Multifunctional magnetic-responsive hydrogels to engineer tendon-to-bone interface

doi:10.1016/j.nano.2017.06.002

Nanomedicine: Nanotechnology, Biology and Medicine

Volume 14, Issue 7, October 2018, Pages 2375-2385

https://doi.org/10.1016/j.nano.2017.06.002 Get rights and content

Abstract

Photocrosslinkable magnetic hydrogels are attracting great interest for tissue engineering strategies due to their versatility and multifunctionality, including their remote controllability ex vivo, thus enabling engineering complex tissue interfaces. This study reports the development of a photocrosslinkable magnetic responsive hydrogel made of methacrylated chondroitin sulfate (MA-CS) enriched with platelet lysate (PL) with tunable features, envisioning their application in tendon-to-bone interface. MA-CS coated iron-based magnetic nanoparticles were incorporated to provide magnetic responsiveness to the hydrogel. Osteogenically differentiated adipose-derived stem cells and/or tendon-derived cells were encapsulated within the hydrogel, proliferating and expressing bone- and tendon-related markers. External magnetic field (EMF) application modulated the swelling, degradation and release of PL-derived growth factors, and impacted both cell morphology and the expression and synthesis of tendon- and bone-like matrix with a more evident effect in co-cultures. Overall, the developed magnetic responsive hydrogel represents a potential cell carrier system for interfacial tissue engineering with EMF-controlled properties.

Graphical Abstract

A versatile magnetic responsive hydrogel of methacrylated chondroitin sulfate (MA-CS) incorporating iron-based superparamagnetic nanoparticles (MA-CS MNPs) and enriched with platelet lysate was successfully developed. The modulation of intrinsic properties like swelling, degradation and release of platelet lysate-origin growth factors were controlled upon the actuation of an external magnetic field (EMF). Tendon-to-bone magnetic responsive hydrogels units were assembled encapsulating tendon derived cells (TDCs), osteogenically differentiated adipose derived stem cells (O-ASCs) and a co-culture of both under EMF stimulation aiming at interfacial approaches. The developed magnetic system represents a potential cell carrier system for interfacial tissue engineering with EMF-controlled properties.

Section snippets

Production and characterization of methacrylated chondroitin sulfate magnetic nanoparticles (MA-CS MNPs)

MNPs were prepared by co-precipitation of Fe²⁺ and Fe³⁺ with ammonium hydroxide (NH₄OH) (05002, Sigma) as previously described.¹⁷ Briefly, FeCl₃.6H₂O (31,232, Sigma) and FeCl₂.4H₂O (220,299, Sigma) were mixed using a 2:1 molar ratio (Fe³⁺: Fe²⁺) under magnetic stirring in a nitrogen environment. The mixture was heated up to 80 °C and 33% (v/v) of NH₄OH was added to the salt solution until pH 10.5; being then cooled at room temperature for 15 min and a permanent magnet (0.6 T) was used to

Production and characterization of MA-CS MNPs

MA-CS MNPs were successfully developed. NMR confirmed the methacrylation of CS (Figure S1) and TGA analysis revealed 5.9 ± 0.25% of MA-CS in the produced MA-CS MNPs. After the coating the net surface charge at pH = 7 became negative (−26.73 ± 1.03), contrarily to uncoated MNPs (0.20 ± 0.86), as assessed by the electrokinetic potential measurements.

TEM images revealed a typical round shape of the MA-CS MNPs with an average size of 6.9 ± 1.9 nm (Figure 2, A).

The absence of reminiscence and coercive forces

Discussion

The addition of functionalities enabling the modulation of specific components of a construct ex vivo would strongly improve the design of smart biomaterials for tissue interfaces.²⁰ Based on that assumption, we proposed the development of a magnetic responsive hydrogel composed of a methacrylated chondroitin sulfate (MA-CS) matrix enriched with platelet lysate (PL). To provide magnetic responsiveness, magnetic nanoparticles (MNPs) were incorporated within the hydrogel matrix, envisioning the

Conclusions

A multifunctional system was successfully developed incorporating MA-CS MNPs in a MA-CS hydrogel that can be further manipulated by the application of EMF, which enables the control of intrinsic properties of the construct, including the modulation of growth factors release from platelet lysate.

The biological response of different cell types indicates the suitability of the magnetic system to be used as a cell carrier. Furthermore, the physical bond of different units in the proposed hydrogel

Acknowledgments

Authors thank CEMUP and Histology and Electron Microscopy Service of I3S for the cryo-SEM and TEM analysis, respectively. Authors also thank Serviço de Imuno-Hemoterapia, Centro Hospitalar São João, EPE (Porto, Portugal) for providing human platelet concentrate samples and to Hospital da Prelada (Porto, Portugal) for providing lipoaspirate and tendon tissue samples, respectively. Authors also acknowledge Margarida Silva Miranda for the TGA analysis.

References (37)

A. Seidi et al.
Gradient biomaterials for soft-to-hard interface tissue engineering
Acta Biomater
(2011)
J. Klein-Nulend et al.
Mechanobiology of bone tissue
Pathol Biol (Paris)
(2005)
L.J. Santos et al.
Harnessing magnetic-mechano actuation in regenerative medicine and tissue engineering
Trends Biotechnol
(2015)
B. Mattix et al.
Biological magnetic cellular spheroids as building blocks for tissue engineering
Acta Biomater
(2014)
J.J. Lim et al.
The effect of desulfation of chondroitin sulfate on interactions with positively charged growth factors and upregulation of cartilaginous markers in encapsulated MSCs
Biomaterials
(2013)
I.Y. Tóth et al.
Magnetic hyaluronate hydrogels: preparation and characterization
J Magnet Magnetic Mater
(2015)
K.J. Livak et al.
Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method
Methods
(2001)
S. Font Tellado et al.
Strategies to engineer tendon/ligament-to-bone interface: biomaterials, cells and growth factors
Adv Drug Deliv Rev
(2015)
R.D. LeBoeuf et al.
Human fibrinogen specifically binds hyaluronic acid
J Biol Chem
(1986)
R. Barbucci et al.
Biohydrogels with magnetic nanoparticles as crosslinker: characteristics and potential use for controlled antitumor drug-delivery
Acta Biomater
(2012)

R.M. Domingues et al.

Development of injectable hyaluronic acid/cellulose nanocrystals Bionanocomposite hydrogels for tissue engineering applications

Bioconjug Chem

(2015)

Y. Yeo et al.

In situ cross-linkable hyaluronic acid hydrogels prevent post-operative abdominal adhesions in a rabbit model

Biomaterials

(2006)

C. Silva et al.

Following the enzymatic digestion of chondroitin sulfate by a simple GPC analysis

Anal Chim Acta

(2015)

N. Mori et al.

The role of osteopontin in tendon tissue remodeling after denervation-induced mechanical stress deprivation

Matrix Biol

(2007)

L.-F. Wang et al.

Synthesis and characterization of chondroitin sulfate–methacrylate hydrogels

Carbohydr Polym

(2003)

J.E. Phillips et al.

Engineering graded tissue interfaces

Proc Natl Acad Sci

(2008)

M.T. Galloway et al.

The role of mechanical loading in tendon development, maintenance, injury, and repair

J Bone Joint Surg Am

(2013)

A.I. Goncalves et al.

Exploring the potential of starch/Polycaprolactone aligned magnetic responsive scaffolds for tendon regeneration

Adv Healthc Mater

(2016)

Cited by (69)

Untethered: using remote magnetic fields for regenerative medicine
2023, Trends in Biotechnology
Citation Excerpt :
Additionally, advances in tomographic additive manufacturing (also termed as volumetric printing) [85] and photoclick chemistry [86] have reduced fabrication times to <10 s. Since most magnetic nano- or microparticle formulations are optically opaque, the photocrosslinking-based biofabrication of magnetically responsive tissues has been underexplored and mostly limited to thin films [87,88]. Future research into the incorporation and optimization of optically transparent magnetic materials [89] could expand the scope of light-based biofabrication toward magnetically responsive tissue engineering.
Magnetic fields are increasingly being used for the remote, noncontact manipulation of cells and biomaterials for a wide range of regenerative medical (RM) applications. They have been deployed for their direct effects on biological systems or in conjunction with magnetic materials or magnetically tagged cells for a targeted therapeutic effect. In this work, we highlight the recent trends on the broad use of magnetic fields for the homing of therapeutic cells and particles at targeted tissue sites, biomimetic tissue fabrication, and control of cell fate and proliferation. We also survey the design and control principles of magnetic manipulation systems, including their capabilities and limitations, which can guide future research into developing more effective magnetic field-based regenerative strategies.
Three-dimensional printing of hyaluronate-based self-healing ferrogel with enhanced stretchability
2023, Colloids and Surfaces B: Biointerfaces
Hydrogels have been frequently employed for three-dimensional (3D) printing, which is a promising tool for fabricating sophisticated structures useful in many biomedical applications. Ferrogels prepared by combining magnetic nanoparticles with hydrogels also have potential in biomedical engineering because of the responsiveness to a magnetic field and remotely controllable properties. However, typical ferrogels, especially those prepared from natural polysaccharides, have limitations concerning their mechanical properties and the fabrication method of complex structures owing to their rigid and brittle properties. In this study, 3D printable and stretchable ferrogel was designed and prepared to overcome these limitations. Hyaluronic acid (HA) derivatives such as hydrazide-modified HA (hHA) and oxidized HA (oHA) were used as the base materials for gel preparation. Self-healing oHA/hHA hydrogels were prepared by the addition of adipic acid dihydrazide (ADH). Self-healing ferrogels with 3D printability were prepared by adding superparamagnetic iron oxide nanoparticles (SPIONs) to oHA/hHA/ADH hydrogels, which improved the stretchability owing to the double network formation (2.1 times its original length). Various 3D constructs were fabricated by an extrusion-based printing method using ferrogel (structural integrity = 94.3 ± 1.5%). The potential to fabricate hydrogel/ferrogel hybrid constructs for tissue engineering was also investigated. This approach for developing customized 3D constructs using magnetic field-responsive and 3D printable hydrogel systems may find useful applications in tissue engineering approaches.
Smart/stimuli-responsive hydrogels: State-of-the-art platforms for bone tissue engineering
2022, Applied Materials Today
Repair of bone defects is a time-consuming, self-healing process based on tissue modeling and remodeling. However, the capacity of this method is limited, especially for critical-sized bone defects where bone augmentation is necessary. Hence, bone tissue engineering (BTE) with the execution of curative strategies that merge biomolecules, biomimetic scaffolds, and cells plays a decisive role in this track. Smart/stimuli-responsive hydrogels (SSRHs) are superb three-dimensional (3D) biomaterials intended for tissue engineering and other biomedical implementations. They can imitate the diverse mechanical, biological and physicochemical nature of the natural tissues. Besides, they provide 3D configuration, afford ample aqueous circumstances, and support mechanical constancy necessary for cell growth.
Furthermore, they function as competent delivery platforms for various biomolecules. Different nature-derived and synthetic polymers were largely exploited to produce smart scaffolds with distinctive characters and customized functionalities matching BTE applications. In the current review, we outlined the fundamentals of bone biology and the existing strategies for BTE. Moreover, we discussed different SSRHs, their synthesis scheme, their present situation, and future perspectives for BTE. In conclusion, recent progress in the assembly of SSRHs reinforces their potential to serve as smart and intricate platforms for regenerating injured bone tissues.
Magnetic-responsive polysaccharide hydrogels as smart biomaterials: Synthesis, properties, and biomedical applications
2022, Carbohydrate Polymers
This review reports recent advances in polysaccharide-based magnetic hydrogels as smart platforms for different biomedical applications. These hydrogels have proved to be excellent, viable, eco-friendly alternative materials for the biomedical field due to their biocompatibility, biodegradability, and possibility of controlling delivery processes via modulation of the remote magnetic field. We first present their main synthesis methods and compare their advantages and disadvantages. Next, the synergic properties of hydrogels prepared with polysaccharides and magnetic nanoparticles (MNPs) are discussed. Finally, we describe the main contributions of polysaccharide-based magnetic hydrogels in the targeted drug delivery, tissue regeneration, and hyperthermia therapy fields. Overall, this review aims to motivate the synthesis of novel composite biomaterials, based on the combination of magnetic nanoparticles and natural polysaccharides, to overcome challenges that still exist in the treatment of several diseases.
3D printed magnetic polymer composite hydrogels for hyperthermia and magnetic field driven structural manipulation
2022, Progress in Polymer Science
Magnetic hydrogels and soft composites have fueled the development of next generation biomimetic soft robotics due to their precise control and non-cytotoxic nature. Bare magnetic nanoparticles are difficult to regulate via remote controlling whereas, when these nanoparticles are arrested inside polymeric matrices, the whole system become an artificial soft mussel like integrated system. Concurrently, these polymeric magnetic soft materials are also prone to response of external magnetic field (static or oscillatory). Additive manufacturing via spatial assembly of polymeric precursors followed by actuation like behavior is quite a new manufacturing technique to fabricate magnetic soft materials. In this review, we focused on the magnetic nanoparticles and their entrapment into polymeric matrices and assessing their applicability in clinical (hyperthermia) as well as shape morphing behaviours. Both the behaviors are also amassed together to form dual responsive soft microbots. The techniques and their applications are elaborated in this review.
3D bioprinting of multilayered scaffolds with spatially differentiated ADMSCs for rotator cuff tendon-to-bone interface regeneration
2022, Applied Materials Today
Regeneration of the gradient structure of the tendon-to-bone interface is still a significant clinical challenge. This study reports a novel therapeutic method combining three-dimensional (3D) bioprinting and melt electrospinning writing techniques to regenerate a functional tendon-to-bone interface. We generated biomimetic multilayered scaffolds with 3D-bioprinted pre-differentiated autologous adipose-derived mesenchymal stem cells (ADMSC), which recapitulated compositional and cellular structures of the interface. The hydrogel-based bioinks offered high cell viability and proliferative capability for rabbit ADMSCs. The hydrogels with pre-differentiated (into tenogenic, chondrogenic, and osteogenic lineages) or undifferentiated rabbit ADMSCs were 3D-bioprinted into zonal-specific constructs to mimic the structure of the tendon-to-bone interface. These scaffolds were tested in a rabbit rotator cuff injury model and the histological, radiological, and biomechanical changes were analyzed. The in vivo studies demonstrated that the scaffold with spatially differentiated autologous ADMSCs had a superior histological score and improved collagen organization when compared to acellular scaffolds and similar T2 value as the normal interface tissue. The biomechanical characterization demonstrated that the application of multilayered scaffolds improved the biomechanical properties of the tendon-to-bone interface at 12 weeks after rotator cuff reconstruction surgery, but the incorporation of autologous ADMSCs within the multilayered scaffolds showed a limited contribution. Thus, our work provides a 3D-bioprinting-based strategy with the application of autologous ADMSCs to reconstruct massive rotator cuff tendon tears.

View all citing articles on Scopus

: Disclosures: The authors declare no conflict of interest.

: Part of the content of this manuscript was previously presented in a poster communication at the TermStem2016 meeting in Guimarães, Portugal.

: Financial support information: The authors wish to acknowledge the financial support from the Portuguese Foundation for Science and Technology (FCT) for the PhD/post-doctoral fellowships of R.C-A (SFRH/BD/96593/2013), R.M.A.D (SFRH/BPD/112459/2015), M.T.R (SFRH/BPD/111729/2015), and career development grant of M.E.G. (IF/00685/2012) and Recognize project (UTAP-ICDT/CTM-BIO/0023/2014). Authors acknowledge the financial support from FCT/MCTES (Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia, e Ensino Superior) and the Fundo Social Europeu through Programa Operacional do Capital Humano (FSE/POCH), PD/59/2013 by PD/BD/113807/2015. Authors are also grateful to RL3-TECT-NORTE-07-0124-FEDER-000020 project co-financed by ON.2 (NSRF) through ERD.

View full text

Research ArticleSpecial Topic: Two-Dimensional Biomaterials in Regenerative MedicineMultifunctional magnetic-responsive hydrogels to engineer tendon-to-bone interface

Abstract

Graphical Abstract

Section snippets

Production and characterization of methacrylated chondroitin sulfate magnetic nanoparticles (MA-CS MNPs)

Production and characterization of MA-CS MNPs

Discussion

Conclusions

Acknowledgments

Acta Biomater

Pathol Biol (Paris)

Trends Biotechnol

Acta Biomater

Biomaterials

J Magnet Magnetic Mater

Methods

Adv Drug Deliv Rev

J Biol Chem

Acta Biomater

Bioconjug Chem

Biomaterials

Anal Chim Acta

Matrix Biol

Carbohydr Polym

Engineering graded tissue interfaces

Proc Natl Acad Sci

The role of mechanical loading in tendon development, maintenance, injury, and repair

J Bone Joint Surg Am

Exploring the potential of starch/Polycaprolactone aligned magnetic responsive scaffolds for tendon regeneration

Adv Healthc Mater

Research ArticleSpecial Topic: Two-Dimensional Biomaterials in Regenerative Medicine
Multifunctional magnetic-responsive hydrogels to engineer tendon-to-bone interface