Bioactivity and composition of a preserved connective tissue matrix derived from human placental tissue

Joseph Irvin, Carina Danchik, Joseph Rall, Annie Babcock, Matthew Pine, Devin Barnaby, Joseph Pathakamuri, Daniel Kuebler, Joseph Irvin, Carina Danchik, Joseph Rall, Annie Babcock, Matthew Pine, Devin Barnaby, Joseph Pathakamuri, Daniel Kuebler

Abstract

There are a wide variety of extracellular matrices that can be used for regenerative purposes. Placental tissue-based matrices are quickly becoming an attractive option given the availability of the tissue source and the wide variety of bioactive molecules knows to exist in unprocessed placental tissues. As fresh placental tissues are seldom an option at the point of care, we examined both the composition and bioactivity of a commercially packaged flowable placental connective tissue matrix (FPTM) (BioECM® , Skye Biologics, Inc.) that was preserved by the proprietary HydraTek® process. The FPTM contained significant amounts of collagen and various growth factors such as bFGF, EGF, PDGF, KGF, and PIGF. In addition, it contained high levels of tissue inhibitors of metalloproteinases (TIMP-1 and 2) and molecules known to modulate the immune response including TGF-β and IL-4. In terms of its bioactivity, the FPTM displayed the ability (1) to suppress INF-γ secretion in activated T-cells nearly fourfold over control media, (2) to inhibit methicillin resistant Staphylococcus aureus (MRSA) and Staphylococcus saprophyticus proliferation, (3) to increase the migration of adipose-derived stem cells (ASCs) nearly threefold over control media and (4) to adhere to ASCs in culture. When ASCs were exposed to FPTM in culture, the cells maintained healthy morphology and showed no significant changes in the expression of five genes involved in tissue growth and repair as compared to culture in standard growth media. © 2018 The Authors Journal of Biomedical Materials Research Part B: Applied Biomaterials Published by Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2731-2740, 2018.

Keywords: adipose-derived stromal/stem cells; connective tissue; extracellular matrix; growth factors; placenta.

© 2018 The Authors Journal of Biomedical Materials Research Part B: Applied Biomaterials Published by Wiley Periodicals, Inc.

Figures

Figure 1
Figure 1
INF‐gamma secretion following Dynabead T cell activation in the presence or absence of 20% FPTM. INF‐gamma secretion was significantly inhibited by the addition of the placental product. INF‐gamma secretion was consistently reduced across five different lots of FPTM (A–E). The 20% FPTM bar refers to the mean value of the five different lots tested.
Figure 2
Figure 2
ASCs in culture bind to the placental connective tissue in FPTM product. (a) ASCs in control media. (b) ASCs in media supplemented with 10% FPTM. Scale bar = 100 μm.
Figure 3
Figure 3
Changes in gene expression in ASCs cultured in control media versus 10% FPTM supplemented media. The addition of FPTM did not significantly alter the expression of any of the genes tested although the majority of the genes displayed a slight increase in expression levels.
Figure 4
Figure 4
The migratory capacity of ASCs was up‐regulated significantly by the addition of 20% FPTM. The addition of FPTM samples from four different lots (S1–S4) consistently upregulated cell migration nearly threefold over baseline. The 20% FPTM bar refers to the mean value of the four different lots tested.
Figure 5
Figure 5
(a) Effect of FPTM on the growth of MRSA. (b) Effect of FPTM on the growth of Sa.ccharomyces saprophyticus. The relative growth was determined following 10 h at 37°C in nutrient media or nutrient media supplemented with various percentages of FPTM. The FPTM inhibits the growth of both bacterial strains.

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