Harnessing biochemical and structural cues for tenogenic differentiation of adipose derived stem cells (ADSCs) and development of an in vitro tissue interface mimicking tendon-bone insertion graft.

Tendon-bone interface tissue is extremely challenging to engineer because it exhibits complex gradients of structure, composition, biologics, and cellular phenotypes. As a step toward engineering these transitional zones, we initially analyzed how different (topographical or biological) cues affect tenogenic differentiation of adipose-derived stem cells (ADSCs). We immobilized platelet-derived growth factor - BB (PDGF-BB) using polydopamine (PD) chemistry on random and aligned nanofibers and investigated ADSC proliferation and tenogenic differentiation. Immobilized PDGF greatly enhanced the proliferation and tenogenic differentiation of ADSCs; however, nanofiber alignment had no effect. Interestingly, the PDGF immobilized aligned nanofiber group showed a synergistic effect with maximum expression of tenogenic markers for 14 days. We also generated a nanofiber surface with spatially controlled presentation of immobilized PDGF on an aligned architecture, mimicking native tendon tissue. A gradient of immobilized PDGF was able to control the phenotypic differentiation of ADSCs into tenocytes in a spatially controlled manner, as confirmed by analysis of the expression of tenogenic markers and immunofluorescence staining. We further explored the gradient formation strategy by generation of a symmetrical gradient on the nanofiber surface for the generation of a structure mimicking bone-patellar-tendon-bone with provision for gradient immobilization of PDGF and controlled mineralization. Our study reveals that, together with biochemical cues, favorable topographical cues are important for tenogenic differentiation of ADSCs, and gradient presentation of PDGF can be used as a tool for engineering stem cell-based bone-tendon interface tissues.

[1]  L. Soslowsky,et al.  Decorin regulates assembly of collagen fibrils and acquisition of biomechanical properties during tendon development , 2006, Journal of cellular biochemistry.

[2]  Stavros Thomopoulos,et al.  Functional attachment of soft tissues to bone: development, healing, and tissue engineering. , 2013, Annual review of biomedical engineering.

[3]  K. Char,et al.  Delivery of a therapeutic protein for bone regeneration from a substrate coated with graphene oxide. , 2013, Small.

[4]  S. Oh,et al.  Porous membrane with reverse gradients of PDGF-BB and BMP-2 for tendon-to-bone repair: in vitro evaluation on adipose-derived stem cell differentiation. , 2014, Acta biomaterialia.

[5]  R. K. Peterson,et al.  Allograft versus autograft patellar tendon anterior cruciate ligament reconstruction: A 5-year follow-up. , 2001, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[6]  Zi Yin,et al.  The regulation of tendon stem cell differentiation by the alignment of nanofibers. , 2010, Biomaterials.

[7]  Hyun Suk Jung,et al.  Graded functionalization of biomaterial surfaces using mussel-inspired adhesive coating of polydopamine. , 2017, Colloids and surfaces. B, Biointerfaces.

[8]  Rui L Reis,et al.  Engineering tendon and ligament tissues: present developments towards successful clinical products , 2013, Journal of tissue engineering and regenerative medicine.

[9]  Helen H. Lu,et al.  Advances in biologic augmentation for rotator cuff repair , 2016, Annals of the New York Academy of Sciences.

[10]  H. Mizuno,et al.  Tendon regeneration and repair with adipose derived stem cells , 2010 .

[11]  C. Wilkinson,et al.  Grooved substrata facilitate in vitro healing of completely divided flexor tendons , 1995 .

[12]  Franz Pfeiffer,et al.  The microstructure and micromechanics of the tendon-bone insertion. , 2017, Nature materials.

[13]  James Chang,et al.  Tissue engineering of flexor tendons: optimization of tenocyte proliferation using growth factor supplementation. , 2006, Tissue engineering.

[14]  Chih-Yu Wu,et al.  Multifunctional and Continuous Gradients of Biointerfaces Based on Dual Reverse Click Reactions. , 2016, ACS applied materials & interfaces.

[15]  V. Vogel,et al.  Bioactive, Elastic, and Biodegradable Emulsion Electrospun DegraPol Tube Delivering PDGF-BB for Tendon Rupture Repair. , 2016, Macromolecular bioscience.

[16]  Heungsoo Shin,et al.  Controlled Retention of BMP-2-Derived Peptide on Nanofibers Based on Mussel-Inspired Adhesion for Bone Formation. , 2017, Tissue engineering. Part A.

[17]  Esther J. Lee,et al.  Materials from Mussel-Inspired Chemistry for Cell and Tissue Engineering Applications. , 2015, Biomacromolecules.

[18]  B. Harley,et al.  The effect of anisotropic collagen-GAG scaffolds and growth factor supplementation on tendon cell recruitment, alignment, and metabolic activity. , 2011, Biomaterials.

[19]  S. Oh,et al.  Dual growth factor-immobilized asymmetrically porous membrane for bone-to-tendon interface regeneration on rat patellar tendon avulsion model. , 2018, Journal of biomedical materials research. Part A.

[20]  D. Zeugolis,et al.  The biophysical, biochemical, and biological toolbox for tenogenic phenotype maintenance in vitro. , 2014, Trends in biotechnology.

[21]  Haeshin Lee,et al.  Facile Conjugation of Biomolecules onto Surfaces via Mussel Adhesive Protein Inspired Coatings , 2009, Advanced materials.

[22]  Heungsoo Shin,et al.  Polydopamine-mediated immobilization of multiple bioactive molecules for the development of functional vascular graft materials. , 2012, Biomaterials.

[23]  Benjamin B. Rothrauff,et al.  The Rotator Cuff Organ: Integrating Developmental Biology, Tissue Engineering, and Surgical Considerations to Treat Chronic Massive Rotator Cuff Tears. , 2017, Tissue engineering. Part B, Reviews.

[24]  Changsheng Liu,et al.  Facilitated receptor-recognition and enhanced bioactivity of bone morphogenetic protein-2 on magnesium-substituted hydroxyapatite surface , 2016, Scientific Reports.

[25]  Younan Xia,et al.  "Aligned-to-random" nanofiber scaffolds for mimicking the structure of the tendon-to-bone insertion site. , 2010, Nanoscale.

[26]  D. Mcallister,et al.  Current tissue engineering strategies in anterior cruciate ligament reconstruction. , 2014, Journal of biomedical materials research. Part A.

[27]  H. Seeherman,et al.  Tendon‐selective genes identified from rat and human musculoskeletal tissues , 2010, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[28]  R. Gelberman,et al.  Growth factors and canine flexor tendon healing: initial studies in uninjured and repair models. , 1995, The Journal of hand surgery.

[29]  William D Middleton,et al.  The outcome and repair integrity of completely arthroscopically repaired large and massive rotator cuff tears. , 2004, The Journal of bone and joint surgery. American volume.

[30]  N. Maffulli,et al.  Tendon healing: can it be optimised? , 2002, British journal of sports medicine.

[31]  Zhiyong He,et al.  Rho/Rock signal transduction pathway is required for MSC tenogenic differentiation , 2015, Bone Research.

[32]  L. Soslowsky,et al.  Tendon to bone healing: differences in biomechanical, structural, and compositional properties due to a range of activity levels. , 2003, Journal of biomechanical engineering.

[33]  Younan Xia,et al.  Nanofiber scaffolds with gradations in mineral content for mimicking the tendon-to-bone insertion site. , 2009, Nano letters.

[34]  Richard O. Hynes,et al.  The Extracellular Matrix: Not Just Pretty Fibrils , 2009, Science.

[35]  C. Tabin,et al.  Analysis of the tendon cell fate using Scleraxis, a specific marker for tendons and ligaments. , 2001, Development.

[36]  J. Hollinger,et al.  Regenerative tendon and ligament healing: opportunities with recombinant human platelet-derived growth factor BB-homodimer. , 2012, Tissue engineering. Part B, Reviews.

[37]  Heungsoo Shin,et al.  Effects of Immobilized BMP-2 and Nanofiber Morphology on In Vitro Osteogenic Differentiation of hMSCs and In Vivo Collagen Assembly of Regenerated Bone. , 2015, ACS applied materials & interfaces.

[38]  N. Voelcker,et al.  Materials Displaying Neural Growth Factor Gradients and Applications in Neural Differentiation of Embryoid Body Cells , 2015 .

[39]  B. Harley,et al.  Composite growth factor supplementation strategies to enhance tenocyte bioactivity in aligned collagen-GAG scaffolds. , 2013, Tissue engineering. Part A.

[40]  Guanbin Song,et al.  Effect of Focal Adhesion Kinase on the Regulation of Realignment and Tenogenic Differentiation of Human Mesenchymal Stem Cells by Mechanical Stretch , 2011, Connective tissue research.

[41]  Marilisa Quaranta,et al.  Collagen Structure of Tendon Relates to Function , 2007, TheScientificWorldJournal.

[42]  C. Albigès-Rizo,et al.  Presentation of BMP‐2 from a Soft Biopolymeric Film Unveils its Activity on Cell Adhesion and Migration , 2011, Advanced materials.

[43]  Oh Soo Kwon,et al.  Platelet-derived growth factor-BB-immobilized asymmetrically porous membrane for enhanced rotator cuff tendon healing , 2016, Tissue Engineering and Regenerative Medicine.

[44]  Choongsoo S. Shin,et al.  Effective immobilization of BMP-2 mediated by polydopamine coating on biodegradable nanofibers for enhanced in vivo bone formation. , 2014, ACS applied materials & interfaces.

[45]  F. Berenbaum,et al.  Tendon injury: from biology to tendon repair , 2015, Nature Reviews Rheumatology.

[46]  L. Galatz,et al.  In Vivo Evaluation of Adipose-Derived Stromal Cells Delivered with a Nanofiber Scaffold for Tendon-to-Bone Repair. , 2015, Tissue engineering. Part A.

[47]  D. Nicolella,et al.  Platelet-derived growth-factor-releasing aligned collagen-nanoparticle fibers promote the proliferation and tenogenic differentiation of adipose-derived stem cells. , 2014, Acta biomaterialia.

[48]  F R Noyes,et al.  Revision Anterior Cruciate Surgery with Use of Bone-Patellar Tendon-Bone Autogenous Grafts , 2001, The Journal of bone and joint surgery. American volume.

[49]  Evren U Azeloglu,et al.  The guidance of stem cell differentiation by substrate alignment and mechanical stimulation. , 2013, Biomaterials.

[50]  G. Genin,et al.  The development and morphogenesis of the tendon-to-bone insertion - what development can teach us about healing -. , 2010, Journal of musculoskeletal & neuronal interactions.

[51]  Kurt P Spindler,et al.  Clinical practice. Anterior cruciate ligament tear. , 2008, The New England journal of medicine.

[52]  D. Mcallister,et al.  Evaluation of polycaprolactone scaffold with basic fibroblast growth factor and fibroblasts in an athymic rat model for anterior cruciate ligament reconstruction. , 2015, Tissue engineering. Part A.

[53]  F. Guilak,et al.  Aligned multilayered electrospun scaffolds for rotator cuff tendon tissue engineering. , 2015, Acta biomaterialia.

[54]  Younan Xia,et al.  Nanofiber Scaffolds with Gradients in Mineral Content for Spatial Control of Osteogenesis , 2014, ACS applied materials & interfaces.

[55]  Zefeng Zheng,et al.  Alignment of collagen fiber in knitted silk scaffold for functional massive rotator cuff repair. , 2017, Acta biomaterialia.

[56]  Changlian Lu,et al.  PDGF-BB-induced MT1-MMP expression regulates proliferation and invasion of mesenchymal stem cells in 3-dimensional collagen via MEK/ERK1/2 and PI3K/AKT signaling. , 2013, Cellular signalling.

[57]  O. Akkus,et al.  Heparinized collagen sutures for sustained delivery of PDGF-BB: Delivery profile and effects on tendon-derived cells In-Vitro. , 2016, Acta biomaterialia.

[58]  Mark D. Miller,et al.  Controversies in ACL Reconstruction: Bone-patellar Tendon-bone Anterior Cruciate Ligament Reconstruction Remains the Gold Standard , 2009, Sports medicine and arthroscopy review.