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.

Summary: We have analyzed a number of collagen‐derived biomaterials for the matrix‐ induced and assisted bone and cartilage tissue regeneration. These include the Small intestine submuosa (SIS) Restor ™, ACI‐Maix collagen membrane, Chondro‐ Gide collagen membrane, Permacol collagen Ossix and lycoll collagen membrane and five types of collagen‐based marine sponge skeletons. Certain characteristics of different scaffold materials with comparable chemical composition may vary significantly. This variation may have a relevant impact on the suitability of the scaffolds for bone and cartilage regeneration. It suggests that the ACI‐Maix® membrane is the best available collagen‐derived material for an MACI®/MACT® application. In addition, the study of marine sponge indicates that the collagenous fibre skeleton of marine sponges provides a suitable bioscaffold for bone regeneration, as it supports the adhesion, migration and proliferation of osteoblasts in vitro.