Commitment of Autologous Human Multipotent Stem Cells on Biomimetic Poly-L-Lactic Acid-Based Scaffolds Is Strongly Influenced by Structure and Concentration of Carbon Nanomaterial

Abstract

Nanocomposite scaffolds combining carbon nanomaterials (CNMs) with a biocompatible matrix are able to favor the neuronal differentiation and growth of a number of cell types, because they mimic neural-tissue nanotopography and/or conductivity. We performed comparative analysis of biomimetic scaffolds with poly-L-lactic acid (PLLA) matrix and three different p-methoxyphenyl functionalized carbon nanofillers, namely, carbon nanotubes (CNTs), carbon nanohorns (CNHs), and reduced graphene oxide (RGO), dispersed at varying concentrations. qRT-PCR analysis of the modulation of neuronal markers in human circulating multipotent cells cultured on nanocomposite scaffolds showed high variability in their expression patterns depending on the scaffolds’ inhomogeneities. Local stimuli variation could result in a multi- to oligopotency shift and commitment towards multiple cell lineages, which was assessed by the qRT-PCR profiling of markers for neural, adipogenic, and myogenic cell lineages. Less conductive scaffolds, i.e., bare poly-L-lactic acid (PLLA)-, CNH-, and RGO-based nanocomposites, appeared to boost the expression of myogenic-lineage marker genes. Moreover, scaffolds are much more effective on early commitment than in subsequent differentiation. This work suggests that biomimetic PLLA carbon-nanomaterial (PLLA-CNM) scaffolds combined with multipotent autologous cells can represent a powerful tool in the regenerative medicine of multiple tissue types, opening the route to next analyses with specific and standardized scaffold features.

Graphene-Based Scaffolds for Regenerative Medicine

“…Its presence in a poly(lactic-co-glycolic acid (PLGA) film increased its hydrophilicity and enhanced neuronal differentiation of neuronal stem cells (NSCs) [58]. In our lab we [47,59] designed composite poly-L-lactic acid (PLLA) scaffolds with different carbon Figure 3. Assembly of RGO-cellulose hybrid paper through deposition of GO followed by in situ reduction [55].…”

“…Its presence in a poly(lactic-co-glycolic acid (PLGA) film increased its hydrophilicity and enhanced neuronal differentiation of neuronal stem cells (NSCs) [ 58 ]. In our lab we [ 47 , 59 ] designed composite poly-L-lactic acid (PLLA) scaffolds with different carbon nanostructures (CNS) as filler—namely RGO, carbon nanohorns (CNH) and CNT—covalently functionalized with p-methoxyphenyl (PhOMe) groups in order to improve biocompatibility, and the electrical and mechanical properties of materials. RGO- and CNH-based scaffolds (RGO-PhOMe and CNH-PhOMe respectively) showed promising activity in enhancing the expression of myogenic markers during human circulating multipotent stem cell (hCMCs) differentiation.…”

“…These materials trigger reduced biological responses without the impairment of the GBMs capability to cross cell membranes and deliver therapeutic species [ 239 ]. As our lab has recently shown, organic-functionalized carbon nanofillers dispersed in a polymeric poly(L-lactic) acid exhibited enhanced cell viability (~90%) and supported cell growth [ 59 ], while having interest effects on the differentiation of neuronal precursors [ 50 ] and human circulating multipotent stem cells [ 47 , 159 ].…”

“… Examples of two-dimensional and three-dimensional scaffolds. ( a ) CVD graphene on Si/SiO 2 chip and AFM images of graphene transferred on different polymeric substrates [ 45 ]; ( b ) graphene transferred on nanopatterned substrate and AFM image [ 46 ]; ( c ) PLLA-RGO film obtained as reported in [ 47 ]; ( d ) graphite oxide paper [ 48 ]; ( e ) GO foams and SEM images [ 49 ]; ( f ) PLLA-RGO electrospun fibers obtained as reported in [ 50 ]; ( g ) peptide–GO hybrid hydrogels and TEM images [ 51 ]. …”

Advances on Graphene-Based Nanomaterials and Mesenchymal Stem Cell-Derived Exosomes Applied in Cutaneous Wound Healing

“… 111 Graphene could act as a biocompatible and conductive substrate for human induced pluripotent stem cells (hiPSCs) and mimic the biomimetic conductive cardiogenic niche to promote the self-renewal and cardiac differentiation of hiPSCs. 112 , 113 It is reported that highly conductive GBNs, such as the carbon nanotubes, could enhance the neuronal differentiation of multipotent autologous cells in most cases. On the contrary, several GBNs-conjugated composite scaffolds which are less conductive appeared to boost the expression of myogenic-lineage marker genes.…”