Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo

Abstract

Frutapin is a lectin with affinity for glucose/mannose found in the seeds of Artocarpus altilis (breadfruit) and shares structural similarity with artocarpin, which is known for its immunostimulatory and wound-healing properties. Given its low natural extraction yield, the expression of its recombinant form in Escherichia coli was standardized, thereby increasing its production and enabling studies on its pharmacological potential. This study aimed to investigate the recombinant form of frutapin (rFTP) for its wound-healing potential in vitro and in vivo by evaluating the viability of fibroblasts (L929), keratinocytes (HaCaT), and macrophages (RAW 264.7) using the resazurin assay, as well as nitric oxide production (Griess assay) and cytokines TNF-α, IL-1β, IL-6, and IL-10 (ELISA) in RAW 264.7 cell supernatants. Subsequently, rFTP (0.1%, 0.5%, and 1%) was incorporated into xyloglucan-based biomembranes and tested in excisional wounds in BALB/c mice (n = 6 animals/group), compared with untreated controls (sham) and biomembrane without the active compound. The animals were euthanized on days 3, 7, and 14 post-surgery. rFTP (0.15–1 mg/mL) increased the proliferation of L929 and HaCaT cells and modulated the inflammatory response in RAW 264.7 macrophages compared to the CTRL group, significantly enhancing proliferation and IL-10 production while reducing the levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β, both in the presence and absence of LPS. The xyloglucan biomembrane containing 0.1% rFTP accelerated wound contraction and reduced inflammatory cell infiltration on days 3 and 7. Moreover, epithelial thickness was significantly higher in the rFTP-treated groups (0.1%) compared to the sham and control groups, highlighting rFTP’s ability to stimulate tissue regeneration and extracellular matrix remodeling. Levels of IL-1β, TNF-α, and IL-6 were reduced, while IL-10 was considerably increased, suggesting that rFTP modulates inflammation. These results indicate that rFTP is a promising candidate for wound management by promoting inflammatory control and regenerative stimulation. Thus, this work encourages further studies to elucidate the specific molecular mechanisms of rFTP and to assess its efficacy and safety in clinical trials.