Histologic Evaluation of the Effects of Folinic Acid Chitosan Hydrogel and Botulinum Toxin A on Wound Repair of Cleft Lip Surgery in Rats

Abstract

The aim of the present study was to compare the effects of folinic acid chitosan hydrogel and botulinum toxin A on the wound repair of cleft lip surgery in rat animal models. Cleft lip defects were simulated by triangular incisions in the upper lip of 40 Wistar rats. Then, the rats were randomly assigned to four groups: control (CTRL), chitosan hydrogel (CHIT), and folinic acid chitosan hydrogel (FOLCHIT), in which the wounds were covered by a gauze pad soaked in normal saline, chitosan hydrogel, and folinic acid chitosan hydrogel, respectively for 5 min immediately after closure; and botulinum toxin A (BOT) with the injection of 3 units of botulinum toxin A in the wound region. Fibroblast proliferation, collagen deposition, inflammatory cell infiltration, neovascularization, and epithelial proliferation and each parameter were rated on days 14 and 28. Statistical analysis was performed by Kolmogorov-Smirnov test, Shapiro-Wilk test, Kruskal-Wallis, and post-hoc tests (α = 0.05). The mean score for fibroblast proliferation was significantly higher in the FOLCHIT group compared with the BOT group at days 14 and 28 (p < 0.001, p = 0.012, respectively). At day 28, collagen deposition was significantly higher in the FOLCHIT group compared with the BOT group (p = 0.012). No significant difference was observed between the inflammatory infiltration of the study groups at the two time points (p = 0.096 and p = 1.000, respectively). At day 14, vascular proliferation of group FOLCHIT was significantly higher than groups CTRL and CHIT (p = 0.001 and p = 0.006, respectively). The epithelial proliferation in the FOLCHIT group was significantly higher than groups CHIT and CTRL at day 14 (p = 0.006 and p = 0.001, respectively) and day 28 (p = 0.012). In simulated lip cleft defects, topical application of folinic acid induces faster initial regeneration by higher inflammation and cellular proliferation, at the expense of a higher tendency for scar formation by slightly higher fibroblast proliferation and collagen deposition. While injection of botulinum toxin A provides less fibroblast proliferation and collagen deposition, and thus lower potential for scar formation compared with the folinic acid group. Therefore, in wounds of the esthetic zone, such as cleft lip defects, the application of botulinum toxin A shows promising results.

In‐vivo assessment of application of folinic acid and botulinum toxin A in cleft lip surgical defects

“…Botolinum toxin A also showed positive effects on neo‐vascularization and epithelial healing. The increase in neo‐vascularization by botulinum toxin A was not observed in the previous study by Namdar et al 22 . The reason can be attributed to different dosages of the administered botulinum toxin A in the two studies.…”

“…In a previous study, folinic acid loaded in chitosan hydrogel led to relatively similar findings with the present study in terms of inflammatory response, fibroblast proliferation, epithelial healing and collagen deposition. However, in that study folinic acid chitosan hydrogel did not show similar angiogenic properties with botulinum toxin 22 . Beneficial effects of folic acid on the skin fibroblasts and collagen neosynthesis has been previously reported by promoting DNA repair in dermal fibroblasts 34 .…”

“…However, in that study folinic acid chitosan hydrogel did not show similar angiogenic properties with botulinum toxin. 22 Beneficial effects of folic acid on the skin fibroblasts and collagen neosynthesis has been previously reported by promoting DNA repair in dermal fibroblasts. 34 The effects of a topical folic acid and creatine formulation on skin firmness, collagen genes expression, procollagen synthesis, collagen fibril organization, and collagen density has been evaluated in the literature.…”

“…Folinic acid is a 5‐formyl derivative of tetrahydrofolic acid and is one of the forms of folate naturally found in foods, as opposed to folic acid (a synthetic form of folate), which can be converted to all other forms of folate and has the full properties of folic acid in the body 21 . Nonetheless, the role of folinic acid in skin wound healing needs further investigation 22 . Since postoperative scars play a critical role in the psychological and social effects of cleft surgery in childhood, this study aimed to compare the effects of folinic acid and botulinum toxin A on wound healing after cleft lip surgery.…”

“…21 Nonetheless, the role of folinic acid in skin wound healing needs further investigation. 22 Since postoperative scars play a critical role in the psychological and social effects of cleft surgery in childhood, this study aimed to compare the effects of folinic acid and botulinum toxin A on wound healing after cleft lip surgery.…”