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. 2023;30(2):222-233.
doi: 10.15388/Amed.2023.30.2.15. Epub 2023 Jul 31.

Regeneration of Rabbit Auricular Cartilage After the Intravenous Stem Cell Injection

Pavlo Virych  1 , Nadiia Shuvalova  1 , Anton Karas  1 , Galina Karas  1 , Svitlana Chaika  1 , Tetiana Kucherenko  1 , Ganna Minina  1 , Marina Timchenko  1 , Oleg Melnykov  1 , Yurii Minin  1
Affiliations

Regeneration of Rabbit Auricular Cartilage After the Intravenous Stem Cell Injection

Pavlo Virych et al. Acta Med Litu. 2023.

Abstract
in English, Italian

Background: The restoration of auricular cartilage is a major problem of otolaryngology. The low regenerative capacity of cartilage requires alternative approaches such as cell and tissue engineering. Stem cells are one of the ways to repair auricular cartilage damages. The aim of the investigation was the regeneration of an artificial defect of the auricular cartilage of rabbits after the intravenous injection of stem cells.

Materials and methods: The study was carried out on rabbits. A narrow strip of auricular cartilage was surgically removed. A previously prepared suspension of homologous mesenchymal stem cells (5 million) in 0.5 ml physiological solution was injected into the vein of the opposite ear. Tissue samples from the site of the injury were collected after 1, 2, and 3 months. Histological examinations of the tissues were carried out after staining with fuchsin-eosin, azure II-eosin, and according to Weigert. In addition, the amount of interleukin-6 (IL-6) and the transforming growth factor β1 (TGF-β1) in the blood serum were determined.

Results: The main method of healing is the formation of a connective tissue scar. Yret, an increase of the number of fibroblasts and single islands of the newly formed auricular cartilage was found, which indicates the migration of the injected stem cells to the site of the damage and settling there. The intravenous injection of stem cells did not affect the secretion of pro-inflammatory IL-6, but significantly increased the amount of TGF-β1.

Conclusions: We assume that regenerative processes were stimulated. Nevertheless, they were aimed at quickly restoring the tissue integrity through the typical stages of scar formation. The restoration of cartilage integrity requires additional regulatory factors which will determine the chondrogenic differentiation of stem cells.

Kontekstas: Ausies kremzliᶙ atkūrimas yra svarbi otolaringologijos problema. Kadangi kremzliᶙ regeneracijos pajėgumas yra nedidelis, būtini alternatyvūs sprendimai – pavyzdžiui, ląsteliᶙ ir audiniᶙ inžinerija. Kamieninės ląstelės yra vienas iš būdᶙ atkurti ausᶙ kremzlėms padarytą žalą. Šio tyrimo tikslas buvo dirbtinai suformuoto triušio ausies žalos defekto regeneracija po intraveninės kamieniniᶙ ląsteliᶙ injekcijos.

Medžiagos ir metodai: Šis tyrimas atliktas su triušiais. Chirurginiu būdu buvo pašalinta siaura triušio ausies kremzliᶙ juostelė. Iš anksto buvo paruošta homologizuota mezenchiminiᶙ kamieniniᶙ ląsteliᶙ (5 milijonai) suspensija su 0,5 ml fiziologinio tirpalo. Šios suspensijos buvo įšvirkšta į kitos ausies veną. Sužeidimo vietos audiniᶙ mėginiai imti po 1, 2 ir 3 mėnesiᶙ. Surinktᶙ audiniᶙ histologinis tyrimas buvo atliktas ištepus juos su fuchsinu ir eozinu, azūro II-eozinu bei naudojant Weigerto metodiką. Be to, krau jo serume buvo nustatytas interleukino-6 (IL-6) ir transformuojančio augimo faktoriaus β1 (TGF-β1) kiekis.

Rezultatai: Pagrindinis gijimo metodas yra jungiamojo audinio rando susidarymas. Buvo nustatytas fibroblastᶙ skaičiaus padidėjimas. Taip pat nustatyta naujai susidariusiᶙ ausies kremzliᶙ saleliᶙ. Tai rodo, kad įšvirkštos kamieninės ląstelės migruoja į sužeidimo vietą ir ten įsikuria. Intraveninė kamieniniᶙ ląsteliᶙ injekcija nepaveikė prouždegiminio IL-6 išskyrimo, tačiau reikšmingai padidino TGF-β1 kiekį.

Išvados: Darome prielaidą, kad buvo stimuliuojami regeneraciniai procesai. Tačiau jais siekta greitai atkurti audinio integralumą įprastiniais rando susidarymo etapais. Kremzliᶙ integralumui atkurti reikia papildomᶙ reguliuojamᶙjᶙ veiksniᶙ, kurie paskatintᶙ chondrogeninę kamieniniᶙ ląsteliᶙ diferenciaciją.

Keywords: auricular cartilage; inflammation; intercellular matrix; regeneration; stem cells.

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Figures

Figure 1
Figure 1
Surface antigen expression by mesenchymal stem cells (MSCs) at passage 2: A – CD34, B – CD45, C – CD73, D – CD105. The number of living cells was about 75% of the total population (E); (flow cytometry by FACSAria).
Figure 2
Figure 2
Morphological characteristics of artificially damaged rabbit auricular cartilage after one (1) and three months (2) after surgery. A – without MSCs injection, B – intravenous MSCs injection (staining by hematoxylin-eosin, magnification x100).
Figure 3
Figure 3
The density of the elastin mesh of the connective tissue of the skin (C), auricular cartilage (1) and tissue at the point of cartilage damage (2) for 1, 2 and 3 months after surgery (tissue staining by Weigert, grayscale value, medians and quartiles).
Figure 4
Figure 4
The concentration of IL-6 (A) and TGF-β1 (B) in the blood plasma of rabbits after 1, 2 and 3 months after the artificial auricular cartilage damage and the injection of stem cells. Control – animals without stem cells injection, MSC – intravenous injection of 5•106 of homologous mesenchymal stem cells in saline (M±SD, *p<0.05).
See this image and copyright information in PMC

References

    1. Bichara DA, O’Sullivan NA, Pomerantseva I, et al. The tissue-engineered auricle: past, present, and future. Tissue Eng Part B Rev. 2012;18(1):51–61. doi: 10.1089/ten.TEB.2011.0326. - DOI - PubMed
    1. de Chalain T, Phillips JH, Hinek A. Bioengineering of elastic cartilage with aggregated porcine and human auricular chondrocytes and hydrogels containing alginate, collagen, and kappa-elastin. J Biomed Mater Res. 1999;44(3):280–288. doi: 10.1002/(sici)1097-4636(19990305)44:3<280::aid-jbm6>3.0.co;2-h. - DOI - PubMed
    1. García-López J, Garciadiego-Cázares D, Melgarejo-Ramírez Y, et al. Chondrocyte differentiation for auricular cartilage reconstruction using a chitosan based hydrogel. Histol Histopathol. 2015;30(12):1477–1485. doi: 10.14670/HH-11-642. - DOI - PubMed
    1. García-Martínez L, Campos F, Godoy-Guzmán C, et al. Encapsulation of human elastic cartilage-derived chondrocytes in nanostructured fibrin-agarose hydrogels. Histochem Cell Biol. 2017;147(1):83–95. doi: 10.1007/s00418-016-1485-9. - DOI - PubMed
    1. Li L, Yu F, Zheng L, et al. Natural hydrogels for cartilage regeneration: Modification, preparation and application. J Orthop Translat. 2018;17:26–41. doi: 10.1016/j.jot.2018.09.003. - DOI - PMC - PubMed

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