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Rotary cell culture systems enhance engineered nerve construction by promoting adipose-derived mesenchymal stem cell proliferation and neural differentiation, leading to improved peripheral nerve repair.

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Area of Science:

  • Biomaterials Science
  • Regenerative Medicine
  • Stem Cell Biology

Background:

  • Tissue-engineered nerves (TENs) using mesenchymal stem cells (MSCs) often suffer from poor cell proliferation and migration.
  • This limits the microstructure of TENs and hinders effective nerve defect repair.
  • Rotary cell culture systems (RCCSs) show promise for improving MSC-based TENs by enhancing differentiation and transport.

Purpose of the Study:

  • To construct TENs using adipose-derived MSCs (ADSCs) and acellular nerve grafts (ANG) within an RCCS.
  • To compare the efficacy of RCCS-based TENs with those prepared using a static 3D culture method.
  • To investigate the mechanisms underlying RCCS-driven improvements in ADSC behavior and neural differentiation.

Main Methods:

  • Construction of TENs using ADSCs and ANG in both RCCS and static 3D culture.
  • Assessment of ADSC proliferation using MTT assays and cell counts.
  • Analysis of cell cycle, apoptosis, gene expression (qPCR), and protein levels (ELISA).
  • Evaluation of neural differentiation via Notch1 signaling pathway inhibition (DAPT).
  • In vivo assessment of TENs in a sciatic nerve defect model using TEM, axon immunostaining, and retrograde labeling.

Main Results:

  • RCCS significantly increased ADSC proliferation compared to static 3D culture, attributed to enhanced mass/nutrient transport, not microgravity.
  • RCCS promoted ADSC neural differentiation, evidenced by increased expression of relevant genes, which was mediated by the Notch1 signaling pathway.
  • In vivo studies demonstrated superior regenerative capacity of RCCS-prepared TENs in peripheral nerve repair.

Conclusions:

  • RCCS is a viable method for constructing improved MSC-based TENs.
  • The system enhances ADSC proliferation and neural differentiation through improved transport and Notch1 signaling modulation.
  • RCCS-generated TENs show greater potential for peripheral nerve regeneration compared to static methods.