Fibrin promotes proliferation and matrix production of intervertebral disc cells cultured in three-dimensional poly(lactic-co-glycolic acid) scaffold

Abstract

Previously, we have proven that fibrin and poly(lactic-co-glycolic acid) (PLGA) scaffolds facilitate cell proliferation, matrix production and early chondrogenesis of rabbit articular chondrocytes in in vitro and in vivo experiments. In this study, we evaluated the potential of fibrin/PLGA scaffold for intervertebral disc (IVD) tissue engineering using annulus fibrosus (AF) and nucleus pulposus (NP) cells in relation to potential clinical application. PLGA scaffolds were soaked in cells-fibrin suspension and polymerized by dropping thrombin-sodium chloride (CaCl(2)) solution. A PLGA-cell complex without fibrin was used as control. Higher cellular proliferation activity was observed in fibrin/PLGA-seeded AF and NP cells at each time point of 3, 7, 14 and 7 days using the MTT assay. After 3 weeks in vitro incubation, fibrin/PLGA exhibited a firmer gross morphology than PLGA groups. A significant cartilaginous tissue formation was observed in fibrin/PLGA, as proven by the development of cells cluster of various sizes and three-dimensional (3D) cartilaginous histoarchitecture and the presence of proteoglycan-rich matrix and glycosaminoglycan (GAG). The sGAG production measured by 1,9-dimethylmethylene blue (DMMB) assay revealed greater sGAG production in fibrin/PLGA than PLGA group. Immunohistochemical analyses showed expressions of collagen type II, aggrecan core protein and collagen type I genes throughout in vitro culture in both fibrin/PLGA and PLGA. In conclusion, fibrin promotes cell proliferation, stable in vitro tissue morphology, superior cartilaginous tissue formation and sGAG production of AF and NP cells cultured in PLGA scaffold. The 3D porous PLGA scaffold-cell complexes using fibrin can provide a vehicle for delivery of cells to regenerate tissue-engineered IVD tissue.

A review of the application of reinforced hydrogels and silk as biomaterials for intervertebral disc repair

“…Material Publication Annulus Fibrosus Enhanced with genipin Likhitpanichkul et al, 2014;Likhitpanichkul et al, 2015;Long et al, 2016a;Schek et al, 2011 Nucleus Pulposus Combined with hyaluronic acid Li et al, 2014;Park et al, 2012 3D Culture Combined with poly(lactic-co-glycolic acid) Sha'ban et al, 2008;Stern et al, 2000;Stich et al, 2015 Correia et al, 2013). It has yet to be shown whether Gellan gum could withstand compressive forces in long-term IVD repair experiments.…”

Scaffolds and tissue regeneration: An overview of the functional properties of selected organic tissues

“…The increased mechanical resistance of the scaffolds for application in intervertebral discs is made possible by the use of composites. 2,5,48,57 Meniscus. The meniscus has a semilunar shape and extends over the upper tibial surface, improving its congruence with the femoral condyles.…”

Stem Cell-assisted Approaches for Cartilage Tissue Engineering

“…Fibrin possesses chemically active gelling property, which typically occurs during blood coagulation in addition to the advantages described previously. Several studies investigating fibrinbased hybrid scaffold demonstrated its potential for the promotion of homogeneous cell distribution and cartilaginous tissue formation (33)(34)(35). PNIPAAm was also used for this purpose, and it possessed the merit of a synthetic thermoresponsive polymer for tissue engineering with versatile manipulation of its chemical structure and molecular weight as well as easy conjugation with other components.…”