A leptin-loaded poly-ϵ-caprolactone 3D printing scaffold for odontoblastic differentiation in human dental pulp cells

Abstract

This study investigated the effects on odontoblast differentiation of a 3D-printed poly-ε-caprolactone (PCL) scaffold that incorporated leptin. Material extrusion-type 3D printing with a 43,000-molecular weight PCL material was used to fabricate a PCL scaffold with a 6 mm diameter, 1 mm height, and 270 to 340 µm pore size. The experimental groups were PCL scaffolds (control group), PCL scaffolds with aminated surfaces (group A), and PCL scaffolds with leptin on the aminated surface (group L). The aminated surface was treated with 1,6-hexanediamine and verified by ninhydrin analysis. Leptin loading was performed using Traut’s reagent and 4-(N-Maleimidomethyl)cyclohexane-1-carboxylic acid 3-sulfo-N-hydroxysuccinimide ester sodium salt (Sulfo-SMCC). Groups A and L showed significantly higher surface wettability, pulp cell adhesion, and proliferation than the control group. Group L exhibited increased alkaline phosphatase (ALP), calcification deposits, and mRNA and protein expression of dentin sialophosphoprotein (DSPP) and dentin matrix acidic phosphoprotein 1 (DMP1) compared with the control group. In this study, a 3D-printed PCL scaffold containing leptin enhanced odontoblast differentiation towards and dental pulp cells adhesion and proliferation.

The potential bone regeneration effects of leptin- and osteolectin-coated 3D-printed PCL scaffolds: an in vivo study

“…Therefore, in this study, bone regeneration was promoted by coating the scaffold with leptin and osteolectin, which are growth factors that stimulate osteogenic differentiation [38][39][40][41][42][43][44]. Adhesion and distribution of leptin and osteolectin were performed as previously described [51].…”

“…It is mainly produced in adipose tissue and acts on the hypothalamus of the brain to regulate appetite and metabolism. The main function of leptin is related to the regulation of energy homeostasis, but it also has various effects on other physiological processes, including bone metabolism [38][39][40][41]51]. Leptin receptors are present in osteoblasts, the cells responsible for bone formation.…”

“…Amination of the PCL scaffolds was done as previously described (figure 1) [46][47][48][49][50][51]. The PCL scaffolds were placed in a 96-well plate, followed by the addition of 100 µl of 10% w/v 1,6-hexanediamine solution in isopropanol to each PCL scaffold and incubation at 37 • C for one hour.…”

Effect of 3D-printed polycaprolactone/osteolectin scaffolds on the odontogenic differentiation of human dental pulp cells

“…As a result of measurement with a stereoscopic microscope, the pore size of the PCL scaffolds was 270-340 µm [30]. A previous study using SEM reported that untreated PCL specimens have smooth surfaces, whereas crystalline deposits were confirmed on the surface of both aminated PCL scaffolds and aminated PCL scaffolds treated with Arg-Gly-Asp (RGD) [32].…”

“…PCL (Mn = 43 000 g mol −1 ; Polysciences, USA) was put into a syringe attached to a Dr INVIVO 4D6 (Rokit Healthcare, Seoul, Korea) printer and melted at 75 • C. The PCL was sprayed with a 0.4 mm diameter nozzle to prepare a lattice-shaped specimen with a pore density of 65%, diameter of 30 mm, and height of 1 mm [29]. The pore size of the PCL specimen was measured using a stereomicroscope (EGVM-452M, EG Tech, Ansan, Korea) [30]. Then, PCL scaffolds were made into disks with a 6 mm diameter using a puncher.…”