Sains Malaysiana 48(10)(2019): 2229–2237

http://dx.doi.org/10.17576/jsm-2019-4810-19

Neurogenic Differentiation Potential of Human Nasal Mucosa Obtained from the Middle and Inferior Turbinates

(Potensi Pembezaan Neurogen Mukosa Hidung Manusia Diperoleh daripada Turbinat Tengah dan Inferior)

TAN SHI NEE¹, YOGESWARAN LOKANATHAN², ROHAINA CHE MAN², AMINUDDIN SAIM^1,3 & RUSZYMAH BT HJ IDRUS⁴*

¹KPJ Healthcare University College, 71800 Nilai, Negeri Sembilan Darul Khusus, Malaysia

²Tissue Engineering Centre, Universiti Kebangsaan Malaysia, 56000 Kuala Lumpur, Federal Territory, Malaysia

³Ear, Nose and Throat Consultant Clinic, Ampang Puteri Specialist Hospital, 68000 Ampang, Selangor Darul Ehsan, Malaysia

⁴Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, 56000 Kuala Lumpur, Federal Territory, Malaysia

Diserahkan: 8 Jun 2018/Diterima: 15 Ogos 2019

ABSTRACT

Olfactory bulb and nasal mucosa are one of the sources for neural stem cell, including the superior and middle turbinates (MT). The middle and inferior turbinates (IT) provides the largest area of nasal mucosa which is technically easier to harvest the stem cell for future transplantation. The ability of nasal respiratory epithelial cells (RECs) and nasal fibroblasts (NFs) from both middle and inferior turbinates to differentiate into neural lineage (NL) cells were compared in this study. Six redundant human MT and IT from post-sinus surgery were digested and cultured. The RECs and NFs were separated and induced with neurotrophic factors of forskolin, human basic fibroblast growth factor (bFGF), platelet-derived growth factor-AA (PDGF-AA) and heregulin-β1-EGF-domain. Based on immunocytochemistry and quantitative PCR, the NL induced NFs of MT expressed GFAP, Nestin and P75 receptor. NL induced RECs from MT and IT expressed GFAP and Nestin but did not express the P75 receptor protein. Regarding the control, the non-induced RECs and fibroblasts expressed Nestin only. This study demonstrated that nasal mucosa cells from both IT and MT have the potential to differentiate into neural lineage cells even though the fibroblasts of MT are superior in term of quality. Hopefully, these tissues will provide better donor area with less morbidity for autologous or allograft transplantation in future neural regenerative medicine.

Keywords: Nasal epithelial cells; nasal mucosa; neural induction; neural lineage; turbinate

ABSTRAK

Salah satu sumber sel stem saraf adalah bebuli olfaktori dan mukosa hidung, terutamanya dalam turbinat superior dan tengah (MT). Walau bagaimanapun, turbinat tengah dan turbinat inferior (IT) menyediakan kawasan terbesar untuk mukosa hidung dan secara teknikal, lebih mudah untuk memperoleh sel untuk pemindahan pada masa hadapan. Dalam kajian ini, keupayaan sel epitelium pernafasan (RECs) dan fibroblas hidung (NFs) daripada kedua-dua turbinat untuk membezakan sel-sel seketurunan saraf (NL) telah dibandingkan. Enam sampel MT dan IT manusia yang berlebihan daripada pasca pembedahan sinus telah dicerna dan sel-sel yang diperoleh dikultur. RECs dan NFs dipisahkan dan diaruh dengan faktor neurotropik, iaitu forskolin, faktor pertumbuhan fibroblas asas manusia, faktor pertumbuhan yang diperoleh daripada platelet dan domain heregulin-β1-EGF. Berdasarkan imunositokimia dan tindak balas berantai polimerase kuantitatif, NL disebabkan MT yang diaruh mengekspreskan GFAP, Nestin dan P75 reseptor. Pengaruhan NL menyebabkan RECs daripada MT dan IT mengekspreskan GFAP dan Nestin tetapi tidak mengekspreskan protein P75 reseptor. RECs dan NFs yang tidak teraruh sebagai kawalan hanya mengekspreskan Nestin. Kajian ini menunjukkan bahawa sel mukosa hidung daripada kedua-dua IT dan MT mampu diaruh kepada sel-sel seketurunan saraf walaupun fibroblas MT didapati lebih sesuai daripada segi kualiti. Diharapkan tisu-tisu ini dapat memberikan kawasan penderma yang lebih baik dengan kurang morbiditi untuk transplantasi autologus atau alograf dalam perubatan regeneratif saraf pada masa depan.

Kata kunci: Aruhan saraf; mukosa pernafasan; seketurunan saraf; sel epitelium hidung; turbinat

RUJUKAN

Au, E. & Roskams, A.J. 2002. Culturing olfactory ensheathing glia from the mouse olfactory epithelium. In Neural Stem Cells: Methods and Protocols. Methods in Molecular Biology, Volume 198, edited by Zigova, T., Sanberg, P.R. & Sanchez- Ramos, J.R. Totowa: Humana Press.

Ball, S.L., Mann, D.A., Wilson, J.A. & Fisher, A.J. 2016. The role of the fibroblast in inflammatory upper airway conditions. The American Journal of Pathology 186(2): 225-233.

Bas, E., Van De Water, T.R., Lumbreras, V., Rajguru, S., Goss, G., Hare, J.M. & Goldstein, B.J. 2013. Adult human nasal mesenchymal-like stem cells restore cochlear spiral ganglion neurons after experimental lesion. Stem Cells and Development 23(5): 502-514.

Bing Cang Li, Chuan Xu, Jie Yuan Zhang, Yue Li & Zhao Xia Duan. 2012. Differing Schwann cells and olfactory ensheathing cells behaviors, from interacting with astrocyte, produce similar improvements in contused rat spinal cord’s motor function. Journal of Molecular Neuroscience 48(1): 35-44.

Delorme, B., Nivet, E., Gaillard, J., Häupl, T., Ringe, J., Devèze, A., Magnan, J., Sohier, J., Khrestchatisky, M. & Roman, F.S. 2009. The human nose harbors a niche of olfactory ectomesenchymal stem cells displaying neurogenic and osteogenic properties. Stem Cells and Development 19(6): 853-866.

Dong Chang Lee, Hyunsu Choi, Jeong-Min Oh, Yupyo Hong, Su Hee Jeong, Choung Soo Kim, Dong-Kee Kim, Won- Kyung Cho, Sung Won Kim, Soo Whan Kim, Jin Hee Cho & Joohyung Lee. 2018. The effect of urban particulate matter on cultured human nasal fibroblasts. International Forum of Allergy & Rhinology 8(9): 993-1000.

Fang, F., Ni, K., Cai, Y., Ye, Z., Shang, J., Shen, S. & Xiong, C. 2017. Biological characters of human dermal fibroblasts derived from foreskin of male infertile patients. Tissue and Cell 49(1): 56-63.

Fulcher, M.L. & Randell, S.H. 2013. Human nasal and tracheo-bronchial respiratory epithelial cell culture. In Methods in Molecular Biology. New Jersey: Springer.

Goldstein, B.J., Hare, J.M., Lieberman, S. & Casiano, R. 2013. Adult human nasal mesenchymal stem cells have an unexpected broad anatomic distribution. International Forum of Allergy & Rhinology 3(7): 550-555.

Hauser, S., Widera, D., Qunneis, F., Müller, J., Zander, C., Greiner, J., Strauss, C., Lüningschrör, P., Heimann, P. & Schwarze, H. 2011. Isolation of novel multipotent neural crest-derived stem cells from adult human inferior turbinate. Stem Cells and Development 21(5): 742-756.

Jae Woong Hwang, Jae Hyung Kim, Hyun Jung Kim, In Hak Choi, Hye Min Han, Ki Jung Lee, Tae Hoon Kim & Sang Hag Lee 2019. Neutrophil extracellular traps in nasal secretions of patients with stable and exacerbated chronic rhinosinusitis and their contribution to induce chemokine secretion and strengthen the epithelial barrier. Clinical & Experimental Allergy 49(10): 1306-1320. https://doi. org/10.1111/cea.13448.

Kato, T., Honmou, O., Uede, T., Hashi, K. & Kocsis, J.D. 2000. Transplantation of human olfactory ensheathing cells elicits remyelination of demyelinated rat spinal cord. Glia 30(3): 209-218.

Khairunnisa Ramli, Ifasha Aminath Gasim, Amir Adham Ahmad, Shariful Hassan, Zhe Kang Law, Geok Chin Tan, Azmi Baharuddin, Amaramalar Selvi Naicker, Ohnmar Htwe & Nor Hazla Mohammed Haflah. 2019. Human bone marrow- Derived MSCs spontaneously express specific Schwann cell markers. Cell Biology International 43(3): 233-252.

Lane, A.P. 2009. The role of innate immunity in the pathogenesis of chronic rhinosinusitis. Current Allergy and Asthma Reports 9(3): 205-212.

Lima, C., Escada, P., Pratas-Vital, J., Branco, C., Arcangeli, C.A., Lazzeri, G., Maia, C.A.S., Capucho, C., Hasse-Ferreira, A. & Peduzzi, J.D. 2010. Olfactory mucosal autografts and rehabilitation for chronic traumatic spinal cord injury. Neurorehabilitation and Neural Repair 24(1): 10-22.

Lin, R.J., Krall, R., Westerberg, B.D., Chadha, N.K. & Chau, J.K. 2012. Systematic review and meta-analysis of the risk factors for sudden sensorineural hearing loss in adults. The Laryngoscope 122(3): 624-635.

Lokanathan Yogeswaran, Min-Hwei Ng, Shariful Hasan, Anuar Ali, Mazzre Mahmod, Ohnmar Htwe, Sharifah Ahmad Roohi, Ruszymah Hj Idrus, Shalimar Abdullah & Amaramalar Selvi Naicker. 2014. Olfactory ensheathing cells seeded muscle-stuffed vein as nerve conduit for peripheral nerve repair: A nerve conduction study. Journal of Bioscience and Bioengineering 118(2): 231-234.

Meng Wang, Lu Chengliang & Fred Roisen. 2012. Adult human olfactory epithelial-Derived progenitors: A potential autologous source for cell-Based treatment for Parkinson’s Disease. Stem Cells Translational Medicine 1(6): 492-502.

Mingliang Tang, Xiaoqian Yan, Qilin Tang, Rongrong Guo, Peng Da & Dan Li. 2018. Potential application of electrical stimulation in stem cell-based treatment against hearing loss. Neural Plasticity 2018: 9506387.

Mohd Heikal Mohd Yunus, Kan Chan Siang, Nurul Izzati Hashim, Ng Pei Zhi, Nur Fathurah Zamani, Primuharsa Putra Sabri, Mohd Fauzi Busra, Shiplu Roy Chowdhury & Ruszymah Haji Idrus. 2014. The effects of human serum to the morphology, proliferation and gene expression level of the respiratory epithelium in vitro. Tissue and Cell 46(4): 233-240.

Mohd Heikal, M.Y., Aminuddin, B.S., Jeevanan, J., Chen, H.C., Sharifah, S.H. & Ruszymah, B.H.I. 2010. Autologous implantation of bilayered tissue-engineered respiratory epithelium for tracheal mucosal regenesis in a sheep model. Cells Tissues Organs 192(5): 292-302.

Müller, J., Ossig, C., Greiner, J.F.W., Hauser, S., Fauser, M., Widera, D., Kaltschmidt, C., Storch, A. & Kaltschmidt, B. 2015. Intrastriatal transplantation of adult human neural crest-derived stem cells improves functional outcome in parkinsonian rats. Stem Cells Translational Medicine 4(1): 31-43.

Murrell, W., Wetzig, A., Donnellan, M., Féron, F., Burne, T., Meedeniya, A., Kesby, J., Bianco, J., Perry, C. & Silburn, P. 2008. Olfactory mucosa is a potential source for autologous stem cell therapy for Parkinson’s disease. Stem Cells 26(8): 2183-2192.

Nur Adelina Ahmad Noruddin, Aminuddin B Saim, Kien Hui Chua & Ruszymah Idrus. 2007. Human nasal turbinates as a viable source of respiratory epithelial cells using co-culture system versus dispase dissociation technique. The Laryngoscope 117(12): 2139-2145.

Nur Hidayah Hassan, Ahmad Fadzli Sulong, Min-Hwei Ng, Ohnmar Htwe, Ruszymah Idrus, Sharifah Ahmad Roohi, Amaramalar Selvi Naicker & Shalimar Abdullah. 2012. Neural-differentiated mesenchymal stem cells incorporated into muscle stuffed vein scaffold forms a stable living nerve conduit. Journal of Orthopaedic Research 30(10): 1674- 1681.

Rabiatul, A.R., Lokanathan, Y., Rohaina, C.M., Chowdhury Shiplu Roy, Aminuddin, B.S. & Ruszymah, B.H.I. 2015. Surface modification of electrospun poly (methyl methacrylate)(PMMA) nanofibers for the development of in vitro respiratory epithelium model. Journal of Biomaterials Science, Polymer Edition 26(17): 1297-1311.

Rabinowitz, P.M., Slade, M.D., Galusha, D., Dixon-Ernst, C. & Cullen, M.R. 2006. Trends in the prevalence of hearing loss among young adults entering an industrial workforce 1985 to 2004. Ear and Hearing 27(4): 369-375.

Raponi, E., Agenes, F., Delphin, C., Assard, N., Baudier, J., Legraverend, C. & Deloulme, J.C. 2007. S100B expression defines a state in which GFAP-Expressing cells lose their neural stem cell potential and acquire a more mature developmental stage. Glia 55(2): 165-177.

Ruszymah Idrus, Nur Adelina, Ahmad Noruddin, Chen Hui Cheng, Yogeswaran Lokanathan & Aminuddin Bin Saim. 2014. Titanium mesh with expanded respiratory epithelial cells in tracheal reconstruction. Journal of Biomaterials and Tissue Engineering 4(5): 367-372.

Ruszymah, B., Izham, B.A., Heikal, M.M., Khor, S.F., Fauzi, M.B. & Aminuddin, B.S. 2011. Human respiratory epithelial cells from nasal turbinate expressed stem cell genes even after serial passaging. Medical Journal of Malaysia 66(5): 440-442.

Tan, C.W., Ng, M.H., Ohnmar, H., Lokanathan, Y., Nur-Hidayah, H., Roohi, S.A., Ruszymah, B.H.I., Nor-Hazla, M.H., Shalimar, A. & Amaramalar, S.N. 2013. Sciatic nerve repair with tissue engineered nerve: Olfactory ensheathing cells seeded poly (lactic-co-glygolic acid) conduit in an animal model. Indian Journal of Orthopaedics 47(6): 547-552.

Xiaodong Zhang, Kathleen M Klueber, Zhanfang Guo, Jun Cai, Chengliang Lu, Welby I Winstead, Mengsheng Qiu & Fred J Roisen 2006. Induction of neuronal differentiation of adult human olfactory neuroepithelial-derived progenitors. Brain Research 1073-1074: 109-119.

Xingjia Wu, William E Bolger & Juanita J. Anders. 2013. Fibroblasts isolated from human middle turbinate mucosa cause neural progenitor cells to differentiate into glial lineage cells. PLoS ONE 8(10): e76926.

Yang Li, Ying Sheng, Jian Min Liang, Xiao Yong Ren & Yan Cheng. 2017. Glial differentiation of human inferior turbinate-derived stem cells: A new source of cells for nerve repair. NeuroReport 28(5): 235-241.

*Pengarang untuk surat-menyurat; email: ruszyidrus@gmail.com