Cytotoxicity difference of 316L stainless steel and titanium reconstruction plate

• Main Authors: Ni Putu Mira Sumarta, Coen Pramono Danudiningrat, Ester Arijani Rachmat, Pratiwi Soesilawati
• Format: Article
• Language: English
• Published: Universitas Airlangga 2011-03-01
• Series: Dental Journal: Majalah Kedokteran Gigi
• Subjects: 316L Stainless steel plate; titanium plate; cyototoxicity; MTT assay; Plat stainless steel; plat titanium; sitotoksisitas
• Online Access: http://e-journal.unair.ac.id/index.php/MKG/article/view/880

<p><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Background: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Pure titanium is the most biocompatible material today and used as a gold standard for metallic implants. However, </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>stainless steel is still being used as implants because of its strength, ductility, lower price, corrosion resistant and biocompatibility. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Purpose: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>This study was done to revealed the cytotoxicity difference between reconstruction plate made of 316L stainless steel and of </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>commercially pure (CP) titanium in baby hamster kidney-21 (BHK-21) fibroblast culture through MTT assay. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Methods: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Eight samples </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>were prepared from reconstruction plates made of stainless steel type 316L grade 2 (Coen’s reconstruction plate</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>®</em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>) that had been </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>cut into cylindrical form of 2 mm in diameter and 3 mm long. The other one were made of CP titanium (STEMA Gmbh</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>®)</em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>) of 2 mm in </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>diameter and 2,2 mm long; and had been cleaned with silica paper and ultrasonic cleaner, and sterilized in autoclave at 121° C for </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>20 minutes.</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>9 </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Both samples were bathed into microplate well containing 50 μl of fibroblast cells with 2 x 10</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>5 </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>density in Rosewell Park </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Memorial Institute-1640 (RPMI-1640) media, spinned at 30 rpm for 5 minutes. Microplate well was incubated for 24 and 48 hours in </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>37° C. After 24 hours, each well that will be read at 24 hour were added with 50 μl solution containing 5mg/ml MTT reagent in </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>phosphate buffer saline (PBS) solutions, then reincubated for 4 hours in CO</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>2 </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>10% and 37° C. Colorometric assay with MTT was used </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>to evaluate viability of the cells population after 24 hours. Then, each well were added with 50 μl dimethyl sulfoxide (DMSO) and </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>reincubated for 5 minutes in 37° C. the wells were read using Elisa reader in 620 nm wave length. Same steps were done for the wells </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>that will be read in 48 hours. Each data were tabulated and analyzed using independent T-test with significance of 5%. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Results: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>This </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>study showed that the percentage of living fibroblast after exposure to 316L stainless steel reconstruction plate was 61.58% after 24 </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>hours and 62.33% after 48 hours. And after exposure to titanium reconstruction plate, the percentage of living fibroblast was 98.69% </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>after 24 hours and 82.24% after 48 hours. Based on cytotoxicity parameter (CD</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>50%</em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>)</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>, </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>both reconstruction plate made of 316L stainless </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>steel or titanium showed as a non-toxic materials to fibroblast. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Conclusion: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Both reconstruction plate made of stainless steel and CP </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>titanium were non-toxic to fibroblast, although the stainless steel plate showed lower cytotoxicity level compared to titanium. Therefore </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>a reconstruction plate made from stainless steel type 316L can be used as a safe material for mandibular reconstruction.</em></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></p><p><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em> <span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Latar belakang: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Titanium murni adalah bahan yang paling biokompatibel saat ini dan digunakan sebagai standar emas implan </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>logam. Saat ini stainless steel masih digunakan karena kekuatan, ductility, harganya yang murah, tahan terhadap korosi dan cukup </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>biokompatibel. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Tujuan: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Penelitian ini dilakukan untuk mengetahui perbedaan sitotoksisitas antara plat rekonstruksi yang terbuat </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>dari titanium murni komersial dan plat rekonstruksi yang terbuat dari stainless steel pada kultur sel fibroblas baby hamster kidney-</em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>21 (BHK-21) menggunakan MTT assay. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Metode: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Delapan sampel yang masing-masing tipe 316L terbuat dari stainless steel 316L </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>grade 2 (Coen’s reconstruction plate</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>®</em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>) yang dipotong berbentuk silinder diameter 2 mm dan panjang 3 mm, serta yang terbuat dari </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>titanium murni komersial (STEMA Gmbh</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>®</em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>) diameter 2 mm dan panjang 2,2 mm; dan dibersihkan dengan kertas silika dan pembersih </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>ultrasonik serta disterilkan dengan autoclave pada suhu 121° C selama 20 menit. Kedua sampel dimasukkan ke dalam sumur mikroplat </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>yang mengandung 50 μl sel fibroblas dengan kepadatan 2 </em><span style="font-family: TimesNewRomanPS-ItalicMT; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>× </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>10</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>5 </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>dalam media Rosewell Park Memorial Institute-1640 (RPMI-1640), </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>diputar dengan kecepatan 30 rpm selama 5 menit. Sumur mikroplat diinkubasi selama 24 dan 48 jam pada suhu 37° C. Setelah 24 "&gt; <span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>jam, pada tiap sumur yang akan dibaca pada jam ke 24 ditambahkan 50 μl cairan yang mengandung 5mg/ml MTT dalam phosphat </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>buffer saline (PBS), kemudian diinkubasi kembali selama 4 jam dalam CO</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>2 </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>10% pada suhu 37° C. Assay kolorimetri dengan MTT </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>digunakan untuk mengetahui viabilitas populasi sel setelah 24 jam. Setiap sumur ditambahkan pelarut dimetil sulfoksida (DMSO) dan </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>diinkubasi kembali selama 5 menit pada suhu 37° C. sumur-sumur tersebut kemudian dibaca dengan Elisa reader dengan panjang </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>gelombang 620 nm. Langkah yang sama dilakukan pada sumur-sumur yang akan dibaca pada jam ke 48. Data kemudian ditabulasi dan </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>dianalisis dengan menggunakan independent T-test dengan signifikansi 5%. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Hasil: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Penelitian ini menunjukkan presentase fibroblas </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>hidup setelah terpapar plat rekonstruksi yang terbuat dari stainless steel adalah 61,58% setelah 24 jam dan 62,33% setelah 48 jam. </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Dan setelah paparan dengan plat rekonstruksi yang terbuat dari titanium murni adalah 98,69% setelah 24 jam dan 82,24% setelah </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>48 jam. Berdasarkan pada parameter sitotoksisitas (CD</em><span style="font-family: TribuneItalic; font-size: 7pt; color: #231f20; font-style: normal; font-variant: normal;"><em>50%</em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>) kedua plat rekonstruksi baik yang terbuat dari titanium murni maupun </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>yang terbuat dari stainless steel tipe 316L merupakan bahan yang tidak bersifat toksik terhadap fibroblas. </em><span style="font-family: TribuneBold; font-size: 10pt; color: #231f20; font-style: normal; font-variant: normal;"><strong>Kesimpulan: </strong><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>Kedua plat </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>rekonstruksi baik yang terbuat dari stainless steel maupun CP titanium tidak bersifat toksik terhadap fibroblas, walaupun plat stainless </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>steel menunjukkan level sitotoksisitas yang lebih rendah daripada titanium murni. Dengan demikian plat rekonstruksi yang terbuat </em><span style="font-family: TribuneItalic; font-size: 9pt; color: #231f20; font-style: normal; font-variant: normal;"><em>dari stainless steel 316 L aman digunakan sebagai bahan untuk rekonstruksi mandibula.</em></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span><br /></em></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></em></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></p>