Parathyroid hormone (1-34) promotes the effects of 3D printed scaffold-seeded bone marrow mesenchymal stem cells on meniscus regeneration

Abstract

Background Cell-based tissue engineering represents a promising management for meniscus repair and regeneration. The present study aimed to investigate whether the injection of parathyroid hormone (PTH) (1-34) could promote the regeneration and chondroprotection of 3D printed scaffold seeded with bone marrow mesenchymal stem cells (BMSCs) in a canine total meniscal meniscectomy model. Methods 3D printed poly(e-caprolactone) scaffold seeded with BMSCs was cultured in vitro, and the effects of in vitro culture time on cell growth and matrix synthesis of the BMSCs–scaffold construct were evaluated by microscopic observation and cartilage matrix content detection at 7, 14, 21, and 28 days. After that, the tissue-engineered meniscus based on BMSCs–scaffold cultured for the appropriate culture time was selected for in vivo implantation. Sixteen dogs were randomly divided into four groups: PTH + BMSCs–scaffold, BMSCs–scaffold, total meniscectomy, and sham operation. The regeneration of the implanted tissue and the degeneration of articular cartilage were assessed by gross, histological, and immunohistochemical analysis at 12 weeks postoperatively. Results In vitro study showed that the glycosaminoglycan (GAG)/DNA ratio and the expression of collagen type II (Col2) were significantly higher on day 21 as compared to the other time points. In vivo study showed that, compared with the BMSCs–scaffold group, the PTH + BMSCs–scaffold group showed better regeneration of the implanted tissue and greater similarity to native meniscus concerning gross appearance, cell composition, and cartilage extracellular matrix deposition. This group also showed less expression of terminal differentiation markers of BMSC chondrogenesis as well as lower cartilage degeneration with less damage on the knee cartilage surface, higher expression of Col2, and lower expression of degeneration markers. Conclusions Our results demonstrated that PTH (1-34) promotes the regenerative and chondroprotective effects of the BMSCs–3D printed meniscal scaffold in a canine model, and thus, their combination could be a promising strategy for meniscus tissue engineering.

The Roles of Different Multigene Combinations of Pdx1, Ngn3, Sox9, Pax4, and Nkx2.2 in the Reprogramming of Canine ADSCs Into IPCs

“…A total of 50 μg protein was separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) as previously described 24 . Primary antibodies against the targeted proteins included an anti-beta-actin antibody (ab8227, 1:2,000; Abcam, Cambridge, UK), anti-PDX1 antibody (ab227586, 1:1,000; Abcam), anti-NGN3 antibody (ab176124, 1:1,000; Abcam), anti-NKX2.2 antibody (ab86024, 1:1,000; Abcam), anti-PAX4 antibody (ab101721, 1:1,000; Abcam), and anti-SOX9 antibody (ab26414, 1:1,000; Abcam).…”

Mesenchymal Stem Cells From Different Sources in Meniscus Repair and Regeneration

“…As early as in 2005, implantation of rat BMSCs together with acellular meniscus scaffolds was reported, showing comparable results to native meniscus chondrocytes and a better morphology than a deep-frozen allogeneic cell-free implant ( Yamasaki et al, 2005 ). Thereafter, animal studies were conducted to implant BMSCs on artificial synthetic or modified natural scaffolds, and different kinds of cytokines or small molecule compounds added in trials performed well; mechanical stimulation also revealed good experimental results ( Yuan et al, 2017 ; Stuckensen et al, 2018 ; Liu et al, 2019 ; Zhao et al, 2020 ).…”

Parathyroid Hormone 1 Receptor Signaling in Dental Mesenchymal Stem Cells: Basic and Clinical Implications

“…A recent study using a canine total meniscal meniscectomy model suggested PTH (1–34) with a BMMSCs-loaded scaffold stimulated meniscus regeneration and alleviated cartilage damage. This implies that PTH could promote the regenerative and chondroprotective function of a BMMSCs scaffold ( Zhao et al, 2020 ). These data yield insight into the efficacy of combining PTH and stem cells in TMJ tissue engineering in the future.…”