Biphasic Biological Response of Ocimum basilicum Loaded Hydrogels with Dose dependent Teratogenic Effects

Abstract

Plant-derived bioactive hydrogels have attracted growing attention in regenerative medicine due to their biocompatibility and multifunctional properties. However, limited studies have explored the integration of Ocimum basilicum extract into crosslinked hydrogels and its effects on angiogenesis and embryonic development. This study aimed to fabricate GPTMS-crosslinked chitosan–polyvinyl alcohol (CP) hydrogels incorporated with different concentrations of O. basilicum extract and to evaluate their physicochemical behavior, angiogenic potential, and embryotoxicological impact using the chick CAM model. Hydrogels were prepared with extract concentrations of 0, 300, and 600 µL (CP-0, CP-3, and CP-6). Swelling and biodegradation properties were assessed under in vitro conditions. In vivo evaluation included angiogenesis quantification using the chick CAM assay, morphological and morphometric analysis of embryos, amniotic fluid biochemical profiling, and histological examination of the liver. The CP-6 hydrogel exhibited the highest swelling and faster degradation, whereas CP-3 showed stable physicochemical behavior. CAM analysis revealed that CP-3 significantly enhanced vessel formation, while CP-6 suppressed angiogenesis. Morphometric and morphological evaluations confirmed normal development in control, CP-0, and CP-3 groups, but CP-6 embryos displayed teratogenic defects, including malformed limbs, abnormal curvature, and abdominal wall defects. Amniotic fluid biomarkers indicated reduced AST (31.8 U/L) and ALT (28.5 U/L) levels in the CP-3 group, demonstrating hepatoprotective effects, whereas the CP-6 group exhibited elevated AST (48.6 U/L) and ALT (42.3 U/L) values, confirming dose-dependent hepatic and renal toxicity.. Histological findings further corroborated these outcomes, with CP-3 preserving hepatic structure and CP-6 causing vacuolation and tissue disorganization. O. basilicum–loaded hydrogels demonstrated a biphasic, dose-dependent response, with CP-3 showing pro-angiogenic and biocompatible potential, while CP-6 induced systemic toxicity. These findings underscore the need for dose optimization to harness herbal extract–based hydrogels for wound healing, bone regeneration, and tissue engineering applications.uminants, offering actionable targets for precision breeding in tropical agricultural economics.