Background Angiogenesis is essential for the replacement of cartilage by bone during skeletal growth and regeneration. Vascular endothelial growth factor-A (VEGF-A) is a key regulator of angiogenesis whereas endostatin, a potent inhibitor of endothelial cell proliferation and migration, is a natural antagonist of VEGF-A The regulatory role of these peptides in angiogenesis and bone formation was investigated using adenoviral gene delivery of VEGF-A and endostatin in a mouse ectopic ossification model.Methods Bone formation was induced in the hamstring muscles of adult mice with native bone morphogenetic protein (BMP) extract implemented in gelatine gel together with VEGF-A and endostatin recombinant adenoviral vectors. The mice were sacrificed 1, 2, and 3 weeks after the operation and ectopic bone formation was followed radiographically and histologically.Results Significant bone formation was induced by BMP extract in all treatment groups. VEGF-A stimulated and endostatin prevented the formation of FVIII-related antigen-positive vessels as well as the number of cartilage-resorbing chondroclasts/osteoclasts. Endostatin alone or in conjugation with VEGF-A reduced bone formation. Excess of VEGF-A stimulated and endostatin reduced bone formation, respectively, at the 3-week time point.Conclusions Our findings indicate that endostatin retards the cartilage phase in endochondral ossification which subsequently reduces bone formation in our experimental model. We conclude that bone growth and healing, which share features with ectopic bone formation, may be regulated by endostatin. Copyright (C) 2007 John Wiley & Sons, Ltd.

A cDNA sequence of bone morphogenetic protein 3b (BMP-3b) of reindeer antler was produced with degenerative homology primers in polymerase chain reaction (PCR). An in situ hybridization study of BMP-3b mRNA in 1-month-old antler showed expression in most differentiated cells in the antler center. In addition, the bone-inductive capacity of the reindeer antler matrix was evaluated. Decalcified and powdered antler matrix of different stages of antler maturity was implanted in gelatin capsules under the rat dorsal muscle fascia for two implantation periods: 3 and 8 weeks. Allogenic matrix prepared from rat long bones was used as a positive control implant. Heterotopic ossification was evaluated histomorphometrically and densitometrically. Allogenic bone matrix induced rapid osteogenesis and mineral accumulation. Both endochondral and intramembranous ossification was evident, endochondral ossification being the dominant form. Mineral density in the induced ossicle was 115 +/- 48 mg/cm(3) as early as at 3 weeks and 350 +/- 69 mg/cm(3) at 8 weeks. The proportional areas of von Kossa-stained mineral were 3.67 +/- 2.1% and 11.6 +/- 0.07%, respectively. The antler preparations induced mineralization, but significantly less than the allogenic bone matrix. At 8 weeks, mineral density was significantly lower in the cast antler preparation than in the allogenic implants. The morphology of the mineralized areas of the antler preparations showed no ossification.

Advances in Skeletal Reconstruction Using Bone Morphogenetic Proteins, pp. 53-61 (2002) No AccessDetermination of the Biological Activity of BMPs in Cell Cultures is UnreliableM. Ulmanen, E. Birr, O. Hietala and T. S. LindholmM. UlmanenBone Transplantation Research Group, University of Oulu, Oulu, Finland, E. BirrBone Transplantation Research Group, University of Oulu, Oulu, Finland, O. HietalaBone Transplantation Research Group, University of Oulu, Oulu, Finland and T. S. LindholmDepartment of Oral and Maxillofacial Surgery, University Central Hospital of Turku, Finlandhttps://doi.org/10.1142/9789812706577_0004Cited by:2 PreviousNext AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsRecommend to Library ShareShare onFacebookTwitterLinked InRedditEmail Abstract: The most reliable means of proving the osteoinductivity of bone morphogenetic proteins (BMPs) has been a bioassay based on cartilage and bone formation in rodent muscle. Bone formation induced by BMPs can usually be detected, at the earliest, a few weeks from implantation, and this makes the bioassay an inconveniently timeconsuming approach. The present study was undertaken with an eye to developing a more rapid method called microassay for detecting the activity of BMPs. Three different cell lines (C3H10T½, M2-10-B4 and SR-4987) were chosen and their responses to BMPs were tested by measuring changes in the alkaline phosphatase (ALP) activity. It emerged that the cell lines C3H10T½ and M2-10-B4 are not suitable for the microassay, but that SR-4987 could possibly be utilised for this purpose. A great deal more research will be required before the microassay can be considered a reliable test of BMP activity. FiguresReferencesRelatedDetailsCited By 2A mesenchymal stromal cell line resistant to paclitaxel that spontaneously differentiates into osteoblast-like cellsAugusto Pessina, Francesca Sisto, Valentina Coccè, Loredana Cavicchini and Emilio Ciusani et al.29 December 2010 | Cell Biology and Toxicology, Vol. 27, No. 3Growth Promoter: General Principles and Experimental Studies on BMPT. Sam Lindholm1 Jan 2004 Advances in Skeletal Reconstruction Using Bone Morphogenetic ProteinsMetrics History PDF download

Structural comparison of human and rat prostate-specific acid phosphatase (hPAP and rPAP) genes indicate that the exon number is different between these species. The hPAP gene contains 10 exons, whereas the rPAP gene was 11 exons. However, exons 2-9 of the genes are identical in size. The 5' regions of the two genes show 71% identity in the most homologous region +1 to +340. The 5' untranslated regions of the human and rat genes are 50 and 49 nucleotides long, respectively. An Alu sequence is present upstream from the proximal promoter of the hPAP gene. Five putative androgen response elements altogether were localized in both the human and rat gene, one of which is conserved in location and sequence between the two genes. Two of these elements in both genes, the conserved one in the proximal promoter region and another one in intron 1, were shown to bind androgen receptor efficiently in vitro.

Streptomyces coelicolor “Müller” DSM3030 excretes a lysozyme comprising both β-1,4-N-acetyl-and β-1,4-N,6-O-diacetyl muramidase activities. The lysozyme is named Cellosyl. Gene libraries have been established using genomic DNA from the wild-type strain, S. coelicolor DSM3030, and from an overproducing mutant, S. coelicolor HP1, which exhibits about a twofold increase in lysozome production. The lysozyme-encoding genes (cel) from both strains were detected by oligodeoxynucleotide hybridization. The nucleotide sequence of the cel genes isolated from both strains was shown to be identical. The different levels of lysozyme production could not be correlated with any mutations at the cel gene locus. The cel gene isolated from the wild-type strain could not be expressed in some other species of Streptomyces. However, self-cloning of the cel gene into S. coelicolor DSM3030 and HP1 resulted in a 2.5-fold increase in lysozyme production.