P040 - Influence of Type I Collagen-Based Meniscus Scaffold and PRP on Adipose-Derived Stem Cells Differentiation Under Static Culture Condition
- D. Grande (Manhasset, US)
- D. Grande (Manhasset, US)
- W. Chang (Franklin Lakes, US)
- H. Andriamifidy (Manhasset, US)
- J. Bondi (Manhasset, US)
- S. Li (Franklin Lakes, US)
Abstract
Purpose
The overall goal is to develop a type I collagen-based meniscus regeneration implant applying tissue engineering strategy. In the current study, platelet-rich plasma (PRP) was used to induce chondrogenic differentiation of human adipose-derived mesenchymal stem cells (hASC) in a novel type I collagen-based scaffold.
Methods and Materials
Briefly, 0.7% of purified type I collagen fibers in 0.07M lactic acid were swollen and homogenized. Aliquots of dispersion were adjusted to pH4.8 to reconstitute fibers. Fibers were oriented, molded, lyophilized, crosslinked, rinsed, and sterilized. High (HD) and low (LD) density scaffolds were produced.
Effects of human PRP (RegenLab) on hASCs (ATCC) proliferation and differentiation into chondrogenic cells were investigated. 2 × 106 hASCs were seeded onto scaffolds (n=5), i.e., LD, HD, LD+PRP and HD+PRP. 1% PRP was added to the media. Cell viability (Alamar Blue assay) was measured. Differentiation (primers: Collagen I and Aggrecan) was performed with quantitative real-time polymerase chain reaction (qRT-PCR). Histology was used to evaluate cell distribution and ECM synthesis.
Data were analyzed using Analysis of Variance (ANOVA). Statistical significance was set to value ≤ 0.05.
Results
Fig.1A shows that the cell growth kinetics were similar. Fig. 1B shows PRP and scaffold density had a positive effect on type I collagen gene expression, where the highest effect was observed at day 21. Fig. 1C shows the effect of PRP on gene expression of aggrecan. The amount of aggrecan expression was significantly elevated in the presence of PRP by day 21, a sign of chondrogenic differentiation of hASCs.
Fig. 2 shows a cellular migration within the interior of the scaffolds in all groups at day 21.
Conclusion
This study shows that scaffold properties and PRP can play an important role in directing hASC towards chondrogenic differentiation in vitro. The effect of biomechanical stress produced by dynamic loading on the chondrogenic differentiation of hADSC is currently being investigated.