KARAKTERISASI MAKROSKOPIK, MIKROSTRUKTUR, DAN KEKUATAN MEKANIS SARAF TEPI DOMBA DAN BABI KONDISI SEGAR DAN PASCA DESELULARISASI

FAJAR SHODIQ PERMATA, dr. Rina Susilowati, Ph.D.

2013 | Tesis | S2 IKD/Rekayasa Biomedis

Latar Belakang: Domba dikembangkan sebagai donor xenograft saraf tepi pengganti babi. Jaringan xenograft pasca deselularisasi yang baik memiliki karakter mirip karakter jaringan segar. Tujuan penelitian ini adalah melakukan karakterisasi makroskopik, mikrostruktur dan kekuatan mekanis saraf tepi asal domba dan babi kondisi segar dan pasca deselularisasi. Metode: Sebanyak masing-masing 14 sampel n. ischiadicus domba dan babi dibagi menjadi 2 kelompok yaitu saraf segar dan saraf deselularisasi. Deselularisasi menggunakan metode freezing-thawing dilanjutkan perendaman dalam 0,1% Sodium dodecyl sulphate (SDS)-ethylene diamine tetra acetic (EDTA) selama 14 hari untuk saraf domba dan 25 hari untuk saraf babi. Karakterisasi dilakukan secara makroskopik, mikrostruktur pada irisan blok parafin, dan kekuatan mekanis dengan uji tarik. Data kuantitatif disajikan sebagai rerata ± SD. Uji statistik menggunakan independent T-test dan Mann Whitney Test. Hasil dan Pembahasan: Hasil penelitian menunjukkan bahwa saraf domba pasca deselularisasi memiliki diameter makroskopik lebih kecil, jumlah sisa komponen serabut saraf lebih sedikit, namun ultimate tensile stress lebih besar (p<0,05) dibandingkan dengan saraf babi pasca deselularisasi. Saraf domba pasca deselularisasi tidak berbeda (p≥0,05) dengan saraf segarnya pada karakter makroskopik, diameter dan jumlah fasikulus, dan kekuatan mekanis yang diuji. Adanya jaringan lemak di saraf babi menyebabkan larutan deselularisasi sulit masuk ke dalam jaringan dan menyebabkan saraf mudah putus. Mayoritas diameter pori yang dihasilkan pada saraf pasca deselularisasi baik domba dan babi adalah 3-12μm dan >12 μm. Pori yang dihasilkan memungkinkan untuk dikombinasikan dengan growth factor dan sel guna penanganan saraf tepi yang terputus. Kesimpulan: Kesimpulan dari penelitian ini adalah saraf tepi domba memiliki karakter yang berbeda dengan saraf tepi babi baik kondisi segar atau pasca deselularisasi, namun berdasarkan hasil penelitian domba dapat menjadi kandidat donor saraf tepi xenograft acellular pengganti babi di Indonesia.

Introduction: Sheep are developed as peripheral nerve xenograft donor to replace swine. Decellularized xenograft tissue is good if it has same character with fresh tissue. The research purposes are conducting characterization macroscopic, microstuctural, and mechanical properties of sheep and swine peripheral nerve in fresh and decellularized condition Method: Fourteen sciatic nerves of sheep and swine respectively were divided to be 2 groups as fresh nerves group and decellularized nerves group. Decellularization used freezing-thawing method and continued immersion in 0.1% (w/v) sodium dodecyl sulphate (SDS)-ethylene diamine tetra acetic (EDTA) during 14 days for sheep nerves and during 25 days for swine nerves. Characterizations were conducted macroscopically, microstucturally by paraffin block section, and mechanically by tensile test. Quantitative data were showed as mean±SD. Statistic test were used independent T-test and Mann Whitney Test. Result and Discussion: Results showed that decellularized sheep nerves have smaller macroscopic diameter, fewer remnant nerve fiber component, but higher ultimate tensile stress (p<0.05) than those of decellularized swine nerves. Decellularized sheep nerves were insignificant different (p≥0.05) with its fresh nerves in macroscopic character, fascicles diameter and number, mechanical properties which been tested. Because swine nerves have much adipose tissue, decellularization solution was difficult to infiltrate into nerves and both fresh and decellularized nerves were easily broken. Resulted diameter pores in decellularized nerves both of sheep and swine were 3-12μm and >12 μm. The pores are possible to be combined with growth factor and cells to treat injured peripheral nerves in human. Conclusion: The conclusion is that sheep peripheral nerves have different character to swine peripheral nerve both in fresh and decellularized condition. However, based on result, in Indonesia, sheep can be candidate as acellular peripheral nerve xenograft acellular donor substituting swine.

Kata Kunci : domba, babi, saraf tepi deselularisasi, kekuatan mekanis, makroskopik mikrostruktur