Functionalized Electrospun Scaffold–Human-Muscle-Derived Stem Cell Construct Promotes In Vivo Neocartilage Formation

Abstract

Polycaprolactone (PCL) is a non-cytotoxic, completely biodegradable biomaterial, ideal for cartilage tissue engineering. Despite drawbacks such as low hydrophilicity and lack of functional groups necessary for incorporating growth factors, it provides a proper environment for different cells, including stem cells. In our study, we aimed to improve properties of scaffolds for better cell adherence and cartilage regeneration. Thus, electrospun PCL–scaffolds were functionalized with ozone and loaded with TGF-β3. Together, human-muscle-derived stem cells (hMDSCs) were isolated and assessed for their phenotype and potential to differentiate into specific lineages. Then, hMDSCs were seeded on ozonated (O) and non-ozonated (“naïve” (NO)) scaffolds with or without protein and submitted for in vitro and in vivo experiments. In vitro studies showed that hMDSC and control cells (human chondrocyte) could be tracked for at least 14 days. We observed better proliferation of hMDSCs in O scaffolds compared to NO scaffolds from day 7 to day 28. Protein analysis revealed slightly higher expression of type II collagen (Coll2) on O scaffolds compared to NO on days 21 and 28. We detected more pronounced formation of glycosaminoglycans in the O scaffolds containing TGF-β3 and hMDSC compared to NO and scaffolds without TGF-β3 in in vivo animal experiments. Coll2-positive extracellular matrix was observed within O and NO scaffolds containing TGF-β3 and hMDSC for up to 8 weeks after implantation. These findings suggest that ozone-treated, TGF-β3-loaded scaffold with hMDSC is a promising tool in neocartilage formation.

Cartilage regeneration using improved surface electrospun bilayer polycaprolactone scaffolds loaded with transforming growth factor-beta 3 and rabbit muscle-derived stem cells

“…This material and technique already examined in different studies created a proper environment for our cells-rabbit MDSCs and chondrocytes. We did show its potential in the previous experimental study as well (Jankauskaite et al, 2022). The proliferation capacity was increased within the first 14 days.…”

“…The subchondral layer polymer solution was prepared similarly to the chondral layer, except that hydroxyapatite (HA, <15 µm particle size, CAS: 1306-06-5) powder was added. The fabrication procedure of both layers and electrospinning and ozonation techniques (treatment with O 3 ) were described previously (Dabasinskaite et al, 2021;Dabasinskaite et al, 2022). PCL pellets and CA powder (2:1, w/w) were dissolved in acetone and N.N-dimethylformamide mixture at 2:3 (v/v) to obtain 30% (w/v) polymer solution.…”

“…In brief, scaffolds were incubated with 10 ng/ml TGFβ3. After the most efficient binding duration was clarified, samples for in vitro and in vivo experiments were covered with 100 μl of 10 ng/ml TGFβ3 for 24 h, providing sufficient time for protein binding to unmodified and modified scaffolds (Dabasinskaite et al, 2022). Subsequently, the unbound protein was washed with PBS, and scaffolds were submitted for further experiments.…”

Autologous Stem Cells Transplants in the Treatment of Temporomandibular Joints Disorders: A Systematic Review and Meta-Analysis of Clinical Trials

“…Due to the properties of adipose-derived stem cells, such as immunomodulatory and anti-inflammatory activity, autologous transfer is promising [ 19 , 20 , 21 ]. Targeting MSCs differentiation at the formation of cartilage tissue is currently the subject of experimental research [ 22 , 23 ]. Favorable conditions can stimulate the differentiation of MSCs towards chondroblasts which is promising for the treatment of cartilage degradation [ 22 , 23 , 24 ].…”

“…Targeting MSCs differentiation at the formation of cartilage tissue is currently the subject of experimental research [ 22 , 23 ]. Favorable conditions can stimulate the differentiation of MSCs towards chondroblasts which is promising for the treatment of cartilage degradation [ 22 , 23 , 24 ].…”