The effect of two point mutations in GDF-5 on ectopic bone formation in a β-tricalciumphosphate scaffold

Kasten, Philip; Beyen, Ingo; Bormann, Dirk; Luginbühl, Reto; Plöger, Frank; Richter, Wiltrud

doi:10.1016/j.biomaterials.2010.01.109

Cited by 23 publications

(20 citation statements)

References 36 publications

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“…Due to the presence of a flexible spring-loaded latch in MBPR-IB, GDF5 shows a 10-to 20-fold higher affinity for MBPR-IB than MBPR-IA, which does not contain the spring and has a higher energy requirement for the GDF5: MBPR-IA complex [Nickel et al, 2005;Kotzsch et al, 2009]. Furthermore, compared to BMP2, point mutations introduced in GDF5 at the sites that are involved in binding to MBPR-I lead to an enhanced ectopic osteogenesis as BMP2 in a ␤ -tricalcium phosphate ( ␤ -TCP) ceramic scaffold [Kasten et al, 2010]. Whether the GDF5 mutant would show altered MBPR-IA and MBPR-IB binding affinity and whether other biological effects, such as angiogenic properties, might exist was not investigated.…”

Section: Discussionmentioning

confidence: 99%

Overexpression of GDF5 through an Adenovirus Vector Stimulates Osteogenesis of Human Mesenchymal Stem Cells in vitro and in vivo

Cheng

Yang

Meng

et al. 2012

Cells Tissues Organs

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The use of stem cells combined with gene therapy could be an important way to facilitate bone regeneration. In this study, the aim was to investigate the potential of growth and differentiation factor-5 (GDF5) to genetically manipulate human mesenchymal stem cells (hMSCs) for bone regeneration. Recombinant adenovirus Ad-GDF5 and Ad-GFP were constructed and identified, and the titer of both were determined. Third-passage hMSCs were infected with adenovirus, and the expression of GDF5 was confirmed by detection of GFP-positive cells, GDF5 mRNA levels, Western blotting, and enzyme-linked immunosorbent assay (ELISA). hMSCs at passage 3 were divided into four groups: (1) an experimental group infected with Ad-GDF5, (2) a positive control group cultured with osteogenic differentiation medium, (3) a control group infected with Ad-GFP cultured with standard medium, and (4) a blank control group cultured with standard medium. Evaluation of cell morphology and proliferation, analysis of the expression of genes related to osteogenic differentiation, von Kossa staining, and immunofluorescent staining of collagen I were used to investigate the osteogenesis of cells among the groups. After culturing the cells for 2 days under each corresponding condition, the cells were detached and subcutaneously injected into the backs of nude mice to evaluate bone formation. Samples were collected for histological staining, protein Western blotting, and micro-computer tomography. When infected with Ad-GDF5, hMSCs could overexpress GDF5 for a prolonged period in vitro and reach a concentration of 160 ng/ml. Cells infected with Ad-GDF5 or cultured in osteogenic medium displayed osteogenic differentiation based on their histological and cellular properties and on their gene and protein expression patterns. Furthermore, Ad-GDF5 showed a better ability to upregulate the expression of collagen I, alkaline phosphatase, and osteocalcin mRNA than the osteogenic medium. Furthermore, Ad-GDF5 expression was associated with enhanced bone formation in vivo. Our findings suggest that hMSCs infected with Ad-GDF5 can differentiate in an osteogenic direction and may be a promising cell source for bone regeneration.

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Section: Discussionmentioning

confidence: 99%

Overexpression of GDF5 through an Adenovirus Vector Stimulates Osteogenesis of Human Mesenchymal Stem Cells in vitro and in vivo

Cheng

Yang

Meng

et al. 2012

Cells Tissues Organs

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“…As autogenously harvested bone transplants are limited and callus distraction is not feasible in every defect site, alternative strategies involving the use of bone substitutes, bone marrow stromal cells (BMSC), and growth factors have successfully been explored [Niemeyer et al, 2009a, b;Janicki et al, 2010;Kasten et al, 2010]. It is known that vascularization is important and tightly regulated in bone healing [DePalma et al, 1972;Burchardt and Enneking, 1978].…”

Section: Introductionmentioning

confidence: 99%

Comparison of Platelet-Rich Plasma and VEGF-Transfected Mesenchymal Stem Cells on Vascularization and Bone Formation in a Critical-Size Bone Defect

Kasten

Beverungen

Lorenz

et al. 2012

Cells Tissues Organs

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Both platelet-rich plasma (PRP) and vascular endothelial growth factor (VEGF) can promote regeneration. The aim of this study was to compare the effects of these two elements on bone formation and vascularization in combination with bone marrow stromal cells (BMSC) in a critical-size bone defect in rabbits. The critical-size defects of the radius were filled with: (1) a calcium-deficient hydroxyapatite (CDHA) scaffold + phVEGF<sub>165</sub>-transfected BMSC (VEGF group), (2) CDHA and PRP, or (3) CDHA, autogenous BMSC, and PRP. As controls served: (4) the CDHA scaffold alone and (5) the CDHA scaffold and autogenous BMSC. The volume of new bone was measured by means of micro-CT scans, and vascularization was assessed in histology after 16 weeks. Bone formation was higher in the PRP + CDHA, BMSC + CDHA, and PRP + BMSC + CDHA groups than in the VEGF group (p < 0.05). VEGF transfection significantly promoted vascularization of the scaffolds in contrast to BMSC and PRP (p < 0.05), but was similar to the result of the CDHA + PRP + BMSC group. The results show that VEGF-transfected BMSC as well as the combination of PRP and BMSC improve vascularization, but bone healing was better with the combination of BMSC and PRP than with VEGF-transfected BMSC. Expression of VEGF in BMSC as a single growth factor does not seem to be as effective for bone formation as expanded BMSC alone or PRP which contains a mixture of growth factors.

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“…growth and restoration comprise a well-orchestrated interplay47 of various factors and mediators responsible for cellular recruitment,48 proliferation and differentiation. Secreted growth factors, in particular 49 Bone Morphogenetic Proteins (BMPs) and Growth and Differentiation 50 Factors (GDFs), play a key role as they exert their effects on mesenchy-51 mal stem cells by promoting their differentiation into the chondro-or 52 osteogenic lineage [1,2].…”

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confidence: 99%

Improved bone defect healing by a superagonistic GDF5 variant derived from a patient with multiple synostoses syndrome

Degenkolbe

Schwarz

Ott

et al. 2015

Bone

Self Cite

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Multiple synostoses syndrome 2 (SYNS2) is a rare genetic disease characterized by multiple fusions of the joints of the extremities, like phalangeal joints, carpal and tarsal joints or the knee and elbows. SYNS2 is caused by point mutations in the Growth and Differentiation Factor 5 (GDF5), which plays an essential role during skeletal development and regeneration. We selected one of the SYNS2-causing GDF5 mutations, p.N445T, which is known to destabilize the interaction with the Bone Morphogenetic Protein (BMP) antagonist NOGGIN (NOG), in order to generate the superagonistic GDF5 variant GDF5(N445T). In this study, we tested its capacity to support regeneration in a rat critical-sized defect model in vivo. MicroCT and histological analyses indicate that GDF5(N445T)-treated defects show faster and more efficient healing compared to GDF5 wild type (GDF5(wt))-treated defects. Microarray-based gene expression and quantitative PCR analyses from callus tissue point to a specific acceleration of the early phases of bone healing, comprising the inflammation and chondrogenesis phase. These results support the concept that disease-deduced growth factor variants are promising lead structures for novel therapeutics with improved clinical activities.

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The effect of two point mutations in GDF-5 on ectopic bone formation in a β-tricalciumphosphate scaffold

Cited by 23 publications

References 36 publications

Overexpression of GDF5 through an Adenovirus Vector Stimulates Osteogenesis of Human Mesenchymal Stem Cells in vitro and in vivo

Overexpression of GDF5 through an Adenovirus Vector Stimulates Osteogenesis of Human Mesenchymal Stem Cells in vitro and in vivo

Comparison of Platelet-Rich Plasma and VEGF-Transfected Mesenchymal Stem Cells on Vascularization and Bone Formation in a Critical-Size Bone Defect

Improved bone defect healing by a superagonistic GDF5 variant derived from a patient with multiple synostoses syndrome

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