Enhanced cellular infiltration of human adipose-derived stem cells in allograft menisci using a needle-punch method

Abstract

BackgroundThe meniscus plays a crucial role in knee joint stability, load transmission, and stress distribution. Meniscal tears are the most common reported knee injuries, and the current standard treatment for meniscal deficiency is meniscal allograft transplantation. A major limitation of this approach is that meniscal allografts do not have the capacity to remodel and maintain tissue homeostasis due to a lack of cellular infiltration. The purpose of this study was to provide a new method for enhanced cellular infiltration in meniscal allografts.MethodsTwenty medial menisci were collected from cadaveric human sources and split into five experimental groups: (1) control native menisci, (2) decellularized menisci, (3) decellularized menisci seeded with human adipose-derived stem cells (hASC), (4) decellularized needle-punched menisci, and (5) decellularized needle-punched menisci seeded with hASC. All experimental allografts were decellularized using a combined method with trypsin EDTA and peracetic acid. Needle punching (1-mm spacing, 28 G microneedle) was utilized to improve porosity of the allograft. Samples were recellularized with hASC at a density of 250 k/g of tissue. After 28 days of in vitro culture, menisci were analyzed for mechanical, biochemical, and histological characteristics.ResultsMenisci maintained structural integrity and material properties (compressive equilibrium and dynamic moduli) throughout preparations. Increased DNA content was observed in the needle-punched menisci but not in the samples without needle punching. Histology confirmed these results, showing enhanced cellular infiltration in needle-punched samples.ConclusionsThe enhanced infiltration achieved in this study could help meniscal allografts better remodel post-surgery. The integration of autologous adipose-derived stem cells could improve long-term efficacy of meniscal transplantation procedures by helping to maintain the meniscus in vivo.

Tissue-Engineered Grafts from Human Decellularized Extracellular Matrices: A Systematic Review and Future Perspectives

“…Minehara et al [51] used incubations in acetone, hydrogen peroxide, and osmotic solutions. Nordberg et al [52] adopted 24 h enzymatic digestion in 0.05% trypsin EDTA and 48 h incubation in 2% Triton X-100 and 1.5% peracetic acid, apart from other steps in deionized water and neutralization medium. Homogenizing approaches have also been proposed [53].…”

“…Repopulation of decellularized menisci has been performed with rat chondrocytes [51], human adipose-derived stem cells [52], and human synovial fluid-derived mesenchymal stem cells [53]. Various authors have stressed the difficulties in the recellularization of menisci, due to the dense structure of the extracellular matrix with very few interstitial spaces [51,52].…”

“…Repopulation of decellularized menisci has been performed with rat chondrocytes [51], human adipose-derived stem cells [52], and human synovial fluid-derived mesenchymal stem cells [53]. Various authors have stressed the difficulties in the recellularization of menisci, due to the dense structure of the extracellular matrix with very few interstitial spaces [51,52]. Consistently, some authors proposed chemotactic cell seeding techniques (recombinant human bone morphogenetic protein-2) [51] or needle punching (1-mm spacing, 28G microneedle) [52] to facilitate cell invasion.…”

“…Various authors have stressed the difficulties in the recellularization of menisci, due to the dense structure of the extracellular matrix with very few interstitial spaces [51,52]. Consistently, some authors proposed chemotactic cell seeding techniques (recombinant human bone morphogenetic protein-2) [51] or needle punching (1-mm spacing, 28G microneedle) [52] to facilitate cell invasion. Exogenous growth factor supplementation (TGF-β 3 and IGF-1) was required to chondrogenic differentiation of synovial fluid-derived mesenchymal stem cells [53].…”

Patient-specific meniscus prototype based on 3D bioprinting of human cell-laden scaffold

“…5,6 However, there are multiple drawbacks to meniscal allograft transplantation, such as limits in tissue availability, the risk of mismatch and immunoreactivity, impaired cellular infiltration and remodelling capacity, and suboptimal prognosis with results worsening over time. 7,8 To overcome the limitations of allografts, several scaffolds have been developed, 9 but, despite promising preclinical findings, only two cell-free meniscal substitutes have reached clinical practice and been used to address partial meniscus defects: one collagen-based meniscal implant and one consisting of polyurethane and polycaprolactone. 10 Safety and positive clinical results have been reported for both scaffolds, with imaging and, in some cases, arthroscopic evaluation confirming the potential of these bio-engineered devices to stimulate meniscal tissue regeneration, albeit not completely.…”