Hybrid complexes of high and low molecular weight hyaluronan delay in vitro replicative senescence of mesenchymal stromal cells: a pilot study for future therapeutic application

Abstract

Mesenchymal stem cells, a subpopulation of mesenchymal stromal cells (MSCs), are present in the stroma of several tissues. MSC in vitro cultivation for clinical treatments may greatly affect MSC properties. A primary handicap is replicative senescence that impairs MSC functions. Hyaluronan (HA) is present in the extracellular matrix that composes the stem cell niche environment and is under investigation as a key factor for in vitro stem cell growth. We evaluated the effect on MSC cultivation of HA hybrid cooperative complexes (HCC) that are obtained from high (H) and low (L) weight molecules (NAHYCO™). We compared this HCC with H-HA and L-HA. We investigated the effects of these HAs on proliferation, cell cycle, apoptosis, senescence, and differentiation following the addition of the polymer solutions in the culture media at concentrations that did not drastically modify the medium viscosity. Interestingly, 0,16% HCC significantly delayed the senescence compared with the controls. This occurred without alteration of the cell cycle, cytotoxicity, or apoptosis. HCCs also promoted adipogenic and chondrogenic differentiation. Our finding could suggest a potential functional role of HCC above the updated scientific reports of its effects and pave the way to optimization of MSC cultivation for therapeutic application.

Hyaluronan Induces a Mitochondrial Functional Switch in Fast-Proliferating Human Mesenchymal Stem

“…These results suggest that hyaluronan-coated surfaces induce PMDSCs into a resting/quiescent state that preserved their stemness properties. Other recent reports have confirmed the ability of hyaluronan to promote adipogenic and chondrogenic differentiation and induce senescence delay (4). Previous studies have demonstrated a crucial function of hyaluronan on stem cell metabolism (5,6).…”

Hyaluronic Acid (HA), Platelet-Rich Plasm and Extracorporeal Shock Wave Therapy (ESWT) promote human chondrocyte regeneration in vitro and ESWT-mediated increase of CD44 expression enhances their susceptibility to HA treatment

“…Interestingly, the possible correlation between inflammation, hyaluronan fragmentation, CD44 expression on chondrocytes, cartilage disruption and protective homeostatic tissue remodeling during OA[18, 41, 44–46] have already been reported although with contradictory results. In accordance with previous works[19, 20, 24], which revealed the synergistic properties of both low and high ranges of MW for in vitro treatment on human cultured chondrocytes, we used an innovative formulation of HA, consisting in a hybrid complex of combined low (LMW) and high molecular weight (HMW)-HA[24].…”

“…For HA experiments, we used hyaluronic acid hybrid complex commercialized by IBSA Farmaceutici (Italy), (ratio 1:1) with H-HA (MW: 1200 ±100 kDa) and L-HA (MW: 100 ± 10 kDa) prepared as reported[24]. HA stock solution was mixed with culture media and the final concentration of hyaluronan mixture was 0.5%.…”

“…Furthermore, one of the major limitations in healing the damaged cartilage is represented by cellular senescence associated with OA and characterized by an advanced age (or trauma-dependent) decline of tissue regeneration capacity[1, 2, 4, 7, 24, 25]. This chondrosenescence is intimately linked to several factors and may be measured in primary cultured human chondrocytes by different biomarkers of cell senescence, such as the p16 INK4a protein expression (also known as Cdkn2a)[26–28].…”

An in vitro study to assess the effect of hyaluronan-based gels on muscle-derived cells: Highlighting a new perspective in regenerative medicine

“…Besides, the biological properties of HCC have already been tested on several cell cultures used as representative models for tissue regeneration [ 22 – 24 ]. Recently, HCC has been proved to be effective on cell co-culture models subjected to stressful conditions to mimic injuries [ 25 ]. Furthermore, it is known that HA is the subject of the ROS attack, which leads to changes in its structure modulating oxidative condition [ 26 – 28 ].…”