Tension Stimulation of Tenocytes in Aligned Hyaluronic Acid/Platelet-Rich Plasma-Polycaprolactone Core-Sheath Nanofiber Membrane Scaffold for Tendon Tissue Engineering.

Chih-Hao Chen,Andy Deng-Chi Chuang,Banendu Sunder Dash,Jyh-Ping Chen,Dai-Ling Li

doi:10.3390/ijms222011215

Abstract

To recreate the in vivo niche for tendon tissue engineering in vitro, the characteristics of tendon tissue underlines the use of biochemical and biophysical cues during tenocyte culture. Herein, we prepare core-sheath nanofibers with polycaprolactone (PCL) sheath for mechanical support and hyaluronic acid (HA)/platelet-rich plasma (PRP) core for growth factor delivery. Three types of core-sheath nanofiber membrane scaffolds (CSNMS), consisting of random HA-PCL nanofibers (Random), random HA/PRP-PCL nanofibers (Random+) or aligned HA/PRP-PCL (Align+) nanofibers, were used to study response of rabbit tenocytes to biochemical (PRP) and biophysical (fiber alignment) stimulation. The core-sheath structures as well as other pertinent properties of CSNMS have been characterized, with Align+ showing the best mechanical properties. The unidirectional growth of tenocytes, as induced by aligned fiber topography, was confirmed from cell morphology and cytoskeleton expression. The combined effects of PRP and fiber alignment in Align+ CSNMS lead to enhanced cell proliferation rates, as well as upregulated gene expression and marker protein synthesis. Another biophysical cue on tenocytes was introduced by dynamic culture of tenocyte-seeded Align+ in a bioreactor with cyclic tension stimulation. Augmented by this biophysical beacon from mechanical loading, dynamic cell culture could shorten the time for tendon maturation in vitro, with improved cell proliferation rates and tenogenic phenotype maintenance, compared to static culture. Therefore, we successfully demonstrate how combined use of biochemical/topographical cues as well as mechanical stimulation could ameliorate cellular response of tenocytes in CSNMS, which can provide a functional in vitro environmental niche for tendon tissue engineering.

Highlights

Tendinopathy is a common condition negatively affecting the life quality of laborers, athletes and physically active individuals, accounting for up to 30% of musculoskeletal consultation in general practice [1]
No significant difference was noticed in fiber size, reduced mean fiber diameter for Align+ coincide with faster solvent evaporation rate, generated from forced convection air flow around a rotating collector to collect aligned nanofibers
Tenocytes were exposed to a combination of stimuli, including a biochemical stimulus provided by growth factors in platelet-rich plasma (PRP), a topographical cue presented by aligned nanofibers, and mechanical induction by applying uni-axial tension stimulation

Summary

Introduction

Tendinopathy is a common condition negatively affecting the life quality of laborers, athletes and physically active individuals, accounting for up to 30% of musculoskeletal consultation in general practice [1]. While adjunct treatment options such as growth factor and gene therapy are available, current clinically available options for tendon ruptures are mostly confined to tendon prosthesis, allografts, mosaicplasty and autografts [3]. Tissue processing methods such as fresh-freezing, cryo-preservation, ethylene oxide treatment, and gamma irradiation can decrease the risks, detrimental effects on biomechanical properties of the grafts cannot be ruled out [5]. To address these pitfalls, tissue engineering approaches by seeding tendon-derived cells on scaffolds with unique characteristics have yielded promising results, which can provide optimal growth environment while facilitating cellular proliferation and extracellular matrix (ECM) deposition [6,7]

Methods

Results

Conclusion