Cellular Response of Limbal Stem Cells on PHBV/Gelatin Nanofibrous Scaffold for Ocular Epithelial Regeneration

Abstract

The aim of this study was to develop a blend of nanofibrous poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)=gelatin substrate for limbal stem cell (LSC) expansion that can serve as a potential alternative substrate to replace human amniotic membrane. The human Limbus stem cell was used to evaluate the biocompatibility of substrates (nanofibrous scaffold, and human amniotic membrane) based on their phenotypic profile, viability, proliferation, and attachment ability. Biocompatibility results indicated that the all substrates were highly biocompatible, as LSCs could favorably attach and proliferate on the nanofibrous surface. Microscopic figures showed that the human LSCs were firmly anchored to the substrates and were able to retain a normal corneal stem cell phenotype. Microscopic analyses illustrated that cells infiltrated the nanofibers and successfully formed a three-dimensional corneal epithelium, which was viable for two weeks. Immunocytochemistry (ICC) and real time-PCR results revealed no change in the expression profile of LSCs grown on nanofibrous substrate when compared to those grown on human amniotic membrane. In addition, electrospun nanofibrous PHBV substrate provides not only a milieu supporting LSCs expansion, but also serves as a useful alternative carrier for ocular surface tissue engineering and could be used as an alternative substrate to amniotic membrane.

In vivo assessment of a nanofibrous silk tube as nerve guide for sciatic nerve regeneration

“…Because, the structural and physical properties of electrospun nanofibres resemble those of ECM, electrospun nanofibers of synthetic or regenerated natural polymers promote cell proliferation, enhance nutrition and enable waste transfer. Thus, 3D electrospun nanofibres scaffolds have been widely used as scaffolds for various tissue engineering applications [27][28][29][30][31][32][33]. One work demonstrated that nonwoven silk nanofibrous nets were able to support the adhesion, proliferation and cell-cell interactions of variable human cell lines including osteoblasts, fibroblasts, keratinocytes and endothelial cells [28].…”

Electrospun Fibrous Architectures for Drug Delivery, Tissue Engineering and Cancer Therapy

“…Most common clinical substrate for ocular surface repair is human amniotic membrane owing to its biocompatibility, anti‐inflammatory efficacy, and angiogenesis ability 137. However, it also has several disadvantages, such as poor mechanical strength, semitransparent appearance, difficulty of handling, and the potential risk of disease transmission, and therefore, alternative choices should be further explored. Compared to bulk material, electrospun patches with micro/nanosize fiber dimensions and high surface area were considered advantageous in topical application or treating the anterior segment eye diseases.…”

The effect of the carbodiimide cross-linker on the structural and biocompatibility properties of collagen-chondroitin sulfate electrospun mat

“…It can produce the ultra fine fibers with special orientation, high surface-to-volume ratio, surface morphology, pore geometry, and three-dimensional architecture. [6][7][8][9][10][11][12][13][14][15][16][17][18] Thus, the choice of electrospinning technique takes into account the properties of the selected biomaterial, for instance, utilization of natural ECM proteins, to achieve a well-designed nanofiber scaffold, which imitates the desired natural ECM. It is obvious that collagen (CO) is the main abundant ECM protein, and it provides flexibility and structural strength to the ECM tissues.…”