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ترمیم آسیب‌های تاندونی با استفاده از پلاسمای غنی شده از پلاکت در مدل حیوانی خرگوش: بررسی هیستوپاتولوژیکی و بیومکانیکی

ترمیم آسیب‌های تاندونی با استفاده از پلاسمای غنی شده از پلاکت در مدل حیوانی خرگوش: بررسی هیستوپاتولوژیکی و بیومکانیکی

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چکیده 

 تاندون بافت همبند رابط بین استخوان و عضله است که دچار آسیب‌های متعدد می‌گردد. در این میان به دلیل خون‌رسانی ضعیف و احتمال چسبندگی در حین‌ترمیم، مدیریت آسیب تاندون از اهمیت ویژه‌ای برخوردار است. بنابراین یافتن روش سریع در جهت ترمیم بافت تاندون از اهدافی است که طی سال‌ها دنبال می‌شده است. در برخی از مطالعات، استفاده از مواد محرک در ترمیم تاندون گزارش شده است. در این مطالعه به بررسی اثر PRP زنولوگ و مقایسه آن با PRP اتولوگ بر روند ترمیم تاندون خرگوش پرداخته شده است. 45 خرگوش بالغ با وزن تقریبی 5/1 کیلوگرم در محدوده سنی 7 ماه تا 1 سال انتخاب شد. ژل پلاکتی، با اضافه کردن ترومبین به پلاسمای غنی از پلاکت تهیه شد. بعد از قطع تاندون آشیل و بخیه به روش بانل مایر، 5/0 سی سی ژل پلاکتی زنولوگ در پای یک گروه و 5/0 سی سی ژل پلاکتی اتولوگ در گروه دیگر تزریق شد و ارزیابی بیومکانیکی و هیستوپاتولوژی بعد از گذر 7، 14 و 28 روز انجام گرفت.

فاکتورهای بیومکانیکیStrain   Ultimate strength,و حداکثر مقاومت کششی (peak stress) با استفاده از دستگاه Instron مورد ارزیابی قرار گرفت. بلافاصله بعد از ارزیابی بیومکانیکی، نمونه‌ها در فرمالین بافر 10% به آزمایشگاه  منتقل شد و بعد ز آن، با رنگ آمیزی H&E ومیسون تری کروم مورد ارزیابی هیستوپاتولوژی قرار گرفتند. پس از مطالعه با میکروسکوپ نوری و اسکوربندی مورد آنالیز آماری با نرم افزار Graphpad prism 5 و با سطح معنی‌داری 05/0 p<قرار گرفتند.

در این بررسی اختلاف آماری معنی‌داری در فاکتورهای بیومکانیکیStrain  ، Ultimate strengthو Stress سه گروه اتولوگ و زنولوگ و شاهد در هفته اول و چهارم مشاهده شد .این فاکتورها  در گروه اتولوگ و زنولوگ که PRP تزریق شده بود، بهتر مشاهده شد. لازم به ذکر است که این فاکتورها در گروه اتولوگ بهتر از گروه زنولوگ دیده شد.در ارزیابی هیستوپاتولوژی، فاکتورهای شدت التهاب، میزان بلوغ فیبروبلاست‌ها، میزان انباشت رشته‌های همبندی، جهت‌ یابی رشته‌های همبندی و میزان عروق‌زایی مورد بررسی قرار گرفتند. اختلاف آماری معنی دار بین گروه‌های اتولوگ و شاهد و همچنین اتولوگ و زنولوگ در میزان بلوغ فیبروبلاست‌ها در هفته‌های اول، دوم و چهارم، در میزان جهت‌یابی کلاژن در هفته دوم و از نظر میزان انباشت رشته‌های همبندی در هفته دوم و چهارم مشاهده شد. در مقایسه هفته به هفته از نظر انباشت رشته‍‌های همبندی و جهت یابی رشته‌های همبندی در دو گروه اتولوگ و زنولوگ، اختلاف آماری معنادار بین هفته اول و چهارم مشاهده شد. در مطالعه حاضر ژل پلاکتی در برخی فاکتورهای بیومکانیکی اختلاف معنی داری ایجاد کرده و باعث استحکام بافت شده است. از نظر فاکتورهای هیستوپاتولوژی، میزان بلوغ فیبروبلاست‌ها، جهت یابی رشته‌های همبندی و انباشت رشته‌های همبندی را افزایش داده است.

واژگان کلیدی: ترمیم، تاندون، PRP

دسته: , برچسب: , ,

فهرست مطالب

 

مقدمه- 5

کلیات- 8

2-1- ساختار تاندون- 8

2-2- کالبد شناسی تاندون- 8

2-3- بافت‌شناسی تاندون- 9

2-4- خون رسانی تاندون: 13

2-5- عصب رسانی تاندون- 13

2-6- ترمیم تاندون- 14

2-6-1- فاز التهابی ترمیم تاندون- 14

2-6-2- فاز فیبروپلازی- 15

2-6-3- فاز بازآرایی- 17

2-7- مشکلات ترمیم تاندون و مدیریت آن- 18

2-8- انتقال و جذب نیروها در تاندون- 18

2-9- منحنی نیرو-تغییر طول تاندون(force- elongation): 20

2-10- منحنی stress- strain  تاندون: 22

2-11- پلاکت‌ها و PRP- 23

2-11-1- پلاکت‌ها: منشا و ترشحات آن‌ها 24

2-11-2- پلاکت‌ها به عنوان حامل فاکتورهای رشد: 24

2-11-3- تولید PRP و ویژگی‌های آن: 25

2-11-4- PRP به عنوان سیستم تحویل فاکتورهای رشد: 26

2-11-5- پلاسمای غنی از پلاكت خودی (autologous PRP ) 28

2-11-6- کاربردهای کنونی PRP: 29

2-12- بکارگیری ژل پلاکتی زنولوگ– 31

مواد و روش کار 34

3-1- حیوانات، مواد و دستگاه‌های مورد استفاده: 34

3-2- گروه بندی حیوانات: 35

3-3- روش کار 35

3-3-1- تهیه‌ی ترومبین گاوی- 35

3-3-2- تهیه ژل پلاکتی- 36

3-3-3- روش جراحی- 36

3-3-4- نگه‌داری بعد از عمل- 37

3-3-5- نمونه‌ گیری- 37

3-4- روش بررسی و آنالیز داده‌ها 38

3-4-1- تست بیومکانیک و ارزیابی آماری آن- 38

3-4-2- تهیه مقاطع بافتی جهت ارزیابی هیستوپاتولوژیک– 38

3-4-3- آماده سازی نمونه‌ها جهت مطالعه هیستوپاتولوژیک– 38

3-4-3-1- رنگ‌ آمیزی هماتوکسیلین و ائوزین- 39

3-4-3-2- رنگ آمیزی میسون- تری کروم 40

3-4-4- درجه بندی و ارزیابی آماری داده‌های هیستوپاتولوژیک– 40

نتایج- 41

4-1- یافته‌های هیستوپاتولوژیک– 41

4-2- یافته های بیومکانیکی- 50

بحث- 52

5-1- پیشنهادات- 58

منابع- 59

چکیده…………………………………………………………………………………………………………………………………………………..74

 

فهرست جداول

جدول شماره 2-1 عوامل رشد موجود در PRP و عملکرد آنها 30

جدول شماره 4-1- نتایج حاصله از ارزیابی هیستوپاتولوژی بر اساس Median (Min-Max) 42

جدول شماره 4-2- نتایج حاصل از ارزیابی بیومکانیکی فاکتور(Ultimate strength) 50

جدول شماره 4-3- نتایج حاصل از ارزیابی بیومکانیکی فاکتورهای Strain) و (Stress 51

فهرست شکل ها

شکل 2-1- منحنی نیرو-تغییر طول تاندون- 21

شکل 2-2- منحنی Stress-Strain تاندون- 23

شکل 3-1- مرحله ای از مجموع مراحل تهیه ژل پلاکتی- 36

شکل3-2- تاندون آشیل خرگوش– 37

شکل3-3 بخیه تاندون آشیل- 37

شکل 3-4- بافت تاندون در حال کشش در دستگاه تست کشش– 38

شکل شماره 4-1- 43

شکل شماره 4-2- 43

شکل شماره 4-3- 44

شکل شماره4-4- 44

شکل شماره4-5- 45

شکل شماره 4-6- 45

شکل شماره4-7- 46

شکل شماره 4-8- 46

شکل شماره 4-9- 47

شکل شماره 4-10- 47

شکل شماره 4-11- 48

شکل شماره 4-12- 48

شکل شماره 4-13- 49

 شکل شماره 4-14- 49

شکل شماره 4-15- 49

 

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