WOUND HEALING EVALUATION OF NOVEL CISSUS QUADRANGULARIS, BIOCERAMICS AND TENDON EXTRACELLULAR MATRIX INCORPORATED SCAFFOLDS FOR PERIODONTAL BONE REGENERATION WITH ZEBRAFISH MODELS

Background: Wound healing is a complex biological process involving, proliferation, and tissue remodelling,
requiring advanced biomaterials to facilitate effective tissue regeneration. This study evaluated the wound healing
potential of composite scaffolds incorporating Cissus quadrangularis extract, carrageenan, tendon extracellular
matrix (ECM) with bioceramics such as silver hydroxyapatite and silver tricalcium phosphate using a zebrafish
model.
Materials and Methods: The scaffolds were extensively characterized using Scanning Electron Microscopy
(SEM), Fourier Transform Infrared Spectroscopy (FTIR), and X- ray diffraction (XRD) to determine their
physiochemical properties and their correlation with biological efficacy. The wild- type zebrafish were wounded
and treated as following negative control (Group 1), Cissus quadrangularis extract, carrageenan, tendon
extracellular matrix (TEM) (Group 2), Group 3 (contained silver hydroxyapatite + Group 2 components), Group 4
(Silver tricalcium phosphate + Group 2 components) respectively for 21 days. The wound closure ratio was
assessed on 0, 7 and 14 days with histological observations.
Results: As the results, SEM analysis revealed a highly porous architecture with interconnected pores, enhancing
cellular adhesion, proliferation, and migration at the wound site. The surface roughness provided an optimal
microenvironment for fibroblast infiltration and re-epithelization, accelerating the healing process. FTIR analysis
confirmed the presence of hydroxyl, carboxyl, amide, and phosphate groups indicating strong biomaterial
interactions, hydration, and bioactivity. The observed phosphate peaks validated the incorporation of carrageenan,
essential for calcium ion release and osteogenic differentiation. XRD patterns exhibited well- defined crystalline
peaks corresponding to hydroxyapatite and bioactive glass, ensuring stability, controlled degradation, and
prolonged bioactivity at the wound site. Zebrafish wound healing assays demonstrated significant improvements in
wound closure, reduced inflammation, and enhanced tissue remodelling in scaffold treated groups. The histology
observations revealed that the Group 4 treatment induced rapid epidermal formation, immune cell activity, and
vascularization. Faster wound contraction and new scale formation were observed 4 to 21 days post injury,
highlighting the scaffold’s effectiveness compared to the slower recovery in untreated controls. The combined
effects provided a synergistic approach for effective wound healing and tissue regeneration.
Conclusion: The study demonstrated that the application of silver tricalcium phosphate, C. quadrangularis, and
TEM effectively promoted wound healing. Future research should focus on optimizing scaffold composition and
validating efficacy in mammalian models for translational applications.