Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Aug 30;12(9):1411.
doi: 10.3390/jpm12091411.

Regenerative Medicine to Improve Outcomes of Ventral Hernia Reconstruction (REPAIR Study) Phase 1: Find the Best Decellularization Protocol for the Human Dermis

Michele Altomare  1   2 , Luca Ferrario  3 , Laura Benuzzi  3 , Marta Cecilia Tosca  4 , Marta Gipponi  4 , Imam Khodri  4 , Giovanni Sesana  4 , Stefania Cimbanassi  2   5 , Stefano Piero Bernardo Cioffi  2 , Andrea Spota  2 , Roberto Bini  2 , Osvaldo Chiara  2   5 , Andrea Mingoli  6
Affiliations

Regenerative Medicine to Improve Outcomes of Ventral Hernia Reconstruction (REPAIR Study) Phase 1: Find the Best Decellularization Protocol for the Human Dermis

Michele Altomare et al. J Pers Med. .

Abstract

Background: Tissue regeneration is a complex process that allows wounds to heal. Many options are currently available to help human skin repair and to reduce the recurrence of hernias. The aim of this study is to analyze the best decellularization protocol for allogenic human dermal tissues. Methods: Dermal flaps from donors were used and compared with a control group. Each flap was subjected to seven different decellularization protocols and washed with a sequence of five solutions. The samples were then subjected to four control tests (such as Nile Red), and long-term contacts were analyzed to assess whether the decellularized dermis samples could support the growth of human fibroblasts. Results: All the samples had an average residual viability of 60%. Except for one sample, the decellularization treatments were able to reduce cell viability significantly. The Nile Red test showed a significant reduction in phospholipid content (mean 90%, p-value < 0.05) in all treatments. The cell growth increased in a linear manner. As described in the literature, sodium-dodecyl-sulfate (SDS) caused an interference between the test and the detergent. Conclusions: This paper shows the first step to finding the best decellularization protocol for allografting human dermal tissues. Further biocompatibility tests and DNA quantification are necessary.

Keywords: allogeneic tissue; decellularization protocol; fibroblasts; regenerative medicine; viability index; wound healing.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
REPAIR study protocol.
Figure 2
Figure 2
One-way ANOVA test. Mean of viability expressed as OD/mg for each protocol. NT: not treated. CTRL: control samples.
Figure 3
Figure 3
Two-way ANOVA analysis: mean of OD/mg according to donor and treatment.
Figure 4
Figure 4
One-way ANOVA analysis: mean of µg/mg for each treatment.
Figure 5
Figure 5
Two-way ANOVA analysis: mean of µg/mg according to donor and treatments.
Figure 6
Figure 6
Two-way ANOVA analysis: mean of µg/mg of donor-decellularization treatments.
Figure 7
Figure 7
Box plot: percentage of phospholipid residue after treatment application.
Figure 8
Figure 8
H&E dermis MI 49-21 treated and before treatment. (A) Before treatment, (B) C1.5TE, (C) C2TE, (D) C1.5K, (E) C2K, (F) CS, (G) S0.5, (H) S1, (I) MTF.
Figure 9
Figure 9
H&E dermis MI51-21 treated and before treatment. (A) Before treatment, (B) C1.5TE, (C) C2TE, (D) C1.5K, (E) C2K, (F) CS, (G) S0.5, (H) S1, (I) MTF.
See this image and copyright information in PMC

References

    1. Enoch S., Leaper D.J. Basic science of wound healing. Surgery. 2005;23:37–42.
    1. Järbrink K., Ni G., Sönnergren H., Schmidtchen A., Pang C., Bajpai R., Car J. Prevalence and incidence of chronic wounds and related complications: A protocol for a systematic review. Syst. Rev. 2016;5:152. doi: 10.1186/s13643-016-0329-y. - DOI - PMC - PubMed
    1. Kirsner R.S. The wound healing society chronic wound ulcer healing guidelines update of the 2006 guidelines—Blending old with new. Wound Repair Regen. 2016;24:110–111. doi: 10.1111/wrr.12393. - DOI - PubMed
    1. Gainza G., Villullas S., Pedraz J.L., Hernandez R.M., Igartua M. Advances in drug delivery systems (DDSs) to release growth factors for wound healing and skin regeneration. Nanomed. Nanotechnol. Biol. Med. 2015;11:1551–1573. doi: 10.1016/j.nano.2015.03.002. - DOI - PubMed
    1. Sen C.K., Gordillo G.M., Roy S., Kirsner R., Lambert L., Hunt T.K., Gottrup F., Gurtner G.C., Longaker M.T. Human skin wounds: A major and snowballing threat to public health and the economy. Wound Repair Regen. 2009;17:763–771. doi: 10.1111/j.1524-475X.2009.00543.x. - DOI - PMC - PubMed