Biological evaluation of micro-patterned hyaluronic acid hydrogel for bone tissue engineering

Abstract

Design of micro-patterning of hydrogel is of critical importance in both understanding cellular behaviors and mimicking controlled microenvironments and architectures of diverse well-organized tissues. After micro-patterning of hyaluronic acid (HA) hydrogel on a poly(dimethyl siloxane) substrate, its physical and biological properties have been compared with those of a non-patterned hydrogel for its possible applications in bone tissue engineering. The micro-patterned morphologies of HA hydrogel in both swollen and dehydrated forms have been observed with light microscope and scanning electron microscope, respectively, before and after in vitro cell culture. When MC3T3 bone cells were in vitro cultured on both HA hydrogels, the micro-patterned one shows excellence in cell proliferation and lining for 7 days along the micro-pattern paths over those of the non-patterned one, which have shown less cell-adhesiveness. The cytotoxicity of the micro-patterned HA hydrogels was in vitro evaluated by the assays of MTT, BrdU and Neutral red. The viability and morphology of MC3T3 cells on both HA hydrogels were observed with a fluorescence microscope by the live & dead assay, where their viability was confirmed by staining of F-actin development. The results of their H&E staining showed that both micro-patterned and non-patterned hydrogels induced development of tissue regeneration as observed by cell attachment, proliferation, and survivability, but the micro-patterned one induced distinctive patterning of both better initial cells adhesion on the micro-patterns and subsequently development of their proliferation and extracellular matrix, which were considered as important characteristics in their applications to tissue engineering.

Evaluation of MC3T3 Cells Proliferation and Drug Release Study from Sodium Hyaluronate-1,4-butanediol Diglycidyl Ether Patterned Gel

“…Fabrication of sodium hyaluronate-BDDGE patterned gel (HA-BDDGE) was performed using the method reported in our previous paper [ 24 ]. Briefly, sodium hyaluronate (HA, 0.18 g) was homogeneously mixed in 1 mL of 1% NaOH solution ( w / v %) using centrifuge at room temperature with 10,000 rpm speed for 2 h. Then, 72 µL of BDDGE was added and mixed with a spatula.…”

“…Our aim was to develop a shape-specific, hyaluronate-based patterned hydrogel for its application to bone regeneration. Sodium hyaluronate was chosen owing to its unique properties like abundant as natural ECM in human body, and its physico-chemical and immune-neutral characteristics [ 24 , 25 ]. The valuable properties led to development of various hyaluronic acid (HA) hydrogels for biomedical applications such as dermal fillers [ 26 , 27 ], cartilage regeneration [ 28 , 29 ], nucleus pulposus regeneration [ 30 ], and wound healing [ 31 , 32 ].…”

“…Soft lithography includes stamps fabricated from an elastomer or soft material like PDMS [ 6 ]. The PDMS stamp can mark ECM, self-assembled monolayer (SAM), and hydrogel to print on materials [ 6 , 24 ]. The well-defined ECM micro-patterns showed a significant effect on numerous imperative cell behaviors, such as cell adhesion and spreading [ 37 ], cell proliferation and differentiation [ 38 ], cell polarity [ 39 ], and migration [ 40 ].…”

A facile construction of gradient micro-patterned OCP coatings on medical titanium for high throughput evaluation of biocompatibility

“…In contrast, the cells on the OCP-3 micro-units displayed a polygonal shape, while cells on OCP-4 spread much better with a polygonal shape and prominent filopodia and lamellipodia protrusions. 85,86 The expression of F-actin on OCP-4 was significantly better than that on OCP-1, OCP-2 and OCP-3. After 24 h incubation, most cells were well spread and began to proliferate, with a long fusiform shape on the micro-units with OCP-1 and with a polygonal shape on OCP-2, most cell bodies on the microunits and some prominent filopodia on the superhydrophobic gaps.…”

Research trends in biomimetic medical materials for tissue engineering: 3D bioprinting, surface modification, nano/micro-technology and clinical aspects in tissue engineering of cartilage and bone

“…Applications of biomimetics to hydrogel fabrication in bioprinting were possible by chemical modifications of biocompatible polymers such as poly(ethylene glycol), polycarbonates and poly(vinyl alcohol) among many synthetic medical polymers. Functional cross-linking of the polymers were possible through click chemistries such as photo- and thermal chemistry, Michael type addition reactions, pHs and others [ 17 – 21 ]. Inorganic biomaterials have been also employed for 3D printing by controlling the properties of 3D printers such as fabrication methods and their nozzle sizes and surface properties [ 22 , 23 ].…”