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Updated: Apr 30, 2026
An Improved Method for the Preparation of Type I Collagen From Skin
Published on: January 21, 2014
Identification of An Optimized Collagen Extraction Protocol forRegenerative Medicine Application: A Comparative
Tayebeh Sadat Tabatabai1, Arian Ehterami2, Tahereh Sadat Tabatabai1
1Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran.
Objective:
Collagen plays a critical role in tissue engineering and regenerative medicine due to its biocompatibility and structural features. Identifying an optimal extraction method that preserves both functional and structural integrity remains challenging. This study aimed to compare eleven collagen extraction protocols to determine the most effective method for producing high-quality collagen.
Materials And Methods:
In this a comparative experimental study, collagen was extracted using eleven protocols and characterized using scanning electron microscope (SEM), Fourier-transform infrared spectroscopy (FTIR), XRD, Raman spectroscopy, 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 gel electrophoresis (SDS-PAGE), and solubility evaluations at varying pH and salt concentrations. The most promising sample was further evaluated In vivo via subcutaneous transplantation in mice, followed by histological analysis at 30 days.
Results:
Among the evaluated protocols, protocol 4 consistently exhibited superior performance, yielding approximately 35 mg collagen per g tissue. SEM, FTIR, XRD, and Raman analyses confirmed preserved triple-helical structure and fiber morphology. Protocol 4 showed high cell viability, low cytotoxicity, optimal blood compatibility, significant antioxidant 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 samples exhibited incomplete structural features, reduced yield, or increased cytotoxicity.
Conclusion:
Protocol 4 provides a robust approach for extracting collagen with high structural integrity, functional stability, and excellent biocompatibility. This optimized collagen may be suitable for biomedical applications and could support further translational research for collagen-based therapeutic products.
