Rezazadeh Mahboubeh, Akbari Vajihe, Sadeghi Azam, Tavakoli Naser
Department of Pharmaceutics and Novel Drug Delivery System Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.
Department of Pharmaceutical Biotechnology and Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.
Res Pharm Sci. 2025 Aug 25;20(4):580-589. doi: 10.4103/RPS.RPS_30_23. eCollection 2025 Aug.
LL37 peptide is a human antimicrobial peptide with potential application in bone tissue engineering through the stimulation of cell proliferation and osteogenesis. The current study aimed to fabricate chitosan/gelatin/glycerophosphate (CTS/G/GP) and chitosan/collagen/glycerophosphate (CTS/C/GP) thermosensitive hydrogels loaded with LL37 and compared their ability to support cell growth, proliferation, and osteogenesis.
The hydrogel systems were prepared by the physical mixture of chitosan, gelatin, collagen, and GP at the concentrations of 2.5, 1, 1, and 10% w/v, respectively. LL37 was added at a fixed concentration of 1 μg/mL of the hydrogels. The viscosity, friability, release properties, and biological experiments were evaluated based on standard procedures.
FINDINGS/RESULTS: The viscosity of CTS/C/GP increased to 7000 cP at 35 °C in 100-120 s, while for CTS/G/GP, the viscosity and gelation time were recorded as 14000 cP and 30 s, respectively. The friability percent for CTS/G/GP after 72 h was reported as 28%, which was significantly lower than that of 38% for CTS/C/GP. LL37 was released during 8 h from both scaffold systems, and it did not demonstrate any significant differences between the hydrogel systems. Cell viability and alkaline phosphatase activity revealed that the incorporation of LL37 in the hydrogels could accelerate cell proliferation compared to empty scaffolds, and it was higher in gelatin-containing scaffolds.
LL37 was successfully loaded into both hydrogel systems and demonstrated the ability to accelerate cell proliferation and differentiation compared to the empty scaffold.
LL37肽是一种人类抗菌肽，通过刺激细胞增殖和成骨作用在骨组织工程中具有潜在应用价值。本研究旨在制备负载LL37的壳聚糖/明胶/甘油磷酸酯（CTS/G/GP）和壳聚糖/胶原蛋白/甘油磷酸酯（CTS/C/GP）热敏水凝胶，并比较它们支持细胞生长、增殖和成骨的能力。
通过壳聚糖、明胶、胶原蛋白和甘油磷酸酯分别以2.5%、1%、1%和10% w/v的浓度进行物理混合来制备水凝胶体系。以1 μg/mL的固定浓度向水凝胶中添加LL37。基于标准程序评估粘度、易碎性、释放特性和生物学实验。
CTS/C/GP在35 °C下100 - 120 s内粘度增加到7000 cP，而CTS/G/GP的粘度和凝胶化时间分别记录为14000 cP和30 s。CTS/G/GP在72 h后的易碎百分比报告为28%，显著低于CTS/C/GP的38%。LL37在8 h内从两种支架体系中释放，且水凝胶体系之间未显示出任何显著差异。细胞活力和碱性磷酸酶活性表明，与空支架相比，水凝胶中掺入LL37可加速细胞增殖，且在含明胶的支架中更高。
LL37成功负载到两种水凝胶体系中，与空支架相比，显示出加速细胞增殖和分化的能力。
