مقايسه تاثير دو نوع داربست تجاري بر رفتار زيستي سلول هاي اندوتليال در ترميم لثه (آزمايشگاهي)

Comparison of two commercial dermal matrix on biological behavior of endothelial cells (in vitro)

سابقه و هدف: رفتار زيستي سلول هاي اندوتليال به لحاظ رشد و اتصال روي داربست (ADM) Acellular Dermal Matrix اهميت دارد. اين مطالعه با هدف بررسي رفتار زيستي سلول هاي اندوتليال، روي دو نوع داربست تجاري داخلي در سال 1397 انجام گرفت ﻣﻮاد و روش ﻫﺎ: تحقيق به روش تجربي انجام گرفت ﺳﻠﻮل ﻫﺎي اندوتليال اﻧﺴﺎﻧﯽ مشتق شده از سياهرگ نافي انساني به صورت آزمايشگاهي ﮐﺸﺖ داده ﺷﺪﻧﺪ. دو نوع (ADM ) شامل گروه 1: همانندساز بافت كيش و گروه 2 شركت دانش بنيان فرآورده بافت ايرانيان) در مطالعه به كار رفت. ﺗﮑﺜﯿﺮ ﺳﻠﻮﻟﻬﺎ در ﻧﻤﻮﻧﻪ دارﺑﺴﺖ ﻫﺎ و ﮔﺮوه ﮐﻨﺘﺮل در 12 ﺳﺎﻋﺖ، 24 ﺳﺎﻋﺖ، 84 ﺳﺎﻋﺖ و 7 روز ﺑﻌﺪ از ﮐﺸﺖ ﺑﺎ آزﻣﻮن ﮐﻠﺮيﻤﺘﺮيﮏ MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ) ارزيﺎﺑﯽ شد. ﺑﻪ ﻣﻨﻈﻮر ارزيﺎﺑﯽ ﻣﻮرﻓﻮﻟﻮژي سلولي، ﺳﻠﻮل ﻫﺎ ﺑﻌﺪ از 24 ﺳﺎﻋﺖ اﻧﮑﻮﺑﺎﺳﯿﻮن ﺑﻪ وﺳﯿﻠﻪ اﺗﺎﻧﻮل و Hexamethyldisilazane (HMDS) دﻫﯿﺪراﺗﻪ ﺗﻮﺳﻂ ﻗﻄﺮات ﭘﻼﺗﯿﻨﯿﻮم ﭘﻮﺷﺶ داده ﺷﺪه و ﺑﻪ ﻟﺤﺎظ ﻣﺴﺎﺣﺖ و ﻣﺪور ﺑﻮدن ﺗﻮﺳﻂ ﻣﯿﮑﺮوﺳﮑﻮپ اﻟﮑﺘﺮوﻧﯽ ﻣﻮرد ارزيﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. يافته ها: ﻣﯿﺰان اﺗﺼﺎل ﺷﺎﺧﺺ MTT و DAPI (4′,6-diamidino-2-phenylindole) در هر ﺳﻪ ﮔﺮوه ﻣﺸﺎﺑﻪ ﺑﻮد(0/4P< ) در پيگيري 1 هفته اي ميزان رشد در گروه كنترل 0/001 ± 2/66، داربست 1، 0/192±2/09 و داربست 2، 0/17±1/64 بود كه اين اختلاف لحاظ آماري بين سه گروه معنادار بود. (0/01>P) ﻧﺘﯿﺠﻪ ﮔﯿﺮي: به نظر مي رسد در ﺷﺮايﻂ آزﻣﺎيﺸﮕﺎﻫﯽ هر دو داربست مورد مطالعه از ﻟﺤﺎظ اﺗﺼﺎل ﺳﻠﻮﻟﻬﺎي اندوتليال مشابه مي باشند اﻣﺎ رﺷﺪ ﺳﻠﻮل ﻫﺎي اندوتليال ﺑﺮ روي دارﺑﺴﺖ 1 ﻧﺴﺒﺖ ﺑﻪ دارﺑﺴﺖ 2 ﭘﺲ از 7 روز ﺑﯿﺸﺘﺮ بود.

Aim and background: Behavior of endothelial cells can have significant role regarding their proliferation and attachment on acellualr dermal grafts in regeneration procedures.The purpose of this study is to evaluate the effects of two dermal scaffolds on attachment and proliferation of human umbilical vein endothelial cells. Methods and Materials: This study was conducted on experimental approach. Human umbilical vein endothelial cells were seeded, experimentally. Two commercially available types of ADM were used. 1. Kish tissue regeneration co. 2. Knowledge Based Company of Persian Textile Products. The proliferation rate in matrix and control groups were evaluated 12,24, 84 hours and 7 days after culture using MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ) test. The morphologic evaluation was made after 24 hours incubation with Ethanol and Dehydrated Hexamethyldisilazane (HMDS) using Platinium coated drops. The surface and circularity of cells was examined by electronic microscope. Results: The results for DAPI and MTT index was similar in 3 groups(P<0.4).After 1 week the control group rate was 2.66 ± 0.001, fist group, 2.09 ±0.001 and second group, 1.64±0.17. The result was statistically signi􀏐icant.(P<0.1) Conclusion: It seems, in experimental status, both matrix was similar in endothelia cell adhesion. But, proliferation rate of endothelial cells was more in matrix 1 v.s matrix 2 after 7 days.