Papers by Keyword: Bone Morphogenetic Protein

Abstract: Introduction: Marrow mesenchymal cells contain stem cells and can regenerate tissues. We previously reported the clinical application of autologous cultured bone to regeneration therapy. However, in cases with low numbers of active cells, culture is often unsatisfactory. If frozen marrow cells retain their osteogenic potential, we could clinically use them in regeneration therapy as alternatives to high active cells obtained from youngsters. Here, we examined osteogenic potential of frozen human mesenchymal stem cells in combination with recombinant human bone morphogenetic protein (rhBMP) using biochemical and histological analyses. Method: Marrow fluid was aspirated from the human iliac bone of a 46-year-old man with lumbar canal stenosis during surgery. Two weeks after primary culture in standard medium, bone marrow mesenchymal stem cells (BMSCs) were trypsinized for the preparation of a cell suspension, and cells were concentrated to 106 cells/ml by centrifugation. Cells were kept at – 80 °C until use. To impregnate porous hydroxyapatite (HA) with rhBMP, 1 3g rhBMP/20 3l 0.1 % trifluoroacetic acid was applied on HA, and then desiccated under vacuum. In the present study, we used 4 subgroups: BMSC/rhBMP/HA, BMSC/HA, rhBMP/HA, and HA only. HA constructs from the 4 subgroups were implanted at subcutaneous sites on the back of 5-week-old nude mice (BALB/cA Jcl-nu). Eight weeks after implantation, implanted HA constructs were harvested, and biochemical and histological analyses were performed. Alkaline phosphatase activity (ALP) and human osteocalcin (hOs) levels were measured. Results and Discussion: ALP activity and hOs in the BMSC/BMP/HA subgroup were 2 or 3 times that in the BMSC/HA subgroup. Histological analysis showed that significant bone formation was observed in these two subgroups, and supported biochemical data. However, in the BMP/HA and HA only subgroups, significant bone formation could not be detected histologically nor biochemically. These results indicated that a combination of rhBMP and BMSCs, and only with a minimal amount of 1 3g rhBMP, allowed successful generation of human bone. In the human body, rhBMP in the order of milligrams is necessary for bone formation. However, by combining BMSCs, HA and rhBMP, only a small amount of rhBMP was needed to dramatically enhance osteogenic potential. As we reported here, cryopreserved BMSCs also showed high osteoblastic activity. In conclusion, this study provided histological and biochemical evidence that combination of cryopreserved BMSCs, BMP, and porous HA could enhance osteogenic potential.

Abstract: The objective of this study is to examine feasibility of biodegradable gelatin hydrogels incorporating bone morphogenetic protein (BMP)-2 in inducing bone regeneration at a bone defect of non-human primates and rabbits considering their BMP-2 release profiles. As a result, controlled release by the hydrogel system enabled BMP-2 to induce successful bone regeneration in non-human primates even at the BMP-2 dose as low as that for rabbit case (0.034 mg of BMP-2/cm3 of hydrogel).

Abstract: b-glucan, an immunomodulator, can selectively enhance the immunobiological activities of neutrophils and macrophages without stimulating proinflammatory cytokine production. Biologic response modifiers, like beta-glucan, will modulate immunity, modify neoplastic disease and increase resistance to microbial challenge. Therefore, beta-glucan polymers can be applied in bone induction and regeneration model and have a possibility of association with bone morphogenetic protein (BMP) because of tissue-regenerative and antimicrobial effects of those polymers. In this report, we studied an E. coli expression system for BMP-7 production and the biological activities of b-glucan associated with BMP-7. The proliferation of MC3T3-E1 osteoblastic cells was enhanced by treatment with Aureobasidium b-glucan, while neither mushroom b-glucan nor barley b-glucan increased the cell proliferation. Mushroom b-glucan alone or associated with BMP-7 increased alkaline phosphatase (ALP) activity of MC3T3-E1 cells, one of the osteoblast phenotype markers, but the other b-glucans did not affect ALP activity of the cells. In mineralization assay, a highly significant increase in nodular staining was observed in cultures treated with both mushroom and Aureobasidium b-glucans in the presence of BMP-7 compared with nontreated controls, while barley b-glucan showed a significant decrease in nodule number compared with cultures treated only with BMP-7.