fa تمایز سلول‌های بنیادی مشتق از پالپ دندان انسانی با استفاده از نوروسفر به نورون‌های دوپامینرژیک فیزیولوژی Physiology مقاله پژوهشی Research Article <span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="FA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">سابقه و هدف</span></span></b><span lang="FA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">:</span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> سلول&shy;های </span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">بنیادی پالپ دندان انسانی</span></span><span dir="LTR" new="" roman="" style="font-family:" times="">(hDPSc) </span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">&nbsp;با توجه &shy;به قرابت جنین شناختی که با سلول&shy;های اکتومزانشیمی دارند، می&shy;توانند در حضور فاکتور&shy;های نوروتروفیک مناسب و با استفاده از تکنیک نوروسفر به نورون&shy;های دوپامینرژیک تمایز یابند. هدف از این مطالعه القای تمایز </span></span><span dir="LTR" new="" roman="" style="font-family:" times="">hDPSc</span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> به نورون&shy;های دوپامینرژیک با استفاده از تکنیک نوروسفر<b> </b>است.</span></span><span dir="LTR" style="font-family:"Times New Roman","serif""></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="FA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">مواد و روش &shy;ها</span></span></b><span lang="FA" style="font-size:18.0pt"><span b="" nazanin="" style="font-family:">: </span></span><span dir="LTR" new="" roman="" style="font-family:" times="">hDPScs</span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> به روش مکانیکی_آنزیمی جدا و پس از کشت با استفاده از روش تمایزی کشت نوروسفر به سلول&shy;های بنیادی/پیش سازعصبی و سپس تحت تأثیر</span></span><b> </b><b><span dir="LTR" new="" roman="" style="font-family:" times="">(</span></b><span dir="LTR" new="" roman="" style="font-family:" times="">Glial cell-derived Neurotrophic Factor,</span><span dir="LTR" new="" roman="" style="font-family:" times=""> GDNF</span><span dir="LTR" new="" roman="" style="font-family:" times="">)</span> <span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">و</span></span><span dir="LTR" new="" roman="" style="font-family:" times="">(</span><span dir="LTR" new="" roman="" style="font-family:" times="">Brain-derived</span><b> </b><span dir="LTR" new="" roman="" style="font-family:" times="">Neurotrophic Factor</span><span dir="LTR" new="" roman="" style="font-family:" times="">,BDNF)</span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> به</span></span> <span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">سلول&shy;های دوپامینرژیک تمایز داده شدند. سلول&shy;ها با ایمنوسایتوشیمی و</span></span><span dir="LTR" new="" roman="" style="font-family:" times="">RT&ndash;PCR </span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">&nbsp;مورد ارزیابی قرار گرفتند. هم&shy;چنین میزان رهاسازی دوپامین با دستگاه</span></span><span dir="LTR" new="" roman="" style="font-family:" times="">HPLC </span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">&nbsp;اندازه&shy;گیری شد. تجزیه و تحلیل داده&shy;ها توسط نرم&shy;افزار </span></span><span dir="LTR" new="" roman="" style="font-family:" times="">SPSS</span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> انجام گرفت.</span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="FA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">یافته&shy; ها:</span></span></b><b> </b><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">در گروه&shy; تحت تأثیر القاگرهای تمایزی، سلول&shy;ها به لحاظ فنوتیپی تغییرهای قابل ملاحظه&shy;ای داشتند و سلول&shy;های با ظاهر شبه نورونی مشاهده شدند. یافته&shy;های مولکولی حاکی از این است که ژن&shy;های پرتوان نظیر </span></span><i><span dir="LTR" new="" roman="" style="font-family:" times="">Nestin </span></i><i>&nbsp;</i><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">و<i> </i></span></span><i><span dir="LTR" new="" roman="" style="font-family:" times="">Sox2</span></i><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> با شروع روند تمایز به&shy;تدریج کاهش یافته&shy;اند ولی طی افزودن القاگرهای عصبی میزان بروز ژن&shy;های دخیل در فرآیند نوروژنزیس/ دوپامینرژیک مثل</span></span> <i><span dir="LTR" new="" roman="" style="font-family:" times="">DAT,</span></i><span dir="LTR" new="" roman="" style="font-family:" times=""> ,</span><i><span dir="LTR" new="" roman="" style="font-family:" times="">PITX3,</span></i><i> </i><i><span dir="LTR" new="" roman="" style="font-family:" times="">&nbsp;,Nurr1</span></i><i><span dir="LTR" new="" roman="" style="font-family:" times=""> ,TH</span></i><i> </i><i><span dir="LTR" new="" roman="" style="font-family:" times="">PITX3</span></i><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> افزایش قابل ملاحظه&shy;ای نسبت&shy;به گروه کنترل نشان داد. ارزیابی ایمونوسایتوشیمی نشان داد که سلول&shy;های بیان کننده تیروزین هیدروکسیلاز (</span></span> <span dir="LTR" new="" roman="" style="font-family:" times="">Tirozin hidroksilaz</span><span dir="LTR" new="" roman="" style="font-family:" times="">,TH</span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">) نیز در گروه تیمار مشاهده گردید. علاوه&shy;بر این، میزان ترشح دوپامین در گروه تیمار بطور قابل توجهی افزایش یافته </span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">است (05/0</span></span><span dir="LTR" new="" roman="" style="font-family:" times="">.(</span><span dir="LTR" new="" roman="" style="font-family:" times=""><span style="color:black">P <</span></span><span style="font-family:"B Zar"" lang="FA"></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="tab-stops:right 463.5pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="FA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">نتیجه</span></span></b><b><span dir="LTR" style="font-size:16.0pt"><span new="" roman="" style="font-family:" times="">&shy;</span></span></b><b><span lang="FA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">گیری:</span></span></b><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> نتایج نشان می&shy;دهد که سلول&shy;های بنیادی مشتق از پالپ دندان انسان با استفاده از تکنیک نوروسفر و افزودن القاگرهای مناسب قابلیت تمایز به نورون&shy;های دوپامینرژیکی را دارند.</span></span></span></span></span></span></span></span><br> &nbsp; <span style="font-size:11pt"><span style="line-height:normal"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Aim and Background:</span></span></b> <span new="" roman="" style="font-family:" times="">Human dental pulp stem cells </span><span new="" roman="" style="font-family:" times="">(hDPSCs) </span><span new="" roman="" style="font-family:" times="">that closely common origin with ectomesenchymal cells. </span><span new="" roman="" style="font-family:" times="">could be applied as the source of stem cells for treatment of some neurodegenerative diseases. T</span><span new="" roman="" style="font-family:" times="">he hDPSCc are capable of differentiating into dopaminergic neurons in presence of appropriate neurotrophic factors and </span><span style="font-size:13.0pt"><span new="" roman="" style="font-family:" times="">neurosphere</span></span><span new="" roman="" style="font-family:" times=""> technique with regard to fact This study was carried out to assess the differentiation potential of hDPSCc into dopaminergic neurons using </span><span style="font-size:13.0pt"><span new="" roman="" style="font-family:" times="">neurosphere</span></span><span new="" roman="" style="font-family:" times=""> technique.</span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman","serif""></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Materials and Methods:</span></span></b> <span new="" roman="" style="font-family:" times="">In this study, hDPSCs were isolated from the third molar using mechanical-enzymatic methods. The cells were harvested in a proper condition medium and differentiated into precursor neuronal stem cells and then consequently differentiated into dopaminergic cells in the presence of neuronal induction media including BDNF and GDNF. Cells in each groups were evaluated by immunocytochemistry and RT-PCR assays. In addition, the amount of dopamine released was measured by HPLC. </span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman","serif""></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Results:</span></span></b> <span new="" roman="" style="font-family:" times="">Cells treated with differentiation inducer factors exhibited remarkable changes in phenotypic characteristics and neuron like cells were observed. Molecular results showed that pluripotent genes such as </span><i><span new="" roman="" style="font-family:" times="">Nestin &nbsp;, Sox2</span></i><span new="" roman="" style="font-family:" times=""> expression level gradually reduced in the early stages of differentiation process. However, after treatment with neuronal inducers, genes expression level involved in neurogenesis/dopaminergic like </span><i><span new="" roman="" style="font-family:" times="">DAT</span></i><span new="" roman="" style="font-family:" times=""> ,</span><i><span new="" roman="" style="font-family:" times="">PITX3,</span></i><i> </i><i><span new="" roman="" style="font-family:" times="">&nbsp;,Nurr1</span></i><i><span new="" roman="" style="font-family:" times=""> ,TH </span></i><i><span new="" roman="" style="font-family:" times="">PITX3</span></i><span new="" roman="" style="font-family:" times=""> in treated group increased significantly compared to control group. Moreover, immune cytochemistry assay indicated that cells expressing tyrosine hydroxylase (TH) were also detected in treated group. In addition, dopamine release was significantly increased in treated group </span><span new="" roman="" style="font-family:" times="">(</span><span new="" roman="" style="font-family:" times="">P < 0.05).</span> &nbsp;&nbsp;<span style="font-size:12.0pt"><span style="font-family:"Times New Roman","serif""></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="tab-stops:20.15pt 63.0pt"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Conclusion:</span></span></b> <span new="" roman="" style="font-family:" times="">These research findings confirm that the hDPSCs can be differentiated into dopaminergic neurons via </span><span new="" roman="" style="font-family:" times="">neurosphere technique and appropriate inducers. </span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman","serif""></span></span></span></span></span></span><br> &nbsp; سلول‌های بنیادی پالپ دندان انسان, نوروسفر, سلول‌های پیش‌ساز عصبی, سلول‌های دوپامینرژیک, تمایز, Iau Science Human dental pulp stem cells, neurosphere, neuroprogenitors, dopaminergic neurons, differentiation, Iau Science. 103 114 http://ncmbjpiau.ir/browse.php?a_code=A-10-1881-2&slc_lang=fa&sid=1 Maryam Nazm Bojnordi مریم نظم بجنوردی 100319475328460013864 100319475328460013864 No Department of Anatomy & Cell biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran گروه علوم تشریح و بیولوژی سلولی، دانشکده پزشکی، دانشگاه علوم پزشکی مازندران، ساری، ایران Mazaher Gholipourmalekabadi مظاهر قلی پور ملک آبادی 100319475328460013865 100319475328460013865 No Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran گروه بیوتکنولوژی پزشکی، دانشکده پیراپزشکی، دانشگاه علوم پزشکی ایران، تهران، ایران Hatef Ghasemi Hamidabadi هاتف قاسمی حمیدآبادی hatefdr@gmail.com 100319475328460013866 100319475328460013866 No Immunogenetic Research Center, Department of Anatomy & Cell biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran مرکز تحقیقات ایمونوژنتیک، گروه علوم تشریح و بیولوژی سلولی، دانشکده پزشکی، دانشگاه علوم پزشکی مازندران، ساری، ایران Hassan Keshavarz حسن کشاورز 100319475328460013867 100319475328460013867 Yes Emergency medicine department, Rasool-e-akram hospital, Iran University of Medical Sciences (IUMS), Tehran, Iran گروه آموزشی طب اورژانس، بیمارستان رسول اکرم، دانشکده پزشکی، دانشگاه علوم پزشکی ایران، تهران، ایران