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Int J Biol Sci 2011; 7(7):968-977. doi:10.7150/ijbs.7.968

Research Paper

In vitro Evaluation of Natural Marine Sponge Collagen as a Scaffold for Bone Tissue Engineering

Zhen Lin1.2#, Kellie L. Solomon2#, Xiaoling Zhang3#, Nathan J. Pavlos2, Tamara Abel2, Craig Willers2, Kerong Dai3, Jiake Xu2, Qiujian Zheng1✉, Minghao Zheng2✉

1. Division of Orthopaedic Surgery, Guangdong Academy of Medical Sciences, Guangdong General Hospital, Guangzhou, Guangdong, 510080, P.R. China
2. Centre for Orthopaedic Research, School of Surgery, University of Western Australia, Western Australia, 6009, Australia
3. Laboratory of Orthopaedic Cellular and Molecular Biology, Institute of Health Sciences, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences and Shanghai JiaoTong University, Shanghai 200025, P.R. China
# These authors contributed equally to this paper.

Abstract

The selection of a suitable scaffold matrix is critical for cell-based bone tissue engineering. This study aimed to identify and characterize natural marine sponges as potential bioscaffolds for osteogenesis. Callyspongiidae marine sponge samples were collected from the Fremantle coast of Western Australia. The sponge structure was assessed using scanning electron microscopy (SEM) and Hematoxylin and eosin. Mouse primary osteoblasts were seeded onto the sponge scaffold and immunostained with F-actin to assess cell attachment and aggregation. Alkaline phosphatase expression, von Kossa staining and real-time PCR were performed to examine the osteogenic potential of sponge samples. SEM revealed that the sponge skeleton possessed a collagenous fibrous network consisting of interconnecting channels and a porous structure that support cellular adhesion, aggregation and growth. The average pore size of the sponge skeleton was measured 100 to 300 μm in diameter. F-actin staining demonstrated that osteoblasts were able to anchor onto the surface of collagen fibres. Alkaline phosphatase expression, a marker of early osteoblast differentiation, was evident at 7 days although expression decreased steadily with long term culture. Using von Kossa staining, mineralisation nodules were evident after 21 days. Gene expression of osteoblast markers, osteocalcin and osteopontin, was also observed at 7, 14 and 21 days of culture. Together, these results suggest that the natural marine sponge is promising as a new scaffold for use in bone tissue engineering.

Keywords: natural marine sponge, bone tissue engineering

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How to cite this article:
Lin Z, Solomon KL, Zhang X, Pavlos NJ, Abel T, Willers C, Dai K, Xu J, Zheng Q, Zheng M. In vitro Evaluation of Natural Marine Sponge Collagen as a Scaffold for Bone Tissue Engineering. Int J Biol Sci 2011; 7(7):968-977. doi:10.7150/ijbs.7.968. Available from http://www.ijbs.com/v07p0968.htm