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Article
Comparison between three different protocols for isolation and culture of mouse bone marrow derived mesenchymal stem cells
دراسة مقارنة لثلاثة طرق لعزل وزراعة الخلايا الجذعية اللحمية المشتقة من نخاع عظم الفأر

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Abstract

Bone marrow derived mesenchymal stem cells (BM-MSCs) represent a promising source for cell therapy, As they can differentiate into bone, cartilage, fat, tendon and many other organ progenitor cells. Although the culture of MSCs has been studied for over 30 years. The identification of standard protocol for isolation and characterization have yet to be developed. A comparison study was made on three different isolation techniques, which include: direct plating, red blood lysis buffer strategies, and density gradient (DG) method (using two different gradient media: Ficoll and Percoll) to find the optimal isolation and culture condition for adequate amount of MSCs for clinical use.The results demonstrate that direct plating of whole bone marrow (BM) suspension provides a suitable alternative protocol for isolation of BM-MSCs with minimal requirements, which mainly based on the frequent medium change in primary culture. The BM crud cell suspension were isolated by aspiration from albino mice and cultured in fresh complete isolation media (CIM) RPMI-1640/10%FBS. Adherent cells from BM cells suspension were MSCs which then expanded by complete expansion media (CEM) α-MEM/ 10% FBS. By concluding, direct plating of whole bone marrow represent the alternative method for isolating of mesenchymal stem cells from small specimen of bone marrow. By concluding, the present study was designed to investigate the optimal technique for the isolation of BM-MSCs for use in tissue engineering and regenerative medicine.

تعد الخلايا الجذعية اللحمية المشتقة من نخاع العظم مصدرا واعدا في العلاج الخلوي. لقابليتها على التمايز الى خلاياانسجة العظم، الغضروف،الخلايا الدهنية والوتر والكثير من الخلايا المولدة للأعضاء. رغم ان زراعة الخلايا الجذعية اللحمية درست خلال الثلاثين السنة الماضية الا انها لم توفر نمط قياسي لعزل وتمايز هذه الخلايا لحد الان لذلك تم في هذه الدراسة مقارنه ثلاث طرق مختلفة لعزل الخلايا الجذعية المشتقة من نخاع العظم وهي: العزل بواسطة الطريقة المباشرة، استعمال الدارئ المحلل لكريات الدم الحمر، وباستعمال الوسط المتدرج الكثافة وذلك لإيجاد الطريقة والظروف المثلى للعزل والزراعة لحصد كميات ملائمة من الخلايا الجذعية اللحمية. أوضحت النتائج ان الطريقة المباشرة لزراعه معلق نخاع العظم الخام تمثل أفضل طريقه لعزل الخلايا اللحمية، والتي تقوم أساسا على التغيير المتكرر للوسط الزرعي في المزرعة الأولية. تم جمع عالق نخاع العظم الخام من الفئران البيض وانمائها في الوسط الزرعي العازل(RPMI-1640/10%FBS). كانت الخلايا الملتصقة من معلق نخاع العظم في اوعية الزرع هي الخلايا الجذعية اللحمية والتي تم ادامتها بالوسط الزرعي (MEM/ 10% FBS). وعلية صممت الدراسة الحالية لايجاد الطريقة المثلى لعزل الخلايا الجذعية اللحمية من نخاع العظم المستخدمة في هندسة الانسجة والطب التجديدي.


Article
Immunohistochemical study of CD34 in tooth eruption by using amniotic stem cells

Authors: Lubna K. Jassim لبنى جاسم --- Athraa Y. Al- Hijazi عذراء الحجازي
Journal: Journal of baghdad college of dentistry مجلة كلية طب الاسنان بغداد ISSN: 16800087 Year: 2013 Volume: 25 Issue: 2 Pages: 47-53
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Tooth eruption is a more general process, however, which includes certain posteruptive toothmovements. There are two fundamental requirements for both tooth eruption to occur:(1) Require soft tissue, intervening between tooth structure and alveolar bone, which plays an important role inregulating the remodeling of adjacent tissues.(2) Require bone turnover that is temporally and spatially regulated to facilitate specific translocations of teeththrough alveolar boneThese amniotic stem cells are multipotent and able to differentiate into various tissues, which may be useful forhuman application and recently it used in many medical branches. CD34 is an endothelial marker that is extensivelyused in immunohistochemistry and most vascular endothelial cells. Expression of the stem cell antigen CD34 is adefining hallmark of hemopoietic stem cells and progenitors. This study aimed to study the expression of CD34 bydental cells involved in tooth eruption after administration of amniotic stem cellMaterials and Methods: forty eight albino Swiss mice of one day old age injected with isolated amniotic stem cells inthe anterior region of maxilla (incisors area) other 16 mice injected with saline represents control. Sacrifice 4 mice foreach period (4, 7, 10, and 13) day old age. The result were studied histologically and immunohistochemistry.Results: Immunohistochemical result revealed positive expression of CD34 in pulp (Vascular, Paravascular),Mesenchymal cell and in the Dental sac of different groups. Coincidence test of expression marker CD34 in variousstudied group shows that Chorion application affected on CD34 expression in pulp while Amniotic fluid affected ondental sac.Conclusion Immunohistochemical study of expression marker CD34 in various studied groups show that chorionapplication affected on CD34 in pulp .While amniotic fluid affected on dental follicle


Article
Review Article: Stem cells a healing effect in cancer therapy

Journal: Muthanna Medical Journal مجلة المثنى الطبية ISSN: 2226146x Year: 2017 Volume: 4 Issue: 1 Pages: 50-52
Publisher: Al-Muthanna University جامعة المثنى

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Abstract

Cancer is the DNA replications mistakes, where it's enhance by many chemical factors, environmental factors, radiation, in addition to, the viral and immunological diseases. It has been diagnosed many types of cancers in human being, called by the organs and classified to breast cancer, lung cancer, prostate cancer, and colon cancer …..etc. Oncology is the science of genetic materials disease after birth in humans, concern with all metabolic steps guide to abnormal DNA multiplications within somatic cells In order to stop these abnormal multiplications, the researchers tend to find genetic drugs within telomerase line of research; chemical drugs used to kill the cancer cells at early stages of disease; immunological drugs that used in immune system stimulation materials.

Keywords

Stem cells --- regenerations --- cancer --- drug


Article
Isolationand Identification of mouse bone marrow derived mesenchymal stem cells
عزل وتشخيص الخلايا الجذعية اللحمية المشتقة من نخاع عظم الفئران

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Abstract

Bone marrow derived mesenchymal stem cells (BM-MSCs) were successfully used in regenerative medicine. The purpose of this study was to isolate and characterize mesenchymal stem cells from mouse bone marrow for their subsequent use in researches. Previous data suggest that BM-MSCs are typically enriched by plastic adherent cultures, fibroplastoid cell fraction. However, Identification of MSCs achieved through their morphology, phenotypic characteristics and their biological behavior. The cellular morphology played a major part in identifying MSCs in vitro. In general, immature MSCs appeared as small, spindle-shaped cells, whereas mature MSCs was displayed as larger cells with a flat, polygonal morphology. Cells also tended to be locally confluent, growing in distinct colonies. Immunophenotypic analysis demonstrated that mouse BM-MSCs at passage three uniformly positive for CD44, CD90, CD105 and CD106. However, MSCs were always found to be negative for heamatopoietic specific markers CD34, CD45, and endothelial marker CD31. By concluding, mesenchymal stem cells can be successfully derived from mouse bone marrow by direct plating method. These cells represent a valuable source of stem cells for restoring the damaged organs.

استخدمت الخلايا الجذعية اللحمية المشتقة من نخاع العظم بنجاح في الطب التجديدي. هدفت الدراسة الحالية الى عزل وتوصيف الخلايا الجذعية اللحمية من نخاع عظم الفأر لغرض استخدامها في المجالات البحثية.أوضحت معطيات البحوث السابقة بأن الخلايا الجذعية اللحمية المشتقة من نخاع العظم يمكن اغناؤها فقط بواسطة التصاقها بالمزارع البلاستيكية.وفصل الخلايا الليفية المولدة. ان تشخيص الخلايا الجذعية اللحميةيتم من خلال الشكل، والصفات الوراثية المظهرية، والسلوك البيولوجي لها. ويلعب الشكل الخلوي دورا مهما لتشخيص الخلايا الجذعة اللحمية في المزرعة. وبشكل عام، تظهر الخلايا الجذعية اللحمية الفتية صغيرة الحجم ومغزليه بينما تظهر الناضجة منها بشكل خلايا كبيرة الحجم، مسطحة ومتعددة الاضلع. وتميل الخلايا الى التجمع موضعيا والنمو في مستعمرات محددة.أظهرت نتائج تحليل الكيمياء النسيجية المناعية للخلايا الجذعية اللحمية بانها ذات استجابة موجبة لل CD44، CD90، CD105، CD106، وبنسبه منخفضه لل nestin. وذات استجابة سالبة للمعلمات المتخصصة بالخلايا الدميةCD34، CD45، والمعلم الظهاريCD31. ويمكن الاستنتاج بإمكانية عزل الخلايا الجذعية اللحمية وبنجاح من نخاع عظم الفئران البيض بالطريقة المباشرة والتي تعتبر مصدرمهم للخلايا الجذعية لتعويض الأعضاء المتضررة.


Article
Osteogenesis of Mesenchymal Stem Cells Derived from Bone Marrow in vitro

Authors: Shalal M. Hussein --- Shahlaa M. Salih --- Sally T. Younis
Journal: Al-Nahrain Journal of Science مجلة النهرين للعلوم ISSN: (print)26635453,(online)26635461 Year: 2015 Volume: 18 Issue: 4 Pages: 118-123
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

This study was designed to evaluate osteogenic potential of mesenchymal stem cells (MSCs) isolated from mouse bone marrow in vitro. MSCs were isolated through collecting the thigh bone (femur and tibia). Cells were flushed from bones and MSCs isolated based on the ability of adherence to plastic surfaces. Reactivity MSCs to CD105 and CD34 were tested by immunocytochemistry. Isolated MSCs exhibited positive reactivity to CD105 and negative for the haematopoietic surface marker CD34. Differentiation of isolated MSCs into osteoblast was induced by osteogenic medium consisting of high glucose- Dulbecco's Modified Eagle Medium (DMEM) supplemented with 50 µg /ml Ascorbic acid, 1nM dexamethasone and10 Mm of beta glycerophosphate disodium salt hydrate after 21 days. Osteoblast activity was monitored by evaluation alkaline phosphatase (ALP) activity in osteogenic medium by Reflotron at (0, 7, 14 and 21) days of differentiation. Results recorded that a significant increase in Alkaline phosphatase activity in osteogenic medium at 7 and 14 day culture in comparison with zero day and decreased at 21 day.

صممت الدراسة الحالية لتقييم إمكانية تحول الخلايا الجذعية الوسيطة المعزولة من نخاع عظم الفأر الى خلايا مولدة لخلايا العظم في المختبر. عزلت الخلايا من عظم فخذ 50 فأرة على أساس قدرة التصاقها بالسطوح البلاستيكية. اختبرت الخلايا الجذعية الوسيطة للمعلمات CD105 الخاص بالخلايا الجذعية الوسيطة و CD34 الخاص بالخلايا الجذعية المولدة لخلايا الدم بواسطة كيمياء الخلوية المناعية. أظهرت الخلايا الجذعية الوسيطة المعزولة تفاعلا ايجابيا أتجاه CD105 سلبيا للمعلم على سطح الخلايا الجذعية المولدة لخلايا الدم CD34. حفزت الخلايا الجذعية الوسيطة المعزولة على التمايز لخلايا مولدة لخلايا العظم بواسطة وسط مكون يتألف من DMEMمع 50 ميكروغرام/ مل حمض الأسكوربيك، 1نانو مولاري ديكساميثازون و10 ملي مولاري بيتا كليسروفوسفيت هيدرات الصوديوم بعد 21 يوما. درست فعالية الخلايا المولدة لخلايا العظم عن طريق تقييم فعالية انزيم الفوسفاتيزالقاعدي (ALP) في الوسط المكون لخلايا العظم بواسطة Reflotron بعد فترات مختلفة من التمايز (0، 7، 14، 21) يوما. سجلت النتائج زيادة معنويةP≤0.05 في فعالية ALP في وسط التمايز في اليوم 7 و 14 مقارنة مع اليوم صفر (32.13 ± 0.46 و 23.33 ± 0.88 مقابل 5.22 ± 1.76 وحــــــدة دوليــــة/ لتـــــــر) وانخفضـــــت فـــي اليــــــــوم 21(15.33±1.76 وحدة دولية/لتر. تم التأكد من وجود جينات بيتا الأكتين وأوستيوكالسين في الخلايا المتمايزة من قبل (RT-PCR) بعد 14 يوما من التمايز. اكدت النتائج ظهور حزمتين بحجم الجزيئي (200) زوج قاعدي لبيتا الأكتين و (169) زوج قاعدي لأوستيوكالسين.


Article
Potential Healing Effect of Topical Stem Cell Transplantation and Methandrostenoloneon in Induced Cutaneous Wounds in Dogs

Author: Oday K. Luaibi 1 , Laith K.T.AL-Ani 2 , Zena, M. Fahmi 3
Journal: Iraqi Journal of Biotechnology المجلة العراقية للتقانات الحياتية ISSN: 18154794 Year: 2016 Volume: 15 Issue: 1 Pages: 12-21
Publisher: Baghdad University جامعة بغداد

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Abstract

This study was done to explore the influence of stem cells on cutaneous wounds in comparison with treatment with methandrostenoloneon drug. Cutaneous wound was induced on the back of each dog using disposable dermal biopsy punches . Twenty five dogs were divided into five groups: 1stgroup (T1) was control. 2nd group (T2) was induced without any treatment. 3rd group (T3) was induced wound and treated with phosphate buffer saline, 4rth group (T4) was induced and treated with 5mg/kg/B.W. of Methandrostenolone, 5th group (T5) was induced and treated with stem cells 1x106cell/250 µL of phosphate buffer saline. After 7 and 15 days post treatment level of Hb and RBCs count in T2 and T3 group were decline as compared with control group. The WBCs, Alkaline phosphatase , acid phosphataseas in T2 and T3 showed enhance as contrast with T1. After 7 days, Hb and RBCs in T4 and T5 showed important increase as compared with T2. Moreover the WBCs, alkaline phosphatase, acid phosphatase and total protein levels significantly elevated in the T4 and T5 as compared with T1. After 15 days of treatment the Hb, RBCs, levels of Alkaline phosphatase, acid phosphataseas and total protein levels in T4 showed significant increase as compared with T1, The Hb, RBCs, levels of Alkaline phosphatase, acid phosphataseas and total protein in T5 showed no significant changes as compared with control group (T1). The histopathological section in (T2) explained less collagen and more inflammatory cells, the section of wound tissue in (T4) appeared incomplete healing wound tissue and presence little inflammatory cells, the histopathological section of wound treated in (T5) showed return the wound to normal tissue. From this result it can be concluded that the MSCs have ability to access healing of the wound in comparison with methandrostenolone without any side effect.


Article
Comparative study between the effect of stem cells and Metacam® on induced arthritis in dogs

Author: Oday K. Luaibi
Journal: Kufa Journal For Veterinary Medical Sciences مجلة الكوفة للعلوم الطبية البيطرية ISSN: 20779798 Year: 2015 Volume: 6 Issue: 1 Pages: 85-96
Publisher: University of Kufa جامعة الكوفة

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Objective of this project was to study the effect of stem cells on the induced arthritis in the dogs as compared with treatment Metacam®. Incomplete fruend's adjuvant was used to induce arthritis. Twenty five dogs aged 12 month were divided into five groups: 1st group (T1) was negative control, while 2nd group (T2) was induced without any treatment (positive control), 3rd group (T3) was induction arthritis and treated with 0.2 mg/kg B.W. of Metacam® intravenously (I/V) injection, 4th group (T4) was induced and treated with 1ml/kg of B.W of suspension of culture media injected (I/V), and 5th group (T5) was treated with stem cells suspension in a dose of 2.5x106 cells / ml / Kg B.W. (I/V) injection. Typical clinical signs of arthritis appeared on all induction groups. The results of Hb and RBCs showed clear reduction in all induction groups as compared with (T1), the WBCs, Erythrocyte sediment rates (ESR) and C- reactive protein (CRP) in (T2) showed clear elevation as compared with (T1), After 15 and 30 days of treatment the RBCs count and HB level showed significant decline in T2, T3 and T4 as compared with T5 and T1, the WBCs ,ESR and CRP significantly elevated in the T2 and T4 as compared with T1. The T3 and T5 showed clear depletion in levels of WBCs, ESR and CRP as compared with T2 after 15 and 30 days of treatment, while the T5 revealed no significant difference as compared with T1 after 30 days of treatment. The histopathological changes in T2 and T4 showed erosion and infiltration of inflammatory cells, T3 showed hypertrophy of synovial lining and mild inflammatory cells in synovium, T5 explained return the bone tissue to normal. In this experiment concluded a successful use of stem cells in treatment of arthritis without side effect as comparison with Metacam®.


Article
IDENTIFICATION OF CANCER STEM CELLS IN PAEDIATRIC BRAIN TUMOUR GLIOMAS
الكشف عن خلايا عصبية سلالية في السرطانات الدماغية للاطفال

Authors: BETH COYLE بيث كولي --- DEEMA HUSSEIN ديما حسين --- RAMADHAN T. OTHMAN رمضان عثمان
Journal: Duhok Medical Journal مجلة دهوك الطبية ISSN: ISSN: 20717334 (online)/ ISSN: 20717326 (Print) Year: 2008 Volume: 2 Issue: 1 Pages: 54-70
Publisher: University of Dohuk جامعة دهوك

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Abstract

Background Brain tumours are the leading cause of cancer mortality in children and remain difficult to cure despite advances in surgery and adjuvant therapy. Objective To identify cancer stem-like cells in established cell lines from three paediatric brain tumour (PBT) gliomas. Setting Queen’s Medical Centre/ Nottingham city/ UK. Methodology Three glioma cell lines were studied including one high grade glioblastoma multiforme (BT4), one well differentiated oligodendroglioma (Olig1), and one recurrent ependymoma (EPN1). A control cell line of mouse neural stem cells (C17.2) was also included for comparison. Results Established tumour cell lines maintain stem cell marker (nestin and Sox2) expression when grown as monolayers in 15% foetal bovine serum/Dulbecco’s modified Eagle’s medium. Cells derived from these cell lines are able to form neurospheres when cultured in serum-free stem cell media containing basic fibroblast growth factor and human epidermal growth factor. These neurospheres are self-renewable and re-form new neurospheres when dissociated and cultured in fresh medium supplemented with growth factors. The percentages of neural stem cell marker CD133 positive cells were determined by flow cytometry analysis of neurospheres from the three cultured cell lines, BT4 47.2% ± 10.5, EPN1 40.4% ± 8.9 and Olig1 48.7% ± 2.9 (mean ± standard error). Under conditions promoting differentiation, cells derived from neurospheres were multipotent giving rise to neurons, astrocytes, and oligodendrocytes, at different levels for each tumour cell line. Conclusion Paediatric gliomas contain cancer stem-like cells that are able to self-renew and differentiate into three neural lineages

الهدف: تمّيز خلايا سلالية عصبية في ثلاثة أنواع من سرطانات الدماغ في الاطفال. المكان: المجمع الملكي الطبي مدينة نوتنكهام بريطانيا.المنهجية: شملت الدراسة ثلاثة أنواع من سرطانات الدماغ في الاطفال و شملت كل من (high grade glioblastoma multiforme BT4, one well differentiated oligodendroglioma olig1 and one recurrent ependymoma EPN1).و قد شملت الدراسة خلايا عصبية معيارية من الفئران للمقارنة(C17.2).النتائج: أظهرت النتائج أن الخلايا السرطانية تبقي علامات الخلايا السلالية عندما تنمو على شكل طبقة واحدة في 15% من مصل الدم للبليد الجنيني (foetal bovine serum/Dulbecco’s modified Eagle’s medium) هذه الخلايا لها القابلية على ان تنقسم و تتكاثر لتكوين كريات عصبية(neurosphere) عند زرعها في وسط خالي من مصل الدم و ذلك بأضافة محفزات النمو (FGF and EGF). يمكن لهذه الخلايا ان تنقسم و تعيد بناء كريات جديدة عندما يتم فصلها وزرعها في وسط مماثل مع اضافة هذه المحفزات. تم تحديد نسبة الخلايا الايجابية لعلامة خلايا السلالية العصبية (CD133)باستعمال التحليل فلوسايتوميتر (flowcytometry) في جميع الخلايا المزروعة و كانت النسب كالتالي: BT4 47.2%±10.5, EPN1 40.4%±8.9, Olig1 48.7%±2.9 (المتوسط,الخطأ المعياري). يمكن لهذه الخلايا ان تتخصص الى خلايا عصبية وخلايا مساندة neuron, astrocyte, and oligodendrocyte تحت ظروف تساعد على تخصص الخلايا السلالية. ولكن على مستويات مختلفة لكل خط خلية سرطانية.الأستنتاجات: السرطنات الدماغية للأطفال تحتوي على خلايا مشابه للخلايا السرطانية السلالية التي لها القابلية على الانقسام و تجديد نفسها و لها القابلية ع لىالتحول الى ثلاثة انساب عصبية.


Article
Isolation and characterization of bone marrowmesenchymal stem cells from rat and rabbit; a modified method
عزل وتوصيف خلايا نخاع العظم ألجذعيه الميزنكيميه من الجرذانوالأرانب .طريقة محورة

Authors: Limes HusamAlmanseekanaa --- Ali MansoorJasim --- RaedHamzah Mohammed
Journal: karbala journal of pharmaceutical sciences مجلة كربلاء للعلوم الصيدلانية ISSN: 70272221 Year: 2012 Issue: 3 Pages: 44-51
Publisher: Kerbala University جامعة كربلاء

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Bone marrow MSCs were obtained from New Zealand White rabbits (~3 kg, male) and also from white rat (Ratusratus) by the Guideline for Animal Care and use Committee for Teaching and Research using a modified procedure (1,2) Briefly, after the animals was sacrified with carbon monoxide cabinet, tibias and femurs were excised and the bone marrow was extracted under sterile conditions.The collected marrow was mixed and dispersed with PBS. After that the (MSCs) were separated by density-gradient centrifugation over ficoll. Viability of the separated MSCs was determined via trypan blue staining technique. Then, cells were seeded into ten tissue culture flasks containing complete culture medium suplimented with 10% fetal bovine serum (FBS) and antibiotics, incubated at 370C in a humidified atmosphere with 5% CO2. Once the colonies reached 80–90% confluence, they were ready to be detached with trypsin/EDTA and suspended in medium for continuous culture

تضمنت الدراسة عزل و تنمية الخلايا الجذعية الميزنكيمية من نقي العظم للارنب الابيض الذكر والذي يزن حوالي 3 كغم والجرذ الابيض المختبريين طبقا لقوانين هيئة العناية بالحيوان واستعماله للأغراض البحثية و التعليمية وذلك باتباع طريقة محورة لهذا الغرض. بأيجاز, بعد قتل الحيوان داخل كابينة غاز أحادي أوكسيد الكاربون تم استئصال عظمي الفخذ وعظمي القصبة وبعدها تم استخراج نقي العظم منها والذي خلط مع محلول الفوسفات الملحي ( (PBS. ثم تم فصل الخلايا الجذعية الميزنكيمية من الخليط بوساطة السنترفيوج مع فارق الكثافة وذلك بأضافة مادة الفيكول. بعد ذلك تم تحديد حيوية الخلايا المفصولة عن طريق تقنية التصبيغ بصبغة التريبان الزرقاء. بعدها تم استنبات الخلايا في عشرة اطباق زرعية نسيجية يحوي كل منها على الوسط الزرعي النسيجي المتكامل مدعما بالمصل البقري والمضادات الحيوية حيث تم الحضن بدرجة 37 مئوية في محيط رطب يحوي 5% غاز ثاني أوكسيد الكاربون. عندما أصبحت درجة اتصال المستعمرات الخلوية 80-90% كانت جاهزة لفصلها بواسطة محلول أنزيم التربسين مع EDTA واستنباتها في الوسط الزرعي الدائمي


Article
Enhancement of tooth eruption by using amniotic stem cells (Immunohistochemical study of VEGF marker)

Authors: Athraa Y. Al- Hijazi عذراء الحجازي --- Lubna K. Jassim لبنى جاسم
Journal: Journal of baghdad college of dentistry مجلة كلية طب الاسنان بغداد ISSN: 16800087 Year: 2013 Volume: 25 Issue: 2 Pages: 80-88
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Tooth eruption is a localized process in the jaws which exhibits precise timing and bilateral symmetry.Develop within the jaws and their eruption is a complex infancy process during which they move through bone totheir functional positions within the oral cavity. For species with more than one set of teeth, eruption of the second setalso accomplishes. The key to the successful clinical management of tooth eruption consists of understanding thatthis process consists largely of the local regulation of alveolar bone metabolism to produce bone resorption in thedirection of eruption and shift and formation of bone at the opposite side.The amniotic sac contains a considerablequantity of stem cells. These amniotic stem cells are able to differentiate into various tissues, which used in many field.Vascular endothelial growth factor (VEGF) is an important angiogenic factor reported to induce migration andproliferation of endothelial cells, enhance vascular permeability, and modulate thrombogenicity. VEGF expression incultured cells (smooth muscle cells, macrophages, endothelial cells) is controlled by growth factors and cytokines.The aim of this study was to study the administration of cell molecules of (Chorion, Amnion and Amniotic fluid)around developing mouse tooth and studying the expression of VEGF marker.Materials and Methods: forty eight albino Swiss mice of one day old age injected with isolated amniotic stem cells inthe anterior region of maxilla (incisors area) other 16 mice injected with saline represents control. Sacrifice 4 mice foreach period (4, 7, 10, and 13) day old age. The result were studied histologically and immunohistochemistry.Results: VEGF marker localized and identified in 3 areas; pulp, P.D.L, and Bone. In pulp. The mean value of positiveVEGF expression showed to be highest in Amnion group in comparison to the other studied groups. The marginalmean value of all periods reported to be highest in Amnion groups followed by Chorion group. The period 10 dayshowed highest marginal means value for positive VEGF expression for all groups. In P.D.L. area Amniotic fluid recordsthe highest mean and marginal mean value specifically at day-10 in comparison to other studied groups. In Bonearea Amniotic fluid records the highest mean and marginal mean value among the studied groups followed byChorion group. Period 7-day and 10-day shows high mean value for VEGF expression. Coincidence test for VEGFmarker illustrates to be affected by Amniotic fluid application in P.D.L. and in bone area while Amnion and Chorionapplication showed to be concerned with pulp.Conclusion. It reported that amniotic fluid application affected on expression of VEGF in P.D.L and bone whileamnion and chorion showed to affect on expression of VEGF in pulp.The present study highlighted on clinical andresearcher application of Amniotic fluid and Chorion for supplement of stem cell in dental tissue engineering or evenin other body tissues.

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