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Bachtiar EW
Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia
Dentistry

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Endothelial cell cultured on HA/TCP/chitosan scaffold for bone tissue engineering Bachtiar EW; Amir LR; Abbas B; Utami S
Dental Journal (Majalah Kedokteran Gigi) Vol. 44 No. 2 (2011): June 2011
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (469.41 KB) | DOI: 10.20473/j.djmkg.v44.i2.p67-71

Abstract

Background: Angiogenesis is crucial for the success of bone reconstruction through tissue engineering. Currently, is still not known the activity of endothelial cells that is responsible for blood vessel formation, cultured in HA/TCP/chitosan scaffold. The ability of the scaffold to facilitate the proliferation and migration of endothelial cell to form blood vessel is essential for cell survival especially in the inner area of the scaffold that is susceptible for cell death if adequate vascularization is not occurred. Purpose: The purpose of this study was to evaluate the porosity of HA/TCP/chitosan scaffold and the biocompatibility of HA/TCP/chitosan scaffold to endothelial cells. Methods: Endothelial cells were isolated from umbilical vein (human umbilical vein endothelial cells/ HUVEC). HA/ TCP/chitosan scaffold was made from two gelling agents and various basic washing solutions. The characteristic of scaffold was examined by scanning electron microscopy. The activity of HUVEC was evaluated by MTT assay. Results: Initial average scaffold porosity size range from 68 μm and increased up to 134 μm after 7 days incubation with 10 mg/L lysozyme. There was no significant difference in the viability of HUVEC incubated with the scaffold compared to control. Conclusion: HA/TCP/chitosan has a good biocompatibility for HUVEC. This condition supports the activity of HUVEC in the scaffold for angiogenesis process, to provide oxygen and nutrient necessary for osteoblast.Latar belakang: Angiogenesis merupakan proses yang penting untuk keberhasilan rekonstruksi tulang melalui rekayasa jaringan. Saat ini, aktifitas sel endotel pembentuk dinding pembuluh darah pada HA/TCP/chitosan scaffold belum diketahui. Kemampuan scaffold sebagai tempat proliferasi dan migrasi sel endotel untuk membentuk pembuluh darah penting untuk kelangsungan hidup sel osteoblast terutama di bagian dalam scaffold. Tujuan: Mengevaluasi porositas HA/TCP/chitosan scaffold serta sifat biokompatibilitas scaffold terhadap sel endotel. Metode: Sel endotel diisolasi dari vena tali pusat (human umbilical vein endothelial cells/HUVEC). HA/TCP/ chitosan scaffold dibuat dengan variasi gelling agents dan dicuci dengan berbagai larutan basa. Karakter scaffold dievaluasi dengan scanning electron microscope. Aktifitas HUVEC dievaluasi dengan MTT assay. Hasil: Pada tahap awal, rata-rata ukuran porus 68 μm dan meningkat menjadi 134 μm setelah inkubasi dengan 10 mg/mL lysosyme selama 7 hari. Kultur HUVEC pada scaffold selama 24 jam tidak menunjukkan tingkat viabilitas yang berbeda dibandingkan dengan kontrol. Kesimpulan: HA/TCP/chitosan memiliki sifat biokompatibilitas yang baik terhadap sel HUVEC. Kondisi ini memberikan dukungan terhadap aktifitas sel HUVEC pada scaffold untuk proses angiogenesis yang akan memberikan oksigen dan nutrisi untuk osteoblas.
Biocompatibility and osteoconductivity of injectable bone xenograft, hydroxyapatite and hydroxyapatite-chitosan on osteoblast culture Bachtiar EW; Bachtiar BM; Abas B; Harsas NA; Sadaqah NF; Aprilia R
Dental Journal (Majalah Kedokteran Gigi) Vol. 43 No. 4 (2010): December 2010
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (239.252 KB) | DOI: 10.20473/j.djmkg.v43.i4.p176-180

Abstract

Background: Bone graft in the form of injectable paste gives several advantages over the powder form as it could be placed in the defect area that has limited accessibility. Purpose: The purpose of this study was to assess biocompatibility and osteoconductivity of an injectable bone xenograft (IBX), injectable hydroxyapatite (IHA) and injectable hydroxyapatite-chitosan (IHA-C) on osteoblastic cell line (MG-63). Methods: Three concentrations (0.25%, 0.5% and 1.0%) of IBX, IHA and IHA-C were supplemented with DMEM culture medium. The viability cells were measured by MTT assay 4 hour after incubation. ALP activity was measured at day 1, 3, 5 and 7. Calcium deposition was tested at day 3 and day 7 by means of Von Kossa staining. Results: MTT assay showed that the viability cells of all the test groups were above 100% compared to the control group. The cell viability of the 0.25% IHA paste was significantly higher (115.02% ± 4.37%, p < 0.05) compared with IBX paste and IHA-C in all concentrations tested. The highest level of ALP secretion of all test groups was found on the fifth day of exposure. The highest level of ALP in the IBX paste group was 0.25% concentration while the highest level of ALP in the IHA-C and IHA paste group was 1% and 0.25%, respectively. In addition, the highest calcium deposition was shown on IHA 1% at day 7 (p > 0.05). Conclusion: It was suggested that adequate biocompatibility and osteoconductivity was evident for all injectable pastes tested.Latar belakang: Bahan tandur tulang dalam bentuk pasta injeksi memiliki kelebihan dibandingkan bila bahan tersebut berupa bubuk, karena lebih mudah diaplikasikan pada daerah yang sulit dijangkau. Tujuan: Penelitian ini bertujuan untuk mengamati sifat biokompatibilitas dan osteokonduktifitas biomaterial tandur tulang dalam bentuk injectable bone xenograft (IBX), injectable hydroxyapatite (IHA) dan injectable hydroxyapatite-chitosan (IHA-C) pada galur sel osteoblas (MG-63). Metode: Bahan tandur tulang IBX, IHA and IHA-C masing-masing dengan konsentrasi 0,25%, 0,5% dan 1,0% dipaparkan dalam larutan medium kultur sel DMEM yang telah disebari sel MG 63. Selanjutnya setelah 4 jam inkubasi maka viabilitas sel diukur dengan cara uji MTT, sedangkan aktifitas fosfatase alkali (ALP) diukur pada hari ke-1 (24 jam), hari ke-3, 5 dan 7. Deposisi kalsium diukur pada hari ke-3 dan ke-7 dengan metoda pewarnaan Von Kossa. Hasil: Uji MTT menunjukkan bahwa pemberian semua jenis bahan pasta injeksi tandur tulang meningkat di atas 100% dibandingkan kontrol. Viabilitas sel pada pemberian 0,25% pasta IHA tampak paling tinggi dibandingkan pasta IBX dan IHA-C pada semua konsentrasi yang diuji. Sekresi ALP tertinggi pada semua kelompok eksperimen terjadi pada hari ke lima setelah paparan bahan injeksi tandur tulang. Sekresi ALP tertinggi pada tiap jenis pasta terjadi pada pemberian IBX 0,25%, IHA-C 1% dan IHA 0.25%. Sedangkan deposisi kalsium tertinggi terjadi pada pemberian 1% IHA setelah 7 hari kultur sel. Kesimpulan: Semua bahan injeksi tandur tulang yang diuji pada kultur osteoblas bersifat biokompatibel dan berpotensi osteokonduktif
Co-Authors Abas B Abbas B Amir LR Aprilia R Bachtiar BM Harsas NA Sadaqah NF Utami S