A Comparison of Cell Micro-encapsulation Efficiency of a Manually Produced and Capsulation System by Depending on the Cell Isolation Type and the Alginate Percentage

Abstract

Objective: Many industrial sectors in the health sciences are using polymeric materials for different processes. One of these processes is the encapsulation system mainly preferred in therapeutic applications including cell transplantation. Depending on the main approach, the polymer type and sphere properties differ by the encapsulation method. Alginate is one of the natural polymers that show structural changes depending on different polymer ratios. In this study, our aim is to determine the ideal micro-encapsulation procedures by evaluating different alginate concentrations for the capsulation process of parathyroid cells.

Material and Method: In this study, cell isolation of single parathyroid hyperplasia tissue from a patient with secondary hyperparathyroidism was performed by using two methods including mechanical and enzymatic isolation. Two different alginate percentages were used for micro-encapsulation which created manually, and a capsulation system made with two different flow rates. Parathormone levels of micro-encapsulated cells were followed for 64-79 days in vitro.

Results: Morphological stability was observed for 2% alginate concentration with a flow rate of 2 mL/min used group from the micro-encapsulation system. However, parathyroid hormone release showed similar results between cell isolation type and alginate percentages.

Conclusion: Increasing the long-term micro-encapsulation efficiency, certain criteria should be determined from structural and functional aspects. In this study, the microencapsulation of parathyroid cells ob-tained by the enzymatic isolation method formed a more stable structure at increasing flow rate by using the capsulation system.

Keywords

• Parathyroid
• micro-encapsulation
• ultra-pure alginate
• cap-sulation system
• cell isolation

Hücre Mikroenkapsülasyonunda Manuel ve Kapsülasyon Sisteminin Hücre İzolasyon Tipi ve Aljinat Yüzdesine Bağlı Verimliliğinin Karşılaştırılması

Abstract

Amaç: Birçok endüstriyel sektörde kullanılan polimer malzemeler sağlık bilimlerinde de farklı işlemlerde kullanılmaktadır. Bu işlem-lerden biri olan enkapsülasyon sistemi hücre nakli gibi terapötik uygulamalarda tercih edilmektedir. Enkapsülasyon çalışmalarında uygulanacak yaklaşıma göre kapsül yapısında kullanılacak polimer malzeme ve oluşan kapsül boyutu değişmektedir. Aljinat, kahverengi alglerden elde edilen, içeriğindeki farklı polimerik blok oranlarına bağlı olarak değişiklik gösteren doğal polimerlerden biridir. Bu çalışmada, kapsülasyon aşaması için kullanılacak olan değişik aljinat yüzdeleri uygulanarak paratiroid hücrelerinde ideal mikroenkapsülasyon prosedürlerinin belirlenmesi amaçlanmaktadır.

Gereç ve Yöntem: Çalışmada sekonder hiperparatiroidi hastasın-dan alınan bir adet paratiroid hiperplazi dokusundan mekanik ve enzimatik izolasyon yöntemleriyle hücre eldesi gerçekleştirilmiş-tir. İki farklı aljinat yüzdesi kullanarak hem manuel olarak hem de kapsülasyon cihazında otomatize olarak iki farklı akış hızı değerlendirilmiştir. Mikroenkapsüle edilen hücreler 64-79 gün boyunca in vitro olarak parathormon miktarları ölçülerek takip edilmiştir.

Bulgular: Değerlendirilen aljinat yüzdelerinden %2’lik konsant-rasyona sahip mikroenkapsüllerin oluşturulmasında kapsülasyon cihazında kullanılan 2 mL/dk akış hızıyla morfolojik stabilite gözlenmiştir. Ayrıca parathormon salınımı açısından hücre izolasyon tipi ve aljinat yüzdeleri arasında benzer sonuçlar elde edilmiştir.

Sonuç: Uzun süreli mikroenkapsülasyon verimliliğinin arttırılması için yapısal ve fonksiyonel açıdan birçok parametrenin belirlenmesi gerekmektedir. Bu çalışma ile enzimatik izolasyon metoduyla elde edilen paratiroid hücrelerinin kapsülasyon sistemi kullanılarak artan akış hızında daha stabil bir yapı oluşturdukları belirlenmiştir.

Keywords

• Paratiroid
• mikroenkapsülasyon
• ultra saf aljinat
• kapsülasyon sistemi
• hücre izolasyonu

References

1. 1. Candlin JP. Chapter 3 Polymeric Materials: Composition, Uses and Applications. In: Chalmers JM, Meier RJ, editors. Comprehensive Analytical Chemistry 53: Elsevier; 2008. p. 65-119. [CrossRef] google scholar
2. 2. Yücesan E, Başoğlu H, Göncü B, Kandaş N, Ersoy Y, Akbaş, F, et al. In-vitro optimization of microencapsulated parathyroid cells. Dicle Medical Journal 2017; 44(4): 373-80. [CrossRef] google scholar
3. 3. Goncu B, Yucesan E. Microencapsulation for Clinical Applications and Transplantation by Using Different Alginates. IntechOpen; 2021.4. [CrossRef] google scholar
4. 4. Lu Q, Yang Z, Meng X, Yue Y, Ahmad MA, Zhang W, et al. A Review on Encapsulation Technology from Organic Light Emitting Diodes to Organic and Perovskite Solar Cells. Advanced Functional Mate-rials 2021; n/a(n/a): 2100151. [CrossRef] google scholar
5. 5. Güven ŞY. Biyouyumluluk Ve Biyomalzemelerin Seçimi. Suleyman Demirel University Journal of Engineering Sciences and Design 2014; 2(3): 303-11. google scholar
6. 6. Carvalho DN, Inacio AR, Sousa RO, Reis RL, Silva TH. Chapter 18 - Seaweed polysaccharides as sustainable building blocks for bio-materials in tissue engineering. In: Torres MD, Kraan S, Dominguez H, editors. Sustainable Seaweed Technologies: Elsevier; 2020. p. 543-87. [CrossRef] google scholar
7. 7. Omami M, McGarrigle JJ, Reedy M, Isa D, Ghani S, Marchese E, et al. Islet Microencapsulation: Strategies and Clinical Status in Dia-betes. Curr Diab Rep 2017; 17(7): 47. [CrossRef] google scholar
8. 8. Woodhams L, Al-Salami H. The roles of bile acids and applicati-ons of microencapsulation technology in treating Type 1 diabetes mellitus. Ther Deliv 2017; 8(6): 401-9. [CrossRef] google scholar

9. 9. Yücesan E, Goncu B, Başoğlu H, Ozten KN, Kanımdan E, Akbaş F, Ersoy YE, Experimental Alternative Method for the Treatment of Hypothyroidism: Capsulation of the Thyroid Tissue. Akdeniz Med J 2018; 5(3): 417-23. [CrossRef] google scholar
10. 10. Hasse C, Zielke A, Klock G, Schlosser A, Barth P, Zimmermann U, et al. Amitogenic alginates: key to first clinical application of microencapsulation technology. World J Surg 1998; 22(7): 65965. [CrossRef] google scholar
11. 11. Yucesan E, Basoglu H, Goncu B, Akbas F, Ersoy YE, Aysan E. Mic-roencapsulated parathyroid allotransplantation in the omental tissue. Artif Organs 2019; 43(10): 1022-7. [CrossRef] google scholar
12. 12. Aysan E, Kilic U, Gok O, Altug B, Ercan C, Kesgin Toka C, et al. Pa-rathyroid Allotransplant for Persistent Hypocalcaemia: A New Te-chnique Involving Short-Term Culture. Exp Clin Transplant 2016; 14(2): 238-41. google scholar
13. 13. Aysan E, Altug B, Ercan C, Kesgin Toka C, Idiz UO, Muslumanoglu M. Parathyroid Allotransplant With a New Technique: A Prospecti-ve Clinical Trial. Exp Clin Transplant 2016; 14(4): 431-5. google scholar
14. 14. Brown WH. I. Parathyroid Implantation in the Treatment of Tetania Parathyreopriva. Ann Surg 1911; 53(3): 305-17. [CrossRef] google scholar
15. 15. Hasse C, Zielke A, Klock G, Barth P, Schlosser A, Zimmermann U, et al. First successful xenotransplantation of microencapsulated human parathyroid tissue in experimental hypoparathyroidism: long-term function without immunosuppression. J Microencap-sul 1997; 14(5): 617-26. [CrossRef] google scholar
16. 16. Yucesan E, Goncu B, Basoglu H, Ozten Kandas N, Ersoy YE, Akbas F, et al. Fresh tissue parathyroid allotransplantation with short-term immunosuppression: 1-year follow-up. Clin Transplant 2017; 31(11). [CrossRef] google scholar
17. 17. de Vos P, Lazarjani HA, Poncelet D, Faas MM. Polymers in cell en-capsulation from an enveloped cell perspective. Adv Drug Deliv Rev 2014; 67-68: 15-34. [CrossRef] google scholar
18. 18. Poojari R, Srivastava R. Composite alginate microspheres as the next-generation egg-box carriers for biomacromolecules deli-very. Expert Opin Drug Deliv 2013; 10(8): 1061-76. [CrossRef] google scholar
19. 19. Lopes M, Abrahim B, Veiga F, Seiça R, Cabral LM, Arnaud P, et al. Preparation methods and applications behind alginate-based particles. Expert Opin Drug Deliv 2017; 14(6): 769-82. [CrossRef] google scholar
20. 20. Hasse C, Schrezenmeir J, Stinner B, Schark C, Wagner PK, Neu-mann K, et al. Successful allotransplantation of microencapsula-ted parathyroids in rats. World J Surg 1994; 18(4): 630-4. [CrossRef] google scholar
21. 21. Hasse C, Klock G, Zielke A, Schlosser A, Barth P, Zimmermann U, et al. Transplantation of parathyroid tissue in experimental hypo-parathyroidism: in vitro and in vivo function of parathyroid tissue microencapsulated with a novel amitogenic alginate. Int J Artif Organs 1996; 19(12): 735-41. [CrossRef] google scholar
22. 22. Hasse C, Klock G, Schlosser A, Zimmermann U, Rothmund M. Pa-rathyroid allotransplantation without immunosuppression. Lan-cet 1997; 350(9087): 1296-7. [CrossRef] google scholar
23. 23. Hasse C, Schlosser A, Klock G, Barth P, Stinner B, Zimmermann U, et al. [Heterologous transplantation of human parathyroid glands after microencapsulation with clinically suitable algi-nate : long-term function without immunosuppression in the animal model]. Langenbecks Arch Chir Suppl Kongressbd 1998; 115(Suppl I): 713-8. google scholar
24. 24. Hasse C, Bohrer T, Barth P, Stinner B, Cohen R, Cramer H, et al. Parathyroid xenotransplantation without immunosuppression in experimental hypoparathyroidism: long-term in vivo function following microencapsulation with a clinically suitable alginate. World J Surg 2000; 24(11): 1361-6. [CrossRef] google scholar
25. 25. Hasse C, Brune M, Lorenz W, Barth P, Metzler W, Breves G, et al. Fun-ctional parameters before and after parathyroidectomy: a prospe-ctive, randomized long-term trial on different rat strains. Exp Clin Endocrinol Diabetes 2004; 112(7): 364-72. [CrossRef] google scholar
26. 26. Zimmermann U, Cramer H, Jork A, Thürmer F, Zimmermann H, Fuhr G, et al. Microencapsulation-Based Cell Therapy. Biotechno-logy 2001: 547-71. [CrossRef] google scholar
27. 27. Tibell A, Rafael E, Wennberg L, Nordenstrom J, Bergstrom M, Gel-ler RL, et al. Survival of macroencapsulated allogeneic parathyroid tissue one year after transplantation in nonimmunosuppressed humans. Cell Transplant 2001; 10(7): 591-9. [CrossRef] google scholar
28. 28. Ulrich F, Klupp J, Thürmer F, Rayes N, Seehofer D, Tullius S, et al. Allotransplantation of Encapsulated Human Parathyroid Tissue in Patients With Permanent Hypoparathyroidism. Transplantation 2004; 78: 79. [CrossRef] google scholar
29. 29. Cabane P, Gac P, Amat J, Pineda P, Rossi R, Caviedes R, et al. Allot-ransplant of microencapsulated parathyroid tissue in severe post-surgical hypoparathyroidism: a case report. Transplant Proc 2009; 41(9): 3879-83. [CrossRef] google scholar
30. 30. Khryshchanovich V, Ghoussein Y. Allotransplantation of macroen-capsulated parathyroid cells as a treatment of severe postsurgical hypoparathyroidism: case report. Ann Saudi Med 2016; 36(2): 1437. [CrossRef] google scholar