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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.

Medicinski Glasnik, Tarik Kapidžić Zenica, Herzegovina Bosnia, Selma Uzunović Zenica, Filip Čulo, Bosnia And Herzegovina, A. Redžić, Bosnia Sarajevo et al.

Original article 1 Mean platelet volume predicts the glycemic control deterioration in diabetes mellitus type 2 patients 36 Can a finding of cervical vestibular evoked myogenic potentials contribute to vestibular migraine diagnostics? 50 Does odor and taste identification change during hyperemesis gravidarum? 62 Factors affecting mortality in emergency surgery in cases of complicated colorectal cancer ABSTRACT Aim To investigate association of mean platelet volume (MPV) and glycemic control markers, and whether MPV could be used as a predictor of deterioration of glucoregulation in Diabetes mellitus type 2 (DMT2) patients. Methods The cross-sectional study included 106 DMT2 patients, treated at the Primary Health Care Centre in Zenica, distributed into groups according to glycated haemoglobin (HbA1c) values: A (n=44, HbA1c ≤7.0%) and B (n=62, HbA1c>7.0%). Spearman's correlation coefficients were calculated to evaluate the relationships between MPV and glycemic control markers. Binomial logistic regression analysis was performed to estimate the relationship between glycemic control, as dichotomous outcome, and MPV as the main predictor. Diagnostic value of MPV as a marker for poor glucoregulation was estimated by using ROC analysis. Results Mean platelet volume was significantly higher in the group B compared to the group A (p<0.0005). Significant positive correlations of MPV with fasting blood glucose and HbA1c were found in the total sample (rho=0.382, p<0.0005; rho=0.430, p<0.0005, respectively). Mean platelet volume was positively associated with the risk of inadequate glycemic control, with 2 times increased odds of inadequate glycemic control per femtoliter The area under ROC curve for MPV was 0.726 (95% CI: =0.628-0.823, p <0.0005). At the best cutoff value 9.55 fL, MPV showed sensitivity of 82% and specificity of 54.5%. Conclusion Mean platelet volume correlates with glycemic control markers in DMT2 patients. It could be used as a simple and cost-effective predictor of deterioration of glucoregulation.

M. Soliman, H. El Zenati, A. Smajilagić, S. Ibrahim, Kiran Saeed, Osama Kokach

Trismus is the inability to normally open the mouth. Inflammation of soft tissue around impacted third molar tooth is the most common cause of trismus. Other causes include tetanus, inflammation of muscles of mastication, peritonsillar abscess, temporomandibular joint disorders, as a temporary side effect of many stimulants of the sympathetic nervous system and some recreational drugs. Trismus is an anesthetic challenge particularly for airway management. If general anesthesia is induced, difficult ventilation and/or intubation may lead to morbidity or mortality 1-5. We present the case of a patient with a trismus who was in need of incision and drainage and tooth extraction. The case report has been approved for publication from the IRB (HMC, DOHA, QATAR).

A. Smajilagić, M. Aljičević, A. Redžić, S. Filipović, A. Lagumdžija

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.

A. Redžić, A. Smajilagić, M. Aljičević, L. Berberović

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.

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).

A. Smajilagić, Moustafa Al-Khalil, Amira Redjic, S. Filipović, Besima Hadjihasanovic, Sami Lappalainen

OBJECTIVE To obtain information on the feasibility of employing recombinant human bone morphogenetic protein-7 (rhBMP-7), mixed with bone marrow as a substitute for autologous bone graft. METHODS We carried out the study in the University Clinic Center, Sarajevo, Bosnia and Herzegovina, from February 2002 to January 2004. Six New Zealand rabbits underwent hemiresection of the mandible. We placed rhBMP-7 in a concentration of 100 micrograms and collagen (ACS) as carrier mixed with bone marrow in the defects in 3 animals. In another group of 3, we restored the defect by placement of autologous bone graft harvested from the iliac crest. We evaluated the results by the activity of the alkaline phosphatase enzyme (ALP), CT assessment with bone mineral density (BMD) analysis, and clinical and histologic examinations. RESULTS The ALP activity was significantly higher after 14 days, in the rhBMP7/ACS sites than the bone graft sites (30th day). Mean BMD of tissue induced by rhBMP-7/ACS on the 30th day was 510 mg/cm3 in caudal, and 553 mg/cm3 in sagittal plane, and bone graft tissue was 510 mg/cm3 in caudal and 530 mg/cm3 in sagittal plane. Clinical inspection and histologic examination on the 60th day showed complete bridged defects with abundant woven bone in 2 of 3 rhBMP-7/ACS sites (67%), and no incorporation of the autologous bone graft into the other groups treated sites. CONCLUSION The rhBMP-7/ACS mixed with bone marrow was very effective in establishing complete bone regeneration into the defects, indicating that it could be an adequate alternative for autologous bone transplantation.

A. Smajilagić, A. Redžić, S. Filipović, B. Hadžihasanović

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.

A. Smajilagić, F. Dilberović

The great diffusion of the surgical techniques in jaws surgery and the progress of the radiological imagining procedures expressed many interest in clinical anatomy of the mental foramen (MF). The study goal was to determine the precise location of the MF and the surrounding anatomical landmarks. Measurements of the MF position relative to the surgical landmarks and related posterior teeth were made on 20 dry mandibles with complete dentition and intact alveolar bridge obtained from the Institute of Anatomy, School of Medicine, University in Sarajevo. The measurements were made by anthropometric methods on the booth sides of the mandible, and compared with measurement made on the orthopantomogram radiographs of the same mandibles. The most common position of the MF was in line with the longitudinal axis of the second premolar. In the vertical plane on the skulls the MF lays in the midpoint of the distance between the lower border of the mandible and the alveolar margin, however on the orthopantomogram MF appeared slightly bellow the midpoint. In the horizontal plane it lays approximately one third of the distance between the mandibular symphysis and the posterior border of the ramus of the mandible measurement from A-P projection and one quarter of that distance measurement from the profile projection. There were no significant differences between distances MF from posterior border of the ramus of the mandible measurement from A-P and profile projection and the one obtained on orthopantomogram and their ratio is constant value determine das 1,065. The MF was on average 25 mm lateral to the mandibular skeletal midline and symmetrical, and symmetry was preserved on the orthopantomogram. The measurement showed significant differences in distances of M from superior border mandible measurement on dry mandible and orthopantomogram radiographs while distance bellow the MF was not significantly different. The constant values of MF distance to the posterior border of the ramus of the mandible measured as 1,065 and the distance to the medial skeletal line of 2,11 made it possible to also determine average angle of 43 degrees stream of the cor-pus of the mandible behind MF. These values in combination with ratios of MF to the different anatomic landmarks designated as relative horizontal and relative vertical position, would be of importance not only from anatomical but also from practical point of view for estimation of alveolar bridge resorption and preoperative analysis in orthognat postresection or implant surgery in the mandible.

Irma Ramović, T. Masic, A. Smajilagić, E. Imamović, H. Piranić

Education is a process of accepting knowledge and skills or manners modification throught training. One of the most important characteristics in surgery education are classical education program through adequate literature and observation in operation room. Lip and palate clefts are relative rare diseases and classical knowledge acquirement of surgery methods in this field is sometimes defficient. Progress in computer technologies have potential and capacity to improve continuate and self-education training. We used this advancement in additional education in field of lip and palate clefts surgery, thanks to our partnership relations with Smile Train International Organisation. Accent of our cooperation was put in straight of new technology of virtual surgery--The Smile Train Virtual Surgery Videos.

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