Bioscaffold developed with decellularized human amniotic membrane seeded with mesenchymal stromal cells: assessment of efficacy and safety profiles in a second-degree burn preclinical model

Abstract

Therapies to deep burn injuries remain a global challenge. Human amniotic membrane (hAM) is a biomaterial that has been increasingly explored by the field of regenerative medicine. A decellularized hAM (DhAM) can be used as scaffold for mesenchymal stromal cells (MSCs) to grow without the loss of their stemness potential, allowing its application as cell therapy for wound healing. In this work, we associated DhAM with adipose-derived mesenchymal stromal cells (DhAM+AD-MSC), as a therapy strategy for second-degree burns in a preclinical model . Animals with induced second-degree burns were divided into 4 groups: control, which consists of a non-adherent gauze; a synthetic commercial dressing as the positive control (control+); DhAM; and DhAM plus rat AD-MSCs (DhAM+AD-MSCs), followed by detailed and long term analysis (5 weeks). The macroscopical analysis showed the healing improvement in the wound area after the DhAM+AD-MSC treatment. Histological analysis also showed no alteration in the animal organs and a regular epithelial progression in comparison to the control. This observation was also confirmed by the analysis of suprabasal layers in the neoepidermis with CK10, showing a stratified and differentiated epithelium, when compared to Control and Control+. A strong CD73 (ecto-5′-nucleotidase) labeling was observed in the first two weeks postburn in dermis and epidermis. The expression in dermis was stronger in the second week in the middle of the wound, when comparing the Control+ with DhAM+AD-MSCs (p=0.0238). In the epidermis the expression of CD73 was increased in all regions when compared to the control. This data suggests the involvement of this protein on wound healing. A low CD11b labeling was observed in DhAM+AD-MSCs treatment group mainly in the last treatment week, in comparison to Control and Control+ (p<0.0001), which indicates a reduction in the inflammatory process. MSCs through CD73 can release high concentrations of adenosine, an immunosuppressive molecule, suggesting that this could be the mechanism by which the inflammation was better modulated in the DhAM+AD-MSCs group. The results obtained with this preclinical model confirm the effectiveness and safety of this low-cost and highly available dressing for future clinical application as a therapy for burn treatments.

Assessment of the proteome profile of decellularized human amniotic membrane and its biocompatibility with umbilical cord‐derived mesenchymal stem cells

“…We analyzed the cytocompatibility of both, the epithelial and basal sides of D-hAM with hUC-MSC separately. Consistent with earlier studies, 23,25,61 hUC-MSC remained viable and proliferated actively on D-hAM for up to 15 days in culture. Moreover, the present study showed that hUC-MSC seeded on the basal side of D-hAM presented significantly higher proliferation on the basal surface than the epithelial surface, which is a relatively novel finding.…”

“…Earlier studies have reported that preserving the ECM components of decellularized bio-scaffolds provides a suitable microenvironment for allogeneic cells for attachment, proliferation, and differentiation along various lineages. 59,60 Moreover, numerous studies have suggested the combined use of MSCs with a D-hAM, [61][62][63] (i) to enhance healing capacity by accelerating the reepithelialization, modulating inflammation, promoting granulation tissue formation and regulating remodeling of ECM 64 (ii) to increase its anti-infective effects resulting from the increased production of anti-inflammatory proteins like interleukin-10 (IL-10) and M2 macrophages and the reduced expression of pro-inflammatory cytokines like interleukin-1 (IL-1), interleukin-6 (IL-6), and transforming growth factor-β (TGF-β) 65 (iii) to induce and maintain multilineage differentiation in response to various stimulus from three dimensional system, 66 and (iv)to direct angiogenesis by secreting pro-angiogenic factors such as VEGF, insulin like growth factor (IGF), and angiogenin. 67 Hence, we hypothesized that D-hAM would support the attachment and proliferation of hUC-MSC while proving to be a biocompatible scaffold for future tissue engineering and regenerative applications.…”