Bio-Fabrication of Human Amniotic Membrane Zinc Oxide Nanoparticles and the Wet/Dry HAM Dressing Membrane for Wound Healing
Date
2021-07-28
Journal Title
Journal ISSN
Volume Title
Publisher
Frontiers Media
Abstract
The preparation of unique wet and dry wound dressing products derived from
unprocessed human amniotic membrane (UP-HAM) is described. The UP-HAM was
decellularized, and the constituent proteins were cross-linked and stabilized before
being trimmed and packed in sterile Nucril-coated laminated aluminium foil pouches
with isopropyl alcohol to manufacture processed wet human amniotic membrane (PWHAM). The dry type of PD-HAM was prepared by decellularizing the membrane, UV
irradiating it, lyophilizing/freeze-drying it, sterilizing it, and storing it at room temperature.
The UP-HAM consists of a translucent yellowish mass of flexible membranes with an
average thickness of 42 µm. PW-HAM wound dressings that had been processed,
decellularized, and dehydrated had a thinner average thickness of 30 µm and lacked
nuclear-cellular structures. Following successful decellularization, discrete bundle of
fibrous components in the stromal spongy layers, microvilli and reticular ridges were
still evident on the surface of the processed HAM, possibly representing the location of
the cells that had been removed by the decellularization process. Both wet and dry HAM
wound dressings are durable, portable, have a shelf life of 3–5 years, and are available
all year. A slice of HAM dressing costs 1.0 US$/cm2
. Automation and large-scale
HAM membrane preparation, as well as storage and transportation of the dressings,
can all help to establish advanced technologies, improve the efficiency of membrane
production, and reduce costs. Successful treatment of wounds to the cornea of the
eye was achieved with the application of the HAM wound dressings. The HAM protein
analysis revealed 360 µg proteins per gram of tissue, divided into three main fractions
with MWs of 100 kDa, 70 kDa, and 14 kDa, as well as seven minor proteins, with the
14 kDa protein displaying antibacterial properties against human pathogenic bacteria.
Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 1 July 2021 | Volume 9 | Article 695710
fbioe-09-695710 July 22, 2021 Time: 16:39 # 2
Ramasamy et al. HAMP-ZnO Nanoparticles HAM Wound Dressing
Wet and dry wound dressings were
produced. HAM proteins were
purified and analysed. The zinc
oxide nanoparticles (HAMP-ZnO
NP) made from HAM proteins
were characterised and tested for
their antibacterial activity.
Wounds to the cornea of the eye
healed easily when treated with
HAM wound dressings.
Fresh human Amniotic
membrane, Serological
screening, selection of
disease-free HAM, reome
stromal layer, preparation of
HAM.
UNPROCESSED HAM
Cuboidal epithelial cells, basement
membrane, compact layer, stromal
and spongy layers containing
scatted fibroblast cells are visible in
hsitological analysis.
The flow chart depicts the
methods for processing, and
preparation of wet (PWHAM) and dry (PD-HAM)
wound healing dressings.
HAM proteins, Nanoparticle
synthesis (HAMP-ZnO NP) and
analysis.
Antibacterial analysis show
Inhibition of growth and biofilm
formation of pathogenic bacteria .
Processed HAM
lacked a nuclear-cellular
epithelium, but it did have a
distinct fibrous elements in
basement membrane, stromal
and spongy layers.
Processed PW-HAM (Light
&SEM) showed smooth
epithelial surface topography
with microvilli,.
HAM dressing, wet/dry, packed,
labelled, sterilised and processed. They
are durable, portable, have long
shelf life . A slice of HAM dressing
costs US$ 1.0 / cm² . The wound
dressings are ready to be applied.
The dermal wounds and conjunctival
surface can be successfully repaired
using processed HAM wound dressings
GRAPHICAL ABSTRACT | Flow chart depicting the methods, preparing, and characterizing, by histological, and scanning electron microscopy, of wet (PW-HAM)
and dry (PD-HAM)of wound healing dressing, and preparation of nanoparticles (HAMP ZnO NP); and application of HAM wound dressing.
A wide range of antibacterial activity was observed after treatment with 75 µg/ml zinc
oxide nanoparticles derived from human amniotic membrane proteins (HAMP-ZnO NP),
including dose-dependent biofilm inhibition and inhibition of Gram-positive (S. aureus,
S. mutans, E. faecalis, and L. fusiformis) and Gram-negative bacteria (S. sonnei, P.
aeruginosa, P. vulgaris, and C. freundii).
Description
Publication history: Accepted - 25 June 2021; Published online - 28 July 2021.
item.page.type
Article
item.page.format
Keywords
amniotic membrane, wound healing dress, wet/dry membrane, proteins, biofabrication, HAMP's-ZnO nanoparticles, antibacterial activities, biofilm
Citation
Ramasamy, P., Krishnakumar, R., Rekha, R., Vaseeharan, B., Saraswathi, K., Raj, M., Hanna, R. E. B., Brennan, G. P., Dayanithi, G. and Vijayakumar, S. (2021) ‘Bio-Fabrication of Human Amniotic Membrane Zinc Oxide Nanoparticles and the Wet/Dry HAM Dressing Membrane for Wound Healing’, Frontiers in Bioengineering and Biotechnology, 9. doi: 10.3389/fbioe.2021.695710.