Research Article
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Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı ve Keratinosit Kültürü

Year 2018, Volume: 45 Issue: 1, 9 - 14, 12.03.2018
https://doi.org/10.5798/dicletip.363931

Abstract

Amaç: Bu çalışmada üç boyutlu biyoyazıcı için uygun bir biyomürekkep (hidrojel - hücre karışımı)
üretilerek keratinosit hücrelerinin biyobaskısı hedeflendi.Elde edilen hidrojel yapılı epidermis doku
kültüründe hücre proliferasyonu, canlılık analizi, hidrojel içinde hücre dağılımı ve morfolojisi
belirlendi.
Yöntemler: Hücre kültürlerinde derinin üst tabakası olan epidermiste bulunan keratinosit hücreleri
kullanılmıştır. Karakterizasyonu daha önce tamamlanmış olan HS2 insan keratinosit hücre hattı, farklı
biyopolimerhidrojeller (jelatin, aljinat, kitosan) ve karışımları içinde süspanse edilip biyobaskı için en
uygun hidrojel bulundu. 3B keratinosit kültürleri deney başlatılmasından 1, 4 ve 7 gün sonra canlılık
analizi için MTT testine ve hücre dağılımı ve morfolojisinin görüntülenmesi için Hematoksilen- Eozin
boyamasına tabi tutuldu.
Bulgular: Elde edilen MTT sonuçlarına göre hücre canlılıkları iki boyutlu (2B) kültürde elde edilen
keratinosit canlılıklarının %50'sinden yüksek çıkmıştır. MTT sonuçları keratinositlerin üretilen hidrojel
yapısı içerisinde tutunarak canlılıklarını sürdürebildiklerini göstermektedir. Elde edilen hücre içerikli
polimerik hidrojelin histoloji için kesit alımına uygun olduğu ve alınan kesitlere uygulanan
hematoksilen/eosin boyaması sonucunda da hücrelerin hidrojel içinde homojen olarak dağıldıkları ve
canlılıklarını korudukları belirlenmiştir.
Sonuç: Bu çalışmada oluşturulan epidermis benzeri doku kesitleri üç boyutlu biyoyazıcı ile üretilmiş ve
keratinositlerinhidrojeller içinde canlılıklarını sürdürüp doku iskelelerine tutundukları belirlenmiştir.
Üretilen hidrojelbiyomürekkeplerin deri doku mühendisliğinde ve özellikle yanıklarda epidermis
tabakasının onarımında hızlı ve kişiye özel tedavi seçenekleri sunma potansiyeli vardır.

References

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  • 3. Akdoğan E, Omay SB. Organ Mühendisliğinde Kök Hücre Uygulamaları. Turkiye Klinikleri J SurgMedSci. 2006;2:63-8.
  • 4. Kim I-Y, Seo S-J, Moon H-S, et al. Chitosan and itsderivativesfortissueengineeringapplications. BiotechnolAdv. 2008;26:1-21.
  • 5. Murphy SV, Atala A. 3D bioprinting of tissues and organs. NatBiotechnol. 2014;32:773-85.
  • 6. Kolesky DB, Truby RL, Gladman A, Busbee TA, Homan KA, Lewis JA. 3D bioprinting of vascularized, heterogeneouscell‐laden tissueconstructs. Adv Mater. 2014;26:3124-30.
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  • 13. Diduch DR, Jordan LC, Mierisch CM, Balian G. Marrowstromalcellsembedded in alginateforrepair of osteochondraldefects. Arthroscopy: Arthroscopy. 2000;16:571-7.
  • 14. Priya SG, Jungvid H, Kumar A. Skin tissueengineeringfortissuerepair and regeneration. TissueEngineeringPart B: Reviews. 2008;14:105-18.
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  • 16. Yücesan E, Başoğlu H, Göncü B, Kandaş NÖ, Ersoy YE, Akbaş F, Ayşan E. Mikroenkapsüle edilen paratiroid hücrelerinin in-vitrooptimizasyonu. Dicle Tıp Derg. 2017;44:373-80.
  • 17. Uslu B, Arbak S. Doku Mühendisliğinde Kitozanın Kullanım Alanları. 2010.
  • 18. Suh J-KF, Matthew HW. Application of chitosan-basedpolysaccharidebiomaterials in cartilagetissueengineering: a review. Biomaterials. 2000;21:2589-98.
  • 19. Kutlu B, Aydın T, Seda R, Akman AC, Gümüşderelioglu M, Nohutcu RM. Platelet‐richplasma‐loadedchitosanscaffolds: Preparation and growthfactorreleasekinetics. Journal of BiomedicalMaterials Research Part B: J Biomed MaterRes B ApplBiomater. 2013;101:28-35.
  • 20. Pandey AR, Singh US, Momin M, Bhavsar C. Chitosan: Application in tissueengineering and skin grafting. J PolymRes. 2017;24:125.
  • 21. Liu X, Smith LA, Hu J, Ma PX. Biomimeticnanofibrousgelatin/apatite compositescaffoldsfor bone tissueengineering. Biomaterials. 2009;30:2252-8.
  • 22. Hori K, Sotozono C, Hamuro J, et al. Controlled-release of epidermalgrowthfactorfromcationizedgelatinhydrogelenhancescornealepithelialwoundhealing. J Control Release. 2007;118:169-76.
  • 23. Lien S-M, Ko L-Y, Huang T-J. Effect of pore size on ECM secretion and cellgrowth in gelatins caffoldforarticular cartilage tissueen gineering. ActaBiomater. 2009;5:670-9.
  • 24. Kang H-W, Tabata Y, Ikada Y. Fabrication of porousgelatinscaffoldsfortissueen gineering. Biomaterials. 1999;20:1339-44.
  • 25. Skardal A, Zhang J, Prestwich GD. Bioprintingvessellikeconstructsusinghyaluronanhydrogelscrosslinkedwithtetrahedralpolyethyleneglycoltetracrylates. Biomaterials. 2010;31:6173-81.
  • 26. Highley CB, Rodell CB, Burdick JA. Direct 3dprinting of shear-thinninghydrogelsinto selfhealinghydrogels. Adv. Mater. 2015;27:5075–9.
  • 27. Hinton TJ, Jallerat Q, Palchesko RN, et al. Three-dimensionalprintingof complexbiologicalstructuresbyfreeformreversibleembedding of suspendedhydrogels. Sci. Adv. 2015; 1:e1500758.
  • 28. Colosi C,Shin SR, Manoharan V, et al. Microfluidicbioprinting of heterogeneous 3d tissueconstructsusinglowviscositybioink. Adv. Mater. 2015,28:677–84.
  • 29. Augst AD, Kong HJ, Mooney DJ. Alginatehydrogels as biomaterials. Macromol. Biosci. 2006;6:623– 33.
  • 30. Duan B, Hockaday LA, Kang KH, Butcher JT. 3D bioprinting of heterogeneousaorticvalveconduitswithalginate/gelatinhydrogels. J Biomed Mater Res A.2013;101:1255-64.
  • 31. Koch L, Deiwick A, Schlie S, et al. Skintissuegenerationbylasercellprinting. BiotechnoBioeng. 2012;109:1855-63.
  • 32. Miller JS. Thebillioncellconstruct: willthree-dimensionalprintingget us there? PLoSBiol. 2014; 12:e1001882.
  • 33. Murphy SV,Atala A. 3D bioprinting of tissuesand organs. Nat. Biotechnol. 2014;32:773–85.
  • 34. Blaeser A, Campos DFD, Puster U, RichteringW,Stevens MM, Fischer H. Controllingshearstress in 3dbioprinting is a keyfactortobalanceprintingresolution andstemcellintegrity. Adv. Healthc. Mater. 2016;5:326–33.
  • 35. Fedorovich NE, Schuurman W, Wijnberg HM et al. Biofabrication of osteochondraltissueequivalentsbyprintingtopologicallydefined, cell-laden hydrogelscaffolds. TissueEng C Methods.2012;18:33–44.
  • 36. Luo Y, Lode A, Gelinsky M. Direct plottingof three-dimensionalhollow fiberscaffoldsbased on concentratedalginatepastesfortissueengineering. Adv Healthcare Mater. 2013;2:777–83.
  • 37. Malda J, Visser J, Melchels FP et al. 25th Anniversary article: engineeringhydrogelsforbiofabrication. Adv Mater. 2013;25:5011–28.
  • 38. Cohen DL, Malone E, Lipson H et al. Direct freeformfabrication of seededhydrogels in arbitrarygeometries. TissueEng. 2006; 12:1325–35.
  • 39. Chaudhari AA, Vig K, Baganizi DR, Futureprospectsforscaffoldingmethods and biomaterials in skin tissueengineering: a review. Int J MolSci. 2016;17:1974.
Year 2018, Volume: 45 Issue: 1, 9 - 14, 12.03.2018
https://doi.org/10.5798/dicletip.363931

Abstract

References

  • 1. Lanza R, Langer R, Vacanti JP. Principles of tissueengineering: Academicpress; 2011.
  • 2. O'brien FJ. Biomaterials&scaffoldsfortissueengineering. Mater Today. 2011;14:88-95.
  • 3. Akdoğan E, Omay SB. Organ Mühendisliğinde Kök Hücre Uygulamaları. Turkiye Klinikleri J SurgMedSci. 2006;2:63-8.
  • 4. Kim I-Y, Seo S-J, Moon H-S, et al. Chitosan and itsderivativesfortissueengineeringapplications. BiotechnolAdv. 2008;26:1-21.
  • 5. Murphy SV, Atala A. 3D bioprinting of tissues and organs. NatBiotechnol. 2014;32:773-85.
  • 6. Kolesky DB, Truby RL, Gladman A, Busbee TA, Homan KA, Lewis JA. 3D bioprinting of vascularized, heterogeneouscell‐laden tissueconstructs. Adv Mater. 2014;26:3124-30.
  • 7. Bell E, Ehrlich HP, Buttle DJ, Nakatsuji T. Livingtissueformed in vitro and accepted as skinequivalenttissue of fullthickness. Science. 1981;211(4486):1052-4.
  • 8. Billiet T, Vandenhaute M, Schelfhout J, Van Vlierberghe S, Dubruel P. A review of trends and limitations in hydrogel-rapidprototypingfortissueengineering. Biomaterials. 2012;33:6020-41.
  • 9. Currie LJ, Sharpe JR, Martin R. Theuse of fibrin glue in skin grafts and tissue-engineered skin replacements. PlastReconstrSurg. 2001;108:1713-26.
  • 10. Auger FA, Berthod F, Moulin V, Pouliot R, Germain L. Tissue‐engineered skin substitutes: from in vitroconstructsto in vivoapplications. BiotechnolApplBiochem. 2004;39:263-75.
  • 11. Stanton M, Samitier J, Sanchez S. Bioprinting of 3D hydrogels. LabChip. 2015;15:3111-5.
  • 12. Mandrycky C, Wang Z, Kim K, Kim D-H. 3D bioprintingforengineeringcomplextissues. BiotechnolAdv. 2016;34:422-34.
  • 13. Diduch DR, Jordan LC, Mierisch CM, Balian G. Marrowstromalcellsembedded in alginateforrepair of osteochondraldefects. Arthroscopy: Arthroscopy. 2000;16:571-7.
  • 14. Priya SG, Jungvid H, Kumar A. Skin tissueengineeringfortissuerepair and regeneration. TissueEngineeringPart B: Reviews. 2008;14:105-18.
  • 15. Fragonas E, Valente M, Pozzi-Mucelli M, et al. Articularcartilagerepair in rabbitsbyusingsuspensions of allogenicchondrocytes in alginate. Biomaterials. 2000;21:795-801.
  • 16. Yücesan E, Başoğlu H, Göncü B, Kandaş NÖ, Ersoy YE, Akbaş F, Ayşan E. Mikroenkapsüle edilen paratiroid hücrelerinin in-vitrooptimizasyonu. Dicle Tıp Derg. 2017;44:373-80.
  • 17. Uslu B, Arbak S. Doku Mühendisliğinde Kitozanın Kullanım Alanları. 2010.
  • 18. Suh J-KF, Matthew HW. Application of chitosan-basedpolysaccharidebiomaterials in cartilagetissueengineering: a review. Biomaterials. 2000;21:2589-98.
  • 19. Kutlu B, Aydın T, Seda R, Akman AC, Gümüşderelioglu M, Nohutcu RM. Platelet‐richplasma‐loadedchitosanscaffolds: Preparation and growthfactorreleasekinetics. Journal of BiomedicalMaterials Research Part B: J Biomed MaterRes B ApplBiomater. 2013;101:28-35.
  • 20. Pandey AR, Singh US, Momin M, Bhavsar C. Chitosan: Application in tissueengineering and skin grafting. J PolymRes. 2017;24:125.
  • 21. Liu X, Smith LA, Hu J, Ma PX. Biomimeticnanofibrousgelatin/apatite compositescaffoldsfor bone tissueengineering. Biomaterials. 2009;30:2252-8.
  • 22. Hori K, Sotozono C, Hamuro J, et al. Controlled-release of epidermalgrowthfactorfromcationizedgelatinhydrogelenhancescornealepithelialwoundhealing. J Control Release. 2007;118:169-76.
  • 23. Lien S-M, Ko L-Y, Huang T-J. Effect of pore size on ECM secretion and cellgrowth in gelatins caffoldforarticular cartilage tissueen gineering. ActaBiomater. 2009;5:670-9.
  • 24. Kang H-W, Tabata Y, Ikada Y. Fabrication of porousgelatinscaffoldsfortissueen gineering. Biomaterials. 1999;20:1339-44.
  • 25. Skardal A, Zhang J, Prestwich GD. Bioprintingvessellikeconstructsusinghyaluronanhydrogelscrosslinkedwithtetrahedralpolyethyleneglycoltetracrylates. Biomaterials. 2010;31:6173-81.
  • 26. Highley CB, Rodell CB, Burdick JA. Direct 3dprinting of shear-thinninghydrogelsinto selfhealinghydrogels. Adv. Mater. 2015;27:5075–9.
  • 27. Hinton TJ, Jallerat Q, Palchesko RN, et al. Three-dimensionalprintingof complexbiologicalstructuresbyfreeformreversibleembedding of suspendedhydrogels. Sci. Adv. 2015; 1:e1500758.
  • 28. Colosi C,Shin SR, Manoharan V, et al. Microfluidicbioprinting of heterogeneous 3d tissueconstructsusinglowviscositybioink. Adv. Mater. 2015,28:677–84.
  • 29. Augst AD, Kong HJ, Mooney DJ. Alginatehydrogels as biomaterials. Macromol. Biosci. 2006;6:623– 33.
  • 30. Duan B, Hockaday LA, Kang KH, Butcher JT. 3D bioprinting of heterogeneousaorticvalveconduitswithalginate/gelatinhydrogels. J Biomed Mater Res A.2013;101:1255-64.
  • 31. Koch L, Deiwick A, Schlie S, et al. Skintissuegenerationbylasercellprinting. BiotechnoBioeng. 2012;109:1855-63.
  • 32. Miller JS. Thebillioncellconstruct: willthree-dimensionalprintingget us there? PLoSBiol. 2014; 12:e1001882.
  • 33. Murphy SV,Atala A. 3D bioprinting of tissuesand organs. Nat. Biotechnol. 2014;32:773–85.
  • 34. Blaeser A, Campos DFD, Puster U, RichteringW,Stevens MM, Fischer H. Controllingshearstress in 3dbioprinting is a keyfactortobalanceprintingresolution andstemcellintegrity. Adv. Healthc. Mater. 2016;5:326–33.
  • 35. Fedorovich NE, Schuurman W, Wijnberg HM et al. Biofabrication of osteochondraltissueequivalentsbyprintingtopologicallydefined, cell-laden hydrogelscaffolds. TissueEng C Methods.2012;18:33–44.
  • 36. Luo Y, Lode A, Gelinsky M. Direct plottingof three-dimensionalhollow fiberscaffoldsbased on concentratedalginatepastesfortissueengineering. Adv Healthcare Mater. 2013;2:777–83.
  • 37. Malda J, Visser J, Melchels FP et al. 25th Anniversary article: engineeringhydrogelsforbiofabrication. Adv Mater. 2013;25:5011–28.
  • 38. Cohen DL, Malone E, Lipson H et al. Direct freeformfabrication of seededhydrogels in arbitrarygeometries. TissueEng. 2006; 12:1325–35.
  • 39. Chaudhari AA, Vig K, Baganizi DR, Futureprospectsforscaffoldingmethods and biomaterials in skin tissueengineering: a review. Int J MolSci. 2016;17:1974.
There are 39 citations in total.

Details

Journal Section Research Articles
Authors

Aylin Şendemir Ürkmez This is me 0000-0003-1818-6651

Umut Doğu Seçkin This is me 0000-0002-0123-0284

Cansu Görgün This is me 0000-0002-0460-2952

Yiğit Uyanıkgil 0000-0002-4016-0522

Publication Date March 12, 2018
Submission Date December 8, 2017
Published in Issue Year 2018 Volume: 45 Issue: 1

Cite

APA Ürkmez, A. Ş., Seçkin, U. D., Görgün, C., Uyanıkgil, Y. (2018). Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı ve Keratinosit Kültürü. Dicle Tıp Dergisi, 45(1), 9-14. https://doi.org/10.5798/dicletip.363931
AMA Ürkmez AŞ, Seçkin UD, Görgün C, Uyanıkgil Y. Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı ve Keratinosit Kültürü. diclemedj. March 2018;45(1):9-14. doi:10.5798/dicletip.363931
Chicago Ürkmez, Aylin Şendemir, Umut Doğu Seçkin, Cansu Görgün, and Yiğit Uyanıkgil. “Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı Ve Keratinosit Kültürü”. Dicle Tıp Dergisi 45, no. 1 (March 2018): 9-14. https://doi.org/10.5798/dicletip.363931.
EndNote Ürkmez AŞ, Seçkin UD, Görgün C, Uyanıkgil Y (March 1, 2018) Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı ve Keratinosit Kültürü. Dicle Tıp Dergisi 45 1 9–14.
IEEE A. Ş. Ürkmez, U. D. Seçkin, C. Görgün, and Y. Uyanıkgil, “Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı ve Keratinosit Kültürü”, diclemedj, vol. 45, no. 1, pp. 9–14, 2018, doi: 10.5798/dicletip.363931.
ISNAD Ürkmez, Aylin Şendemir et al. “Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı Ve Keratinosit Kültürü”. Dicle Tıp Dergisi 45/1 (March 2018), 9-14. https://doi.org/10.5798/dicletip.363931.
JAMA Ürkmez AŞ, Seçkin UD, Görgün C, Uyanıkgil Y. Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı ve Keratinosit Kültürü. diclemedj. 2018;45:9–14.
MLA Ürkmez, Aylin Şendemir et al. “Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı Ve Keratinosit Kültürü”. Dicle Tıp Dergisi, vol. 45, no. 1, 2018, pp. 9-14, doi:10.5798/dicletip.363931.
Vancouver Ürkmez AŞ, Seçkin UD, Görgün C, Uyanıkgil Y. Deri Doku Mühendisliği Amaçlı Üç Boyutlu Biyobaskı ve Keratinosit Kültürü. diclemedj. 2018;45(1):9-14.