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

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.

Enhancing the stiffness of collagen hydrogels for delivery of encapsulated chondrocytes to articular lesions for cartilage regeneration

“…Among many potential natural and synthetic matrices in tissue engineering, collagen has proven to be useful in hydrogel, membrane, and porous scaffold forms. Collagen is naturally occurring in human tissue, including articular cartilage .…”

Marine Natural Products: New Avenue in Treatment of Osteoporosis

“…On the other side, marine-derived compounds like Mycoepoxydiene, Norzoanthamine, and its homologs, Biselyngbyaside, largazole suppresses RANKL-induced osteoclastogenesis via downregulating the expression of osteoclast differentiating factors (RANKL, NFATc1, and TRAP). and osteocalcin and also increases the levels of TAZ and Runx2 for osteoblast differentiation (Lin et al, 2011;Byun et al, 2012). Biselyngbyaside inhibits osteoclastogenesis by suppressing RANKL and NFATc1 expression whereas largazole mediates osteogenic activity through the increased expression of Runx2 and BMPs (Taori et al, 2008;Yonezawa et al, 2012).…”

“…Sponges contain a complex canal system that builds a porous environment ideal for cellular integration when coalesce with cells for tissue engineering. Marine sponge skeletons are increasingly being used as a source of Scaffold's development for tissue engineering and regeneration because of their potential as cell conductive and inductive framework (Lin et al, 2011;Nandi et al, 2015). Nandi and colleagues investigated that the impregnation of converted sponge scaffold by IGF-1 and BMP-2, promoted excellent osseous tissue formation.…”

“…Processed marine sponge Cally spongiidae skeleton consisted of a collagenous fibrous network having porous structure and interconnecting channels. This collagenous fibrous network provides a good framework that supports cellular aggregation, adhesion and expansion of osteoblast cells with expression of osteoblast markers (Lin et al, 2011;Clarke et al, 2016). Folmer et al concludes that marine algae, bacteria, and invertebrates including marine sponges are a source of NF-κB inhibitors which inhibit TNF-α induced Nuclear factor kappa-light chain enhancer of activated B cells (NF-κB) activation (Folmer et al, 2009).…”

Marine Collagen Scaffolds for Nasal Cartilage Repair: Prevention of Nasal Septal Perforations in a New Orthotopic Rat Model Using Tissue Engineering Techniques

“…Such matrices might represent an alternative source for the production of scaffolds for tissue engineering applications. 30,[32][33][34] Marine collagen is harvested by lyophilization of the invertebrate jellyfish species Rhopilema esculentum. Advantages of this collagen source include its reproducible production process, which relies on renewable natural resources, and the absence of any risk for disease transmission by viruses or bacteria.…”