Transgenic silkworm expressing bioactive human ciliary neurotrophic factor for biomedical application.

Area of Science:

• Biomaterials Science
• Neuroscience
• Biotechnology

Background:

• Ciliary neurotrophic factor (CNTF) is a neuroprotective agent crucial for neuronal survival and regeneration.
• CNTF also induces stem cell differentiation into neurons, indicating broad biomedical potential.
• Challenges exist in the large-scale production of bioactive recombinant human CNTF.

Purpose of the Study:

• To develop a large-scale production method for bioactive recombinant human CNTF.
• To utilize a genetically engineered silkworm silk gland bioreactor for CNTF expression.
• To evaluate the bioactivity of CNTF-functionalized silk materials for neural applications.

Main Methods:

• Genetic engineering of silkworms to express human CNTF in the middle silk gland (MSG).
• Extraction and quantification of CNTF secreted into silk.
• Fabrication of CNTF-functionalized silk materials.
• In vitro assessment of neural cell proliferation, migration, and neurite outgrowth using RGC-5 cells.

Main Results:

• Successful expression and secretion of bioactive human CNTF in silkworm silk at 3.2 mg/g cocoons.
• CNTF-functionalized silk significantly enhanced neural cell proliferation and migration compared to natural silk.
• The functional silk material promoted neurite outgrowth in mouse retinal ganglion cells (RGC-5).

Conclusions:

• Silkworm silk glands serve as an effective bioreactor for large-scale production of bioactive recombinant human CNTF.
• CNTF-functionalized silk materials exhibit significant bioactivity for neural tissue engineering and regeneration.
• This approach offers a promising platform for developing advanced biomaterials for neuroregeneration.