Mesenchymal stromal cells (MSCs) promote wound healing by expediting the inflammatory phase. Local injection of MSCs into injured vocal folds (VFs) is effective in animal models, suggesting suitability for clinical translation. Despite their therapeutic potential, MSCs do not persist within the VF. This study evaluates whether hyaluronan (HA) hydrogels offer a safe delivery vehicle for local injection of MSCs into VFs, and increase longevity of the cells within the injured tissue. MSCs ± HA hydrogel were exposed to interleukin (IL)1β, IL8, and chemokine (C-C motif) ligand 4, and evaluated for mRNA expression of matrix remodeling genes and secretion of immunomodulatory/prohealing factors. Chemotaxis/invasion in response to inflammation was evaluated. A lapin model of VF injury evaluated in vivo effects of MSCs ± HA hydrogel on enhancing VF healing. Histological evaluation of inflammation, type I collagen expression, HA hydrogel resorption, and MSC persistence was evaluated at 3 and 25 days after injury. MSCs within HA hydrogel were responsive to their extracellular environment, upregulating immunomodulatory factors when exposed to inflammation. Despite delayed migration out of the gel in vitro, the MSCs did not persist longer within the injured tissue in vivo. MSCs ± HA hydrogel exerted equivalent dampening of inflammation in vivo. The gel was resorbed within 25 days and no edema was evident. HA hydrogels can be safely used in the delivery of MSCs to injured VFs, minimizing leakage of administered cells. MSCs within the HA hydrogel did not persist longer than those in suspension, but did exert comparable therapeutic effects.
Get full access to this article
View all access options for this article.
References
1.
HansenJK and ThibeaultSL. (2006). Current understanding and review of the literature: vocal fold scarring. J Voice, 20:110–120.
2.
HertegardS, DahlqvistA and GoodyerE. (2006). Viscoelastic measurements after vocal fold scarring in rabbits—short-term results after hyaluronan injection. Acta Otolaryngol, 126:758–763.
3.
ThibeaultSL, GraySD, BlessDM, ChanRW and FordCN. (2002). Histologic and rheologic characterization of vocal fold scarring. J Voice, 16:96–104.
4.
WelhamNV, ChoiSH, DaileySH, FordCN, JiangJJ and BlessDM. (2011). Prospective multi-arm evaluation of surgical treatments for vocal fold scar and pathologic sulcus vocalis. Laryngoscope, 121:1252–1260.
5.
PetreyAC and de la MotteCA. (2014). Hyaluronan, a crucial regulator of inflammation. Front Immunol, 5:101.
6.
DePalmaRL, KrummelTM, DurhamLA3rd, MichnaBA, ThomasBL, NelsonJM and DiegelmannRF. (1989). Characterization and quantitation of wound matrix in the fetal rabbit. Matrix, 9:224–231.
7.
ChoiYH, KimSH, KimIG, LeeJH and KwonSK. (2019). Injectable basic fibroblast growth factor-loaded alginate/hyaluronic acid hydrogel for rejuvenation of geriatric larynx. Acta Biomater, 89:104–114.
8.
LiuF, HuS, YangH, LiZ, HuangK, SuT, WangS and ChengK. (2019). Hyaluronic acid hydrogel integrated with mesenchymal stem cell-secretome to treat endometrial injury in a rat model of Asherman's syndrome. Adv Healthc Mater. e1900411.
9.
RotherS, KronertV, HauckN, BergA, MoellerS, SchnabelrauchM, ThieleJ, ScharnweberD and HintzeV. (2019). Hyaluronan/collagen hydrogel matrices containing high-sulfated hyaluronan microgels for regulating transforming growth factor-beta1. J Mater Sci Mater Med, 30:65.
10.
ShamskhouEA, KratochvilMJ, OrcholskiME, NagyN, KaberG, SteenE, BalajiS, YuanK, KeswaniS, et al. (2019). Hydrogel-based delivery of Il-10 improves treatment of bleomycin-induced lung fibrosis in mice. Biomaterials, 203:52–62.
11.
AlemzadehE, OryanA and MohammadiAA. (2019). Hyaluronic acid hydrogel loaded by adipose stem cells enhances wound healing by modulating IL-1beta, TGF-beta1, and bFGF in burn wound model in rat. J Biomed Mater Res B Appl Biomater [Epub ahead of print];, DOI: 10.1002/jbm.b.34411.
WuKC, ChangYH, LiuHW and DingDC. (2019). Transplanting human umbilical cord mesenchymal stem cells and hyaluronate hydrogel repairs cartilage of osteoarthritis in the minipig model. Ci Ji Yi Xue Za Zhi, 31:11–19.
14.
YingH, ZhouJ, WangM, SuD, MaQ, LvG and ChenJ. (2019). In situ formed collagen-hyaluronic acid hydrogel as biomimetic dressing for promoting spontaneous wound healing. Mater Sci Eng C Mater Biol Appl, 101:487–498.
15.
HertegardS, HallenL, LaurentC, LindstromE, OlofssonK, TestadP and DahlqvistA. (2002). Cross-linked hyaluronan used as augmentation substance for treatment of glottal insufficiency: safety aspects and vocal fold function. Laryngoscope, 112:2211–2219.
16.
HertegardS, HallenL, LaurentC, LindstromE, OlofssonK, TestadP and DahlqvistA. (2004). Cross-linked hyaluronan versus collagen for injection treatment of glottal insufficiency: 2-year follow-up. Acta Otolaryngol, 124:1208–1214.
17.
MathenyKE, TsengEY, CarterKBJr., CobbWB and FongKJ. (2014). Self-cross-linked hyaluronic acid hydrogel in ethmoidectomy: a randomized, controlled trial. Am J Rhinol Allergy, 28:508–513.
18.
ParkYB, HaCW, LeeCH, YoonYC and ParkYG. (2017). Cartilage regeneration in osteoarthritic patients by a composite of allogeneic umbilical cord blood-derived mesenchymal stem cells and hyaluronate hydrogel: results from a clinical trial for safety and proof-of-concept with 7 years of extended follow-up. Stem Cells Transl Med, 6:613–621.
19.
SzkielkowskaA, MiaskiewiczB, RemacleM, KrasnodebskaP and SkarzynskiH. (2013). Quality of the voice after injection of hyaluronic acid into the vocal fold. Med Sci Monit, 19:276–282.
ChhetriDK and MendelsohnAH. (2010). Hyaluronic acid for the treatment of vocal fold scars. Curr Opin Otolaryngol Head Neck Surg, 18:498–502.
22.
OguzH, DemirciM, ArslanN and ArslanE. (2013). Long-term voice results of injection with hyaluronic acid-dextranomere in unilateral vocal fold paralysis. Acta Otolaryngol, 133:513–517.
23.
CarlssonPO, SchwarczE, KorsgrenO and Le BlancK. (2015). Preserved beta-cell function in type 1 diabetes by mesenchymal stromal cells. Diabetes, 64:587–592.
24.
Le BlancK, FrassoniF, BallL, LocatelliF, RoelofsH, LewisI, LaninoE, SundbergB, BernardoME, et al. (2008). Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study. Lancet, 371:1579–1586.
25.
Le BlancK, RasmussonI, SundbergB, GotherstromC, HassanM, UzunelM and RingdenO. (2004). Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells. Lancet, 363:1439–1441.
26.
MatasJ, OrregoM, AmenabarD, InfanteC, Tapia-LimonchiR, CadizMI, Alcayaga-MirandaF, GonzalezPL, MuseE, et al. (2019). Umbilical cord-derived mesenchymal stromal cells (MSCs) for knee osteoarthritis: repeated MSC dosing is superior to a single MSC dose and to hyaluronic acid in a controlled randomized Phase I/II trial. Stem Cells Transl Med, 8:215–224.
27.
UccelliA and de RosboNK. (2015). The immunomodulatory function of mesenchymal stem cells: mode of action and pathways. Ann N Y Acad Sci, 1351:114–126.
28.
NagubothuSR, SugarsRV, TudzarovskiN, AndrenAT, BottaiM, DaviesL, HertegardS and Le BlancK. (2019). Mesenchymal stromal cells modulate tissue repair responses within the injured vocal fold. Laryngoscope [Epub ahead of print];, DOI: 10.1002/lary.27885.
29.
SvenssonB, NagubothuRS, CedervallJ, Le BlancK, Ahrlund-RichterL, TolfA and HertegardS. (2010). Injection of human mesenchymal stem cells improves healing of scarred vocal folds: analysis using a xenograft model. Laryngoscope, 120:1370–1375.
30.
SvenssonB, NagubothuSR, CedervallJ, ChanRW, Le BlancK, KimuraM, Ahrlund-RichterL, TolfA and HertegardS. (2011). Injection of human mesenchymal stem cells improves healing of vocal folds after scar excision—a xenograft analysis. Laryngoscope, 121:2185–2190.
31.
DominiciM, Le BlancK, MuellerI, Slaper-CortenbachI, MariniF, KrauseD, DeansR, KeatingA, ProckopD and HorwitzE. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315–317.
32.
KucharzewskiM, RojczykE, Wilemska-KucharzewskaK, WilkR, HudeckiJ and LosMJ. (2019). Novel trends in application of stem cells in skin wound healing. Eur J Pharmacol, 843:307–315.
GoelAN, GowdaBS, VeenaMS, ShibaTL and LongJL. (2018). Adipose-derived mesenchymal stromal cells persist in tissue-engineered vocal fold replacement in rabbits. Ann Otol Rhinol Laryngol, 127:962–968.
35.
KimCS, ChoiH, ParkKC, KimSW and SunDI. (2018). The ability of human nasal inferior turbinate-derived mesenchymal stem cells to repair vocal fold injuries. Otolaryngol Head Neck Surg, 159:335–342.
ThibeaultSL, KlemukSA, ChenX and Quinchia JohnsonBH. (2011). In Vivo engineering of the vocal fold ECM with injectable HA hydrogels-late effects on tissue repair and biomechanics in a rabbit model. J Voice, 25:249–253.
40.
PostlethwaiteAE, SeyerJM and KangAH. (1978). Chemotactic attraction of human fibroblasts to type I, II, and III collagens and collagen-derived peptides. Proc Natl Acad Sci U S A, 75:871–875.
41.
LiY, RahmanianM, WidstromC, LepperdingerG, FrostGI and HeldinP. (2000). Irradiation-induced expression of hyaluronan (HA) synthase 2 and hyaluronidase 2 genes in rat lung tissue accompanies active turnover of HA and induction of types I and III collagen gene expression. Am J Respir Cell Mol Biol, 23:411–418.
42.
YamadaY, ItanoN, HataK, UedaM and KimataK. (2004). Differential regulation by IL-1beta and EGF of expression of three different hyaluronan synthases in oral mucosal epithelial cells and fibroblasts and dermal fibroblasts: quantitative analysis using real-time RT-PCR. J Invest Dermatol, 122:631–639.
43.
MeranS, ThomasD, StephensP, MartinJ, BowenT, PhillipsA and SteadmanR. (2007). Involvement of hyaluronan in regulation of fibroblast phenotype. J Biol Chem, 282:25687–25697.
44.
CsokaAB, FrostGI and SternR. (2001). The six hyaluronidase-like genes in the human and mouse genomes. Matrix Biol, 20:499–508.
45.
MeranS, MartinJ, LuoDD, SteadmanR and PhillipsA. (2013). Interleukin-1beta induces hyaluronan and CD44-dependent cell protrusions that facilitate fibroblast-monocyte binding. Am J Pathol, 182:2223–2240.
46.
PavanM, GalessoD, SecchieriC and GuariseC. (2016). Hyaluronic acid alkyl derivative: a novel inhibitor of metalloproteases and hyaluronidases. Int J Biol Macromol, 84:221–226.
47.
PhilipsN, KellerT and GonzalezS. (2004). TGF beta-like regulation of matrix metalloproteinases by anti-transforming growth factor-beta, and anti-transforming growth factor-beta 1 antibodies in dermal fibroblasts: implications for wound healing. Wound Repair Regen, 12:53–59.
48.
ItoA, MukaiyamaA, ItohY, NagaseH, ThogersenIB, EnghildJJ, SasaguriY and MoriY. (1996). Degradation of interleukin 1beta by matrix metalloproteinases. J Biol Chem, 271:14657–14660.
49.
CaglianiJ, GrandeD, MolmentiEP, MillerEJ and RiloHLR. (2017). Immunomodulation by mesenchymal stromal cells and their clinical applications. J Stem Cell Regen Biol, 3; DOI: 10.15436/2471-0598.17.022.
50.
KingSN, WooJH, TangS and ThibeaultSL. (2017). Macrophage response to allogeneic adipose tissue-derived stromal cells in hyaluronan-based hydrogel in a porcine vocal fold injury model. Ann Otol Rhinol Laryngol, 126:463–477.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
