Inhibitory effect of porcine cartilage-derived extracellular matrix suspension on angiogenesis in vitro

Alternative Title
김선영
Author(s)
Kim, Sun Young
Alternative Author(s)
김선영
Advisor
민병현
Department
일반대학원 분자과학기술학과
Publisher
The Graduate School, Ajou University
Publication Year
2013-02
Language
eng
Keyword
Porcine cartilage ECM derivedwater suspensionAnti-angiogenesis
Alternative Abstract
Background and Object: Articular cartilage is known as an avascular tissue, which has per se anti-angiogenic factors. Several molecules in cartilage extracellular matrix (ECM) are reported to exhibit antiangiogenic activity, some of which already went through clinical trial for harmaceutical purpose such as chondromodulin-I and type XVIII-derived endostatin. We have found previously that the natural cartilage ECM could be a promising treatment with anti-angiogenic activity. However, wide application of drug necessitates various formula especially in soluble phase. We fabricated evenly distributed water suspension-form biomaterial using the cartilage ECM that could be easily deliverable into the body. In this study, we evaluated whether a water-soluble form of the porcine cartilage-derived ECM biomaterial (PCP-ws, porcine cartilage powder – water suspension) still possesses antiangiogenic property. Materials and Methods: The slices of cartilage tissues obtained from 6-weeks-old porcine knees were lyophilized and comminuted into a particulate form, which was then decellularized by hypotonic buffer and DNase-I treatments. The decellularized ECM powder was solubilized by pepsin/HCl. The toxicity of the PCP-ws was evaluated on mouse fibroblast cell line (L929), rabbit chondrocytes and human umbilical vein endothelial cells (HUVEC) by MTT assay. The effect of PCP-ws on the function of endothelial cells was evaluated including cell adhesion, migration, proliferation and tube formation ability were performed. Results: In this study, it is demonstrated that the porcine cartilage powder (PCP) was successfully decellularized and made to a suspension form (PCP-ws) for easily deliverable biomaterial of anti-angiogenic therapy. The viability of L929 and chondrocyte were slightly decreased. In contrast, the viability of HUVEC was severely decreased by PCP-ws in a dose dependent. PCP-ws was prove to be an excellent substrates for inhibiting endothelial cell adhesion, migration and proliferation, particularly for suppression formation of tube-like structure in vitro. In the tube formation assay, the formation of tube network of HUVECs was significantly inhibited by PCP-ws in a dose-dependent manner. Remarkably, the experimental group treated with 5 mg/ml of PCP-ws showed almost no tube-like structure. The length of tube structure decreased approximately by 17 folds, when compared with the untreated group. Discussion and Conclusion: In this study, it is demonstrated that the porcine cartilage powder (PCP) was successfully decellularized and made to a water suspension form (PCPws) for easily deliverable biomaterial of anti-angiogenic therapy. PCP-ws was prove to be an excellent substrates for inhibiting endothelial cell adhesion, migration and proliferation, particularly for suppression formation of tube-like structure in vitro. Supposedly, PCP-ws have a variety of bioactive factors such as chondromodulin-I, endostatin and sulfated glycosaminoglycan side chains that are known to exist in the articular cartilage. Therefore, these results suggest that PCP-ws has an inhibitory effect on angiogenesis in vitro and could be a useful biomaterial for many applications.
URI
https://dspace.ajou.ac.kr/handle/2018.oak/9490
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Graduate School of Ajou University > Department of Molecular Science and Technology > 3. Theses(Master)
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