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Biomedical subjects

R Luginbühl

Publications and source records attributed to R Luginbühl.

4 recordsLinked to original sources

Comparison of human bone marrow stromal cells seeded on calcium-deficient hydroxyapatite, beta-tricalcium phosphate and demineralized bone matrix.

The aim of this study was to compare three resorbable biomaterials regarding seeding efficacy with human bone marrow stromal cells (BMSCs), cell penetration into the matrix, cell proliferation and osteogenic differentiation. Calcium-deficient hydroxyapatite (CDHA), beta-tricalcium phosphate (beta-TCP), and demineralized bone matrix (DBM) were seeded with human BMSCs and kept in human serum and osteogenic supplements for 3 weeks. Morphologic and biochemical evaluations were performed on day 1, 7, 14 and 21. The allograft DBM and CDHA exhibited both an excellent seeding efficacy while the performance of beta-TCP was lower when compared. The total protein content and the values for specific alkaline phosphatase (ALP) increased on all matrices and no significant difference was found for these two markers. BMSCs in monolayer had a significant increase of protein, but not of ALP. Osteocalcin (OC) values increased significantly higher for BMSC in cultures on DBM when compared to CDHA and beta-TCP. The OC levels decreased significantly in the BMSC monolayer culture. BMSCs were found inconsistently within the synthetic materials, whereas in DBM they were found more homogeneously distributed throughout the matrix. All three matrices promoted BMSC proliferation and differentiation to osteogenic cells. DBM allografts seem to be more favorable with respect to cell ingrowth tested by histology, and osteogenic differentiation ascertained by an increase of OC. CDHA with its high specific surface area showed more favorable properties than beta-TCP regarding reproducibility of the seeding efficacy.

Biocompatible Materials↗

Quantitative interrogation of micropatterned biomolecules by surface force microscopy.

Synthetic biomaterials are widely used in medical implants with success in improving and extending quality of life. However, these materials were not originally designed to interact with cells through specific signaling pathways. As a result, the interaction with the body is mediated through passive adsorption of a disorganized protein monolayer. Next generation biomaterials have been proposed to be active in modifying the biological response of the host through the incorporation of specific biorecognition moieties. An important tool in the development of these novel active biomaterials is the scanning force microscope (SFM). The SFM allows for interrogation of bioactive biomaterials in mapping or spectroscopic modes. In this work, micropatterned protein surfaces were prepared using biomolecules implicated in wound healing. The surfaces were imaged via SFM and the specific binding forces between surface associated biomolecules and antibody functionalized tips were quantified.

Antibodies↗

Immunosensing with photo-immobilized immunoreagents on planar optical wave guides.

Immunocomplexation at wave guiding TiO2/SiO2 surfaces was investigated using an integrated optical grating coupler. For extended application of this label-free monitoring system, F(ab')2 fragments of monoclonal antibodies were photo-immobilized by photolinker polymer-mediated procedures that do not require functionalization of either the immunoreagent or the TiO2/SiO2 surface. Covalent, light-dependent binding of photolinker polymer and F(ab')2 fragments was achieved using a single-step photo-reaction. Bovine serum albumin derivatized with aryldiazirines (T-BSA) served as a photolinker polymer. T-BSA suppressed the non-specific adsorption of analytes to wave guide surfaces. Immunoreagent binding and immunological activity were analyzed and modified surfaces were investigated by scanning force microscopy. Apparent immunoreagent surface densities were 16.7 fmol F(ab')2 per mm2 sensor surface. Optical analyses revealed linear, dose-dependent antigen binding with label free analytes. Immunocompetent surfaces were regenerable by treatment at pH 2.3, rendering the immunosensing system applicable for repetitive use.

Antigen-Antibody Reactions↗

[Induction of bone tissue on different matrices: an in vitro and a in vivo pilot study in the SCID mouse].

AIM: Three resorbable biomaterials were evaluated regarding proliferation and osteogenic differentiation of human bone marrow stromal cells (BMSC) in vitro. In a second step, the new biomaterial, calcium-deficient hydroxyapatite (CDHA), was tested in a pilot in vivo study by subcutaneous implantation in the severe combined immunodeficiency (SCID) mouse. METHODS: CDHA, beta-tricalcium phosphate (beta-TCP), and demineralized bone matrix (DBM) were seeded with human BMSC and cultured in osteogenic supplements for 3 weeks. In the pilot in vivo study, CDHA was seeded with BMSC and kept in osteogenic media for 2 weeks (group A) before subcutaneous implantation in 8 SCID mice for 3 and 8 weeks. In addition, CDHA seeded with BMSC without prior osteogenic induction (group B) and empty ceramics were implanted in each mouse. RESULTS: Total protein content and the values for specific alkaline phosphatase (ALP) increased significantly in vitro on all matrices, but no significant difference between the groups was noted. In the pilot in vivo study all ceramics were well penetrated by cells. After 8 weeks 2 of 4 samples in group B and 1 of 4 samples in group A revealed cells resembling hypertrophic chondrocytes. Specific ALP was higher in the group B (p = 0.012, Z = - 2.5) compared to empty ceramics. There were no significant differences between groups A and B. Differences between group A and the empty control did not become significant (p = 0.069, Z = - 1.8). CONCLUSION: All three matrices promoted BMSC proliferation and differentiation to osteogenic cells in vitro. Human BMSC on CDHA showed signs of osteogenic differentiation after subcutaneous implantation into SCID mice.

Adolescent↗