Comparison of the Biological Behaviour of Human Dermal Fibroblasts seeded on 3D Printed Polylactic acid, Polycaprolactone and Polyethylene Terephthalate Scaffolds in vitro

Abstract

Regenerative medicine is a scientific field that improves and repairs diseased and injured tissues. Three-dimensional (3D) printing is an innovative technology that provides a new application field for regenerative medicine. 3D printed scaffolds by programming pore sizes and shapes serve as a temporary basis for cells until the natural extracellular matrix (ECM) is reconstructed. Dermal fibroblasts are mesenchymal cells located in the dermal skin layer that produce and organize ECM components. They play an essential role in skin wound healing and fibrosis. The aim of this study is to analyze the viability, adhesion, distribution, and collagen IV expression of human dermal fibroblasts (HDFs) seeded on 3D printed polylactic acid (PLA), polyethylene terephthalate (PET), and poly-ε-caprolactone (PCL) scaffolds in vitro. HDFs were seeded on scaffolds or tissue culture plastic plates as control and were cultured for 1 and 3 days. 3D PLA, PCL, and PET scaffolds were prepared using a custom made fused deposition modeling printer. The cell viability was measured by WST-1 assay on days 1 and 3. The cell adhesion was evaluated by scanning electron microscopy (SEM).The distribution was analyzed by hematoxylin and eosin (H&E) staining. Collagen IV expression was analyzed by immunohistochemical (IHC) staining.On day 1, the viability of HDFs on the 3D PLA scaffolds was significantly higher than PCL scaffolds. On day 3, the viability of HDFs on 3D PLA and PET scaffolds was significantly higher than PCL scaffolds. SEM images showed that HDFs on 3D PLA scaffolds attached the surfaces, filled the interfiber gaps and maintained their tissue specific morphology on day 3 compared to PCL and PET scaffolds. Histological images stained with H&E demonstrated that the distribution of HDFs on 3D PLA scaffolds was uniform on days 1 and 3. Collagen IV staining was more intense in HDFs on 3D PLA scaffolds on days 1 and 3. This study shows that 3D PLA scaffolds create a appropriate environment for cell viability, adhesion, distribution and may provide a high advantage in skin tissue regeneration.

3D Bioprinting and Its Role in a Wound Healing Renaissance

“…[ 41 ] WST‐1 assay was used to measure cell proliferation after 18 d. [ 41 ] HDF cell proliferation was observed to be ≈64%‐ to ≈83% for 40% pore sized PLA and ≈54% to ≈76% for 35% pore size PLA, with initial cell seeding concentration ranging from 1 × 10 4 to 4 × 10 4 cells per scaffold. [ 41 ] SEM imaging showed good adhesion by the HDF cells to the scaffolds. [ 41 ]…”

“…[28,37] Synthetic polymers, such as: PLA, PLGA, and PCL, can be used to create mechanically sturdy scaffolds to provide architecture onto which cells can attach and begin repopulating the wound bed. [38][39][40][41] Other synthetic polymers, such as PEG and poloxamers, can be used to create hydrogels into which cells and drugs can be directly incorporated prior to the bioprinting process. [42][43][44] Synthetic polymers are advantageous because of their easily tunable properties.…”

“…[98] Karabay et al prepared PLA scaffolds with two different pore sizes, 35% and 40%, via extrusion 3DP. [41] Subsequently, they seeded the scaffolds with HDF cells and observed the in vitro adhesion and proliferation. [41] WST-1 assay was used to measure cell proliferation after 18 d. [41] HDF cell proliferation was observed to be ≈64%-to ≈83% for 40% pore sized PLA and ≈54% to ≈76% for 35% pore size PLA, with initial cell seeding concentration ranging from 1 × 10 4 to 4 × 10 4 cells per scaffold.…”

“…Poly(lactic acid) (PLA) [41] HDF None Larger pore sizes in PLA scaffolds showed greater cell proliferation. HDF showed good adhesion to PLA.…”

“…[80] Poly (lactic-co-glycolic) acid (PLGA), polyglycolic acid (PGA), polylactic acid (PLA), polyethylene glycol (PEG), polyurethane (PU), and polycaprolactone (PCL) are all commonly used for creating scaffolds using 3DP. [38,41,104,150,151] Afghah et al used 3DP to create polycaprolactone-block-poly(1,3-propylene succinate) (PCL-PPSu) scaffolds and incorporated silver particles (AgNO 3 ) as an antimicrobial at concentrations of 1% and 2.5%, observing some effectiveness in reducing populations of Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus. [150] Additionally, at these concentrations of AgNO 3 , the researchers did not observe a significant cytotoxic effect on cultured human dermal fibroblast (HDF) cells.…”