Ekstraksi dan Karakterisasi Kolagen dari Tulang Bovine Berbasis Enzimatik - Ultrasonik

Dhaningsih, Amallia Sri (2026) Ekstraksi dan Karakterisasi Kolagen dari Tulang Bovine Berbasis Enzimatik - Ultrasonik. Masters thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Kolagen merupakan protein struktural utama dalam matriks ekstraseluler yang berperan penting dalam regenerasi jaringan, terutama pada aplikasi penyembuhan luka (wound healing), karena sifat bioaktif serta biokompatibilitasnya yang tinggi. Namun, metode ekstraksi konvensional seringkali menghasilkan rendemen yang rendah dan struktur molekul yang kurang stabil, sehingga dibutuhkan pendeketan alternatif melalui kombinasi metode konvensional dan yang non-konvensional. Penelitian ini bertujuan untuk mengekstraksi kolagen dari tulang bovine berusia 2 tahun dengan variasi anatomi (femur, tibia, cartilage) dan rasio pepsin (0.5%, 1.0%, 1.5%) menggunakan metode kombinasi enzimatik – ultrasonik, serta mengevaluasi karakteristiknya berdasarkan standar ASTM F2212-20. Kolagen diekstraksi menggunakan metode kontrol (pepsin–ultrasonik) dan metode imobilisasi (pepsin terimobilisasi–ultrasonik), kemudian dikarakterisasi menggunakan FTIR, SEM, DSC, dan SDS-PAGE, serta diuji biokompatibilitasnya melalui uji MTT assay pada sel Adipose Rabbit Mesenchymal Stem Cells (AdMSCs). Hasil penelitian menunjukkan bahwa anatomi tulang bovine berpengaruh signifikan terhadap rendemen kolagen pada kedua metode ekstraksi (p < 0.05), dengan cartilage menghasilkan rendemen tertinggi (0.64%) pada metode pepsin–ultrasonik dan tibia (0.045%) pada metode pepsin terimobilisasi–ultrasonik. Analisis FTIR menunjukkan bahwa kolagen dari kedua metode mempertahankan struktur triple helix dengan rasio Amida III ≥ 1. Namun, kolagen kelompok kontrol menunjukkan keteraturan molekuler dan morfologi fibril yang lebih terdefinisi, serta profil molekuler khas kolagen tipe I yang masih terdeteksi, sedangkan kolagen kelompok imobilisasi didominasi struktur lembaran tipis dan agregat granular. Analisis DSC menunjukkan stabilitas termal kolagen kelompok kontrol lebih tinggi dibandingkan kelompok imobilisasi. Uji sitotoksisitas menunjukkan bahwa kolagen kelompok imobilisasi memiliki viabilitas sel yang lebih konsisten, sementara biokompatibilitas kolagen kelompok kontrol dipengaruhi oleh anatomi tulang bovine. Secara keseluruhan, metode pepsin–ultrasonik menghasilkan kolagen dengan kualitas struktural dan termal yang lebih baik, sedangkan metode pepsin terimobilisasi–ultrasonik memberikan biokompatibilitas in vitro yang lebih stabil. Hasil ini menunjukkan bahwa kolagen hasil ekstraksi dengan metode kombinasi enzimatik–ultrasonik berpotensi dikembangkan sebagai biomaterial tahap awal untuk aplikasi medis.
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Collagen is the main structural protein in the extracellular matrix, playing a vital role in tissue regeneration, especially in wound healing applications, due to its bioactive properties and high biocompatibility. However, conventional extraction methods often produce low yields and less stable molecular structures, necessitating alternative approaches through a combination of conventional and non-conventional methods. This study aims to extract collagen from 2-year-old bovine bones with varying anatomy (femur, tibia, cartilage) and pepsin ratios (0.5%, 1.0%, 1.5%) using a combined enzymatic-ultrasonic method, as well as to evaluate its characteristics based on ASTM F2212-20 standards. Collagen was extracted using control methods (pepsin–ultrasonic) and immobilised methods (immobilised pepsin–ultrasonic), then characterised using FTIR, SEM, DSC, and SDS-PAGE, and its biocompatibility was tested via MTT assay on Rabbit Adipose Mesenchymal Stem Cells (AdMSCs). The results showed that bovine bone anatomy significantly affected collagen yield in both extraction methods (p < 0.05), with cartilage producing the highest yield (0.64%) in the pepsin–ultrasonic method and tibia (0.045%) in the immobilised pepsin–ultrasonic method. FTIR analysis indicated that collagen from both methods maintained the triple helix structure with an Amide III ratio of 1. However, control group collagen exhibited more molecular order and fibril morphology, as well as a characteristic molecular profile of type I collagen, whereas immobilised group collagen was dominated by thin sheet structures and granular aggregates. DSC analysis showed that the thermal stability of control group collagen was higher than that of the immobilised group. Cytotoxicity tests revealed that immobilised group collagen had more consistent cell viability, while the biocompatibility of the control group collagen was influenced by bovine bone anatomy. Overall, the pepsin–ultrasonic method produced collagen with better structural and thermal quality, whereas the immobilised pepsin–ultrasonic method provided more stable in vitro biocompatibility. These findings suggest that collagen extracted via the combined enzymatic-ultrasonic method has potential for development as an early-stage biomaterial for medical applications.

Item Type:	Thesis (Masters)
Uncontrolled Keywords:	Kolagen, Tulang Bovine, Ultrasonik, Enzimatik Collagen, Bovine Bone, Ultrasonic, Enzymatic
Subjects:	Q Science > QD Chemistry > QD251.2 Chemistry, Organic. Biochemistry Q Science > QD Chemistry > QD63 Extraction Q Science > QH Biology > QH541.15.T68 Toxicity testing R Medicine > R Medicine (General) > R857.M3 Biomedical materials. Biomedical materials--Testing.
Divisions:	Faculty of Industrial Technology > Material & Metallurgical Engineering > 27101-(S2) Master Thesis
Depositing User:	Amallia Sri Dhaningsih
Date Deposited:	27 Jan 2026 02:47
Last Modified:	27 Jan 2026 02:47
URI:	http://repository.its.ac.id/id/eprint/130569

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