PENGARUH KONSENTRASI SERISIN PADA SCAFFOLD BIOKOMPOSIT SERISIN-HIDROKSIAPATIT-STARCH BIOPLASTIC TERHADAP VIABILITAS DAN PERLEKATAN SEL FIBROBLAS

ABAJAIDUN MAHULAUW, Ir. Alva Edy Tontowi, M.Sc., Ph.D; Dr. drg. Siti Sunarintyas, M.Kes

2015 | Tesis | S2 Bioteknologi

Hidroksiapatit dan starch bioplastic merupakan biomaterial yang diaplikasikan dalam rekayasa jaringan namun belum maksimal dalam mendukung viabilitas dan perlekatan sel dikarenakan kurangnya gugus fungsional dan bersifat hidrofobik. Serisin dari ulat sutera merupakan protein yang telah diketahui mampu meningkatkan interaksi sel dengan material karena memiliki gugus fungsional seperti hidroksil, karboksil dan amina. Tujuan dari penelitian ini adalah mengkaji pengaruh konsentrasi serisin pada scaffold biokomposit serisin-hidroksiapatit-starch bioplastic terhadap viabilitas dan perlekatan sel fibroblas. Subjek penelitian ini adalah scaffold biokomposit yang disintesis dari serisin (Bombyx moriI), hidroksiapatit (Sigma Aldrich), dan tepung tapioka (PT. Budi Starch & Sweetener TBK). Delapan kelompok scaffold yang terdiri dari satu kelompok scaffold HA/bP tanpa serisin dan tujuh kelompok dengan konsentrasi berbeda (0,005; 0,01; 0,02; 0,04; 0,08; 0,16; dan 0,32%) digunakan untuk mengkaji viabilitas sel fibroblas dan empat kelompok scaffold biokomposit (HA/bP (tanpa serisin), SS/HA/bP (SS 0,005%), SS/HA/bP (SS 0,01%), dan SS/HA/bP (SS 0,32%)) digunakan untuk mengkaji perlekatan sel fibroblas. Uji viabilitas dan perlekatan dilakukan dengan memasukkan scaffold ke dalam kultur sel fibroblas gingiva. Viabilitas sel diukur berdasarkan nilai densitas optikal (OD) menggunakan ELISA Plate Reader setelah dilakukan uji MTT (24 jam waktu kultur). Perlekatan sel dihitung setelah 4 jam waktu kultur menggunakan hemositometer. Karakterisasi permukaan dilakukan dengan menggunakan FTIR dan SEM/EDS untuk mendeteksi gugus fungsional dan porositas scaffold sebelum dan sesudah aplikasi serisin. analisis data menggunakan ANOVA satua arah dan LSD. Sel fibroblas menunjukkan viabilitas lebih baik pada konsentrasi serisin sebesar 0,01% dibanding konsentrasi lain dan lebih signifikan daripada scaffold tanpa serisin. Jumlah sel yang melekat pada HA/bP (7830 ± 0,38) berbeda bermakna (p<0,05) dengan SS/HA/bP dengan serisin 0,005% (10500 ± 0,25), SS/HA/bP dengan serisin 0,01% (10920 ± 0,38), dan SS/HA/bP dengan serisin 0,32% (11250 ± 0,25). Perbedaan bermakna antar kelompok scaffold dengan serisin ditemukan pada SS/HA/bP (SS 0,005%) dan SS/HA/bP (SS 0,32%) dengan nilai probabilitas (p) = 0,022. Grafik FTIR menunjukkan terdapatnya gugus fungsional serisin yang telah terimobilisasi pada scaffold. Hasil SEM menunjukkan porositas yang lebih baik setelah aplikasi serisin yang cocok untuk perlekatan sel fibroblas. Kesimpulan dari penelitian ini adalah aplikasi serisin pada scaffold biokomposit SS/HA/bP mampu meningkatkan viabilitas dan perlekatan sel fibroblas.

Hydroxyapatite and starch bioplastic are applied biomaterials in tissue engineering, but not maximized in favor of viability and cell attachment due to lack functional groups and hydrophobic. Sericin of silkworm silk is a protein that has been known to increase the interaction of cells with the material because it has functional groups such as hydroxyl, carboxyl and amine. The aim of this study was to examine the effect of the concentration of sericin on sericin-hydroxyapatite-starch bioplastic biocomposite scaffold on fibroblasts viability and attachments. The Subject of this study was biocomposite scaffold which synthesized from sericin (Bombyx mori), hydroxyapatite (Sigma Aldrich), and tapioca flour (PT. Budi Starch & Sweetener TBK). Delapan kelompok scaffold yang terdiri dari satu kelompok scaffold HA/bP tanpa serisin dan tujuh (sericin-hydroxyapatite-starch bioplastic (SS/HA/bP) with different concentration of sericin. Eight groups of SS/HA/bP biocomposite scaffold which consists of HA/bP scaffold group without sericin and seven groups with different concentrations of sericin (0,005; 0,01; 0,02; 0,04; 0,08; 0.,16; and 0,32%) were used to assess the viability of fibroblasts and four biocomposite scaffold groups (HA/bP (without serisin), SS/HA/bP (SS 0.005%), SS/HA/bP (SS 0.01%), and SS/HA/bP (SS 0.32%) were used to assess fibroblasts attachment. Viability and attachment test were conducted by immersing biocomposite scaffold into gingiva fibroblast cells culture. Cell viability was measured by optical density values (OD) using ELISA Plate Reader after MTT test (24 hours of culture). Cell attachment was calculated after 4 hours of culture using a hemocytometer. Surface characterization performed by using FTIR and SEM/EDS to detect functional group and the porosity of the scaffold before and after application of sericin. The data were analyzed used ANOVA one way and LSD. Fibroblasts showed better viability at concentrations of 0.01% compared to the other concentration and more significant than the scaffold without sericin. Fibroblast cell attachment to the scaffold applied sericin higher that cells attached to the scaffold-based HA/bP without sericin and there were significant differences between the treatment groups. FTIR chart showed the presence of sericin functional groups which have immobilized to the scaffold. SEM results showed better porosity after application of sericin which suitable for the attachment of fibroblast cells. The conclusion from this study was the application of sericin on SS/HA/bP biocomposite scaffold able to improve the viability and attachment of fibroblast cells. the amount of fibroblast cells attached to HA/bP (7830 ± 0,38) was significantly difference (p<0,05) with SS/HA/bP with sericin 0,005% (10500 ± 0,25), SS/HA/bP with sericin 0,01% (10920 ± 0,38), and SS/HA/bP with sericin 0,32% (11250 ± 0,25). The significant difference among scaffold group with sericin was found on SS/HA/bP (SS 0,005%) and SS/HA/bP (SS 0,32%) with probability value (p) = 0,022. In conclusion, sericin concentration in biocomposite scaffold SS/HA/bP increased the viability and the amount of fibroblast cells attachment.

Kata Kunci : Keywords: Sericin, Hydroxyapatite, Starch Bioplastic, Fibroblasts Viability, Fibroblasts Attachment