Immunological and Differentiation Properties of Amniotic Cells Are Retained After Immobilization in Pectin Gel

Abstract

Mesenchymal stromal cells from the human amniotic membrane (i.e., human amniotic mesenchymal stromal cells [hAMSCs]) of term placenta are increasingly attracting attention for their applications in regenerative medicine. Osteochondral defects represent a major clinical problem with lifelong chronic pain and compromised quality of life. Great promise for osteochondral regeneration is held in hydrogel-based constructs that have a flexible composition and mimic the physiological structure of cartilage. Cell loading within a hydrogel represents an advantage for regenerative purposes, but the encapsulation steps can modify cell properties. As pectin gels have also been explored as cell vehicles on 3D scaffolds, the aim of this study was to explore the possibility to include hAMSCs in pectin gel. Immobilization of hAMSCs into pectin gels could expand their application in cell-based bioengineering strategies. hAMSCs were analyzed for their viability and recovery from the pectin gel and for their ability to differentiate toward the osteogenic lineage and to maintain their immunological characteristics. When treated with a purposely designed pectin/hydroxyapatite gel biocomposite, hAMSCs retained their ability to differentiate toward the osteogenic lineage, did not induce an immune response, and retained their ability to reduce T cell proliferation. Taken together, these results suggest that hAMSCs could be used in combination to pectin gels for the study of novel osteochondral regeneration strategies.

Assessment of the in vivo biofunctionality of a biomimetic hybrid scaffold for osteochondral tissue regeneration

“…The intrinsic immune‐modulatory properties of hAMSCs were confirmed by immunohistochemical results that showed no significant activation of the adaptive immunity (CD4 and CD8), thus confirming the inability of these cells to induce a T‐cell response in accordance with Silini et al (2018). Furthermore, significant activation of the innate immunity was observed (CD45 and CD68).…”

“…Pectin microspheres were studied for 3D culture of bone cells, showing interesting properties for cell viability, metabolic activity and differentiation (Munarin et al, 2010(Munarin et al, , 2011. The pectin gel used in the present study was investigated in preliminary experiments with hAMSCs for the first time, confirming increased cell viability, biocompatibility, ability to differentiate toward osteogenic commitment, and retaining the ability to reduce T cell proliferation (Farè et al, 2015;Silini et al, 2018). As the PU/VAC/PECT scaffold showed the pectin degradation after 4 days of incubation in vitro, cells previously embedded in can be release in the defect site, pro- The intrinsic immune-modulatory properties of hAMSCs were confirmed by immunohistochemical results that showed no significant activation of the adaptive immunity (CD4 and CD8), thus confirming the inability of these cells to induce a T-cell response in accordance with Silini et al (2018).…”

“…Human AMSCs are poorly immunogenic and exert their biological function through their ability to modulate the immune response, as demonstrated in different preclinical models of inflammatory‐based diseases (Parolini et al, 2014; Silini et al, 2017). Furthermore, in preliminary experiments, hAMSC seeded with pectin gel onto PU scaffolds (Farè et al, 2015; Silini et al, 2018), or seeded on PU scaffold alone (Bertoldi et al, 2010), showed increased viability, biocompatibility, osteogenic commitment, did not induce an immune response, and retained the ability to reduce T cell proliferation (Silini et al, 2018). Our strategy was to develop a novel hybrid scaffold in which the load‐bearing structure is represented by a 3D porous PU, coated under vacuum with calcium phosphates (PU/VAC).…”

Viability and Morphology of Human Dental Pulp Stem Cells in the Presence of Citrus Pectin

“…Human dental pulp stem cells (hDPSCs) which were isolated and characterized in a previous study [39] were a kind gift from Prof. Dr. Menemşe Gümüşderelioğlu, from Hacettepe University, Turkey. The cells were cultured in 15% (v/v) fetal bovine serum (FBS, Biowest, France) including alpha-Minimum Essential Medium (α-MEM, Biochrom, Germany).…”

“…In a study, human amniotic membrane MSCs were cultured in hydroxyapatite/LM pectin composite gel structure. It was shown that MSCs preserved their osteogenic differentiation capacity and inhibition of T cell proliferation in the presence of LM pectin, also they did not trigger immune response [39]. Pectin/chitosan membrane can provide attachment, adhesion and proliferation of adipose-derived stem cells (ADSCs) [40].…”