Abstract
Unlike humans, salamanders regrow their amputated limbs. Regeneration depends on the presence of regenerating axons which upregulate the expression of newt anterior gradient (nAG) protein. We had the hypothesis that nAG might have an inhibitory effect on collagen production since excessive collagen production results in scarring, which is a major enemy to regeneration. nAG gene was designed, synthesized, and cloned. The cloned vector was then transfected into primary human fibroblasts. The results showed that the expression of nAG protein in primary human fibroblast cells suppresses the expression of collagen I and III, with or without TGF-β1 stimulation. This suppression is due to a dual effect of nAG both by decreasing collagen synthesis and by increasing collagen degradation. Furthermore, nAG had an inhibitory effect on proliferation of transfected fibroblasts. It was concluded that nAG suppresses collagen through multiple effects.
Highlights
Tissue regeneration and wound healing are two different processes [1]
We show that newt anterior gradient (nAG) had an inhibitory effect on proliferation of transfected fibroblasts
Matrix metalloproteinase (MMPs) proteins with gelatinolytic activity were determined in conditioned media deprived from serum for 24 hours for normal fibroblasts and fibroblasts transfected with nAG plasmid and treated with 3 ng/mL TGF-β1. 20 uL of undiluted cell culture supernatant was mixed with an equal volume of nonreducing SDS sample buffer, using MMP-2 (R&D Systems, Minneapolis, USA) as positive control
Summary
Tissue regeneration and wound healing are two different processes [1]. Wound healing is a process of tissue repair mainly by fibrous tissue (collagen) formation [6]. Healing by fibrous tissue is essential for “normal” tissue repair, excessive fibrosis is a harmful pathological process. The amputation stump of the salamander forms a blastema which is a mound of proliferating mesenchymal cells surrounded by wound epithelium. NAG protein expression by schwann cells of regenerating axons peaks at BioMed Research International. At 10–12 days, the protein is expressed in glands in the dermis underlying the wound epithelium [12,13,14,15]. We had the hypothesis that nAG might have an inhibitory effect of collagen production since excessive collagen production results in scarring, which is a major enemy to regeneration. We show that nAG had an inhibitory effect on proliferation of transfected fibroblasts
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