KANDUNGAN KOLAGEN DAN MYELIN PADA N.ISCHIADICUS BABI DAN DOMBA SETELAH PROSES DESELULARISASI “KAJIAN DESKRIPTIF MIKROSKOPIK”

MYA THAT MON, dr. Rina Susilowati, Ph.D

2014 | Skripsi | PENDIDIKAN DOKTER

Pendahuluan: Domba dikembangkan sebagai donor xenograft saraf tepi pengganti babi. Jaringan xenograft pasca deselularisasi sebaiknya memiliki karakteristik yang mirip dengan jaringan sebelum deselularisasi. Metode deselularisasi yang baik akan menghasilkan jaringan acellular dengan komponen residual selular yang sedikit dan kerusakan matriks ekstraselular yang minimal sebagai perancah. Tujuan dari penelitian ini adalah untuk membandingkan kandungan kolagen dan myelin pada n.ischiadicus domba dan babi setelah proses deselularisasi. Metode: Jenis penelitian adalah observasional analitik. Sampel adalah n. ischiadicus domba dan babi. N.ischaidicus kanan digunakan sebagai saraf segar dan n.ischiadicus kiri digunakan sebagai saraf deselularisasi. Deselularisasi dilakukan dengan 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. Spesimen dicat dengan Osmium tetroxide dan dibuat irisan blok parafin dilanjutkan dengan pengecatan Mallory’s trichrome. Hasil dideskripsikan secara kuantitatif dan semi-kuantitatif. Analisis statistik menggunakan Mann Whitney Test untuk membandingkan antara spesiman domba dan babi. Hasil dan pembahasan: Spesimen domba terdeselularisasi secara menyeluruh, kerusakan perineurium sedikit, menunjukkan jaringan kolagen masih tersisa diantara fasikulus dan hanya temukan sedikit jaringan lemak pada spesimen pasca deselularisasi. Sedangkan spesimen babi ditemukan beberapa komponen residual seluler masih tersisa, kerusakan perineurium banyak ditemukan, kolagen antara fasikulus sangan sedikit dan banyak jaringan lemak masih tersisa pada spesimen pasca deselularisasi. Persentase kerusakan matriks ekstraseluler pada saraf babi lebih banyak dibandingkan saraf domba (p<0,05). Persentase sisa kandungan myelin juga ditemukan lebih banyak pada saraf babi dibandingkan saraf domba (p<0,05). Kesimpulan: N.ischiadicus domba memiliki karakter yang berbeda dengan n.ischiadicus 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: Peripheral nerves of sheep are developed as xenograft donor to replace swine nerve. Good decellularization method can results acellular nerve with low cellular residual content and minimal damage of extracellular matrix scaffold. The decellularized nerve need to have similar characteristics of microstructure and macrostructure as fresh tissue. The research purpose is conducting characterization of collagen and myelin content in peripheral nerve of sheep and swine after decellularization process. Method: Type of research is observational analysis. Samples used are sciatic nerve of sheep and swine which is divided into 2 groups as fresh group and decellularized group. Method of decellularization is freezing-thawing. Samples were immersed in 0,1% Sodium dodecyl sulphate (SDS)-ethylene diamine tetra acetic (EDTA) as long as 14 days for sheep and 25 days for swine. Specimens were stained with Osmium tetroxide and made into slices of paraffin which is stained again with Mallory’s trichrome stain to examine the content of collagen and residual myelin. The results were analyzed qualitatively and semi-qualitatively between the groups. Mann Whitney Test was used to analyze statistically. Result and Discussion: Sheep nerve specimens were able to be decellularized evenly, collagen still remain between fascicles and have few perineurium damage and only small amount of adipose tissues can be found after decellularization. Mean while in nerve specimen of swine, some parts of specimen showed remnant of cellular residuals, have few collagen remain between fasicicles and much perineurium damage and adipose tissues remain after decellularization because it have thick fat sheath so that decellularization solution was difficult to infiltrate into nerves. Swine nerve showed higer percentage of damage of extracellular matrix and residual myelin than sheep nerve (p<0,05). Conclusion: 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 : n.ischiadicus deselularisasi, domba, babi, myelin, kolagen.