Journal of Taibah University Medical Sciences · 2026
Development and characterization of an injectable dental follicle stem cell-secretome/nano-hydroxyapatite chitosan-alginate hydrogel: A cytocompatible scaffold for periodontal regeneration.
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Al-Wesabi Mohammed A, Ridwan Rini D, Al-Sanaani Saba A, Al Junaid Mohammed A, Qaid Huda R, Heah Khor G, Hamrun Norlina, Syam Syamsiah, Nugraha Alexander P
Аннотация
OBJECTIVE: To develop and characterize a novel injectable hydrogel containing third molar dental follicle stem cell (DFSC)-derived secretome and nano-hydroxyapatite (nHA) with low molecular weight chitosan to create a bioactive, osteoconductive, and immunomodulatory scaffold for periodontal regeneration.
METHODS: A chitosan/sodium alginate matrix was synthesized via solvent evaporation to prepare three injectable hydrogel formulations: secretome-loaded hydrogel (TG1), nHA-reinforced hydrogel (TG2), and a composite hydrogel incorporating both active agents (TG3). Following sterilization, all formulations were assessed to determine their organoleptic properties, injectability, gelation time, and pH. The morphology and porosity of the samples were characterized using scanning electron microscopy (SEM), mineral content by energy-dispersive X-ray spectroscopy (EDS), and osteoblast cytocompatibility with the MTT assay.
RESULTS: All hydrogels were extruded from fine-gauge needles and maintained a uniform flow rate of up to 30G. The gelation time at 37 °C was approximately 8.5 ± 2.5 min for each hydrogel group. The hydrogels had highly porous and interconnected three-dimensional structures with irregular and varied pore distributions throughout the matrices. Porosity analysis performed by Otsu threshold segmentation of SEM images for the composite formulation (TG3) indicated an overall porosity of approximately 58.3%. EDS detected the presence of calcium and phosphorus (nHA) within the scaffold. The MTT results showed that the cell viability exceeded 90% for the composite formulation (secretome + nHA) and over 75% for all single-agent formulations, with no significant cytotoxicity relative to the control (p < 0.05, one-way analysis of variance with Tukey's HSD).
CONCLUSION: The injectable chitosan/alginate hydrogel loaded with DFSC-derived secretome and nHA exhibited favorable injectability, rapid gelation, high porosity, an interconnected microarchitecture, and excellent cytocompatibility. The successful incorporation of nHA was confirmed by EDS analysis, suggesting that the developed bioactive scaffold has potential uses in periodontal tissue engineering and regenerative applications.
METHODS: A chitosan/sodium alginate matrix was synthesized via solvent evaporation to prepare three injectable hydrogel formulations: secretome-loaded hydrogel (TG1), nHA-reinforced hydrogel (TG2), and a composite hydrogel incorporating both active agents (TG3). Following sterilization, all formulations were assessed to determine their organoleptic properties, injectability, gelation time, and pH. The morphology and porosity of the samples were characterized using scanning electron microscopy (SEM), mineral content by energy-dispersive X-ray spectroscopy (EDS), and osteoblast cytocompatibility with the MTT assay.
RESULTS: All hydrogels were extruded from fine-gauge needles and maintained a uniform flow rate of up to 30G. The gelation time at 37 °C was approximately 8.5 ± 2.5 min for each hydrogel group. The hydrogels had highly porous and interconnected three-dimensional structures with irregular and varied pore distributions throughout the matrices. Porosity analysis performed by Otsu threshold segmentation of SEM images for the composite formulation (TG3) indicated an overall porosity of approximately 58.3%. EDS detected the presence of calcium and phosphorus (nHA) within the scaffold. The MTT results showed that the cell viability exceeded 90% for the composite formulation (secretome + nHA) and over 75% for all single-agent formulations, with no significant cytotoxicity relative to the control (p < 0.05, one-way analysis of variance with Tukey's HSD).
CONCLUSION: The injectable chitosan/alginate hydrogel loaded with DFSC-derived secretome and nHA exhibited favorable injectability, rapid gelation, high porosity, an interconnected microarchitecture, and excellent cytocompatibility. The successful incorporation of nHA was confirmed by EDS analysis, suggesting that the developed bioactive scaffold has potential uses in periodontal tissue engineering and regenerative applications.