Differences in Swelling and Water Content Percentage Between Non-Crosslinked and 0.25% Glutaraldehyde-Crosslinked Chitosan–Gelatin–Carbonate Hydroxyapatite Composite Scaffolds

Togi Ozora Sihaloho¹, Devi Rianti²*, Titien Hary Agustantina²

Abstract

Background: Scaffolds are three-dimensional structures that serve as a site for cell attachment, proliferation, and differentiation in bone tissue engineering. The combination of chitosan, gelatin, and carbonate hydroxyapatite (K-G:CHA) is widely used because of its biocompatibility, bioactivity, and similarity to the natural extracellular matrix of bone. However, its mechanical strength still needs to be improved through a crosslinking process. Crosslinking using 0.25% glutaraldehyde is known to improve the mechanical stability of scaffolds, but it can also affect their physical characteristics, particularly their ability to absorb and retain fluids, which can be evaluated through swelling and water content percentage (WCP). These two parameters are important for supporting nutrient diffusion, metabolite exchange, and providing a suitable physiological environment for cell growth and differentiation. Objective: This study aimed to analyze the differences in swelling and water content percentage of K-G:CHA composite scaffolds with ratios of 30:70 (w/w) and 40:60 (w/w), both non-crosslinked and crosslinked with 0.25% glutaraldehyde, after immersion in simulated body fluid (SBF). Methods: K-G:CHA scaffolds with ratios of 30:70 (w/w) and 40:60 (w/w) were synthesized using deep- freezing and freeze-drying methods. The scaffolds were then divided into non-crosslinked and 0.25% glutaraldehyde-crosslinked groups. The samples were immersed in SBF for 1, 3, and 7 days (24, 72, and 168 hours). Swelling and water content percentage were calculated based on changes in scaffold weight before and after immersion. Statistical analysis was performed using the Shapiro–Wilk test, Levene’s test, one-way ANOVA, and the post hoc Tukey test. Results: Both swelling and water content percentage increased with longer immersion time. The scaffolds crosslinked with 0.25% glutaraldehyde showed higher swelling and water content percentage values compared with the non-crosslinked groups. Statistical analysis showed significant differences between the treatment groups (p < 0.05). Conclusion: Crosslinking K-G:CHA composite scaffolds with 0.25% glutaraldehyde increased their swelling and water content percentage values by improving fluid retention capacity and structural stability. Therefore, these scaffolds have the potential to support their function as biomaterials for bone tissue engineering. 

Keywords

Scaffold; chitosan–gelatin–carbonate hydroxyapatite; 0.25% glutaraldehyde; swelling; water content percentage

Cite This Article

Sihaloho, T. O., Rianti, D., Agustantina, T. H. (2026). Differences in Swelling and Water Content Percentage Between Non-Crosslinked and 0.25% Glutaraldehyde-Crosslinked Chitosan–Gelatin–Carbonate Hydroxyapatite Composite Scaffolds. International Journal of Scientific Advances (IJSCIA), Volume 7| Issue 6: Nov – Dec 2026, Pages 975-982. URL: https://www.ijscia.com/wp-content/uploads/2026/11/Volume7-Issue6-Nov-Dec-No.1120-975-982.pdf

Volume 7 | Issue 6: Nov – Dec 2026