Biotinated bone morphogenetic protein-2: In vivo and in vitro activity

Uludağ, Hasan; Golden, J.; Palmer, R.; Wozney, John M.

doi:10.1002/(sici)1097-0290(19991220)65:6<668::aid-bit7>3.0.co;2-8

Cited by 26 publications

(18 citation statements)

References 14 publications

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“…With a growth factor, it will be important to optimize the conjugation efficiency to strike a balance between the high mineral affinity (positively correlated with the extent of modification) and the preservation of biological activity (inversely correlated with the extent of modification). For biotinylated recombinant human bone morphogenetic protein‐2, for example, the site of biotin coupling was critical for the protein's osteoinductive activity (35). For basic fibroblast growth factor, an optimum degree of biotinylation was observed, beyond which a significant activity loss of was obtained (36).…”

Section: Discussionmentioning

confidence: 99%

Bisphosphonate Conjugation to Proteins as a Means To Impart Bone Affinity

et al. 2000

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Growth factors are endogenous proteins capable of stimulating new bone formation, but their clinical benefit for systemic stimulation of bone mass has not been demonstrated. The critical challenge is to deliver a significant dose of the proteins to bone after intravenous injection. This challenge may be overcome by derivatizing proteins with ligands that exhibit a high bone affinity (e.g., bisphosphonates). To demonstrate the feasibility of this approach, 1-amino-1,1-diphosphonate methane (aminoBP) was conjugated to a model protein, albumin. The conjugation was performed by (1) converting the amino group of aminoBP to a thiol group using 2-iminothiolane, (2) derivatizing the albumin amino groups with a thiol-reactive sulfosuccinimidyl-4-(N-maleimidomethyl)-1-cyclohexane carboxylate, and (3) reacting the derivatized albumin with thiolated aminoBP. Typically, 1-4 aminoBP molecules per albumin were obtained. The conjugated albumin exhibited a high affinity to hydroxyapatite that was proportional to the extent of conjugation. The conjugates were shown to exhibit a high affinity to bone matrix in vitro in a serum-containing medium. Once bound to bone matrix, the conjugates were found to desorb more slowly than the unmodified albumin, especially from bone whose organic matrix was removed by ashing. In conclusion, conjugation of bisphosphonates to albumin was shown to impart a high bone affinity to the protein, and such conjugates can be potentially targeted to bone.

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Section: Discussionmentioning

confidence: 99%

Bisphosphonate Conjugation to Proteins as a Means To Impart Bone Affinity

et al. 2000

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“…Carrier‐free recombinant human BMP‐2 (3 μg) was incubated with Biotin‐NHS (1.5 nmol) in 0.1 M sodium bicarbonate at room temperature for 1.5 h as described previously (26) . Biotinylated BMP‐2 was separated from free biotin‐NHS by centrifugation using Spin Column 30 (Sigma) and reconstituted in PBS.…”

Section: Methodsmentioning

confidence: 99%

αvβ Integrins Play an Essential Role in BMP-2 Induction of Osteoblast Differentiation

Lai

Cheng

2005

Journal of Bone and Mineral Research

127

121

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Both integrins and BMP-2 exert similar effects on osteoblasts. We examined the relationship between the ␣v-containing integrins (␣v␤) and BMP-2 in osteoblast function. BMP-2 stimulates ␣v␤ expression. BMP-2 receptors co-localize/overlap with ␣v␤ integrins, and the intact function of ␣v␤ is essential in BMP-2 activity.Introduction: Bone morphogenetic protein (BMP)-2 not only induces osteoblast differentiation and bone matrix mineralization, but also stimulates osteoblast migration on and adhesion to bone matrix proteins. The ␣v␤-and ␤1-(␣␤1) containing integrins mediate osteoblast interaction with many bone matrix proteins and play important roles in osteoblast adhesion, migration, and differentiation. Because ␣v␤ integrins and BMP-2 share common effects on osteoblasts, we analyzed their relationship in osteoblast function. Materials and Methods:The effects of BMP-2 on integrin expression were determined by surface labeling/ immunoprecipitation and cell adhesion to matrix proteins. Confocal analysis of the immunostained cells and co-immunoprecipitation of cell extracts were used to study the spatial relationship between integrins and BMP-2 receptors. A function-blocking anti-␣v␤ integrin antibody (L230) was employed to investigate the roles of ␣v␤ integrins in BMP-2 function. Results: Human osteoblasts (HOBs) express ␣␤1, ␣v␤3, ␣v␤5, ␣v␤6, and ␣v␤8 integrins at focal adhesion sites. BMP-2 increases the levels of these integrins on osteoblast surface and enhances HOB adhesion to osteopontin and vitronectin. Immunoprecipitation and immunostaining analyses show that BMP-2 receptors co-localize or overlap with ␣v␤ and ␣␤1 integrins. Incubation of HOBs with L230 abolishes the antiproliferative effect of BMP-2 and reduces the capacity of BMP-2 to stimulate alkaline phosphatase activity and the expression of osteocalcin, osteopontin, and bone sialoprotein. Furthermore, L230 prevents BMP-2 induction of matrix mineralization. Although BMP-2 retains its receptor-binding capability in the presence of L230, BMP-2 stimulation of Smad signaling is abolished by L230. Conclusion: BMP-2 upregulates the expression of ␣v␤ integrins, and these integrins, in turn, play a critical role in BMP-2 function in osteoblasts.

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“…Also, the phenol/imidazole‐treated rhBMP‐2 failed to retain the rhBMP‐2 bioactivity . Moreover, Uludag et al showed that protein pI has influence on early retention of rhBMP‐2 …”

Section: Bone Regeneration With Modified Bmp‐2mentioning

confidence: 99%

A review on carrier systems for bone morphogenetic protein‐2

Agrawal

Sinha

2016

J Biomed Mater Res

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Bone morphogenetic protein-2 (BMP-2) has unique bone regeneration property. The powerful osteoinductive nature makes it considered as second line of therapy in nonunion bone defect. A large number of carriers and delivery systems made up of different materials have been investigated for controlled and sustained release of BMP-2. The delivery systems are in the form of hydrogel, microsphere, nanoparticles, and fibers. The carriers used for the delivery are made up of metals, ceramics, polymers, and composites. Implantation of these protein-loaded carrier leads to cell adhesion, degradation which eventually releases the drug/protein at site specific. But, problems like ectopic growth, lesser protein delivery, inactivation of the protein are reported in the available carrier systems. Therefore, it is need of an hour to modify the available carrier systems as well as explore other biomaterials with desired properties. In this review, all the reported carrier systems made of metals, ceramics, polymers, composites are evaluated in terms of their processing conditions, loading capacity and release pattern of BMP-2. Along with these biomaterials, the attempts of protein modification by adding some functional group to BMP-2 or extracting functional peptides from the protein to achieve the desired effect, is also evaluated. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 904-925, 2017.

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Biotinated bone morphogenetic protein-2: In vivo and in vitro activity

Cited by 26 publications

References 14 publications

Bisphosphonate Conjugation to Proteins as a Means To Impart Bone Affinity

Bisphosphonate Conjugation to Proteins as a Means To Impart Bone Affinity

αvβ Integrins Play an Essential Role in BMP-2 Induction of Osteoblast Differentiation

A review on carrier systems for bone morphogenetic protein‐2

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