferred
embryos and have the advantage of being pluripotent (the ability to differentiate into all tissues of theembryo) and able to self-renew indefinitely. However, their clinical application is limited because
they represent an allogeneic resource and thus their use would require high dose immunosuppressive
therapy.
The hope that many diseases can someday be treated with stem cell therapy is inspired by thehistorical success of bone marrow transplants in increasing the survival of patients with leukemia andother cancers, inherited blood disorders, and diseases of the immune system [25]. Nearly 40 years
ago, the cell type responsible for those successes was identified as the hematopoietic stem cell [26].
The ability of hematopoietic stem cells (HSCs) to self-renew continuously in the marrow and todifferentiate into the full complement of cell types found in blood qualifies them as the premier adultstem cells. HSCs are among the few stem cells to be isolated in adult humans. They reside in thebone marrow and under some conditions migrate to other tissues through the blood. HSCs are also
normally found in the fetal liver and spleen and in umbilical cord and placenta blood. There is a
growing body of evidence that HSCs are plastic—that, at least under some circumstances, they areable to participate in the generation of tissues other than those of the blood system. A few studies
have shown that HSCs can give rise to liver cells [11, 24]. It was unexpected that a component ofblood could cross over a developmental separation to form a tissue type that ordinarily has acompletely different embryonic origin [11]. The findings noted above and other reports of cardiac
and muscle tissue formation after bone marrow transplantation in mice [17] and of the developmentof neuron-like cells from bone marrow [2, 6] have raised expectations that HSCs will eventually beshown to be able to give rise to multiple cell types from all three germ layers. One study has, in fact,demonstrated that a single HSC transplanted into an irradiated mouse generated not only blood
components (from the mesoderm layer of the embryo), but also epithelial cells in the lungs, gut,
(endoderm layer) and skin (ectoderm layer) [10]. There is also evidence that transplants derived fromumbilical cord blood are less likely to provoke graft versus host disease, possibly because the cells incord blood are immature and less reactive immunologically [12]. The quantity of HSCs present incord blood and its attached placenta is small, and transplants from cord blood take longer to graft, but
for children, whose smaller bodies require fewer HSCs, cord blood transplants are valuable,
especially when there is no related sibling to donate HSCs [4]. Banks of frozen umbilical cord andplacenta blood (drawn out of the umbilical vein of the cord) are an important source of HSCs
because the histocompatibility markers on the cells in these tissues can be identified and cataloguedin advance of the need for a transplant. If HSCs are truly multipotent, their potential for life-saving
regenerative therapies may be considerably expanded in the future.
Human mesenchymal stem cells (MSCs) - also known as stromal cells - are also used for celltherapies in different conditions associated with organs and tissues disorders. Multipotent
mesenchymal stromal cells are endowed with multilineage differentiative potential andimmunomodulatory properties. It is still a matter of debate whether donor MSCs have sustained
engraftment potential in host bone marrow after allogeneic hematopoietic stem cell transplantation[18]. There are multiple sources of MSCs, some of them common to HSCs sources: bone marrow,peripheral blood adipose tissue, umbilical cord and umbilical cord blood, etc. They are known todifferentiate into at least three cell lineages: osteocyte, adipocyte and chondrocyte, but are likely tobe able to transdifferentiate and dedifferentiate into much more cell types, required for cell basedtherapies. Their potential is still under investigation and the possibilities for differentiation into
different cell types seem to be limitless. Even if the origin and the functions of MSCs are still
unclear, their presence in the adult peripheral blood might relate to adult stem cell biology, such as
the mechanism of systemic migration of bone marrow-derived multi-potent stromal cells and theexistence of a common hematopoietic-mesenchymal precursors.
Specific therapeutic applications of tissue engineering research include, but are not limited to:• Novel cell delivery models and approaches, including delivery of cells in scaffolds to promotehealing for repair, replacement or regeneration of tissues;• Development of scaffolds with appropriate characteristics to promote cell and tissue survival andintegration;• Development of novel animal and culture models for regenerative medicine applications,
including innovative models of acute and chronic injury, aging models, organ cultures and coculture systems;• Molecular and biochemical basis of vascularization and angiogenesis in native and exogenouslytransplanted tissues and organs;• Approaches to minimize cell death and promote cell survival and differentiation in transplants;• Application of tissue-engineered biomaterials as conduits or shunts in tissue regeneration;• Development of important new insights into "normal" structure, function and/or development oftissue and organ systems of interest;• Development of effective new strategies for improving healing, repair, biological replacement orregeneration of tissue and organ systems of interest;• Ethical, legal, social, cultural and economic consequences of regenerative medicine based on
tissue engineering strategies.
There are several teams involved in the study of stem cells and their therapeutic potential.
Their reports were published in acknowledged speciality journals (Nature, Science, PNAS, Blood,
Stem Cells, NEJM, etc.) and are still the most challenging domain in medical research all over theworld.
Cell therapy as base of regenerative medicine can address a number of human conditions
from chronic to life threatening: organ transplant, Parkinson’s disease, Alzheimer’s, arthritis,
osteoporosis, emphysema (smoker’s lung), liver cirrhosis, diabetes, heart disease, trauma, sports
injuries:• Heart diseases: Minguell JJ. and co [15] revealed the benefits of regenerative medicine in heartdiseases. Most attempts for cardiomyoplasty have considered the bone marrow as the source ofthe repair stem cell(s), assuming that the hematopoietic stem cell can be a choice. Since 1995 it
has been known that under in vitro conditions, MSCs differentiate into cells exhibiting features ofcardiomyocytes. Freyman T. [3] proved that intracoronary and endocardial injection of MSCs
post- myocardial infarction resulted in increased engraftment within infarcted tissue whencompared with intravenous infusion. Nagaya N and co [16] showed that MSC transplantation
improved cardiac function in a rat model of dilated cardiomyopathy, possibly through inductionof myogenesis and angiogenesis, as well as by inhibition of myocardial fibrosis.
• Bone and cartilage disorders: In the ortopedic field it was demonstrated the posibility ofautologus human condrocytes or mezenchymal stem cells transplant in order to reconstruct the
injured cartilage, as a result of trauma or some pathologic entities, and to enable the new bone
tissue formation in treatment of skeletal anomalies, trauma and bone tumors. There is aconceptual change regarding hard tissues repair, in the way that there is no need to find materials
that could replace the destroyed part of the bone, but we need to find materials that could help theorganism to the process of auto-repair of the damaged tissues, such as bioactive materials of thethird generation [7].
• Diabetes mellitus: Beta-cell replacement therapy via islet transplantation has received renewedinterest due to the recent improved success. In order to make such a therapy available, newsources of insulin-producing cells must be readily available. The most promising sources arestem cells, with efforts of deriving new beta-cells from both embryonic and adult stem cells [21].
Under strong induction of pancreas regeneration, pancreatic duct cells dedifferentiate to grow,and re-differentiate toward other cell types including islet cells [23].

•

•

•

•
•

Neurodegenerative diseases: New cell therapies will generate different types of neural cells forthe treatment of degenerative diseases such as Alzheimer's and Parkinson's. It would be possiblethat spinal cord injury patients may regain full function of their body. Even some genetic
disorders could be cured, not just treated [27].
Kidney diseases: Stem cells offer an exciting potential for kidney regeneration. Lin F. [13]revealed evidences to support the conclusion that intra-renal cells, including surviving tubularepithelial cells and potential renal stem/progenitor cells, are the main source for renal
regeneration. Future research will be focussed in selecting the type of stem cells, frequency androute of administration of the cells will be fundamental in successful cell replacement therapy inacute renal failure. Recent studies indicate the possibility of collecting autologous bladder cells
from human patients, expanding them in culture, and returning them to the human donor in
sufficient quantities for reconstructive purposes.
Muscle diseases: According to Bach AD [1] there are two principle strategies for the
replacement of impaired muscle tissues: (1) the application of isolated and differentiated cells (invivo tissue engineering), using a transport matrix for the cell delivery; (2) use of in vitrodesigned and pre-fabricated tissue equivalents (in vitro tissue engineering). Duchenne muscular
dystrophy can be treated using bone marrow-derived human MSCs genetically modified with thefull-length Dys-coding sequence [5].
Transplant: MSCs can be used in novel therapeutic strategies designed to improve engraftment
or to suppress graft-versus-host disease in bone marrow transplantation, due to the fact that MSCs
can inhibit NK-cell proliferation [22]Cancer: Both human stem cells and mature stromal cells can play an important role in thedevelopment and growth of human malignancies. In contrast to these tumour-promoting
properties, human MSCs home to sites of tumour-genesis and potently inhibit tumour growth.
MSCs possess intrinsic antineoplastic properties and might be of particular utility for treating
those human malignancies characterized by dysregulated Akt, as Kaposi's sarcoma [9].

The research on cell therapies in Romania: the enormous potential of stem cell research, both innovel therapeutics and in understanding the basic biology of malignancy, has driven some Romanian
research units to initiate basic and clinical research in this field and to provide collaborativeinteractions with other scientists. The most advanced are the following:• The University of Medicine and Pharmacy Victor Babes Timisoara (www.umft.ro) has as
strategy in research to become an excellence centre in stem cell therapies till 2007. It is theleading Regional Centre specialized in stem cell therapies, with an existing Stem Cell Cluster of
9 departments (immunology and physiology, histology, biochemistry, biophysics, haematology,
cardiology, paediatrics, orthopaedics, experimental microsurgery), organized around the
Immune-physiology and Biotechnologies Centre, collaborating on a range of research projects
involving stem cells of adult origin. The University has well equipped facilities for basic
research, as well as for experimental studies and clinical trials and well-trained researchers
involved in the field. It is the first university in Romania who began a clinical trial using stem
cells (autologous transplantation of AC133+ bone marrow-derived stem cells in patients withacute and recent myocardial infarction), and participates to a FP6 integrated project in the field ofstem cell research (GENOSTEM, LSHB 1.1-503161). The university is already partner in two
European projects regarding Quality Management Systems, financed by EU Public Health
Programme, Luxemburg (EU-QMS-BLOODNET and European Quality System for Tissue
Banking - no 2003209). The mobility within EU is sustained by bilateral agreements withrecognized EU universities (Frankfurt, Heidelberg, Montpellier), by Leonardo da Vinci
Mobilities Programme, and by the training programme developed by GENOSTEM.
• The University of Medicine and Pharmacy Carol Davila Bucuresti (www.univermedcdgm.ro) is specialized in the in vitro study of embryonic and human adult stem cell, with strong
collaborative links with Karolinska Institute in this field and just won a Romanian LSH-SSA
within the specific call for ACC/2004 in rheumatic disease study.
• The Institute Nicolae Simionescu Bucharest (www.icbp.ro) is an Excellence Center ofRomanian Academy, specialized in the in vitro study of stem cells and its applications in cardiovascular diseases, mainly in endothelial cells dysfunctions. The Institute has many national grants

•in this field, and participates to FP5 as well as FP6 projects and is also recognized as EU
Excellence Center. The Institute just won the other Romanian LSH-SSA within the specific callfor ACC/2004 - SERA. The fundamental concept of SERA is to act as a support for
reinforcement of our team and our country research capacity. The work is financially supportedby the European Community, in the framework of “Integrating and Strengthening the European
Research Area” – Specific Support Action. The activities are targeted to the Thematic Priority 1of European Community FP6.
The team of the Clinical Institute Fundeni (www.icfundeni.ro) under the lead of Prof. Irinel
Popescu has many accomplishments and national premiers in surgical techniques. The mostimportant of these is the introduction of the liver transplant in Romania, which is a very complex
and laborious surgical procedure. Another accomplishment at the Clinical Institute Fundeni wasin November 2005 when the first stem cell transplant for a liver disease has been performed. The
stem cells have been isolated and prepared at the Cambridge University in the United Kingdom.
This intervention is the 6th out of 25 within a study performed by the Hammersmith Hospital,
London and the Cambridge University.

The current state of the art for stem cell research seeks to: (1) explore adult stem cells in nonhomologous settings, such as the use of HSCs in heart repair; (2) exploit further sources of stem
cells, notably embryonic stem cells, for the treatment of paediatric, heart, pancreatic, liver and brain
impairments; (3) use stem cell lines as tools in drug discovery and development; (4) increase theunderstanding and treatment of cancer by further studies of endogenous adult stem cells; (5) generate
embryonic stem cells with the same nuclear genetic material to that of the patient using therapeuticcloning techniques, to avoid the potential rejection of cell therapies. Another ambitious goal for thefield involves the use of endogenous stem cells, naturally resident in tissues of the human body, todirect the repair of damaged or diseased tissues. In all of the above examples, timescales are
unknown and merely indicative [20].
In this regard, regenerative medicine may become a successful strategy by providing a dynamic,
interactive, and individualized therapeutic approach that responds to the pathophysiologicalcondition of the patient. Humes H.D. [8] revealed the fact that most common disease processes are
not due to the deficiency of a single protein but develop due to alterations in the complex interactionsof a variety of cell components. Cells may provide innovative methods for drug delivery of biologics,
immunotherapy, and tissue regenerative or replacement engineering. The translation of this discipline
to medical practice has tremendous potential, but in many applications technological issues need to
be overcome. Since many cell-based indications are already being evaluated in the clinic, the fieldappears to be on the threshold of a number of successes. Collaborations between biotechnology andpharmaceutical companies with academic institutes and research centres will help overcome thetechnological challenges associated with the development of the stem cell-based therapies. For
pharmaceutical companies, stem cells contribute to drug discovery through their application infinding novel drug targets and through the development of new technology platforms. Therefore,
pharmaceutical companies will increasingly explore the different methods in which stem cells can beused in the drug discovery phase to accelerate the discovery of novel drug molecules. In time,regenerative medicine will also provide a cost-effective, long-term solution to many age-related andlethal conditions.
2. ROMANIAN SCIENTIFIC PROGRESS IN INTERNATIONAL STRESS RESEARCH
AND STRESS - AGING MEDICINE
Petru Derevenco
Romanian Academy of Medical Sciences, Cluj-Napoca, Romania
ABSTRACTThe aim of this paper is to reveal the Romanian scientific progress in stress research and stress-aging
medicine made by the Department of Stress Research and Prophylaxis, headed by Sorin Riga MD,
PhD and Dan Riga, MD, PhD, at the "Al Obregia" Clinical Hospital of Psychiatry, Bucharest. Our
synthesis is justified because the international publications and presentations of Drs Riga's team areinsufficiently well-known in Romania, compared to their significance and impact in bio-medicine
and therapy.
In the background section we describe the stress globalization phenomenon, which is a major
concern of the United Nations specialized structures: WHO, ILO, UNESCO. Advances in stressmedicine and research reveal the quick progress and main directions in this field. Moreover, thepaper includes the long-term contribution (1980-2005) made by the Academy of Medical Sciences
and the University of Medicine from Cluj-Napoca, Romania.
In addition we characterized the new Romanian concepts and excellence in stress-aging research,bio-medicine, and therapeutics, ellaborated by Drs Riga's team, from 1972 until nowadays. Theirconcepts and research work in the field of neuroscience and anti-stress and anti-aging medicine areincluded in the diagnosis-therapy couple, applied at neuro-psycho-biologic level:• anti-adaptive, anti-homeostatic and etio-pathogenic concept: distress ↔ impairment ↔ senescence↔ disease; and• homeostatic and orthomolecular therapy: anti-stress, anti-impairment and anti-aging.
Finally, we emphasized the international recognition of Romanian excellence in science andmedicine owed to Drs Riga's department. Moreover, due to the medico-social actuality and important
impact of stress and senescence, we underline the need to develop this department and its researches.
Key words: stress-aging medicine and therapy; Romanian scientific researches and progress;
Department of Stress Research and Prophylaxis, "Al Obregia" Clinical Hospital of Psychiatry
Bucharest.
3. CHANGES OF SOME OXIDATIVE STRESS MARKERS AND ANTIOXIDANT STATUS
AFTER THE ADMINISTRATION OF VITAMIN E AND RESPECTIVELY SOY
ISOFLAVONE IN FEMALE RATS WITH EXPERIMENTALLY INDUCED MENOPAUSE
Lavinia Sabau, Suciu Soimita, Adriana Muresan, Doina Daicoviciu, Clichici Simona, David Ion,
DoinaColcear
Department of Physiology, University of Medicine and Pharmacy ”Iuliu Hatieganu”, Cluj-Napoca,
Romania.
Address for correspondence: Dr. Lavinia Sabau, Department of Physiology, “Iuliu Hatieganu” University of Medicine
and Pharmacy Cluj-Napoca, Romania, Clinicilor Street

ABSTRACTThe aim of our study was to determine the effect of vitamin E and respectively soy isoflavone onserum levels of reactive oxygen species, antioxidants and nitric oxide in female rats withexperimentally induced menopause. We found that an increased serum level of reactive oxygen
species and decreased levels of antioxidants and nitric oxide accompanies experimentally induced
menopause in rats. Soy isoflavone reduces significantly the level of free radicals in these rats.
Vitamin E has significantly higher antioxidant effect than has soy isoflavone. Both vitamin E and soy
isoflavone has antioxidant protective effects in ovariectomized rats, but the effect of vitamin E isstronger than that of soy isoflavone.
Key words: oxidative stress, menopause, isoflavone, vitamin E.
4. MICROFRACTURES IN FULL THICKNES CARTILAGE DEFECTS OF THE KNEE
R.F.Tatu1 , Carmen Tatu2, R. Bredicianu1
1 st
I Department of Orthopaedics and Traumatology,2
Department of Physiology and Immunology „Victor Babes” University of Medicine and Pharmacy
Timisoara
Address for correspondence: Dr. Romulus Fabian Tatu, Ist Department of Orthopaedics and Traumatology, “Victor
Babes” University of Medicine and Pharmacy Timisoara, Bd. Prof. Dr. Iosif Bulbuca, 300736 Timisoara, e-mail:tatuf@xnet.roABSTRACT
Cartilage lesions in the knee are still awaiting the optimal treatment method. Several techniques,
starting with the old Pridie drilling and ending with themodern and expensive autologus chondrocite
transplant (ACT) are not bringing the optimal results. Based on the stem cell potentials, themicrofracture technique brings them into the defect region realising a compromise between theacceptable functional result and low costs especially in the light of the new studies which areestablishing no difference between ACT and microfracture.
Key words: cartilage lesion, microfracture, autologus cartilage transplant
5. ABSTRACTS PRESENTED AT THE WORKSHOP: NEW STRUCTURES AND
BIOMATERIALS FOR LIFE AND HEALTH. TRENDS AND PRIORITIES IN THE
EUROPEAN RESEARCH AREA
ORGANIZERS:
Victor Babes University of Medicine and Pharmacy Timişoara
National R&D Programme New Materials, Micro and Nanotechnologies – MATNANTECH
Politechnics University Bucharest
April 14, 2006
1. ANGIOGENIC GENE THERAPY IN PATIENTS WITH SEVERE LOWER LIMBISCHAEMIA
A. Andrei, B. Mut-Vitcu, M. Cătălin, E. Şeclăman, Şt. I. Drăgulescu“Victor Babeş” University of Medicine and Pharmacy, Timişoara
Institute for Cardiovascular Medicine, Timişoara
ABSTRACTThe ongoing study focuses on the application of a therapeutical strategy in patients with chronic
severe lower limbs ischaemia by means of plasmid vector encoding VEGF165 gene. The
methodology for angiogenic gene therapy administration has been improved. A 1-year follow-up
study in patients who underwent therapeutic angiogenesis will clinically assess its efficiency as wellas the safety profile.
As part of this study, it has been verified the hypothesis that proposes that injection of a genetic
material able to induce secretion of the angiogenic growth factors (VEGF165) into the criticallyischemic lower limb will promote neovascularization with more efficient blood perfusion of theaffected limb. This result is clinically demonstrated by disappearance of rest pain, healing of
ischaemic ulcers and avoidance of amputation. The primary end-point has been the complete relief ofrest pain and/or healing of ischaemic ulcers. Secondary outcomes have been the prolongation ofwalking time on the rolling carpet assessed by Gardner protocol, prevention of limb amputation,
angiographic demonstration of newly visible collateral vessels as well as the safety of treatment(pathological angiogenesis will be evaluated: proliferative retinopathy, tumour growth, progressionof atherosclerosis).
In the first stage, the vector has been designed to encode the gene of interest. Its purified form has
been synthesized in sufficient quantity. VEGF165 mRNA has been isolated from human blood andused in complementary DNA synthesis by means of RT-PCR technique using random hexamer
primers. VEGF gene amplification has been performed by PCR with specific oligonucleotide primers
for the cDNA template. VEGF gene insert has been ligated into the plasmid mammalian expression
vector – pCMV-Script Vector (Stratagene), and further cloned into E. coli strains.
E. coli colonies which incorporated the plasmid encoding VEGF have been isolated. The plasmid has
been purified by affinity chromatography. Thus, it has been obtained the ultrapurified plasmid vector
which can either be used immediately after suspension in saline or lyofilizated and stored at -80˚C.
The obtained product has been certified as regards its sterility and toxicity.
Up to now, there have been selected 3 patients with chronic ischaemia of the lower limbs (persistentrest pain and/or unhealed ischaemic ulcers) who cannot undergo interventional or surgicalrevascularization. For these patients, who are in so-called near-amputation stage, nowadays, there isno other therapeutical option but amputation. Patients received intramuscular injection of plasmid
encoding VEGF165 at 16 sites of the affected limb separated by a distance of 3-4 cm.
After therapy, the patients were further hospitalized for the observation of both the ischaemic
symptoms evolution and the possible side-effects of therapy (edema of the cured lower limb, fever,increased levels of acute phase reactants). The only side-effect of angiogenic therapy noticed in thesepatients was a temporary increase (within the first 6-10 hours) of the acute phase reactant levels,
which demonstrates the normal evolution after parenteral administration of the plasmid vectors,
resulted from successful penetration into the muscular tissue and the beginning of their local action.
After 1 month, the first 2 patients showed positive evolution with the settlement of rest pain, partial
revitalization of the lower limb’s cyanotic areas and a mean 5-minute walk distance of about 400 m.
This study is supported by a grant from the Academy of Medical Sciences – VIASAN Programme.
2. HEALTH PROMOTION IN ELDERLY PATIENTSElena Ardeleanu Elena, Daniela Gurgus, Răzvan Şuşan, Minodora Baaj“Victor Babes” University of Medicine and Pharmacy Timisoara
ABSTRACTThe purpose of the study was to analize the morbidity of this population and to apreciate the way
how elderly people benefit form screening tests and desease prevention.
Material and methods: The study was performed on a period of 4 years beginning in 2003 at the
Medical Family Departament in collaboration with 7 family medicine doctors, on 3208 patients withage ranging between 65 and 87 years. The study was based on the retrospective revue of the medical
recordings and to fill in a special form.
Results: Morbidity analisis: 706 pateints (22%) presented phisical sufferings, cerebral vasculardeseases 994(31%), utinary tract infections 451(14%), cardiovascular pathology 1605 (51%),dental
problemes 88%,ear and eye disorders 30%,osteophorosis 18%,rheumatic disease 70%.2791 patients(87%) have been instructed regarding the decrease of cardiovascular risc, periodic mesure of bloodpressure was performed in 7o%,for 1027 patients (32%) EKG was performed, 1860 (58%) ofpatients were subjects of glucose blood determination, 481 (15%) were submited to cholesterol
leaveles screening for a period less than three years and 1604 (50%) of these patients were instructed
about a properly administrated medication.Tests for early stages in prostathic cancer were performed
at 21% of men,mamography was prescribed in 0,8% of the wemen involved in this study and forcolorectal cancer were screened 5% of the patients.Eldery peoples,age ranging between 65-75 years
and high school graduated patients have particulary benefit by the majotity of prorphilactic actiones.
Conclusions:The preventive clinical services and screening tests are insufficient promoted on eldery
patients.Health improving and disease prevention stategies have to be implemented on a larger scaleby all factores liable for public health.
3. THE INCIDENCE OF Β HEMOLYTIC GROUP A STREPTOCOCCI FROMPHARYNGEAL SWABS
Delia Berceanu Văduva, Matilda Linţa, Marcela Adămuţ, F. Horhat, M. Berceanu Văduva, Dana
Velimirovici, Maria Rada, Roxana Moldovan“Victor Babes” University of Medicine and Pharmacy Timisoara
ABSTRACT
Purpose: Streptococci can produce primary infections with various locations, wich are severe due toinfective and postinfective complications. The most exposed persons are children, teenagers andyoung people from crowed communities.
In order to detect the incidence of β hemolytic group A streptococcus (Streptococcus pyogenes), in
the pharynx, 4607 pharyngeal swabs have been collected during 2005, in a private laboratory fromTimişoara.
Material and Method: After performing all the bacteriological stages, investigations have been
completed with ASO test (ASLO - serological test), to all patients clinically suspected, including

patients in which β hemolytic streptococcus group A was detected, as well as patients whith negative
swab results, but with typical simptoms.
After final lab results, we studied the incidence of poststreptococcal complications, which included 8
patients that have been identified during 2005.
Results: From 4607 collected and performed samples, 3593 (78%) had normal flora, and 1014
(22%) were positive.
From 1014 pathogen species, 117 (11,53%) strains were identified as β hemolytic group A
streptococci, 76 (7,5%) were β hemolytic group A streptococci in association with other germs, andthe rest of 821 (80,97%) were other pathogenic germs (most frecvent Staphilococcus aureus, othergroups of streptococci and gram-negative bacilli: Escherichia coli, Klebsiella spp., Proteus spp.,
Enterobacter spp., Pseudomonas aeruginosa etc).
Conclusions: This study detected a high number of β hemolytic group A streptococcus strains - 193(19,03%), responsible for upper respiratory tract infections. Prevention of spreading of β hemolytic
group A streptococcus infections needs a good sanitary education of the population.
4. CHANGING THE REMOVABLE PARTIAL DENTURE METALLIC FRAMEWORKTECHNOLOGY USING “LIGHT CURING” WAXES
Cristina Bortun¹, Liliana Sandu¹, Brandusa Ghiban²
1“Victor Babes” University of Medicine and Pharmacy Timisoara, University School of Dentistry,Specialization of Technical Dentistry2
Polytechnic University of Bucharest, Department of Material Science and Physical Metallurgy
ABSTRACTAIM OF STUDY: Changing the removable partial denture metallic framework technology is anachievable objective in order to reduce intermediary working stages and use new materials in wax upof metallic framework.
MATERIALS AND METHOD: Materials used for wax up were “LiWa” and “Ti Light” lightcuring waxes. Were conceived many removable partial denture design choices for different
topographical situations. Then, were achieved metallic framework wax patterns directly on theworking cast without duplicating and achievement of the refractory cast.
RESULTS: Wax up with “LiWa” is more difficult then that with “Ti Light”, first material being
more sticky, adherent to instruments more elastic. After light curing these materials, removal of wax
pattern from the cast is a delicate procedure because the pattern can crack (“Ti Light” is more
breakable then “LiWa”.
CONCLUSIONS: Wax up and light curing duration is 25 minutes, so it is more rapid then classic
technology, witch is achieved in several hours. In future, materials quality can be improved and theprocedure be applied currently.
5. THIN LAYERS FOR IMPROVING THE BIOCOMPATIBILITY OF IMPLANT
MATERIALS
M.Braic1, S.Ivanescu2, C.Trisca-Rusu3, V.Braic1, M.Balaceanu1, A.Kiss1, M. Valeanu4,
A.Vladescu1, M.Albulescu5
1
The National Institute for Development and Research in Optical Electronics- The Research Center
TEHNOPROF, Magurele
2
SC R&D Consulting and Services, Bucharest
3
Polytechnic University, Bucharest, Spl.Independentei 313, Bucharest
4
The National Institute for Materials Physics, Str. Atomistilor 1, Magurele
5
The National Institute for Development and Research in Pharmaceutical Chemistry, Calea Vitan
112, Bucharest
ABSTRACTThe characteristics of the materials employed for implants can be improved by maximizing theirbiocompatibility and their resistance to chemical corrosion and mechanical wear through the coatingwith anorganic thin layers, 0.002-0.01 thick, obtained by PVD vacuum deposition.

The materials used as substrates are: 316L stainless steel, Ti5A12 ,5Fe alloy and shape-memory
alloy NiTiNb. Thin layers of TiN, TiAlN and multilayered TiN/TiAlNb nitrides were vacuum
deposited by the PVD cathodic spring method. Comparative measurement of resistance to corrosionof uncoated and coated materials was performed by measuring the corrosion and critical passivationin physiological solution and artificial saliva currents. Layer composition, microhardness and theadherence to substrate were determined by X-ray difraction. Auger electron spectroscopy, Vickers
microhardness and Rockwell diamond scratching adherence test. The biocompatibility of layers wasassessed by comparison with uncoated materials according to the Tripan Blue rat hepatocyte culture
method.
The objective of this paper is demonstrating the improvement of the biocompatibility, corrosion
resistance and superficial hardness of implant materials by thin layer coatings with TiN and TiAlN
monolayer and TiN/TiAlN multilayer structures.
The results showed that all explored coatings have a good biocompatibility, show no signs ofcytotoxicity and irritating potential. The celular viability was 94.3 % for TiAlN, 03.4% for
TiN/TiAlN and 92.6% for TiN. Multilayered TiN/TiAlN had the highest resistance to corrosion,
followed by samples coated with TiAlN and TiN. There was no significant difference between layer
properties depending on the nature of the substrate.
We conclude that TiN and TiAlN thin layers obtained through vacuum cathodic spring method
improve the biocompatibility of implant materials.
6. HIGH PERFORMANCE TECHNICALLY ADVANCED COMPOSITE MATERIALSDESTINED TO BE USED IN THE MEDICAL INDUSTRY
Irina Carceanu*, Gheorghe Dinescu**, Tascu Gheamalinga**** Metals Research Institute, Bucuresti – irina_carceanu@yahoo.co.uk** National Institute of Laser, Plasma and Radiation Physics, Magurele*** SC BIMETAL SA
ABSTRACT
Objective: Obtaining porous superelastic materials, to be used in the medical field, from NiTi alloys,
and improving the performance of bench-marks obtained by protective and functional covering
techniques – laser ablation.
The structure of the nitinol thinlayers obtained by laser ablation

AFM images of: a) Ti subtrate,b) NiTi thin layer

AFM images of thin layers of
NiTi on microareas:a) si b) – scanning area 10X10µm;

X-ray difraction on NiTi thinlayers/ Ti substrate

Results• Outstanding porosity at the tissue-implant interface;• The tissue-implant contact area is 40% greater for Nitinol than for titanium;• Remarcable biofunctional integration;• Drastic decrease in the risk of restenosis;• Offers favourable conditions for cellular growth
Conclusions• The porous superelastic Nitinol is a highly attractive material in the fields of orthopedics,
maxilo-facial surgery and dental implants• The Nitinol thin layers obtained by laser ablation show homogenous structure and uniform
distribution of their components.
7. INTEGRATED OPEN SOURCE HOSPITAL INFORMATION SYSTEM & EHEALTHENVIROMENT CARDIOPREVENT™
Gabriel Ciorica1, Silvia Mancas1, Lucian Stoica1,Calin Muntean2, D.M. Duda-Seiman1, C.A.
Sarau1, Dan Gaita1, Stefan I. Dragulescu1
1Institute of Cardiovascular Diseases Timisoara – Prevention and Rehabilitation Department,
2
University of Medicine and Pharmacy Timisoara, Medical Informatics Department
ABSTRACTThe e-health project proposed under the label CARDIOPREVENT™ is a complete open-source,
informatic system built around an internet portal providing the necessary web-services, with an open
and easily modifiable architecture. These applications are developed under the provisions set by open
EHR standards, in a open source environment.
We have realised more relational data-base systems (SQL type) using a collaborative work mediumthat allows a more pertinent and complete data analysis. We have also included into the system theexistent forms (medical files) and used a series of formularies adapted to the project.Data collectionis accomplished through the use of the same paramaters of analysis. The developed application iseasily deployable to every hospital due to its inclusion of the therapeutical and diagnostic procedures
codification and classification according to the ICD 10 standard.
This pilot-project is in current use and development at the Institute of Cardiovascular Diseases
Timisoara, Clinical Department of Preventive Cardiology and Cardiovascular Rehabilitation. We
have already included more than 3000 patients in the last 24 months, and we use this database forstudies, grants, research.
From a common entry-level interface the users are then presented with custom forms for data
input or query, according to specific department needs and medical procedures. The DICOM
standard is used for medical imaging techniques (X-Ray, CT, angiography etc), providing
interoperability with other database systems and medical imaging software. Among otherfacilities, the user interface includes data-export features for reporting data to local andnational healthcare institutions (for epidemiology, statistic analysis, DRG reports).
1. CARDIOPREVENT™ can develop generic, standards-based interfaces between cardiology
systems and enterprise systems for scheduling, ordering and results reporting
So far the results we obtained indicate that a multiportal such as CARDIOPREVENT™ canrepresent, as it is, an electronic system of multifactorial evaluation in cardiovascular diseases

2. This project needs time though to quantify its penetrability and aplicability, but can represent amodel for a standard in Romanian e-health. Currently it is being developed horizontally, verticaldevelopment depending on many economical and political decision makers.
3. CARDIOPREVENT™ can providing an implementation framework for open connectivity using
existing standards in preventive cardiology and rehabilitation programs

8. MAGNETOIMPEDANCE-BASED BIOSENSOR PROTOTYPE FOR
BIOMEDICAL APPLICATIONS
H. Chiriac1, D.-D. Herea1, 2
1National Institute of Research and Development for Technical Physics, Iasi, Romania,
2“Al. I. Cuza” University, Iasi, Romania

SPECIFIC

ABSTRACT
Goal: At present, there are known different detection and bio-detection techniques implemented inbiomedical devices such as optical, electrochemical, thermometric, piezoelectric or magnetic. The
last one includes the giant magneto-resistance (GMR) and, relative recently, the giant magnetoimpedance (GMI) effect. These effects can be used in different GMR or GMI magnetic biosensors
that make use of magnetic particles as markers for target biomolecules. In our work we designed andtested a GMI-based biosensor using a special sensing element and in home-prepared or commerciallyavailable magnetic micro/nanoparticles.
Material and method. The sensing element. Co-Fe-Si-B glass-coated amorphous microwires of 35
micrometers in diameter and 30 mm length was prepared by glass-coating melt-spinning method andused as GMI sensing element. The magnetic particles. In home-prepared amorphous microparticles
and cobalt nanoparticles, and commercially magnetic beads were used to test their influence on thesensing element.
The magnetic amorphous microwire was arranged parallel to an external d.c.
magnetic field. The magnetic particles were dispersed around the microwire. An external magnetic
field has been applied on the microwire that is passed by an a.c. current.
Results. Different magneto-impedance variations were obtained when using amorphousmicroparticles with different sizes. Moreover, the sensing element was sensitive even when using
magnetic nanoparticles. As for example an important magneto-impedance variation of about 40 Ω
was obtained when using amorphous magnetic microparticles.
Conclusions. The magnetic biosensor prototype presents high sensitivity and it can detect bothmagnetic micro/nanoparticles and commercially magnetic beads. Therefore, such a magnetic
detection system can be used successfully in magnetic bio-detection applications for specificbiomolecule identification.
This work was partially supported from the contract CEEX no. 42/2005 RO-NANOMED.
9. EMPLOYMENT OF TITANIUM ALLOYS IN THE MANUFACTURE OF SOME TYPESOF SPINAL IMPLANTS
Danut Cojocaru*, Ion Cinca*, Ioan Dan**, Victor Moagăr- Poladian***, A. Trisca*****Polytechnic University, Bucuresti; **SC R@D Consultanta si Servicii;*** National Imstitute for Research and Development of Microtechnologies – Bucharest;**** Academy for Economical Studies - Bucharest
ABSTRACT
Objective. Obtaining and characterizing from a functional, structural, dimansional and material pointof view, Ti and Ti alloy spinal implants
Materials and methods.The Ti6Al4V alloy has been widely used as implant material, but its current
usage is restricted due to Vanadium toxicity. Thus, the Ti6Al7Nb alloy was perfected by replacing V
from the Ti6Al4V alloy with the equivalent quantity of Nb. The resistance of the Ti6Al7Nb alloy is
933 Mpa, a little less than that of the Ti6Al4V alloy. The Ti6Al7Nb alloy is less corrosive and its
resistance to corrosion is close to that of the comercially available pure titanium. The rigidity of theimplant and the characterstics of its fastening on the bone tissue are important factors of bone

remodelling. Consequently, the Ti6Al7Nb alloy conveniently meets both qualities of mechanicalresistance and biocompatibility.
In order to clarify the functional, structural, dimensional and material properties of the implants, thefollowing scientific research activities will be initiated: synthesis of the alloys; structural, mechanicaland biocompatibility tests; designing and obtaining of the implants. To optimize the results
mathematical modelling of structure-properties relations and of implant functionality will be
performed.
Results and conclusions.
This project will be finalized with the obtaining of spinal implants that will satisfy the following
requirements:- Monoaxial pedicle screws, with a diameter from 4,5 mm to 65 mm- Pedicle hooks, laminar hooks, transverse hooks;- Fixing nut, fixing rod of 5 mm with a length of 200 to 650 mm
The range above comprises all necessary dimensions and types of surgical implants for all categories
of patients.
10. MICROSENSORS AND ACTUATORS FOR MICRO-OPTO-ELECTROMECHANICALSYSTEMS MANUFACTURED BY SHAPE MEMORY ALLOY NANOENGINEERING
Corneliu M. Craciunescu1,2, Viorel Serban2, Victor Budau2, Octavian Oanca3, Virgil Paunescu4,
Gabriela Tanasie4, Dorin Jurcau5
1
National R&D Institute for Electrochemistry and Condensed Matter2
Politechnics University Timisoara
3
National R&D Institute for Welding and Material Testing4
University of Medicine and Pharmacy “Victor Babes” din Timisoara
5
LK-ELDUS S.R.L Timisoara
ABSTRACT
Goal: The main goal of the project refers to the fabrication of microsensors and actuators based on
performant shape memory alloys, materialized through secondary objectives like: fabrication of
nanoengineered shape memory alloys and characterization of materials behavior and manufactured
products in various environments.
Materials and method. The focus is on Cu-Zn-Al and Cu-Al-Ni alloys and the biocompatibility
tests have been oriented toward NiTi bulk materials. The basic method used for achieving the goals
was based on the “melt-spinning” technique and the aim was to generate alloys with favorable
texture, thus leading to shape memory effect with larger amplitude.
Results:
Copper based pre-alloys have been manufactured based on Cu-Zn-Al and Cu-Al-Ni families,
following several steps, like:- design and implementation of the technology for rapid solidification and heat treatment;- characterization by optical and X-ray diffraction techniques;- optimization based on analysis of as cast and quenched structures.
A system for ribbons fabrication has been optimized (especially on what concerns the control of thewheel rotation and ejecting temperature) and shape memory alloy ribbons with a width between 2.5
and 4 mm and 30 to 50 μm thickness have been produced.
Assembling technologies for shape memory alloy ribbons and bulk materials have been developed
for similar and dissimilar materials as well as for ribbon joins.
Microsensor and actuator demonstrative products have been designed and produced, based on the
manufactured materials and the NiTi behavior in biocompatible environments has been studied.
Conclusions. The possibility to fabricate better quality ribbons and to use them to design prototypes
for new applications has been demonstrated and a technique to assess the shape memory alloy
biocompatibility has been analyzed.

11. NANOCONTROL AND MULTIFUNCTIONALITY IN MATERIALS, MICROLAYERS
AND ARCHITECTURES WITH SHAPE MEMORY PROPERTIESCorneliu M. Craciunescu1,2, Victor Budau2, Iona Mitelea2, Mihaela Valeanu3, Horia Chiriac4,
Aurel Ercuta5, Valentin Bardean6, Virgil Paunescu7, Dieter Schinle8, Mihaela Stoica91
National R&D Institute Institutul for Electrochemistry and Condensed Matter2
Politechnics University Timisoara
3
National R&D Institute for Materials Physics Bucharest
4
National Physics R&D Institute Iasi
5
Western University Timisoara
6
National R&D Institute National for Welding and Material Testing
7
University of Medicine and Pharmacy Victor Babes Timisoara
8
Machines Building Factories Resita
9
National R&D Institute in the Laser, Plasma and Radiation Physiscs Bucharest
ABSTRACTGoal. The main goal of the project refers to the analysis of the shape memory alloys functionality
with focus on new ferrous and ferromagnetic alloy families, as well as on the use of these materials
in performant structures and architectures.
Material si metoda. The main alloy families cicaonsidered are ferrous shape memory alloys (FeMn-Si, Fe-Ni-Co-Ti as well as FeCrNiMnSiCo stainless alloys), Heusler-type alloys (Co-Ni-Ga, NiMn-Ga, Co-Ni-Fe, Co-Ni-Al, etc.), and NiTi family, respectively. The basic methods proposed arethose through which particular materials will be manufactured as ribbons, glass coated wires andsuction solidified bars. The characterization methods used are complex, based on thermal,
magnetical, magnetoelastic, biological techniques, and the finality is related to the fabrication ofadvanced materials.
Results. Bulk ferous shape memory alloys have been manufactured so far by arc melting andsubsequently rapidly solidified ribbons (FeMn32Si6) have been produced using the melt-spinningtechnique. The ribbons have been vacuum heat treated, and as result the shape memory effect has
been observed above room temperature and as a function of the applied heat treatment they also
becomed magnetic.
Other ribbons (Fe-Ni-Co-Ti) showed the shape memory effect at liquid nitrogen temperature.
Preliminary investigations by electrical analysis showed anomalies in the behavior of Fe-Mn-Si
alloys, in the vicinity of 0- 50 K, that has been attributed to a phase transition.
Conclusions. Promising research results have been obtained on shape memory alloys, based on
thorough analysis and ferrous shape memory alloy bulk and ribbons, with shape memory properties
and – as a function of composition – magnetic or stainless properties.
12. FACTOR ANALYSIS OF PULSE WAVE VELOCITY INCREASE IN ASYMPTOMATIC
SUBJECTS
Laura David, Silvia Mancas, C.A. Sarau, Stela Iurciuc, Livia Barbu, Simina Urseanu, MariaRada, Ciorica G., Stanciu B., Elena Nistor
Preventive Cardiology and Cardiovascular Rehabilitation Clinic, Timisoara, Romania
ABSTRACT
Objectives: Insulin resistance (IR) is an essential feature of the metabolic syndrome, but theassociation IR – endothelial dysfunction is still disputed. There is scientific proof regarding therelation carotid-radial pulse wave velocity (PWV-CR), endothelial function and cardiovascular risk.
The work proposes to verify the hypothesis that PWV-CR is increased in subjects with IR vs. without
IR.
Methods: PWV-CR was assessed using the Complior device in order to observe its behavior inasymptomatic subjects with (n=44) and without IR (n=14).
Results: The association IR and blood hypertension was present in 65.51% from the subjects, IR andleft ventricular diastolic dysfunction in 63,79%, IR and TG/HDL≥3.5 in 56,89%, IR and LDL>100
mg/dL in 46,55%. In the group with IR, PWV-CR had a mean value of 10.83±2.52 m/s. Increased

PWV-CR correlated significantly with IR (p=0.004), with increased TG levels (p=0.01) andextremely significant with the TG/HDL≥3.5 ratio (p=0.0000001). The correlation with LDL was not
significant.
Conclusions: (a) PWV-CR was with 13.85% higher in the IR group and with 8.77% higher insubjects with metabolic syndrome. (b) It is possible that this vascular dysfunction to be present earlyin subjects at risk of developing metabolic syndrome. (c) The TG/HDL≥3.5 ratio was the best IR
predictor.
13. THE OSTEOGENIC DIFFERENTIATION OF CANINE MESENCHYMAL STEMCELLS IN COMPOSITE MATRICES
Erika Deak1, Mihai Palko2, Simona Anghel1, Tiberiu Popescu-Negreanu3, Cornel Igna2, Gabriela
Tanasie1
1Center of immunophysiology and Biotechnology, University of Medicine and Pharmacy Victor
Babes Timisoara,
2
Department of Surgery, University of Agricultural Sciences and Veterinary Medicine Timisoara,
3“Sticla si ceramica PONETI” Ltd
ABSTRACT
Tissue engineering based on stem cells involves the exploration of two main characteristics of thesecells: the high proliferation capacity and the pluripotency.
Aim of the study: Investigation of in vivo proliferative and differentiation potential of canine
mesenchymal stem cells cultured on composite matrices.
Materials and methods: The mesenchymal stem cells were isolated from bone marrow aspirates(harvested by humerus punction) from 5 healthy dogs. The harvesting procedures were performed
safe and sterile according to the approval of university ethics committee. For cell isolation we used
the plastic adherence method. The cells were cultivated on semiconfluence, trypsinised and seeded in
DMEM+10% FCS on a density of 105cells/cm2. At second passage the cells were seeded on 3Dcomposite matrices, 106 cells/matrix. The media was supplemented in osteogenic factors (betaglicerophosphate, ascorbate, dexametasone). We counted the unadherent cells (trypan blue
technique) and after 14 days we checked for bone mineralization using von Kossa staining.
Results: Canine mesenchymal stem cells showed a characteristic, fibroblastoid morphology, similar
with human mesenchymal stem cells. These cells can be relatively easy differentiated into
osteoblasts in vitro, using the appropriate media supplementation. In vitro, the cells had a good
adherence on composite matrices. The ex vivo examination of mixed implants (biovitroceramicscolagen-mesenchymal stem cells) proved the mesenchymal stem cells differentiation into osteoblasts.

Fig. 1. Canine mesenchymal stem cells,
passage 1, ob. 40x

Fig. 2. Von Kossa positive staining

The study was supported by national excellence grant CEEX 45/2005 (CELLART)

14. INFLUENCE OF IMMOBILISATION METHOD ON THE KINETICS OF PROTEASESENTRAPPED IN NANOPOROUS SILICA GELS OBTAINED USING THE SOL-GELTECHNIQUE
Monica Dragomirescu1, Gabriela Preda2, Beatrice Oros2, Rodica Căpriţă1
1
Banatul University of Agricultural Sciences and Veterinary Medicine, Faculty of Animal Scienceand Biotechnology,
ABSTRACT

Aims: The sol – gel process is a method allowing the entrapment of functionally active biomolecules
in inorganic silica at room temperature. The kinetic constants for immobilized enzymes are usuallydifferent from those for the native form, as conformational changes, steric hindrances and partitionand diffusion effects may occur on enzyme immobilization. The conformational effects may alsochange the affinity between enzyme and substrate. This is not always a disadvantage, because theporous silica network may protect proteins against unfolding.
The aim of this work was the immobilisation of two proteases, a commercial product, Alcalase, and aprotease preparation obtained by submerged fermentation of Bacillus licheniformis B 40 local strain.
Two immobilisation methods were used: entrapment in silica gels and entrapment-deposition on ainorganic support. The enzyme-substrate affinity modification was investigated based on the kinetics
of native and immobilised enzymes respectively.
Materials and methods: The general immobilisation method used tetraethoxysilane (TEOS) as gel
precursor. The sol-gel synthesis was done in two steps, by acid (pH 3.5-4.0) and base (pH 7.5-8.0)
catalysis. A local ceramic material (provider Procema SA) was used as co support for entrapmentdeposition. The kinetics of the native and immobilised enzymes was studied in the hydrolysis
reaction of a model substrate, a low molecular mass ester, para-nitrophenyl acetate.
Results and discussion: Alcalase immobilisation provided entrapped products with 10 time’s higher
protease activity and better immobilisation yields, compared with the Bacillus licheniformis B 40
enzyme. For both enzymes, the Michelis constant Km and the maximum rate Vmax of the immobilised
enzymes were higher than those of the soluble ones. The Bacillus licheniformis B 40 enzyme has
kinetic parameters comparable with those of the commercial enzyme.
Conclusions: For both enzymes, the hydrolysis reaction follows a Michaelis-Menten type kinetic.
Immobilisation changes the kinetic parameters. Vmax/Km ratio suggests that, by immobilisation, thecatalytic efficiency increases, thus compensating the partial decrease of the enzyme-substrateaffinity.
15. IMPORTANT STRUCTURAL DESCRIPTORS TO DEFINE BIOLOGICAL ACTIVITY
AND MOLECULAR MODELING TO DESIGN NEW ANTI-HIV DRUGSCorina Duda-Seiman1, D. M. Duda-Seiman2, R. Nuţiu1, D. Ciubotariu3
1Western University of Timişoara, Dep. of Chemistry – Organic Chemistry2
University of Medicine and Pharmacy „Victor Babeş” Timişoara, Medical Ambulatory andEmergencies Dep.
3
University of Medicine and Pharmacy „Victor Babeş” Timişoara, Facultaty of Pharmacy
ABSTRACT
Aim: to perform a QSAR (quantitative structure-activity relationship) study using molecularmodeling methods upon a series of 79 HEPT ligands which act as HIV reverse-transcriptase (RT)inhibitors in order to map the space of the interaction site, as well as to assess structural parameters
important to define biological activity.
Materials and methods: Molecular modeling was performed using the HyperChem 7.01 programand conformational analysis using Conformational Search. The most stable conformations wereidentified in the series with 79 HEPT ligands and they were used to obtain the hypermolecule inorder to perform a classic MTD method. A large number of structural parameters was assessed with
the Dragon program and their predictive power was tested with correlation methods.
Results: Biological active molecules are less symmetrical shapes. The obtained multiparametricmodels with various structural descriptors led to correlation coefficients of 0.9.

Conclusions: Beyond the possibility to identify and to remove from the beginning those molecular
structures which don’t show any therapeutic potentiality, well designed QSAR studies allow tocollect essential structural data necessary for an increased biological activity, suggesting certain
requirements for an increased anti-HIV effect.
16.
CARDIOVASCULARRISK:ENDOTHELIAL
DYSFUNCTION,
INSULINRESISTANCE AND ASSOCIATED FACTORS
Duda-Seiman D.M., Sarau C.A., Iurciuc Stela, Iurciuc M., Barbu Livia, Duda-Seiman Corina,
Velimirovici Dana, Rada Maria, Ciorica G., Petcov Bileana, Mancas Silvia
Preventive Cardiology and Cardiovascular Rehab. Clinic, Timisoara, Romania
ABSTRACT
Objectives: Epidemiological evidence shows that smoking increases the incidence of fatal and nonfatal cardiovascular events through insulin resistance (IR). The work proposes to assess the relation
between smoking, IR and endothelial dysfunction in asymptomatic hypertensive men.
Methods: We assessed the smoking state (pack years), carotid-radial pulse wave velocity (PWV-CR)
with Complior and IR (IRHOMA) in asymptomatic hypertensive men.
Results: 42 asymptomatic hypertensive men, mean age 57.57±16.4 years: 26 (61.9%) with PWVCR≥9.5 m/s and 28 (66.66%) with IRHOMA≥1. Waist circumference has a positive correlation withthe state of IR, especially in those with PWV-CR≥9.5 m/s and LDL>100 mg/dL (R=0.59, R2=0.34,
p=0.004). The odds to have IR are 2.75 if TG/HDL≥3.5 and the relative risk in this case to develop
IR is 1.38. The odds to have PWV-CR≥9.5 m/s are 5.40 in subjects with IRHOMA≥1 and therelative risk to develop increased PWV-CR is 1.85 when IRHOMA≥1. Smoking amplifies
significantly the PWV-CR increasing level (p=0.0003).
Conclusions: (a) Smoking is a public health problem because of the consequences on the vessels
quality. (b) IR plays a key role in cardiovascular risk assessing in middle-aged asymptomatic men.
(c) Adding other modifiable factors to IR, the vascular impact is aggravated. (d) The costeffectiveness of prevention programmes is in favor of a healthy lifestyle adopted as early as possible.
17. IMPAIRED ENDOTHELIAL-DEPENDENT VASORELAXATION IN ISOLATEDHUMAN MAMMARY ARTERIES PREINCUBATED WITH TRIGLICERIDE-RICH
REMNANTS LIPOPROTEIN
O. Fira-Mladinescu1, A. Avram2, L. Noveanu3, C. Fira-Mladinescu1, D. Muntean1, D. Gaita2
1
Research Centre of Preventive Medicine, 2Department of Preventive Cardiology and Rehabilitation,3
Department of Physiology, "Victor Babes" University of Medicine and Pharmacy, Timisoara,
Romania
ABSTRACT
Several studies have now shown an impairment of endothelial function in association withhypertriglyceridemia. Major changes in lipoprotein metabolism occur in these individuals, resulting
in the formation of triglyceride-rich remnant lipoprotein, small and dense LDL (predispose toperoxidation), and decreased HDL-cholesterol. We examined “in vitro” the effect of remnant
lipoproteins on acetylcholine (Ach)-induced endothelium-dependent relaxation, and sodium
nitroprusside (SNP)-induced endothelium-independent relaxation in human mammary arterial rings.
Our results suggest that preincubation with triglyceride-rich remnant lipoproteins inhibited
endothelial-dependent vasorelaxation (maximal relaxation expressed as percentage was 34.15 ± 4.88
in remants group versus 49.03 ± 9.85 in control group, p=0.008) but did not affect endotheliumindependent vasorelaxation (maximal relaxation was 93.33 ± 7.36 in remnants group, versus in 95.86± 3.48 control group, p=0.46). This endothelial dysfunction induced by remnant lipoprotein could
contribute to the pathogenesis of atherosclerosis associated with hypertriglyceridemia. Thisconclusion supports the concept that each of the changes in the plasma lipoproteins associate withelevated triglyceride plasma level contribuite to the increased risk of premature cardiovasculardisease.

18. INTERFACE MECHANICS AND HISTOLOGY OF TITANIUM AND
HYDROXYAPATITE-COATED TITANIUM FOR MEDICAL IMPLANT APPLICATIONS
C. Igna1, A. Sala1, L. Schuszler1, N. Markocsan2
1
USAMVBT, Faculty of Veterinary Medicine Timişoara, ignacornel@gmail.com,
2
INCDSIM Timişoara
ABSTRACT
Objectives: in-vivo testing biocompatibility of implants coated with osteoconductive supports infemoral assembling on canine biological model; clinical, radiological and histological studies toestablish adhesion between biological support and implant and bone contact reaction checking(presence of osteogenic and osteoclastic cells, presence of blood vessels and preosseous matrix
formation).
Experimental model: eight types of titanium lamellar implants (osteosinthesys plate type, PCT, AO
model, dimensions between 10-18 mm made by Aesculap that have the internal face (bone contactface) coated at INCDSIM Timisoara with various concentrations of titanium powders,hydroxyapatite and titanium-hydroxyapatite combinations were fixed into the femoral cortex of fivedogs (common breed, females, three year old and 18 kg bodyweight) using a standard asepticallytechnique.
Methods: Clinical study – behavior changes, central body temperature and body weight were
followed and recorded on all the experimental period.
Radiological examination of the femours with implants was made by cranio-caudal incidence
radiographs, using 65 kVp and 20 mAs.
Macroscopical examination – the femours with implants were prelevated and all
macroscopical changes noted.
Microscopical examination – the pieces were demineralized (immersion in 10% tricloracetic acid)and histological processed by Hematoxylin-Eozin and Mallory tricromic coloration.
Results: The adhesion of bone support to implants is better on those coated with hydroxyapatite thatat the ones with titanium powders. The macroscopical changes reveal the biocompatibility with thecanine bone support of the implants in study. The biological changes radiographic observed aroundthe implants from the second experiment are placed in the normal, physiological evolution of bone
support under these kind of manipulation, which reveals a perfect biocompatibility. The active
biointerface behavior was observed on implants coated with hydroxyapatite and combination of
porous titanium and hydroxyapatite, on which the histological changes observed were osteoinduction
and osteoconduction phenomenons.
Clinical relevancy: The implants coated with hydroxyapatite have biocompatibility with canine
bone support and also have osteoinductive and osteoconductive capacity which makes themrecommended for clinical testing.
19.
FLOWCYTOMETRIC
MEASUREMENTOF
BIOCOMPATIBILITYOF
MULTILAYERED NANOSTRUCTURES DEPOSITED BY MAGNETRON REACTIVEATOMISATION
C. Iordachel1, D. Bratosin1, L. Buia1, M. Braic2, M. Balaceanu2, C.Trisca-Rusu3, A. Kiss2, V.
Braic2, A. Vladescu2
1
The National Institute for Biological Sciences, Splaiul Independentei 269, Bucharest
2
The National Institute for Development and Research in Optical Electronics – Research Centre
TEHNOPROF, Magurele
3
Polytechnics University Bucharest
ABSTRACTThe study of the biocompatibility of implant materials can be accurately performed by using thecalcein-AM flowcytometry.

The materials used as substrates were 316L stainless steel and Ti6A14V alloy. The multilayered
nanostructures (Ti/TiN/TiAlN)n, n being the total number of layers, were vacuum deposited bymagnetron reactive atomisation, as it results in a minimal surface rugosity. Comparative
measurement of the biocompatibility of uncoated and coated materials was performed using theCalcein-AM test, the best indicator of cell viability due to its high retention inside the cell andrelative insensitivity of the fluorescence to pH values within physiological boundaries. Calcein, thehydrolysis product of Calcein-AM, a polianionic derivate of fluoresceine, has 6 negative and 2
positive charges at pH 7.0. The activity of cellular esterases, resulting in an increased fluorescence, isa degree of cellular viability. Layer composition, microhardness and the adherence to substrate were
determined by X-ray difraction. Auger electron spectroscopy, Vickers microhardness and Rockwell
diamond scratching adherence test.
The objective of this paper is demonstrating the improvement of the biocompatibility, corrosion
resistance and superficial hardness of implant materials by thin layer coatings with multilayered
nanostructures of (Ti/TiN/TiAlN)n=100 with reduced surface rugosity.
The results showed that all explored coatings have a good biocompatibility. Calcein, as a hydrolysis
product of Calcein-AM, confers the cells a high degree of fluorescence related to cellular viability as
this compound is only retained by cells that have an intact membrane. Cell viability was 49% for the
Ti6Al4V alloy, 42% for 316L stainless steel, 75% for(Ti/TiN/TiAlN)n=100, 65% for TiN and 63% for
TiAlN.
We conclude that the Calcein-AM flowcytometry test is a fast and sensitive method for assessing thebiocompatibility of implant materials.
20. THEORETICAL ANALYSIS OF SEQUENCE-STRUCTURE RELATIONSHIP FOR
CALCIUM BINDING PROTEINS
Adriana Isvoran1, Laura Unipan2, Liliane Mouawad3, Constantin T. Craescu4
1Western University of Timisoara, Department of Chemistry, Str. Pestalozzi 16, 300115 Timisoara,
Romania2
University of Agricultural Sciences and Veterinary Medicine of Banat, Department of Agriculture,
Cl. Aradului 119, Timisoara, Romania
3
INSERM U759/ Institute Curie-Recherche, Univerity Center Paris-Sud, 110-112, 91405 Orsay,
FranceABSTRACT
Aim : theoretical analysis of intramolecular interactions responsible for tertiary structure folding of
calcium binding proteins belonging to the “E-F hand” family and prediction or tertiary structure ofcentrins.
Method:- sequence alignment for proteins under study ;- prediction of the tertiary structure using Mitaku’s server ;- determination of net charge of entire protein and those of their N and C-terminal domains;- determination of the ratio of charged amino acids and that of hydrophobic amino acids both forthe entire protein and for its domains.
Results. Sequence similarity is high (more than 60%) for the extended structures and quite small forcompact structures (less than 26%). Centrins have a high sequence similarity with extended
structures. Theoretical analysis of physicochemical properties of amino acids belonging to these
proteins shows that extended structures are abundant in charged amino acids (more than 30%) in
both domains. They also have a high negative net charge. Compact structures are abundant inhydrophobic residues, especially in the central linker. Average hydropaticity is higher for compact
structures than for extended ones.
Conclusions. The high ratio of charged residues and high negative net charges of domains confirm
the hypothesis that electrostatic repulsions are responsible for great stability of extended structures.
In the case of compact structures, the high ratio of hydrophobic residues confirms that hydrophobic
interactions contribute significantly to the stability of these proteins. Comparison of thephysicochemical properties of amino acids belonging to centrins sequences show that human centrin

3 and yeast centrin could adopt a compact tertiary structure and human centrins 1 and 2 could adopt
extended tertiary structures.
21. THE IMPORTANCE OF PULSE WAVE VELOCITY IN HYPERTENSIVE PATIENTS
WHICH ARE TO BE INCLUDED IN A REHABILITATION PROGRAM.
Stela Iurciuc, M. Iurciuc, Livia Barbu, S. Ursoniu, Gabriela Otiman, D. Gaita, Simona Dragan,
Avram Adina, D. Duda-Seiman, C. A. Sarau, L Berinde, Dana Velimirovici, Maria Rada, G.
Ciorica, F.I. Cobzariu, Laura Craciun, Silvia Mancas
Department of Cardiovascular Rehabilitation, University of Medicine and Pharmacy Victor Babes
Romania.
ABSTRACTIntroduction: The pulse wave velocity (PWV) represents a method of evaluating the arterial
stiffness. Recent epidemiological studies have proved that the PWV and the pulse pressure (PP)represent independent predictors of cardiovascular mortality in hypertensive old subjects. Thechange of arterial rigidity appears in initial states of arterial hypertension, and this is why the early
identifying of PWV values could allow the identifying of and intervention on the complications ofhypertensive disorder.
Methods: We studied 89 subjects, with average age of 55.22±11,69 years; of which: men: 47.19%
and women: 52.81%, with essential hypertension: normal high, grade 1 and 2 (ESC-ESH 2003). The
subjects have been evaluated from a clinical and paraclinical point of view according to the standard
protocol of inclusion in the recovery program. The PWV of the carotid-radial (PWVc-r) has been
determined for all subjects through automatic monitoring using the Complior device.
Results: We have noticed that the following cardiovascular risk factors (CVRF): male aged over 55
associates prevalently with values of PWVc-r over 10 m/s (bivaried analysis through the chi square
test: OR=3.11; 95%CI: 1.20-8.07), the same correlation has also been obtained for smokers with
PWVc-r values >10 m/s (OR=3.39; 95%CI: 1.13-10.20). There is also a simple linear correlation
between PWV (the dependent variable) and the following independent variables: the systolic bloodpressure (SBP) and the pulsed pressure PP (r=0.24, p=0.027).
Conclusion: Evaluating the arterial stiffness by measuring the PWV may constitute a reference inthe evaluation of the benefits of the cardiovascular rehabilitation program for a hypertensive patient.
Follow up data reviles: the arterial rigidity correlates positively with some CVRF: males aged over
55, as well as with the condition of being smokers. The SBP, as well as the PP values modify thearterial stiffness parameters like PWV.
22. GETTING THROUGH ALTERNATIVE METHODS OF DOPED TiO2
NANOCRYSTALS WITH METALICS IONS.APLICATIONS STUDY FOR HEALTH,BIOLOGY AND ENVIRONMENT
Carmen Lazau, I. Grozescu, Paulina Vlazan, L. Mocanu, Iasmina`Miron 1
National Research-Development Institute for Electrochemistry and Condensed Matter Timisoara
ABSTRACT
Purpose. The purpose to this project is getting the undoped and doped TiO2 nanocrystals with Au
and Ag ions through sol-gel method and hydrothermal method attend ultrasonic/microunde.The
interest crystallization form of nanocrystals is anatas and rutil, and medium dimensions of theparticles are to maximum 30nm.The titan dioxide doped with metallic’s ions get through alternative
methods(sol-gel, hydrothermal) offer an large aria of photocatalitics applications in medicine,environment protection and agriculture, opening new perspectives for technical performances of thedevices with solid body.
Materials and methods. The precursors used in synthesis of TiO2 doped with metallic’s ions weretitan izopropoxid, titan tetrachloride, etilic alcohol, oxalic acid, distillate water, silver azotate andtetracloroauric acid (dopaded precursors). The used methods in titan dioxide synthesis were sol-gel
and hydrothermal method. The field of baking and autoclave temperatures was between 2000C and
6000C.

Results: