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Excised human larynx in N-vinyl-2-pyrrolidone-embalmed cadavers can produce voiced sound by pliable vocal fold vibration

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Abstract

Tissue-hardening effect and health-hazard issue of formaldehyde (FA) have long been a great disadvantage of this conventional fixative in anatomical research. We recently developed a FA-free embalming method for cadavers which utilizes N-vinyl-2-pyrrolidone (NVP) and enables assessment of motion kinetics by maintaining the softness of embalmed tissue. By assessing the feasibility of NVP-embalmed tissue to mimic vocalization, this study aimed to prove the potential of embalmed cadavers, which have previously been used only for the understanding of anatomical morphology, for the assessment of precise motion physiology in the human body. Ten cadavers embalmed in NVP (n = 6) and FA (n = 4) were incorporated in this study. Excised larynges underwent experimental phonation to mimic vocalization with fast and pliable vibration of vocal folds. High-speed digital imaging was utilized for the assessment of vocal fold vibration. Furthermore, acoustic analysis of the voiced sound, and reproducibility examination were also performed. Regular vocal fold vibrations successfully produced voiced sounds during experimental phonation using NVP-embalmed larynges. The vibratory frequency, vibration amplitude, and stretch rate of the vocal folds were comparable to those of living humans. Six months after the first experiment, the vocal parameters were reproduced, to suggest the long-term preservation potential of our NVP-embalming technique. On the other hand, neither voiced sound nor vocal fold vibration were observed in FA-embalmed larynges. This novel embalming technique could pioneer the next era to utilize embalmed cadavers for the examination of motion physiology in the human body.

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References

  • Alipour F, Finnegan EM, Jaiswal S (2013) Phonatory characteristics of the human larynx in comparison to other species. J Voice 27:441–447

    Article  PubMed  PubMed Central  Google Scholar 

  • Balta JY, Cronin M, Cryan JF, O’Mahony SM (2015) Human preservation techniques in anatomy: a 21st century medical education perspective. Clin Anat 28:725–734

    Article  PubMed  Google Scholar 

  • Barton DP, Davies DC, Mahadevan V, Dennis L, Adib T, Mudan S, Sohaib A, Ellis H (2009) Dissection of soft-preserved cadavers in the training of gynaecological oncologists: report of the first UK workshop. Gynecol Oncol 113:352–356

    Article  PubMed  Google Scholar 

  • Blaugrund SM, Taira T, el-Assuooty A, Lin PT, Isshiki N, Gould WJ (1990) Effects of lateral manual compression upon glottic incompetence: objective evaluations. Ann Otol Rhinol Laryngol 99:249–255

    Article  CAS  PubMed  Google Scholar 

  • Blum F (1893) Der Formaldehyd als härtungsmittel. Z Wiss Mikrosk 10:314–315

    Google Scholar 

  • Campbell JW, Margrave JL (1998) Preservative and embalming fluid. Google Patents

  • Conroy ER, Hennick TM, Awan SN, Hoffman MR, Smith BL, Jiang JJ (2013) Effect of variations to a simulated system of straw phonation therapy on aerodynamic parameters using excised canine larynges. J Voice 28:1–6

    Article  PubMed  PubMed Central  Google Scholar 

  • der Berg V (1962) Modern research in experimental phoniatrics. Folia Phoniat 14:81–149

    Article  PubMed  Google Scholar 

  • Enver N, Doruk C, Kara E, Asliyüksek BB (2018) A morphometric analysis of laryngeal anatomy: a cadaveric study. Tr-ENT 28:71–77

    Article  Google Scholar 

  • Fox CH, Johnson FB, Whiting J, Roller PP (1985) Formaldehyde fixation. J Histochem Cytochem 33:845–853

    Article  CAS  PubMed  Google Scholar 

  • Garcia M, Herbst CT (2018) Excised larynx experimentation: history, current developments, and prospects for bioacoustics research. Anthropol Sci 126:9–17

    Article  Google Scholar 

  • Goyri-O’Neill J, Pais D, Freire de Andrade F, Ribeiro P, Belo A, O’Neill A, Ramos S, Neves Marques C (2013) Improvement of the embalming perfusion method: the innovation and the results by light and scanning electron microscopy. Acta Med Port 26:188–194

    PubMed  Google Scholar 

  • Graham CR (2014) Fundamental frequency range in Japanese and English: the case of simultaneous bilinguals. Phonetica 71:271–295

    Article  PubMed  Google Scholar 

  • Haaf F, Sanner A, Straub F (1985) Polymers of N-vinylpyrrolidone: synthesis, characterization and uses. Polym J 17:143–152

    Article  CAS  Google Scholar 

  • Haizuka Y, Nagase M, Takashino S, Kobayashi Y, Fujikura Y, Matsumura G (2018) A new substitute for formalin: application to embalming cadavers. Clin Anat 31:90–98

    Article  PubMed  Google Scholar 

  • Hayashi S, Homma H, Naito M et al (2014) Saturated salt solution method: a useful cadaver embalming for surgical skills training. Medicine 93:e196

    Article  PubMed  PubMed Central  Google Scholar 

  • Hayashi S, Naito M, Kawata S et al (2016) History and future of human cadaver preservation for surgical training: from formalin to saturated salt solution method. Anat Sci Int 91:1–7

    Article  CAS  PubMed  Google Scholar 

  • Holliday MA, Bones VM, Malekzadeh S, Grant NN (2015) Low-cost modular phonosurgery training station: development and validation. Laryngoscope 125:1409–1413

    Article  PubMed  Google Scholar 

  • IARC (1999) N-vinyl-2-pyrrolidone and polyvinyl pyrrolidone. IARC Monogr Eval Carcinog Risks Hum 71(Pt 3):1181–1187

    Google Scholar 

  • IARC (2006) International Agency for Research on Cancer Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. No. 88- Formaldehyde, 2-Butoxyethanol and 1-tert Butoxy-2-propanol

  • Ishikawa CC, Pinheiro TG, Hachiya A, Montagnoli AN, Tsuji DH (2017) Impact of cricothyroid muscle contraction on vocal fold vibration: experimental study with high-speed videoendoscopy. J Voice 31:300–306

    Article  PubMed  Google Scholar 

  • Jansen MM, Hazenberg CEVB, de Ruiter QMB, van Hamersvelt RW, Bleys RLAW, van Herwaarden JA (2020) Feasibility of fresh frozen human cadavers as a research and training model for endovascular image guided interventions. PLoS ONE 15:e0242596. https://doi.org/10.1371/journal.pone.0242596 (eCollection 2020. PMID: 33254200)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kawaida M, Fukuda H, Kano S, Shiotani A, Kohno N (1990) Dynamic movement of air tract fluid in lubrication of the larynx during phonation: a basic study using excised canine larynges and experimental air tract fluid by means of X-ray stroboscope system. Auris Nasus Larynx 16:237–243

    Article  CAS  PubMed  Google Scholar 

  • Kawamura Y, Okamura H, Yumoto E (1986) Multiparametric display of voice dysfunction: use of radar chart. Nihon Jibiinnkoka Gakkai Kaiho 89:448–454

    Article  CAS  Google Scholar 

  • Lagier A, Guenoun D, Legou T, Espesser R, Giovanni A, Champsaur P (2017) Control of the glottal configuration in ex vivo human models: quantitative anatomy for clinical and experimental practices. Surg Radiol Anat 39:257–262

    Article  PubMed  Google Scholar 

  • Macchi V, Munari PF, Brizzi E, Parenti A, Caro RD (2003) Workshop in clinical anatomy for residents in gynecology and obstetrics. Clin Anat 16:440–447

    Article  CAS  PubMed  Google Scholar 

  • Maruyama K, Yokoi H, Nagase M et al (2019) Usefulness of N-Vinyl-2-pyrrolidone embalming for endoscopic transnasal skull base approach in cadaver dissection. Neurol Med Chir 59:379–383

    Article  Google Scholar 

  • Messmer C, Kellogg RT, Zhang Y, Baiak A, Leiweke C, Marcus JR, Levin LS, Zenn MR, Erdmann D (2010) A technique to perfuse cadavers that extends the useful life of fresh tissues: the Duke experience. Anat Sci Educ 3:191–194

    Article  PubMed  Google Scholar 

  • Nagase M, Kimoto Y, Sunami E, Matsumura G (2020) A new human cadaver model for laparoscopic training using N-vinyl-2-pyrrolidone: a feasibility study. Anat Sci Int 95:156–164

    Article  CAS  PubMed  Google Scholar 

  • Nakagawa H, Fukuda H, Kawaida M, Shiotani A, Kanzaki J (1998) Lubrication mechanism of the larynx during phonation: an experimental in excised canine larynges. Folia Phoniatr Logop 50:183–194

    Article  CAS  PubMed  Google Scholar 

  • Nishizawa N (1989) Stereoendoscopic observation of the Larynx -Vocal fold length in respiration and in phonation with vocal pitch change-. Nihon Jibiinkoka Gakkai Kaiho 92(8):1239–1252 (in Japanese. PMID: 2585198)

    Article  CAS  PubMed  Google Scholar 

  • Omori K, Slavit DH, Kacker A, Blaugrund SM, Kojima H (2000) Effects of thyroplasty type I on vocal fold vibration. Laryngoscope 110:1086–1091

    Article  CAS  PubMed  Google Scholar 

  • Pinheiro TG, Ishikawa CC, Imamura R, Frizzarini R, Montagnoli AN, Tsuji DH (2019) Fundamental frequency in excised human larynges after anterior Glottic web formation of various extents. Otolaryngol Head Neck Surg 160:672–678

    Article  PubMed  Google Scholar 

  • Salazar J, Gras JR, Sanchez-Guillen L, Sánchez-Del-Campo F, Arroyo A (2021) Phonosurgery training in human larynx preserved with thiel’s embalming method. ORL. https://doi.org/10.1159/000512725 (Online ahead of print. PMID: 34530430)

    Article  PubMed  Google Scholar 

  • Shiotani A, Fukuda H, Kawaida M, Kanzaki J (1996) Vocal fold vibration in simulated head voice phonation in excised canine larynges. Eur Arch Otorhinolaryngol 253:356–363

    Article  CAS  PubMed  Google Scholar 

  • Svec JG, Schutte HK, Miller DG (1999) On pitch jumps between chest and falsetto registers in voice: data from living and excised human larynges. J Acoust Soc Am 106:1523–1531

    Article  CAS  PubMed  Google Scholar 

  • Thiel W (1992) The preservation of the whole corpse with natural color. Ann Anat 174:185–195

    Article  CAS  PubMed  Google Scholar 

  • Thiel W (2002) Supplement to the conservation of an entire cadaver according to W. Thiel Ann Anat 184:267–269

    Article  PubMed  Google Scholar 

  • Tolhurst DE, Hart J (1990) Cadaver preservation and dissection. Eur J Plast Surg 13:75–78

    Article  Google Scholar 

  • Tsuji DH, de Almeid ER, Sennes LU, Butugan O, Pinho SMR (2003) Comparison between thyroplasty type I and Arytenoid rotation: a study of vocal fold vibration using excised human larynges. J Voice 17:596–604

    Article  PubMed  Google Scholar 

  • Xue C, Pulvermacher A, Calawerts W, Devine E, Jiang J (2017) An excised canine model of anterior glottic web and its acoustic, aerodynamic, and high-speed measurements. J Voice 31:246.e21-246.e32

    Article  Google Scholar 

  • Zhuang P, Sprecher AJ, Hoffman MR et al (2009) Phonation threshold flow measurements in normal and pathological phonation. Laryngoscope 119:811–815

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We would like to thank Editage (www.editage.com) for English language editing. Part of this study was supported by JSPS KAKENHI Grant-in-Aid for Scientific Research(C) Number 20K09737.

Funding

Part of this study was supported by JSPS KAKENHI Grant-in-Aid for Scientific Research(C) Number 20K09737.

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Authors and Affiliations

  1. Department of Otolaryngology-Head and Neck Surgery, Kyorin University School of Medicine, 6-20-2 Shinkawa, Mitaka, Tokyo, 1818611, Japan

    Makoto Miyamoto, Itaru Watanabe, Hideki Nakagawa & Koichiro Saito

  2. Department of Anatomy, Kyorin University School of Medicine, Tokyo, Japan

    Miki Nagase & George Matsumura

  3. Department of Hygiene and Public Health, Kyorin University School of Medicine, Tokyo, Japan

    Kanae Karita

  4. Department of Otolaryngology, University of São Paulo School of Medicine, São Paulo, Brazil

    Domingos Hiroshi Tsuji

  5. Department of Electric Engineering, Federal University of São Carlos, São Paulo, Brazil

    Arlindo Neto Montagnoli

Authors
  1. Makoto Miyamoto
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  2. Miki Nagase
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  3. Itaru Watanabe
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  4. Hideki Nakagawa
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  6. Domingos Hiroshi Tsuji
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  7. Arlindo Neto Montagnoli
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  8. George Matsumura
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  9. Koichiro Saito
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Contributions

Conceptualization: MM, MN, HN, DHT, KS. Data Curation: MM, MN, IW, KK, ANM, GM, KS. Formal analysis: MM, MN, HN, ANM. Funding acquisition: KS. Investigation: MM, IW, KS. Methodology: MM, HN, DHT, KS. Project administration: MN, KS. Resources: MN, DHT, GM. Software: ANM. Supervision: MN, KS. Validation: MM, MN, IW, HN, KK, DHT, GM, KS. Visualization: MM, MN, DHT, ANM, GM, KS. Writing—Original Draft: MM, KS. Writing—Review and Editing: MN, IW, HN, KK, DHT, ANM, GM, KS.

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Correspondence to Koichiro Saito.

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Miyamoto, M., Nagase, M., Watanabe, I. et al. Excised human larynx in N-vinyl-2-pyrrolidone-embalmed cadavers can produce voiced sound by pliable vocal fold vibration. Anat Sci Int 97, 347–357 (2022). https://doi.org/10.1007/s12565-021-00646-3

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