Structural evaluation and animal implantation of porous eggshell wastederived hydroxyapatite graft as bone substitution

Abstract

The development of hydroxyapatite graft with high economically value is needed for orthopedic practice in developing countries. Eggsell waste is well known as natural substance for calcium resource. It has been used as raw material in producing hydroxyapatite. This study was conducted to synthesize porous hydroxyapatite from eggshell waste and evaluate its activity as bone substitution. The porous hydroxyapatite graft was manufactured from eggshell and sugar as a raw material using hydrothermal process. The porous eggshell waste-derived hydroxyapatite (EW-HAP) graft was characterized using X ray difractometer (XRD) and analytical scanning electron microscope (SEM) and compared with commercial hydroxyapatite (HAP) JCPDS 09-432 graft (Bangros ®) as standard. The porous EW-HAP graft obtained was then implanted on critically sized femoral defects surgically created in the right thigh of male Wistar rats (Rattus norvegicus) with Bangros ® as control. Radiological examination using XRD and histological examination using hematoxyline-and-eosin staining of the bone femour were performed at 28 days after implantation. The results showed that the XRD pattern for EW-HAP was likely similar with the HAP standard. However, the SEM examination showed that the pasticle size of EW-HAP graft (2.5-3 µm) was higher than those HAP standard graft (1.5-2 µm). Radiographs according to the International of Limb Salvage (ISOLS) radiological evaluation system between EW-HAP graft (6.1 ± 1.45) and HAP control graft (6.9 ± 2.10) was not significantly different (p>0.05). Moreover, histological examination according to Lane and Shandu scoring system between the both graft (4.0 ± 0.94 versus 4.4 ± 0.92) was also not significantly different (p>0.05). It can be concluded that the structure EW-HAP graft is similar with HAP graft standard. The both grafts have also equal outcome as bone substitution. ABSTRAK Pengembangan graft hidroksiapatit dengan nilai ekonomi tinggi diperlukan dalam bidang ortopedi di negara berkembang. Limbah cangkang terus dikenal luas sebagai bahan alam sumber kalsium. Limbah cangkang telur telah digunakan sebagai bahan untuk menghasilkan hidroksiapatit. Penelitian ini dilakukan untuk mensintesis graft hidroksiapatit berpori dari limbah cangkang telur dan mengkaji aktivitasnya sebagai pengganti tulang. Graft hidroksiapatit berpori dibuat dari bahan dasar cangkang telur (HAP-LCT) dan gula menggunakan proses hidrotermal. Graft yang dihasilkan dikarakterisasi difraktometer X-ray (DXR) dan mikroskop elektron skaning (MES) analitik dan dibandingkan dengan graft JCPDS 09-432 hidroksiapatit komersial (Bangros ®) sebagai standar. Graft berpori HAP-LCT yang diperoleh kemudian diimplantasikan pada critically sized tulang femur yang dibuat melalui operasi pada paha kanan tikus Wistar jantan (Rattus norvegicus) dengan Bangros ® sebagai kontrol. Pemeriksaan radiologi menggunakan DXR dan histologi dengan pengecatan hematosilin-eosin tulang femur dilakukan pada hari ke 28 setelah implantasi. Hasil 177 Sakti & Magetsari, Structural evaluation and animal implantation of porous eggshell waste-derived hydroxyapatite graft as bone substitution penelitian menunjukkan bahwa gambaran DXR graft HAP-LCT sama dengan graft HAP standar. Namun demikian, hasil pemeriksaan MES menunjukkan ukuran partikel HAP-LCT (2,5-3 µm) lebih tinggi dari pada graft HAP standar (1,5-2 µm). Gambaran radiografi menurut sistem evaluasi radiologi International of Limb Salvage (ISOLS) graft HAP-LCT (6,1 ± 1,45) dan and graft HAP kontrol (6,9 ± 2,10) berbeda bermakna (p>0.05). Lebih lanjut dari hasil pemeriksaan histologi menurut sistem penilaian Lane dan Shandu antara kedua graft (4,0 ± 0,94) tidak berbeda bermakna (p>0,05). Dapat disimpulkan bahwa struktur graft HAP-LCT sama dengan graft HAP standar. Kedua graft juga mempunyai luaran sama sebagi pengganti tulang.