فهرست نشریات علمی دسترسی آزاد ایران

Objective: Collagen plays a critical role in tissue engineering and regenerative medicine due to its biocompatibilityand structural features. Identifying an optimal extraction method that preserves both functional and structural integrityremains challenging. This study aimed to compare eleven collagen extraction protocols to determine the most effectivemethod for producing high-quality collagen.Materials and Methods: In this a comparative experimental study, collagen was extracted using eleven protocols andcharacterized using scanning electron microscope (SEM), Fourier-transform infrared spectroscopy (FTIR), XRD, Ramanspectroscopy, MTT and anti-inflammatory assays, lactate dehydrogenase (LDH) cytotoxicity, total antioxidant capacity(TAC), blood compatibility tests, protein quantification (BCA and Bradford), sodium dodecyl-sulfate polyacrylamide gelelectrophoresis (SDS-PAGE), and solubility evaluations at varying pH and salt concentrations. The most promisingsample was further evaluated in vivo via subcutaneous transplantation in mice, followed by histological analysis at 30days.Results: Among the evaluated protocols, protocol 4 consistently exhibited superior performance, yielding approximately35 mg collagen per g tissue. SEM, FTIR, XRD, and Raman analyses confirmed preserved triple-helical structureand fiber morphology. Protocol 4 showed high cell viability, low cytotoxicity, optimal blood compatibility, significantantioxidant activity, and improved protein content (P<0.05). In vivo studies demonstrated normal tissue regeneration,including collagen deposition, angiogenesis, and epidermal restoration, without inflammatory response. Other samplesexhibited incomplete structural features, reduced yield, or increased cytotoxicity.Conclusion: Protocol 4 provides a robust approach for extracting collagen with high structural integrity, functionalstability, and excellent biocompatibility. This optimized collagen may be suitable for biomedical applications and couldsupport further translational research for collagen-based therapeutic products.