การศึกษาการปล่อยเพปไทด์ต้านจุลชีพจากแผ่นเยื่อกั้นชนิดพอลิ (แอล-แลกไทด์-โค-คาร์โปรแลคโตน) ในห้องปฏิบัติการ

Abstract

Guided bone regeneration (GBR) is widely used for bone regeneration around the dental implants which require a barrier membrane to exclude soft tissue from the bone defect site. Electrospun poly (L-lactide-co--caprolactone), PLC, is a polymer membrane which is biocompatibility, biodegradable in an appropriate time and mimic extracellular matrix in human tissues to promote cell adhesion. LL-37 is an antimicrobial peptide found in innate immunity of humans that are against pathogens, promote wound healing and bone regeneration. Currently, there is still no study about adding LL-37 into the barrier membrane. Hence, in this study, we fabricated a polymer membrane for guided bone regeneration which can provide sustained-release antimicrobial peptides. Therefore, the objective of this work was to study the release of LL-37 from the PLC membrane in vitro. reserved In this study, LL-37 was physically adsorbed to collagen scaffold (LL/Col) followed by coated with PLC nanofibrous scaffold to produce the LL/Col/PLC membrane. The LL/Col/PLC and LL/Col membranes were incubated in sterile PBS at 37C for 14 days. Cumulative release of LL-37 in release medium at 4 hours, 1 day, 7 days and 14 days were measured by using enzyme-linked immunosorbent assay. Characterization of the LL/Col/PLC membranes was studied by scanning electron microscope. The effect of LL-37 released from the LL/Col/PLC membranes on the vitality of human monocytes and mouse fibroblasts were studied by MTT assay. According to the result of ELISA, burst release of LL-37 was exhibited in the control group. In contrast, the study group showed sustained release of LL-37 that was significantly less than the control group at 4 hours, 1 day, 7 days and 14 days (P<0.05), which improved wound healing and bone regeneration. Furthermore, the surface morphology of the electrospun PLC mimicked the extracellular matrix for cell adhesion. LL-37 released from the PLC membranes significantly increased the vitality of human monocytes (P<0.05) and had no toxicity to mouse fibroblasts. Based on these results, the LL/Col/PLC membrane developed in this study can sustain release of LL-37 in clinically relevant doses. Therefore, the LL/Col/PLC membrane is a promising material for guided bone regeneration membrane which will further enhance bone regeneration.