Cell and Tissue-based Therapies for Hearing Loss
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1 Cell and Tissue-based Therapies for Hearing Loss Anne L. van de Ven 19 Nov 2003 BioE 492/592 Sensory Neuroengineering Lecture 16
2 Causes of Hearing Loss Age (34%) Loud noise (40 %) Disease/trauma (21%) Congenital (5%) Deafness prevention (gene & drug therapy) Hair cell regeneration (stem cell & growth factor therapy) Reconstructive surgery (tissue engineered substitutes) Each requires different strategies for treatment
3 Tissue Engineering Repair or replacement of tissue function with biological materials generated in labs
4 Tissue Engineering
5 Tissue Engineering of the Ear Cochlear regeneration Outer ear reconstruction Tympanic membrane repair Middle ear ossiculoplasty
6 Ossicles Utricle Cochlea Auricle Tympanic membrane (eardrum)
7 Tissue Engineering in the Cochlea Aimed at regenerating/replacing damaged hair cells Potential cell sources: Embryonic stem (ES) or progenitor cells Adult inner ear stem cells Trans-differentiation of non-hair cells
8 Stem Cell Fate
9 Stem Cell Fate
10 Stem Cell Fate Auditory progenitor cells
11 Generation of hair cells by stepwise differentiation of embryonic stem cells Li H,Roblin G, Liu H, Heller S.
12 Creating Auditory Progenitor Cells ES cell propagation ES cell medium + serum Cell expansion & differentiation as embryoid bodies 10 days EGF, IGF-1 Progenitor cell enrichment 8 days bfgf, EGF, IGF-1 Differentiated progenitor cells
13 10 days 8 days
14 Hair cell markers are co-expressed. % of total cells in vitro
15 Progenitor cells are grafted into chick embryos LacZ+ progenitor cells Cells examined 3 & 11 days post-injection Stage 17
16 Grafting promotes hair cell differentiation in the cochlea.
17 Implications 1. ES cells have the potential to differentiate into hair cells 2. Auditory progenitor cells can be successfully created in vitro 3. Can this work in the adult cochlea?
18 Adult Stem Cells in the Inner Ear Utricle stem cells Are highly proliferative Can self-renew in vitro Myosin IIVA+ / Espin+ hair cells Are pluripotent in vivo Can make hair cells in vivo Espin+ / f-actin+ stereocilia
19 Trans-differentiation in the Inner Ear Transcription factor Math1 is required for hair cell differentiation In vivo transfection of cochlear support cells with Math1 trans-differentiation into hair cells These new hair cells attract growth of new auditory neurons!
20 Outer Ear Reconstruction Performed since 1597 BC Why repair: cosmetic reasons improved hearing Indications for reconstruction: Trauma Cancer removal Congenital abnormalities (1 per 7000 births)
21 Outer Ear Reconstruction Gold standard: autologous rib cartilage unsatisfactory cosmetic results complications at secondary site Shaping rib cartilage Before After
22 Tissue Engineering of Auricle Aimed at creating a pre-molded, autologous cartilage auricle Advantages Reduces/eliminates surgery at 2 site Provides more realistic auricle Less dependence on artistic skills of the prosthetist
23 Key Challenges Obtaining sufficient cartilage cells Creating appropriate anatomical shape Controlling polymer degradation in vivo Reducing immune reactivity
24 A tissue-engineering model for the manufacture of auricular-shaped cartilage implants Haisch A, Klaring S, Groger A, Gebert C, Sittinger M
25 Key Challenges Obtaining sufficient cells - Chondrocyte expansion in culture Creating appropriate shape - Pre-molding of PGLA scaffold Controlling polymer degradation - Co-polymer (PGLA and fibrin) Reducing immune reactivity - Use of autologous cells
26
27 Results after 12 wks implantation Smooth, whitish cartilage-like morphology Some collagen & proteoglycan present Encapsulated in fibrous tissue 6 weeks 12 weeks
28 Customized Ear Reconstruction? CT/MRI scans can be combined with rapid prototyping techniques to create custom reconstructions Research reduced to the university level
29 References Li H,Roblin G, Liu H, Heller S. Generation of hair cells by stepwise differentiation of embryonic stem cells. PNAS 2003; 100: Li H, Liu H, Heller S. Pluripotent stem cells from the adult mouse inner ear. Nature Medicine 2003; 9(10): Kawamoto K, Ishimoto S, Minoda R, Brough DE, Raphael Y. Math1 gene transfer generates new cochlear hair cells in mature guinea pigs in vivo. Journal Neuroscience 2003; 23(11): Haisch A, Klaring S, Groger A, Gebert C, Sittinger M. A tissue-engineering model for the manufacture of auricularshaped cartilage implants. Eur Arch Otorhinolaryngol. 2002; 259(6):
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