Human dental pulp mesenchymal stem cells isolation and osteoblast differentiation.

AIM: This study was focused on the isolation and characterization of mesenchymal stem cells (MSCs) from human dental pulp (DPSC). METHODS: The study was performed in the Department for Oral and Cranio-Maxillo- Facial Surgey Hamad Medical Corporation, Doha, Qatar and Weill Cornell Medical Colleague Doha, Qatar, in period 2010-2011. Dental pulp was extracted from premolars and third molars of 19 healthy patients. The pulp was digested in a solution of 3 mg/mL collagenase type I and 4 mg/mL dispase for 1 hour at 37C. After filtration, cells were cultured in Dulbecco's Modified Eagle Medium (DMEM Low Glucoses) with 20% Fetal Bovine Serum (FBS), 2mM L-glutamine and antibiotics (100 U/mL penicillin, 100 ug/mL streptomycin) at 37 °C under 5% CO2. Cultures were treated with osteoinductive medium for differentiation MSC in to the osteoblast cell line. Staining with Alizarin red were used for the detection of the osteoblast production and calcification new formed tissue. RESULTS: On the total of three out of 19 patients it was possible to isolate DPMSCs after 2 to 3 weeks: in one patient it was not possible to expand MSCs because of infection, and in other two patients positive Alizarin red staining reaction showed osteogenic differentiation capability and strong mineralization in vitro. CONCLUSION: The main advantage of using DPSC is absence of morbidity. MSCs could be isolated noninvasively from teeth, routinely extracted in the clinic and discarded as medical waste. Standardization of clinical and laboratory protocols for DPMSCs isolation and team work coordination could lead to significantly improved result.

Rat bone marrow stem cells isolation and culture as a bone formative experimental system.

Bone marrow mesenchymal cells have been identified as a source of pluripotent stem cells with multipotential potential and differentiation in to the different cells types such as are osteoblast, chondroblast, adipoblast. In this research we describe pioneering experiment of tissue engineering in Bosnia and Herzegovina, of the isolation and differentiation rat bone marrow stromal cells in to the osteoblast cells lineages. Rat bone marrow stromal cells were isolated by method described by Maniatopulos using their plastic adherence capatibility. The cells obtained by plastic adherence were cultured and serially passaged in the osteoinductive medium to differentiate into the osteocytes. Bone marrow samples from rats long bones used for isolation of stromal cells (BMSCs). Under determinate culture conditions BMSCs were differentiated in osteogenic cell lines detected by Alizarin red staining three weeks after isolation. BMSCs as autologue cells model showed high osteogenetic potential and calcification capatibility in vitro. In future should be used as alternative method for bone transplantation in Regenerative Medicine.

Impact of reversionary and other etiological factors on prognosis and course of schizophrenia.

BACKGROUND: To identify the presence of schizophrenia among patients and their relatives, factors affecting duration and prognosis of the disease and other etiological factors related to schizophrenia. METHODS: This retrospective, descriptive, analytical and epidemiological research, which was conducted at the Psychiatric hospital of the Clinical Center of the University of Sarajevo during 2007, covered randomly selected 100 hospitalized patients with schizophrenia according to diagnostic criteria of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). Diagnosis of schizophrenia among relatives was based on anamnesis- Structural Clinical Interview (SCID) and it was applied to confirm DSM-IV diagnosis of schizophrenia. RESULTS: The presence of schizophrenia among patient relatives was the most important in etiology of schizophrenia (62%), and etiological factors were represented in 38 % of examinees (p=0,0001). Among relatives of examinees aged 20 - 30 years, schizophrenia was present in 37 (59.7%) cases. Schizophrenia among relatives caused earlier appearance of the disease. Duration of hospitalization of over 60 days was in the group of examinees which have the relatives with schizophrenia, 18 (29.0%); multiple hospitalizations were noted in the group of relatives in 40 (64.5%) cases; in one case (8.3%) traumatic experience was noted, in three (42.8%) acute stress, and in four 4 cases (28.6%) non-adequate living conditions. CONCLUSIONS: The results of this study show that reversionary factors are responsible for inducing schizophrenia, which leads towards chronic course of the disease and worsened prognosis.

In vivo osteoinductive effect and in vitro isolation and cultivation bone marrow mesenchymal stem cells.

Bone marrow contains cell type termed mesenchymal stem cells (MSC), first recognized in bone marrow by a German pathologist, Julius Cohnheim in 1867. That MSCs have potential to differentiate in vitro in to the various cells lines as osteoblast, chondroblast, myoblast and adipoblast cells lines. Aims of our study were to show in vivo capacity of bone marrow MSC to produce bone in surgically created non critical size mandible defects New Zeland Rabbits, and then in second part of study to isolate in vitro MSC from bone marrow, as potential cell transplantation model in bone regeneration. In vivo study showed new bone detected on 3D CT reconstruction day 30, on all 3 animals non critical size defects, treated with bone marrow MSC exposed to the human Bone Morphogenetic Protein 7 (rhBMP-7). Average values of bone mineral density (BMD), was 530 mg/cm3, on MSC treated animals, and 553 mg/cm3 on control group of 3 animals where non critical size defects were treated with iliac crest autologue bone graft. Activity of the Alkaline Phosphatase enzyme were measurement on 0.5, 14, 21, 30 day and increased activity were detected day 14 on animals treated with bone marrow MSCs compared with day 30 on iliac crest treated animals. That results indicates strong osteoinduction activity of the experimental bone marrow MSCs models exposed to the rhBMP-7 factor Comparing ALP activity, that model showed superiorly results than control group. That result initiates us in opinion that MSCs alone should be alternative for the autolologue bone transplantation and in vitro study we isolated singles MSCs from the bone marrow of rat's tibia and femora and cultivated according to the method of Maniatopoulos et all. The small initial colonies of fibroblast like cells were photo-documented after 2 days of primary culture. Such isolated and cultivated MSCs in future studies will be exposed to the growth factors to differentiate in osteoblast and indicate their clinically potential as alternative for conventional medicine and autologue bone transplantation. That new horizons have potential to minimize surgery and patient donor morbidity, with more success treatment in bone regenerative and metabolism diseases.

[Sociodemographic characteristics of posttraumatic stress disorder (PTSD) patients]. / Sociodemografske karakteritike kod pacijenata sa posttraumatskim stresnim poremecajem (PTSP).

UNLABELLED: Posttraumatic Stress Disorder (PTSD) represents a delayed response to stress events, which are catastrophic and likely to cause pervasive distress to almost everyone. This disorder is frequently associated with other psychiatric disorders. PTSD can be caused by many traumatic events, but the war related events are very important and significant. WHO assess that a million of BiH population suffers from one of the stress related disorder. The aim of this research is to show sociodemographic characteristics of PTSD patients during 2007. MATERIAL AND METHOD: This research was conducted at the Psychiatric Clinic KCU Sarajevo. The study is epidemiological and retrospective analytical-descriptive. The final results show that there were 343 patients with PTSD admitted in Psychiatric Clinic. PTSD is the most common among male population in 45-64 age group. Majority (74%) had high school education. Most of them, that is 206 (60%), were unemployed and 274 (80%) were married.

Influence of some socio-economic factors on growth and development of the boys in the Tuzla region (Bosnia and Herzegovina).

The impact of certain exogenous factor (socio-economic, ecological) has been investigated with special attention paid to the parents' living standard, and number of family members on some anthropometric parameters like: body height, body mass, chest circumference, upper leg circumference, upper arm circumference, sitting height, arm length, leg length, pelvis width, shoulders width, lenght of head and with of head on the sample of 698 boys aged 11 to 16 (17) years in the Tuzla region (the northeastern Bosnia, Western Balkan peninsula). Anthropometric measurements have been carried out using methodology proposed by the International Biological program (IBP). The results of these investigations have shown that there is a certain impact of the socio-economic conditions on the growth and development of boys. Children from families that have better living standard are, as a rule, taller, which is indicated by the statistical significant differences (P > 0.01). This trend indicates also value of Body Mass Index (BMI), which is in younger children from the families with lower living standard 16, while in the same category in the children from the families with better living standard it has value 18.5. The real impact of living conditions on the dynamics of development could be the best seen in the period of puberty. The number of children in the family has negative relationship with anthropometric features. Statistically significant differences (P > 0.001) have been detected in numerous analysed features in families with one or two children in comparison with families with three, four, or five children. Therefore, BMI has been significantly lower (16) in children from families with several children, while in the families with one child in the same growth class (11 years) it was significantly higher (17.4). Similar value of BMI (17.9) have children from the families with five children and which are 17 years old. Besides socio-economic conditions, high level of environmental pollution which is typical for Tuzla region for a long time, has also significant impact on the growth and development of children.

Pharmacokinetics and biological effect of the recombinant human bone morphogenetic protein-7.

BACKGROUND: During Bone Morphogenetic Protein (BMP) induced osteoblastogenesis and bone formation, strong vascularization is observed at the transition of preosteoblasts to mature osteoblasts. The cellular and molecular mechanisms that determine autonomous hematopoietic competence in the BMP induced osteoblastogenesis and angiogenesis have not been characterized yet. AIMS: In the present study we investigated, whether rhBMP-7 in collagen as carrier (ACS) stimulates osteoblastogenesis have effect on the physiologic autonomous hematopoetic system and clinical condition on animal model. MATERIAL AND METHOD: Study was' performed in the Clinic for Maxillofacial Surgery University Clinic Center Sarajevo from 2003 to 2004. Mandible hemiresection non-critical size created defect were treated with rhBMP7/ACS on 7 rabbits, and on other group of 7 defects were treated with autologues bone graft iliac crest. Biochemical and hematologist tests were performed measurement Alkaline Phosphates enzyme (ALP) activity and Erythrocytes (Er), Leukocytes (Le), Hematocrit(HCR) and Hemoglobin(Hg) count extracted from the periphery blood preoperative, 5, 14, 21, 30 day. RESULT: ALP activity showed strong osteoinductive effect rhBMP-7 determinate with day 21 significance increased activity and on bone graft sites day 30. All hematologist values were in physiologic range on both groups. CONCLUSION: Strong osteoinductive response rhBMP/ACS construct determinate by the higher value of the ALP enzyme didn't affect hematopoetic competence determinate by the Er. Le, HCR and Hg. counts parameters. Result indicated that BMP-inducedangiogenesis may thus be the result of BMP-induced secretion some angiogenetic factor locally from osteoblasts named Vascular Endothelial Growth Factor (VEGF).

[Obesity, personal and social problem]. / Gojaznost, osobni i drustveni problem.

Obesity is risk-factor for the most common nowdays diseases, as cardiovascular diseases, diabetes, hypertension, some carcinomas, degenerative diseases of weight bearing joints (spine, hips, knees), and present a huge medical and social problem, as well. It causes not only somatic but mental problems as well, especially in females and younger persons. Some of them undergo risky weight reducing methods, surgical procedures, etc, to reduce their body weight and to release mentally stressing body deformities. Aim of the work is to quantify negative impaction of obesity on functionality of the knee, the key joint in everyday body activities (rising, standing, walking, and climbing). Study has analyzed 22 randomly chosen patients (5 male, 17 female) with strong degenerative alteration of the knee (osteoarthritis), who were treated on the Department for orthopedic and traumatology, Clinical center Sarajevo during 2005 and 2006. Average age was 63.6 +/- 10.6 (54-76) years, with Body Mass Index (level of obesity) 31.1 +/- 3.5 (27-38) points, and average duration of symptoms 9.1 +/- 7.4 (1-25) years. The knee functionality was assessed by Knee Society Knee Score (KSKS). Completely healthy knee has 200 points--50 points for pain free knee, 25 points for stabile knee, and forfull extension-flexion arch, both, 50 points for normal walking and stairs climbing, both. Average value of KSKS was 118.1 +/- 35.0 points. As it was expected, there were strong significant correlation between KSKS and age (r = -0.50, p = 0.015). The duration of disability correlated with KSKS (r = -0.5, p = 0.02), and level of pain (r = 0.60, p=0.00). The obesity has significantly increased level of pain in the knee (r = -0.44, p = 0.04), all patients were obese persons. Persons with reduced functional abilities avoid body activity, it causes increasing of obesity, and one makes other worse. Concerning on incidence of described problem and its impaction on society in general, it is necessary to make every effort in prevention and treatment of obesity, as important risk factor and consequence of reduced functional abilities and risk factor of most common nowadays diseases.

Alcoholism during adolescence.

It is well known that drug abuse is common in early adolescence with almost the same epidemiological characteristics in economically developed and undeveloped countries. Alcohol abuse is a problem worldwide, which destroys ones social, economical and family life. Our country is in a transition period and consists of postwar society. Therefore, all risk factors for alcohol and other substances abuse are present such as social, economical and medical risk factors. Parents and peers seem to have strong impact on adolescent's behavior. Youngsters like to experiment with a risky life style without adequate knowledge about long-term health damages and risks. Finding risk factors related to alcohol abuse among early adolescents is necessary for appropriate prevention approach. Research covers 600 adolescents (400 in rural and urban part of Sarajevo and 200 in urban and rural part of the Canton Tuzla, as well). Adolescents are of equal gender and age distribution. We used Q 2000 as research tool. Study design is prospective, epidemiological and analytical. Results have been compared within groups, between cantons and among groups. Out of total number of adolescents 93 (15.55 %) were abusing alcohol, 62.4 % of them were from urban and 37.6 % were from rural areas. Results show numerous risk factors related to alcohol abuse among adolescents which could be used as base for preventive activities.

Alcohol abuse and risk behavior among adolescents in larger cities in Bosnia and Herzegovina.

OBJECTIVE: To investigate adolescents who abuse drugs and to describe aspects of risk behavior among alcohol abusers. METHOD: study was conducted on 598 adolescents, aged 12-17 years, with equal distribution in rural and urban areas. Study was based on self-reported questionnaire Q 2000. Study designed as prospective, epidemiological, analitical. Results have been compared within groups, between cantons and among groups. Evaluation of data was done using EPI-info software, with standard statistic methods. RESULTS: Out of the total number of 598 adolescents 15.55% had abused alcohol (urban 62.4%, rural 37.6%); 6.69% tobacco (urban 67.5%, rural 32.5%); 3.34% cannabis (urban 70.0%, rural 30.0%). Results show various aspects of risk behavior among adolescents who consume alcohol such as: truancy 44.1%; low success at school 14.0%; suicidal thoughts 36.6%; unprotected sex 17.7%; drunken driving 10.0%; non use of seat belts 24.7%; deliquency (stealing) 22.2%; destructive behavior 18.9%. Adolescents who abuse drugs have planned to continue with similar behavior in the future: to abuse drugs 31.6%; to smoke 52.2%, to drink alcohol 44.4%, to fight 27.8%; to drive without seat belt 36.8%; to have unprotected sex 16.7%. Risk behavior related to cannabis abuse and tobacco smoking is also described. CONCLUSION: Data suggests that major aspects of adolescents' risk behavior is related to alcohol abuse, what should be considered while designing prevention activities and programmes.

[Biological activities of bone morphological protein in bone regeneration]. / Bioloske aktivnosti kostanog morfogenetskog proteina u kostanoj regeneraciji.

Bone matrix contents various development factors which control structuring and absorption and those factors play important role in bone and cartilage development. Bone morphological proteins are members of TGF-beta super family and their activity is certain becoming from the bone. This activity leads to the serial development processes which include chemo taxis, proliferation and differentiation which results in trans resistant formation of cartilage as well as production of life cells of a bone tissue. Biological activities of re combinative human bone morphogenetic protein 7 (rhBMP-7). induction bone formation of non critical size mandible defect of New Zealand rabbits were researched in the study. Markers of osteoblastic differential in the study included ALP specific activity. Histological analysis performed 7, 14, 30, 60 postoperative days, C-T analysis with determination Bone Mineral Density value of new structured tissue within the defect was done 30 days. Results indicate that ectopic bone formation has been inducted with rhBMP-7 and histological analysis shown mature bone with collagen and ostheociti 60th day. Early 7 day granulocyte tissue with angiogenesis was detected, and after 30 days ostheoblastsis shown with a lot of vascular and mezenhimal tissue. Ostheogenetic processes were characteristic for typical inter membraneous ossification without cartilage tissue. ALP activity was significantly increased 21 days. C-T and Bone Mineral Density value shown density of new structured tissue determinate as bone (413 mg/cm3 and 519 mg/cm3). Studies showed that concentration of 100 mg rhBMP-7 in collagen as career had strong ostheo inductive capacity. Conditions which module BMP depend ostheo induction should be considered in the future. Information could lead to improvements of rhBMP as substitution for bone graft in clinical practice.

Genetic examination of children suffering from cystic fibrosis.

CFTR protein (cystic fibrosis trans membrane conductance regulator) is expressed in multiple epithelial tissues, including upper and lower respiratory tracts, pancreas, sweat glands and gastrointestinal tract. More than 800 mutations and 100 polymorphic variants of DNA sequences were identified in patients with CF (Cystic fibrosis) and CFTR-diseases. In this study, genetic CFTR analysis of the children suffering from chronic lung disease (cystic fibrosis) is presented. They are treated and regularly controlled at the Pediatric hospital Sarajevo. CFTR analysis was done in 9 cases, 4 boys (44.4%) and 5 girls (55.55%). There are 3 children (33.3%) in the age group 1 to 3 years, 1 child (11.1%) in the age group 3 to 6 years, 3 children (33.3%) in the age group 6 to 9 years and 2 children (22.2%) in the age group 9 to 12 years. Genetic analysis was conducted at the Medical center for molecular biology, School of Medicine, Ljubljana. PCR method with PAGE and direct sequestration on ABI PRISM 31 was applied. The majority of children (7 children, i.e. 77.77%) had CFTR mutation Delta F 508 whilst one child had G542X mutation and one child R 1174 mutation. The purpose of this study is to emphasize the need for CFTR gene identification in the institutes of our country.

Psycho-social characteristics of cannabis abusing youth.

It is a well known fact that drug abuse is most common in early adolescence. The most popular substances among youth are cannabis products (made from Cannabis sativa L., Cannabaceae). The majority of heroin and cocaine addicts have started with marijuana. The aim of this study is to show some psycho-social characteristics of adolescents who abuse cannabis. Research conducted during the year 2001 was epidemiological and prospective. The study group included 600 adolescents of equal gender and age distribution. Q 2000 questionnaire was used, as a comprehensive tool for all aspects of adolescent life. The results show strong peer impact on one's behavior. Youth who use cannabis had 2-3 friends of the same behavior, compared to others who had none. We found positive correlation between life stressful events and cannabis abuse. We also noticed tendency to delinquent behavior related to cannabis abuse (35%).

[ABO blood groups and haemolytic disease of newborns--population-genetic analysis]. / Krvne grupe ABO sistema i hemoliticka bolest novorodencadi--populaciono-geneticka analiza.

Classical works dealing with the possibility of mother-child incompatibility with regard to basic ABO blood groups give contradictory conclusions (e.g.--Matsunaga et al. 1962, Thompson et al. 1991). Bioreproductive and population-genetic indicators have been studied in a sample of live births and in two pregnancy samples characterised by different "a priori" and "a posteriori" risk assessment. The analysis points out that ABO blood groups can influence fertility of different parental pairs, with a possible impact on the assessment of the individual pregnancy risc.

Impact of ABO blood groups on the fertility of different parental pairs.

Classical works dealing with the possibility of mother-child incompatibility with regard to basic ABO blood groups give contradictory conclusions. Bioreproductive and population-genetic indicators have been studied in a sample of live births and in two pregnancy samples with different "a priori" and "a posteriori" risk assessment. The analysis points out that ABO blood groups can influence fertility of different parental pairs, and consequently--assessment of the individual pregnancy risk.

Birth order influence on the anthropometric properties of the boys from Tuzla region (Bosnia and Herzegovina).

Birth order and its effect on growth and development of children and youths have rarely been studied so far. The objective of this research was an analysis of the birth order effects on some anthropometric properties of the boys 11-16 years old. The sample consisted of 748 boys from the Tuzla region. As the sample included very few boys born as the third, forth, or fifth child, we decided to consider only the differences in the mean values for some anthropometric parameters between the groups of the first- and the second-born. Measurements were taken according to IBP and the following parameters were investigated: body height, body mass, chest circumference, upper arm circumference, upper leg circumference, sitting height, arm length, leg length, pelvis width, shoulders width, length and width of head. We established that in most generations the firstborn boys have larger mean values for most anthropometric variables in comparison to the second-born.

Some aspects of the psychoactive substances abuse in adolescents.

Cannabis is a plant whose consumption causes hallucinations soon after it has been inhaled. This study was conducted with 600 adolescents of age 12 to 17 with even gender distribution from two residence locations (rural and urban). Epidemiological-analytic study was prospectively conducted, whose results were analysed with standard statistical methods. The results showed that the use of cannabis is greater by the adolescents in high school (15-17 years old), than the adolescents in the elementary school (12-14 years old). The x2 test showed great significance. Among the tested adolescents the consumption of cannabis is more present in urban than in rural type of residency. However, gender structure did not show significant differences. It is obvious that the prevention of the use of psychoactive substances should be at its maximum.

Suicidal thoughts related to psychoactive substance abuse among adolescents.

The enquiry of suicide is of particular importance. According to WHO statistics suicide in people aged from 15 to 24 years has shown the greatest increase of relative mortality. In the group of suicidal adolescents, depression, behavioural disorders, abuse of different substances and personality deviations are frequent factors in presence. The aim of our investigation was to determine the correlation between psychoactive substance abuse and the occurrence of suicidal thoughts in adolescents. The specific "Q 2000" test was used to question 600 adolescents, from Tuzla and Sarajevo Cantons, with the same gender and age distribution and rural-urban ambient. The more prevailing suicidal idea occurrence was observed in cannabis abusers (50.0%) and alcohol abusers (36.6%) in comparison to non-abusers regardless gender and/or living ambient (settlement types) (19.5% and 17.6% per each, x2 = 17.184 p = 0.00001). The augmentation in number of suicidal ideas was not observed in tobacco smokers.

[Type I Gaucher's disease--a rare genetic metabolic disease]. / Tip I Gaucherove bolesti--rijetke geneticki uvjetovane metabolicke bolesti.

Morbus Gaucher is a rare disease. It occurs once in 40,000 to 65,000 persons in the whole world. This is the most frequent lysosome disease in the clinics. This is the first lysosome disease used in perinatal diagnostics and the first one where enzyme therapy was implemented. Case of gaucher disease morbus type I was found in girl of age 14 in Cantonal hospital "Dr. Irfan Ljubijankic" in Bihac. The clinical investigations were carried out at the same hospital, and samples necessary for the further detailed biochemical analysis were taken together with all other relevant data for this disease. In this case, taking into account rules of distribution of certain genes responsible for transfer of characteristics and processes, and following genealogical series, it could be concluded that parents of our patient are heterozygotes (healthy) and carriers of Gaucher gene, and that 1/4 of their children are dominant homozygotes (healthy), 2/4 are heterozygotes (healthy) and carriers of Gaucher gene, and 1/4 of children, together with the patient, are recessive homozygotes, and carriers of Gaucher gene with expression of the type I of this disease. For therapy of Gaucher disease there are in the market two enzyme. Aglucerosis obtained from human placenta and imiglucerasis obtained from cells of ovarian of hamster. The second preparation is cheaper and it is in wide use. There are no differences in the activity, as well as in creation of antibodies--the enzyme entered in use for the first time in 1989, and since 1991 is in wide use. In Bosnia and Herzegovina, therapy is not possible due to the financial restrictions.

Simultaneous abuse of different psychoactive substances among adolescents.

Although the drug abuse has been evidenced in every age of the human life, it seems that its occurrence is crucial during adolescence period with its well-known consequences on the further personality development. Adolescents like to experiment with risky lifestyles without adequate knowledge about their possible harmful effects and consequences. International experiences have represented that early onset of the risky behaviour predisposes young persons for serious problems in social, psychological and physical future lives. It has been noticed that adolescents like to combine different psychoactive substances. Therefore the aim of this study was to present the most important and the most actual substances abused by young adolescents together with the substance characteristics. This research included 600 adolescents with the same prevalence regarding age, sex and living zone (rural-urban). Our research showed that simultaneous abuse of various harmful psychoactive substances (polydrug abuse) is the most common (tobacco smoking and cannabis consumption in 75% of cases, alcohol consumption and cannabis consumption in 80% of cases).