Table 3.
Novel bioengineering and regenerative medicine approaches to produce treatments for hypothyroidism.
| Reference | Author | Scaffold Material | Cells Type | Study | Method | Result |
|---|---|---|---|---|---|---|
| (146) | Antonica et al., 2012 | Matrigel | Murine ESCs | In vivo (mice) | Recombinant tetracycline inducible NKX2.1 & PAX8 ESC lines generated, which were differentiated into embryoid bodies in hanging drops and into thyroid follicles in Matrigel-supported 3D culture with addition of rhTSH and Dox. The organoids were grafted under the renal capsule of hypothyroid female mice. | Overexpression of NKX2.1 & PAX8 induced differentiation of ESCs into FTCs. Cells showed iodide organification activity and formed follicular organoid structures in vitro. These rescued thyroid hormone plasma levels in vivo. |
| (147) | Arauchi et al., 2009 | NA | Primary rat thyroid cells | In vivo (rats) | Thyroids were minced & incubated with collagenase type II and IV before culturing on temperature responsive PIPAAm dishes. Cell sheets were implanted subcutaneously into the gluteus after 1-week of culture. | Serum T3 and T4 significantly increased 1 week after transplant and were maintained for 4 weeks. Morphological analysis showed typical follicle organization, microvessel formation and the presence of follicle epithelial cells. |
| (148) | Bulanova et al., 2017 | Collagen hydrogel | Primary murine thyroid cells & Allantoids | In vivo (mice) | Thyroid spheroids and allantoic spheroids were used as a source of thyrocytes and endothelial cells. These were printed in close association within a collagen hydrogel and implanted into mice. | Epithelial cells invaded and vascularized thyrocytes leading to the progressive formation of follicles. These were able to restore body temperature and serum T4 levels in mice. |
| (149) | Gabr et al., 2018 | Decellularized porcine liver organoid | Wharton Jelly derived MSCs | In vivo (mice) | Thyroid islets, produced through addition of Activin A and TSH, were loaded onto decellularized porcine liver organoids supplemented with TSH and iodine, then implanted intraperitoneal or intramuscular into mice. | Organoid corrected the hypothyroid state and showed viability and function at both implantation sites. Intramuscular transplantation showed higher vascularity and iodine uptake. |
| (150) | Kurmann et al., 2015 | Matrigel | Murine iPSCs | In vitro | Nkx2-1(GEP); Pax8(tdTomato trace) iPSCs were generated and differentiation directed towards thyroid with specific factors to induce Nkx2-1 expression. Cells were cultured 2D or in 3D Matrigel for organoid production. | Successfully matured thyroid progenitors derived from mouse iPSCs into thyroid follicular organoids which secrete T3, T4 and TSH. |
| (151) | Martin et al., 1993 | Matrigel | Human follicular thyroid cells | In vivo (mice) | Organoids were constructed from thyrocytes, embedded into a basement membrane preparation, and transplanted into SCID mice. | Widespread neofollicle formation was observed after 4 weeks of transplant. Murine T4 levels were not altered but human Tg was secreted and increased in response to TSH stimulation. |
| (152) | Mastrogiacomo, et al., 2012 | 3.5-5% PLLA in anhydrous dichloromethane/amylene or 3% PCL in anhydrous tetrahydrofuran/BHT | Primary rat thyroid cells | In vitro | The biomaterial scaffolds were prepared on wet glass support and cells seeded, allowing 8 days for growth. | Both biomaterial sheets promoted survival, adhesion, and proliferation of primary thyroid elements. T3 and T4 secretion varied but were a higher volume than in monolayer cultures. |
| (153) | Pan et al., 2019 | Decellularized thyroid gland (Male Lewis Rats) | FRTL-5 or Primary human follicular thyroid & parathyroid cells | In vitro | Thyroid gland decellularized with 1% SDS and recellularized perfusion seeding | Retained ECM and vascular network of native thyroid. Recellularized thyroid-maintained expression of Tg, thyroid peroxidase, and parathyroid hormone. |
| (154) | Risheng Ma, 2015 | Gelatin/Matrigel | Human ESCs | In vitro | Human PAX8 and NKX2-1 were expressed in ESCs with pEZ-lentiviral vectors, followed by differentiation into thyroid cells directed by Activin A and TSH within Gelatin/Matrigel | Double transfected cells expressed thyroid specific genes and the differentiation approach induced thyroid follicle formation. TSH stimulation induced dose-dependent cAMP generation and radioiodine uptake. |
| (155) | Strusi, V., et al., 2012 | Decellularized thyroid gland (Sprague-Dawley Male Rats) | Primary follicular thyroid or ABCG2+ thyroid stem/precursor (S/P) cells | In vitro | Thyroid matrix obtained by freezing/detergent/enzyme processing. Cells were expanded in a monolayer or 3D Matrigel culture before scaffold recellularization. | Thyroid architecture and SVS were maintained. S/P initiated follicle formation. Thyroid hormones were secreted for 7 days (minimum). |
| (156) | Strusi, V., et al., 2011 | Decellularized thyroid gland (Sprague-Dawley Male Rats) | Primary rat thyroid cells | In vitro | Rat thyroids were decellularized through freezing/thawing and sequential washes with 0.02% trypsin/0.05% EDTA, 3% Triton-x100, 4% deoxycholic acid and 0.1% peracetic acid (1% P/S/Fungizone). Cells were seeded in the inner surface of the matrix. | Native 3D architecture and the thyroid SVS were retained. Thyroid-derived cells aggregate, form intracytoplasmic cavities up to follicular coating and undergo secretory de-differentiation. |
| (157) | Weng, et al., 2021 | Decellularized thyroid gland (New Zealand White Rabbits) | Human follicular thyroid cells | In vitro | Thyroid gland decellularized with 1% SDS and recellularized with HFTCs | Maintained biomechanical properties, ECM, and cytokine composition of the native thyroid. Thyroid peroxidase secretion present. No cytotoxicity was induced. |
| (158) | Yang et al., 2016 | Alginate-poly-l-ornithine-alginate (APA) | Primary porcine thyroid cells | In vitro | Porcine primary thyroid cells were obtained through dissociation and collagenase digestions. The cells were suspended in low-viscosity-high-mannuronic acid solution, and the inner alginate core was formed by a microfluidic device. The inner core was coated to form an APA multilayer following cross-linking in a calcium chloride solution | The porcine cells successfully formed thyroid follicles in the microcapsule and displayed viability and proliferation. T4 was released significantly higher in encapsulated cells than unencapsulated cells. |
NKX2.1, NK2 Homebox 1; PAX8, Paired Box 8; ESC, embryonic stem cell; rhTSH, recombinant human thyroid stimulating hormone; FTC, follicular thyroid cell; NA, Not applicable; PIPAAm, Poly(N-isopropylacrylamide); MSC, mesenchymal stem cell; iPSC, induced pluripotent stem cell; SCID, severe combined immunodeficient; Tg= Thyroglobulin; PLL, Poly-L-lactic acid; PCL, Poly-Ꜫ-caprolactone; BHT, butylated hydroxytoluene; FRTL-5, Fischer Rat Thyroid low serum 5%; SDS, Sodium Dodecyl Sulphate; ECM, extracellular matrix; EDTA, ethylenediaminetetraacetic acid; SVS, stromal vascular scaffold; S/P, Stem/Precursor; P/S, Penicillin/Streptomycin; HFTC, human follicular thyroid cell.