Einfluß verschiedener Desinfektions- und Sterilisationsverfahren für allogene Knochentransplantate auf die Osteoblastenfunktion in-vitro
Loading...
Files
Date
Authors
Publisher
Philipps-Universität Marburg
Supervisors
Abstract
One important reason for different clinical
outcome after bone allografting might be the alteration of cell
adherence, viability, and differentiation by different
processing methods. In order to assess the influence of eight
different sterilisation and disinfection methods for bone
allografts on adhesion, proliferation, and differentiation of
bovine periosteal osteoblasts, cells were grown in culture and
then plated onto pieces of human bone allografts. We tested
autoclaved bone (AUT), demineralised and low-temperature-plasma
sterilised bone (D-LTP), ethylene oxide sterilised bone (EtO),
80°C-thermodisinfected bone (80°C), and chemical solvent
disinfected bone (CSD). The seeding efficiency was determined
during the first hour of incubation to detect the number of
attached cells but before mitosis starts. The cell viability
was tested after a culture period of 7 and 21 days using the
MTT-assay and determination of the total DNA content. Tests to
confirm the osteoblastic function and differentiation of the
cells included measuring of the total protein amount,
histochemical alkaline phosphatase staining and quantitative
determination of AP-activity, ELISA for osteocalcin and
SDS-PAGE for collagens. Results showed that heat processing of
bone matrix clearly influenced the adhesion, proliferation and
differentiation of osteoblasts. Further, serum protein mediated
cell adhesion upon bone allograft surfaces was altered by
different processing methods. Most cells adhered to D-LTP-bone.
Highest proliferation rates of periosteal osteoblasts as
measured via MTT-activity and total DNA-content were also found
within the D-LTP-group after one and three weeks. Highest
expression levels of solved and ECM-proteins were found in CLD-
and D-LTP groups, followed by the 80°C-group. Extracellular
matrix proteins (e.g. Collagen Type I) are necessary for the
attachment, migration, proliferation, and differentiation of
osteoblasts. The present study clealy shows that different
processing methods directly influence the gene expression of
collagens Type I and III. The highest expression levels of this
ECM-marker was found in the D-LTP-Group, followed by 80°C, AUT,
CLD, and EtO-groups. Corresponding to this findings, expression
of specific differentiation markers of osteoblastic phenotype
AP and osteocalcin were significantly increased in D-LTP group.
Growth factor activity (TGF?s, BMP?s) of the allograft matrix
may be responsible for these effects. The study reports an
in-vitro model to examine the influence of different processing
methods for allogenic bone grafts on single cell types. For the
first time a direct change of osteoblast growth and funtion
caused by processed bone grafts was shown in the present
work.
Review
Metadata
Contributors
Supervisor:
Dates
Created: 2004Issued: 2004-04-08Updated: 2011-08-10
Faculty
Medizin
Publisher
Philipps-Universität Marburg
Language
ger
Data types
DoctoralThesis
Keywords
Osteoblastallograft, cell culturebone
DFG-subjects
ZellkulturOsteoblastAllogenes Transplantat , Knochentransplantation
DDC-Numbers
610
show more
Hofmann, Alexander (128938307): Einfluß verschiedener Desinfektions- und Sterilisationsverfahren für allogene Knochentransplantate auf die Osteoblastenfunktion in-vitro. : Philipps-Universität Marburg 2004-04-08. DOI: https://doi.org/10.17192/z2004.0215.
License
This item has been published with the following license: In Copyright