3D PCL/Gelatin/Genipin Nanofiber Sponge as Scaffold for Regenerative Medicine

Abstract

Recent advancements in tissue engineering and material science have radically improved in vitro culturing platforms to more accurately replicate human tissue. However, the transition to clinical relevance has been slow in part due to the lack of biologically compatible/relevant materials. In the present study, we marry the commonly used two-dimensional (2D) technique of electrospinning and a self-assembly process to construct easily reproducible, highly porous, three-dimensional (3D) nanofiber scaffolds for various tissue engineering applications. Specimens from biologically relevant polymers polycaprolactone (PCL) and gelatin were chemically cross-linked using the naturally occurring cross-linker genipin. Potential cytotoxic effects of the scaffolds were analyzed by culturing human dermal fibroblasts (HDF) up to 23 days. The 3D PCL/gelatin/genipin scaffolds produced here resemble the complex nanofibrous architecture found in naturally occurring extracellular matrix (ECM) and exhibit physiologically relevant mechanical properties as well as excellent cell cytocompatibility. Samples cross-linked with 0.5% genipin demonstrated the highest metabolic activity and proliferation rates for HDF. Scanning electron microscopy (SEM) images indicated excellent cell adhesion and the characteristic morphological features of fibroblasts in all tested samples. The three-dimensional (3D) PCL/gelatin/genipin scaffolds produced here show great potential for various 3D tissue-engineering applications such as ex vivo cell culturing platforms, wound healing, or tissue replacement.

Bioengineering of fibroblast‐conditioned polycaprolactone/gelatin electrospun scaffold for skin tissue engineering

“…28,53 Different studies indicated the PCL/Gel composites for overcoming PCL deficiencies providing suitable biocompatibility, mechanical, and chemical traits. 36,54 Indeed, our in vitro biocompatibility assays demonstrated no difference between conditioned scaffolds and plasma-treated ones, contrary to in vivo biocompatibility results indicating better biocompatibility of conditioned scaffolds than other groups. Our in vitro and in vivo biocompatibility results were similar to those reported by Samadikuchaksaraei et al 26 They investigated the effects of osteoblast-conditioning on mechanics, biocompatibility, and osteoinductivity of a nanohydroxyapatite/gelatin scaffold.…”

“…Electrospun PCL scaffolds coated by gelatin improved the proliferation and survival of fibroblast cells in vitro and in vivo 28,53 . Different studies indicated the PCL/Gel composites for overcoming PCL deficiencies providing suitable biocompatibility, mechanical, and chemical traits 36,54 . Indeed, our in vitro biocompatibility assays demonstrated no difference between conditioned scaffolds and plasma‐treated ones, contrary to in vivo biocompatibility results indicating better biocompatibility of conditioned scaffolds than other groups.…”

Acrylonitrile and Pullulan Based Nanofiber Mats as Easily Accessible Scaffolds for 3D Skin Cell Models Containing Primary Cells

“…Consequently, the use of other materials, such as chitosan, polycaprolactone (PCL), and poly(ethylene glycol) (PEG) has been investigated [4]. To simulate the inherent ECM architecture of skin, with its nanofibrous structure and porosity, electrospinning of these polymers can be employed [42][43][44][45][46]. Despite the evident success of electrospun scaffolds, there are numerous ongoing challenges, such as achieving suitable cell infiltration [43], cell adhesion [47], or mechanical properties [43].…”

“…Despite the evident success of electrospun scaffolds, there are numerous ongoing challenges, such as achieving suitable cell infiltration [43], cell adhesion [47], or mechanical properties [43]. Research is mainly focused on biopolymers (e.g., hyaluronic acid, alginate, collagen, silk protein, fibrinogen, chitosan, starch, and poly(3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV)) [48] or on synthetic polymers such as poly(lactic acid-coglycolic acid) (PLGA), PCL or PLA [42,46,48]. To promote cell adhesion, proliferation, migration, or morphology, bioactive molecules are commonly added to the scaffolds, in particular collagen, gelatin, or elastin [47], which are however associated with challenges in terms of batch-to-batch variability or adhesion efficiency.…”

Genipin-Crosslinking Effects on Biomatrix Development for Cutaneous Wound Healing: A Concise Review

“…They found that nonmodified PET-CH revealed quick degradation after 3 days of buffer exposure. Several concentrations of GNP (0.5%, 1.0%, and 2.5%) were also added, by Merk et al (2021) , into polycaprolacton/GEL formulation to produce sponges for regenerative purposes. A remaining mass of >92% was found in all GNP-crosslinked samples after 8 weeks of incubation in buffer solution.…”