Collagen-phosphophoryn sponge as a scaffold for bone tissue engineering

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Non-collagenous phosphoproteins that interact with type I collagen are thought to nucleate bone mineral into collagen network of mineralized tissues. Previously phosphophoryn cross-linked to collagen type I was reported to be an effective nucleator of apatite (Saito et al. Bone 21,305 (1997); Saito et al. J. Bone Miner. Res. 13,265 (1998)). On the basis of the above study, we expected that the collagen-phosphophoryn sponge could work as a good scaffold for bone tissue engineering and examined the in vivo bone formation in orthotopically transplanted collagen-phosphophoryn sponge and bone marrow osteoblasts composite in Fisher rats (bone defect model). Osteoblastic primary cells were obtained from the bone shaft of femorae of Fischer 344 male -weeks-old rats, according to the method of Maniatopoulos et al. The cell suspension of marrow cells was distributed through a T-75 flask with 15 ml of standard culture medium and incubated at 37℃ in a humidified atmosphere containing 95% air and 5% CO_2. When culture dishes become near confluent, after 10 days, they were treated with 0.1% trypsin and then concentrated by centrifuge to (10)^6 cells/ml and subcultured onto the synthesized collagen-phosphophoryn sponge and collagen sponge (control). After 14 days culture with standard medium supplemented with 10nM Dexamethasone, 10mM β-glycerophosphate, 50μg/ml ascorbic acid, the composites of collagen-phosphophoryn and osteoblastic cells and control composites were transplanted into bone defect sites with 2mm diameter hole of Fischer rats and then the wounds were sutured. The composites were harvested at 1,2,6,8 weeks after implantation, fixed with 4% paraformaldehide and 0.25% glutaraldehyde, decalcified, embedded in paraffin, and stained with hematoxirin and eosin. Results clearly showed that more hone formation was observed in collagen-phosphophoryn sponge and osteoblasts composites than control composites from 1 week to 8 weeks, suggesting that the collagen-phosphophoryn sponge is a good candidate as a scaffold for bone tissue engineering.
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Collagen-phosphophoryn sponge as a scaffold for bone tissue engineering

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