Biomedical subjects

D P Luxenberg

Publications and source records attributed to D P Luxenberg.

5 recordsLinked to original sources

PubMed1993

Bone morphogenetic protein-2 causes commitment and differentiation in C3H10T1/2 and 3T3 cells.

C3H10T1/2 cells are an established mesenchymal stem cell line which can differentiate into muscle, fat and cartilage cells when treated with azacytidine. Bone morphogenetic protein-2 (BMP-2) caused a dose dependent differentiation of these cells into fat, cartilage and bone cells-low concentrations favoring adipocytes and high concentrations chondrocytes and osteoblasts. The differentiated phenotypes were stable in the absence of BMP-2. Furthermore, the addition of other growth factors during the differentiation process altered the frequency of the differentiated colony formation. Transfection of the C3H10T1/2 cells with a BMP-2 cDNA also induced a phenotypic change from the parental fibroblast to adipocytes and osteoblasts. Our results in this model system indicate that a single protein factor can cause differentiation of a stem cell line to multiple phenotypes, that phenotypes induced can be regulated by factor concentration, and that other factors can also influence BMP-2 induced differentiation.

3T3 Cells↗

PubMed1991

Mandibular reconstruction with a recombinant bone-inducing factor. Functional, histologic, and biomechanical evaluation.

Bone morphogenetic protein-2 (BMP-2) is a human recombinant bone-inducing factor that stimulates bone formation within 14 days. Twenty-six dogs underwent reconstruction of 3-cm full-thickness mandibular defects. After stabilizing the defects with stainless steel reconstruction plates, test implants composed of inactive dog bone matrix carrier and human recombinant BMP-2 were placed in defects of 12 animals (group 1). Control implants (carrier without BMP-2) were used in 10 animals (group 2), and no implants were placed in mandibular defects of four animals (group 3). Animals were killed at 3 and 6 months. The reconstructed segments were evaluated by roentgenography, analysis of functional stability, histology, histomorphometry, and analysis of biomechanical strength using three-point bend testing. In group 1, reconstruction plates were removed at 10 weeks because stiff, noncompressible mineralized bone formed across the defects, allowing the animals to chew a solid diet. The defects from groups 2 and 3 showed minimal, if any, bone formation and remained grossly unstable, prohibiting plate removal or advancement to a solid diet. Histomorphometric analysis at 6 months revealed that 68% of the group 1 implants were replaced by mineralized bone, whereas mineralized bone occupied less than 4% of the implants in groups 2 and 3. Biomechanical testing at 6 months revealed that the average bending strength of the reconstructed hemimandibles (expressed as a percentage of the contralateral hemimandible) was 27% for group 1 and 0% for group 2. The biomechanical strength of the defects reconstructed with BMP-2 increased significantly from 3 to 6 months and was related to degree of mineralization and thickness of bone bridging the defect.

Animals↗

PubMed1988

Purification and characterization of other distinct bone-inducing factors.

We purified a factor that induces bone formation greater than 300,000-fold from guanidinium chloride extracts of demineralized bone. Fifty nanograms of highly purified protein was active in an in vivo cartilage and bone-formation assay. The activity resided in a single gel band, corresponding to a molecular mass of approximately 30 kDa, which yielded proteins of 30, 18, and 16 kDa on reduction. The partial amino acid sequence obtained from these proteins confirmed our identification of specific factors that induce new bone formation in vivo.

Amino Acid Sequence↗

PubMed1985

Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins.

Clones of complementary DNA encoding the human lymphokine known as granulocyte-macrophage colony-stimulating factor (GM-CSF) were isolated by means of a mammalian cell (monkey COS cell) expression screening system. One of these clones was used to produce recombinant GM-CSF in mammalian cells. The recombinant hematopoietin was similar to the natural product that was purified to apparent homogeneity from medium conditioned by a human T-cell line. The human T-cell GM-CSF was found to be 60 percent homologous with the GM-CSF recently cloned from murine lung messenger RNA.

Amino Acid Sequence↗

PubMed1985

Isolation of cDNAs encoding human and gibbon GM-CSF.

We have used a mammalian cell expression cloning system to identify cDNA clones encoding human granulocyte-macrophage colony stimulating factor. The human clone was used as a hybridization probe to identify the corresponding sequence from a cDNA library prepared from a gibbon T-cell line. The human cDNA has been used to produce recombinant GM-CSF in monkey COS-1 cells. The purified protein from COS cells is very similar to the GM-CSF isolated from a continuous human T-cell line.

Amino Acid Sequence↗