The Effect of Blood-Derived Products on the Chondrogenic and Osteogenic Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells Originated from Three Different Locations

Abstract

Background. Adipose-derived mesenchymal stem cells (AD-MSCs) from fat tissue considered “surgical waste” during joint surgery may provide a potent source for regenerative medicine. Intra-articular, homologous fat tissue (Hoffa’s fat pad, pouch fat) might possess a superior chondrogenic and osteogenic differentiation potential in comparison to extra-articular, nonhomologous fat. Blood products might further enhance this potential. Methods. AD-MSCs were isolated from fat tissue of 3 donors from 3 locations each, during total knee replacement. Isolated cells were analyzed via flow cytometry. Cells were supplemented with blood products: two types of platelet-rich plasma (EPRP—PRP prepared in the presence of EDTA; CPRP—PRP prepared in the presence of citrate), hyperacute serum (hypACT), and standard fetal calf serum (FCS) as a positive control. The viability of the cells was determined by XTT assay, and the progress of differentiation was tested via histological staining and monitoring of specific gene expression. Results. Blood products enhance ex vivo cell metabolism. Chondrogenesis is enhanced by EDTA-PRP and osteogenesis by citrate PRP, whereas hyperacute serum enhances both differentiations comparably. This finding was consistent in histological analysis as well as in gene expression. Lower blood product concentrations and shorter differentiation periods lead to superior histological results for chondrogenesis. Both PRP types had a different biological effect depending upon concentration, whereas hyperacute serum seemed to have a more consistent effect, independent of the used concentration. Conclusion. (i) Blood product preparation method, (ii) type of anticoagulant, (iii) differentiation time, and (iv) blood product concentration have a significant influence on stem cell viability and the differentiation potential, favouring no use of anticoagulation, shorter differentiation time, and lower blood product concentrations. Cell-free blood products like hyperacute serum may be considered as an alternative supplementation in regenerative medicine, especially for stem cell therapies.

Optimization of Lyophilized Hyperacute Serum (HAS) as a Regenerative Therapeutic in Osteoarthritis

“…HAS is being tested as a possible therapeutic of OA with the aim of preventing joint degeneration. Previous studies showed that freshly isolated HAS more significantly induced cell proliferation of mesenchymal stromal cells, chondrocytes and in bone explants than PRP supplementation [ 15 , 16 , 17 , 18 ]. Moreover, Vacz et al demonstrated that in ischemia conditions, freshly isolated HAS significantly increased cell proliferation within bone fragments when compared to PRP [ 14 ].…”

“…In contrast, hyperacute serum (HAS) is a cell and fibrin-free solution, obtained after pressing out fibrin clots, which maintains the highly regenerative capacities of PRP. Previous studies of HAS used the freshly isolated formulation, a liquid version, which proved to induce cell proliferation of chondrocytes, bone marrow mesenchymal stem cells, osteoblasts in cell culture and in a bone ischemic model [ 14 , 15 , 16 , 17 , 18 ]. Kardos et al established a more complex biological model consisting of a human explant co-culture of bone, articular cartilage and synovium, which was used to study the effect of HAS under inflammatory conditions showing that HAS reduced inflammation as well as induced cell proliferation [ 19 ].…”

Platelet-Rich Plasma Does Not Inhibit Inflammation or Promote Regeneration in Human Osteoarthritic Chondrocytes In Vitro Despite Increased Proliferation

“…The general opinion is that PRP and PRP derivatives like platelet lysate stimulate proliferation of a variety of cell types. Many studies looked into the effects of PRP on MSCs derived from various sources, [35][36][37][38] where all studies report on increased proliferation when PRP was compared with conventional culture medium. Likewise, studies describing cell types derived from other tissues in the knee also report on increased proliferation.…”

Regenerative Potential of Blood-Derived Products in 3D Osteoarthritic Chondrocyte Culture System

“…Addition of hypACT to adipose-derived stem cells cultured in 3D pellets together with chondrogenic differentiation medium resulted in obtaining pellets with good tissue quality and enhanced expression of cartilage-related genes. However, our previous studies evaluating hypACT and PRP influence on OA chondrocytes behaviour were limited to monolayer culture experiments which may not adequately mimic the in vivo conditions [11][12][13].…”