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Human biomaterials from placenta for tissue engineering and regenerative medicine

Authors:
Johannes Hackethal at THT Biomaterials
Johannes Hackethal
  • THT Biomaterials
Severin Mühleder
Severin Mühleder
Severin Mühleder
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Simone Hennerbichler
Simone Hennerbichler
Simone Hennerbichler
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Heinz Redl at Ludwig Boltzmann Institute Traumatology
Heinz Redl
  • Ludwig Boltzmann Institute Traumatology
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Abstract

Human Extracellular Matrix (ECM) is nature’s perfect cell niche. Currently, many biomaterials extracted from ECM are gold standard products in many clinical fields. Main sources for the isolation of these proteins are still nonhuman tissues, associated with immune reactions and xenogenic disease transmission. In contrast, placenta tissue as a potential source for ECM proteins is consistently available from full-term birth and it fulfills all demands of cost-efficient bioengineering on a large industrial scale. The aim of these studies was to establish effective isolation methods from ECM proteins from human placenta.
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AMBA 2017: Advanced Materials for Biomedical Applications, Ghent, September 27th-29nd 2017 1
Human biomaterials from placenta for tissue engineering and regenerative
medicine
Johannes Hackethal1,2, Severin Mühleder1,2, Simone Hennerbichler2,3, Heinz Redl1,2, Andreas Teuschl2,4
1Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria 2Austrian Cluster for
Tissue Regeneration, Vienna, Austria
3Red Cross Blood Transfusion Service of Upper Austria, Linz, Austria
4Department of Biochemical Engineering, University of Applied Sciences Technikum Wien, Vienna, Austria
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INTRODUCTION
Human Extracellular Matrix (ECM) is nature’s
perfect cell niche. Currently, many biomaterials
extracted from ECM are gold standard
products in many clinical fields. Main sources
for the isolation of these proteins are still non-
human tissues, associated with immune
reactions and xenogenic disease transmission.
In contrast, placenta tissue as a potential
source for ECM proteins is consistently
available from full-term birth and it fulfills all
demands of cost-efficient bioengineering on a
large industrial scale. The aim of these studies
was to establish effective isolation methods
from ECM proteins from human placenta.
MATERIALS AND METHODS
The proteins were biochemically characterized
by determining residual DNA contents,
showing the protein fractions on gel
electrophoresis (SDS-PAGE) analysis combined
with total amino acid quantification,
identifying the isolated ECM proteins by
Western Blot analysis. Finally, the
biocompatibility of the isolated ECM proteins
was demonstrated in 2D and 3D in vitro assays
using NG-108 cell line, HUVEC, primary rat
hepatocytes and Schwann cells.
RESULTS AND DISCUSSION
We have developed effective methods to
isolate ECM proteins such as atelocollagen type
1 and 3 or laminin-111 from human basal
placenta tissue based on enzymatic or non-
enzymatic isolation methods. Mean values of
isolated ECM proteins were increased
compared to current methods published. DNA
residues of the isolates were assessed using a
HOECHST-33342 dye-based DNA quantification
assay. Mean DNA content was significantly
reduced after processing. By using SDS-PAGE
and Coomassie blue staining, we determined
the purity of our extracts. The specific ECM
proteins were assessed by Western blot
analysis. Amino Acid quantification was used to
determine the amino-acid fingerprint of the
isolates. In 2D and 3D cell culture assays, ECM
proteins from human placenta revealed
excellent structural and functional properties.
CONCLUSIONS
Despite the ongoing development and
research in new biomaterials over the last
decades, human-derived ECM proteins are still
regarded as the best option for the generation
of medicinal products.
REFERENCES
Badylak SF., et al, 2009. Extracellular matrix as
a biological scaffold material: Structure and
function. Acta Biomaterialia 5(1-13)
Lobo SE., et al., 2016. The placenta as an organ
and a source of stem cells and extracellular
matrix: a review. Cells Tissues Organs 201,239-
246.
ACKNOWLEDGEMENT
The authors acknowledge the Red Cross Blood
Transfusion Service, Linz, Upper Austria, for
providing the placenta tissues.
... Since bones exhibit superelastic biomechanical properties, the ideal scaffolds should mimic its strength, stiffness, and mechanical behavior, so as to avoid bone resorption and consequent implant failure. Additionally, an ideal scaffold should be non-toxic, non-immunogenic, and biocompatible, and it should be easily functionalized with bioactive proteins and/or chemicals [42]. Another key factor is the biodegradability of the material: the degradation rate of tissue engineering scaffolds should mirror the rate of new tissue formation [42]. ...
... Additionally, an ideal scaffold should be non-toxic, non-immunogenic, and biocompatible, and it should be easily functionalized with bioactive proteins and/or chemicals [42]. Another key factor is the biodegradability of the material: the degradation rate of tissue engineering scaffolds should mirror the rate of new tissue formation [42]. Additionally, the resulting products should be non-toxic and easily metabolized and cleared from the body. ...
... This serves as a physical foundation for the development of new tissues with specific functions. Therefore, one of the prerequisites of practical tissue engineering technology is to obtain 3-D matrices with good biocompatibility [42]. ...
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... Since bones exhibit superelastic biomechanical properties, the ideal scaffolds should mimic its strength, stiffness, and mechanical behavior, so as to avoid bone resorption and consequent implant failure. Additionally, an ideal scaffold should be non-toxic, non-immunogenic, and biocompatible, and it should be easily functionalized with bioactive proteins and/or chemicals [42]. Another key factor is the biodegradability of the material: the degradation rate of tissue engineering scaffolds should mirror the rate of new tissue formation [42]. ...
... Additionally, an ideal scaffold should be non-toxic, non-immunogenic, and biocompatible, and it should be easily functionalized with bioactive proteins and/or chemicals [42]. Another key factor is the biodegradability of the material: the degradation rate of tissue engineering scaffolds should mirror the rate of new tissue formation [42]. Additionally, the resulting products should be non-toxic and easily metabolized and cleared from the body. ...
... This serves as a physical foundation for the development of new tissues with specific functions. Therefore, one of the prerequisites of practical tissue engineering technology is to obtain 3-D matrices with good biocompatibility [42]. ...
View
... water vapor annealing process [5,6]. ...
View
Biomimetic Nanostructures with Compositional Gradient Grown by Combinatorial Matrix-Assisted Pulsed Laser Evaporation for Tissue Engineering
Article
  • Sep 2019
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The Placenta as an Organ and a Source of Stem Cells and Extracellular Matrix: A Review
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  • Apr 2016
  • CELLS TISSUES ORGANS
The placenta is a temporal, dynamic and diverse organ with important immunological features that facilitate embryonic and fetal development and survival, notwithstanding the fact that several aspects of its formation and function closely resemble tumor progression. Placentation in mammals is commonly used to characterize the evolution of species, including insights into human evolution. Although most placentas are discarded after birth, they are a high-yield source for the isolation of stem/progenitor cells and are rich in extracellular matrix (ECM), representing an important resource for regenerative medicine purposes. Interactions among cells, ECM and bioactive molecules regulate tissue and organ generation and comprise the foundation of tissue engineering. In the present article, differences among several mammalian species regarding the placental types and classifications, phenotypes and potency of placenta-derived stem/progenitor cells, placental ECM components and current placental ECM applications were reviewed to highlight their potential clinical and biomedical relevance.
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Extracellular Matrix as a Biological Scaffold Material: Structure and Function
Article
  • Jan 2009
Biological scaffold materials derived from the extracellular matrix (ECM) of intact mammalian tissues have been successfully used in a variety of tissue engineering/regenerative medicine applications both in preclinical studies and in clinical applications. Although it is recognized that the materials have constructive remodeling properties, the mechanisms by which functional tissue restoration is achieved are not well understood. There is evidence to support essential roles for both the structural and functional characteristics of the biological scaffold materials. This paper provides an overview of the composition and structure of selected ECM scaffold materials, the effects of manufacturing methods upon the structural properties and resulting mechanical behavior of the scaffold materials, and the in vivo degradation and remodeling of ECM scaffolds with an emphasis on tissue function. Copyright © 2008. Published by Elsevier Ltd.
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