´Ù¾çÇÑ rhBMP-2 ÄÚÆà ÀÓÇöõÆ® Ç¥¸é¿¡¼ÀÇ Àΰ£ °£¿± Áٱ⼼Æ÷ÀÇ ºÎÂø, Áõ½Ä, ºÐȹÝÀÀ ºÐ¼®
Analysis of attachment, proliferation and differentiation response of human mesenchymal stem cell to various implant surfaces coated with rhBMP-2
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ÀÌÀα¸ ( Lee In-Ku ) - °í·Á´ëÇб³ ÀÇ°ú´ëÇÐ Ä¡ÀÇÇб³½Ç
ÇÑÀÎÈ£ ( Han In-Ho ) - ƯÇãû ÈÇлý¸í°øÇнɻ籹
Ȳ¼±¿í ( Hwang Sun-Wook ) - °í·Á´ëÇб³ ¾È»êº´¿ø ½Å°æ»ý¸®Çаú
·ùÀçÁØ ( Ryu Jae-Jun ) - °í·Á´ëÇб³ ¾È»êº´¿ø Ä¡°úº¸Ã¶°ú
KMID : 0362420120500010044
Abstract
¿¬±¸ ¸ñÀû: º» ½ÇÇèÀº °ÅÄ£ Ç¥¸éÀ¸·Î À¯ÀǼº ÀÖ´Â Áٱ⼼Æ÷¹ÝÀÀÀ» ³ªÅ¸³Â´ø 4 °¡ÁöÀÇ Æ¼Å¸´½ Ç¥¸é À§¿¡ rhBMP-2¸¦ ÄÚÆÃÇßÀ» ¶§ ¾î¶² À¯ÀÇÇÑ Áٱ⼼Æ÷¹ÝÀÀ(¼¼Æ÷ºÎÂø, Áõ½Ä, ºÐÈ)ÀÌ ³ªÅ¸³ª´ÂÁö ºñ±³ ºÐ¼®ÇÔÀ¸·Î rhBMP-2 ÄÚÆÃÀ» À§ÇÑ °¡Àå ÀûÀýÇÑ Ç¥¸éÀ» Æò°¡Çϱâ À§ÇØ ½ÃÇàµÇ¾ú´Ù.
¿¬±¸ Àç·á ¹× ¹æ¹ý: ´ëÁ¶±ºÀÎ ±â°èÀý»èÇ¥¸é(machined surface)°ú ½ÇÇ豺ÀÎ ¾ç±Ø»êÈ(anodized), RBM, SLA Ç¥¸é¿¡ rhBMP-2¸¦ ÄÚÆÃÇÑ ÈÄ ÄÚÆÃÇÏÁö ¾ÊÀº Ç¥¸é°ú °°ÀÌ 8°¡Áö Ç¥¸é À§¿¡ Àΰ£Áٱ⼼Æ÷¸¦ ¹è¾çÇÏ¿´´Ù. ¹è¾ç ÈÄ 24½Ã°£ ÈÄ SEMÀ» ÅëÇØ Áٱ⼼Æ÷ÀÇ ºÎÂøÀ» Æò°¡ÇÏ¿´°í ¹è¾ç 3, 7, 14ÀÏÈÄMTT¿Í ALP °Ë»ç¸¦ ÅëÇØ Áٱ⼼Æ÷ÀÇ Áõ½Ä°ú ºÐȹÝÀÀÀ» Æò°¡ÇÏ¿´´Ù. ±×¸®°í ¹è¾ç 7ÀÏÈÄRT-PCR °Ë»ç¸¦ ÅëÇØ Type I collagen, osteocalcin, osteopontinÀÇ À¯ÀüÀÚ ¹ßÇöÀÇ º¯È¸¦ Æò°¡ÇÏ¿´´Ù.
°á°ú: SEM Æò°¡¿¡¼ 4°¡Áö rhBMP-2 Ç¥¸éÀÌ ÄÚÆÃÇÏÁö ¾ÊÀº Ç¥¸é¿¡ ºñÇØ ¼¼Æ÷ºÎÂø ¸éÀûÀÌ ³Ð°í ±ä¹ÐÇÏ¸ç ¼¼Æ÷µ¹±â°¡ ´õ ¸¹ÀÌ °üÂûµÇ¾ú´Ù. ¾ç±Ø»êÈ rhBMP-2ÄÚÆÃÇ¥¸é¿¡¼ °¡Àå µÎµå·¯Áö°Ô °üÂûµÇ¾ú´Ù. MTT °Ë»ç¿¡¼ Å©°Ô ÀÇ¹Ì ÀÖ´Â Â÷ÀÌ´Â ³ªÅ¸³ªÁö ¾Ê¾Ò´Ù. ALP°Ë»ç¿¡¼ ¾ç±Ø»êÈ rhBMP-2ÄÚÆà ǥ¸éÀº ´ëÁ¶±º°ú ºñ±³Çؼ (3, 14ÀÏ) ¶Ç RBM rhBMP-2 ÄÚÆà ǥ¸é°ú ºñ±³Çؼ (14ÀÏ) À¯ÀǼº ÀÖ´Â ALP È°¼ºµµÀÇ Áõ°¡¸¦ ³ªÅ¸³»¾ú´Ù(P<.05). RT-PCR °Ë»ç¿¡¼ osteocalcin°ú osteopontinÀÇ À¯ÀüÀÚ ¹ßÇöÀº ¾ç±Ø»êÈ rhBMP-2ÄÚÆà ǥ¸é¿¡¼ ³ô°Ô ³ªÅ¸³µ´Ù.
°á·Ð: ¾ç±Ø»êÈ rhBMP-2ÄÚÆÃÇ¥¸éÀÌ Áٱ⼼Æ÷ÀÇ ºÎÂø°ú ºÐȽÇÇè¿¡¼ ´ëÁ¶±ºÇ¥¸é°ú rhBMP-2¸¦ ÄÚÆÃÇÑ ±â°èÀý»èÇ¥¸éÀ̳ª RBM Ç¥¸é¿¡ ºñÇØ À¯ÀǼº ÀÖ´Â Áõ°¡¸¦ ³ªÅ¸³Â´Ù(P<.05).
Purpose: In this paper we tried to evaluate the most appropriate surface for rhBMP-2 coating among 4 rough titanium surfaces.
Materials and methods: We used machined surface as a control group and anodized, RBM and SLA surfaces as test groups. We coated rhBMP-2 on the 4 surfaces and with uncoated surfaces for each case, we cultured human mesenchymal stem cells on all 8 surfaces. 24 hours after we measured the stem cell¡¯ attachment with SEM, and on 3rd, 7th, and 14th days, we checked the cell proliferation and differentiation by using MTT and ALP activity assay. And on the 7th day after the culture, we performed RT-PCR assay to determine whether the expression levels of Type I collagen, osteocalcin, osteopontin were changed.
Results: We observed with SEM that 4 rhBMP-2 coated surfaces exhibited wider and tighter cell attachment and more cell process spreading than uncoated surfaces. The anodized rhBMP-2 surface caused robustest effects. In MTT assay we could not find any meaningful difference. In ALP assay there was a significant increase (P<.05) in the ALP activity of anodized rhBMP-2 coated surface compared with that of the control (3rd and 14th days) and with that of the RBM rhBMP-2 coated surface (14th day). In RT-PCR assay there was increased expressions in the anodized rhBMP-2 coated surface for osteocalcin, and osteopontin.
Conclusion: We found that the anodized rhBMP-2 coated surface were most prominent stem cell attachment and differentiation in compared to control and Machined rhBMP-2 coated, RBM rhBMP-2 coated surface.
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Áٱ⼼Æ÷; ¾ç±Ø»êÈ Ç¥¸é; RBM Ç¥¸é; SLA Ç¥¸é
Stem cell; rhBMP-2; Anodized; RBM; SLA surface
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