The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure

Jin-Ho Chung; Jin Kim; Seong-Ho Choi; Eui-Seong Kim; Jiyong Park; Seung-Jong Lee

doi:10.5395/JKACD.2010.35.4.285

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Basic Research The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure: Jin-Ho Chung¹, Jin Kim², Seong-Ho Choi³, Eui-Seong Kim¹, Jiyong Park⁴, Seung-Jong Lee¹; 2010;35(4):-294.
DOI: https://doi.org/10.5395/JKACD.2010.35.4.285
Published online: July 31, 2010

¹Department of Conservative Dentistry, College of Dentistry, Yonsei University, Seoul, Korea.

²Department of Oral Pathology, College of Dentistry, Yonsei University, Seoul, Korea.

³Department of Periodontology, College of Dentistry, Yonsei University, Seoul, Korea.

⁴Department of Biotechnology, College of Life Science and Biothchnology, Yonsei University, Seoul, Korea.

Corresponding Author: Seung-Jong Lee. Department of Conservative Dentistry, Yonsei University School of Dentistry 134, Shinchon-dong, Sudaemoon-gu, Seoul, 120-752, Korea. Tel: +82-2-2228-3148 Fax: +82-2-313-7575, sjlee@yuhs.ac

• Received: May 29, 2010 • Revised: June 19, 2010 • Accepted: July 3, 2010

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Abstract
REFERENCES

Abstract

The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group.

Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at 0℃ under pressure of 2 MPa), group 8 (low-temperature preservation at 0℃ under no additional pressure), group 9 (low-temperature preservation at -5℃ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37℃ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization.

In both MTT and WST-1 assay, group 7 (0℃/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance).

By the results of this study, low-temperature preservation at 0℃ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.
Keywords: Periodontal ligament cell; MTT; WST-1; Viability; Low temperature preservation under pressure; Rapid freezing; Cryopreservation

Figure 1

Schematic diagram of program freezer with pressure vessel.

a. Oxygen container: 2 or 3 MPa of pressure, b. Program freezer, c. Pressure bottle, d. 2 ml Cryotube: 1 ml F medium + 10% DMSO, e. Tooth treated 10% DMSO, f. Pressure valve, g. Thermometer.

Table 1

Experimental Groups

Table 2

The averages and standard deviations of optical density of MTT

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

Table 3

The averages and standard deviations of optical density of WST-1

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

REFERENCES

1. Schwartz O, Andreasen JO. Cryopreservation of mature teeth before replantation in monkeys(I). Effect of different cryopreservation agents and freezing devices. Int J Oral Surg. 1983;12: 425-436.PubMed
2. Schwartz O, Andreasen JO, Greve T. Cryopreservation before replantation of mature teeth in monkeys(II). Effect of preincubation, different freezing and equilibration rates and endodontic treatment upon periodontal healing. Int J Oral Surg. 1985;14: 350-361.PubMed
3. Kristerson L. Autotransplantation of human premolars. A clinical and radiographic study of 100 teeth. Int J Oral Surg. 1985;14: 200-213.PubMed
4. Lindskog S, Blomlof L, Hammarstrom L. Repair of periodontal tissues in vivo and in vitro. J Clin Periodontol. 1983;10: 188-205.PubMed
5. Melcher AH. Repair of wounds in the periodontium of the rat. Influence of periodontal ligament on osteogenesis. Arch Oral Biol. 1970;15: 1183-1204.Article PubMed
6. Andreasen JO. Interrelation between alveolar bone and periodontal ligament repair after replantation of mature permanent incisors in monkeys. J Periodont Res. 1981;16: 228-235.Article PubMed
7. Andreasen J.O0, Kristerson L. "The effect of limited drying or removal of the periodontal ligament. Periodontal healing after replantation of mature permanent incisors in monkeys". Acta Odontol Scand. 1981;39(1):1-13.PubMed
8. Schwartz O, Rank C.P. "Autotransplantation of cryopreserved tooth in connection with orthodontic treatment". Am J Orthod Dentofacial Orthop. 1986;90(1):67-72.Article PubMed
9. Schwartz O. Cryopreservation of teeth before replantation or transplantation. Atlas of replantation and transplantation of teeth. 1992;Medglobe; 241-256.
10. Rubinsky B. Principles of low temperature cell preservation. Heart Fail Rev. 2003;8(3):277-284.PubMed
11. Pegg DE. The history and principles of cryopreservation. Seminars in reproductive medicine. 2002;20(1):5-13.PubMed
12. Chesne C, Guillouzo A. Crypreservation of isolated rat hepatocytes: a clinical evaluation of freezing and thawing conditions. Cryobiology. 1988;25(4):323-330.Article PubMed
13. Kawasaki N, Hamamoto Y, Nakajima T, Irie K, Ozawa H. Periodontal regeneration of transplanted rat molars after cryopreservation. Archives of Oral Biology. 2004;49: 59-69.Article PubMed
14. Lee YE, Kim ES, Kim J, Han SH, Lee SJ. Efficacy of programmed cryo-preservation under pressure in rat periodontal ligament cells. J Korean Acad Conserv Dent. 2009;34: 356-363.Article
15. Schluter O, Urrutia Benet G, Heinz V, Knorr D. Metastable states of water and ice during pressure-supported freezing of potato tissue. Biotechnol Prog. 2004;20: 799-810.PubMed
16. Arav A, Natan Y. Directional freezing: a solution to the methodological challenges to preserve large organs. Semin Reprod Med. 2009 11;27(6):438-442 Epub Oct 5, 2009.Article PubMed
17. Inuzuka K, Unno N, Yamamoto N, Sagara D, Suzuki M, Nishiyama M, Konno H. Effect of hyperbarically oxygenated-perfluorochemical with university of Wisconsin solution on preservation of rat small intestine using an original pressure-resistant portable apparatus. Surgery. 2007;142(1):57-66.Article PubMed
18. Takahashi T, Kakita A, Takahashi Y, Sakamoto I, Yokoyama K, Fujiu T, Yamashina S, Tamaki T, Takazawa Y, Muratsubaki R. Funtional integrity of the rat liver after subzero preservation under high pressure. Transplantation Proceedings. 2000;32: 1634-1636.Article PubMed
19. Takahashi T, Kakita A, Takahashi Y, Yokoyama K, Sakamoto I, Yamashina S. Preservation of rat livers by supercooling under high pressure. Transplantation Proceedings. 2001;33: 916-919.PubMed
20. Pribenszky C, Molnar M, Cseh S, Solti L. Improving post-thaw survival of cryopreserved mouse blastocysts by hydrostatic pressure challenge. Anim Reprod Sci. 2005;87(1-2):143-150.Article PubMed
21. Pribenszky C, Du Y, Molnar M, Harnos A, Vajta G. Increased stress tolerance of matured pig oocytes after high hydrostatic pressure treatment. Anim Reprod Sci. 2008;106(1-2):200-207.Article PubMed
22. Du Y, Lin L, Schmidt M, Bogh IB, Kragh PM, Sorensen CB, Li J, Purup S, Pribenszky C, Molnar M, Kuwayama M, Zhang X, Yang H, Bolund L, Vajta G. High hydrostatic pressure treatment of porcine oocytes before handmade cloning improves developmental competence and cryosurvival. Cloning and stem cells. 2008;10(3):325-330.Article PubMed
23. Du Y, Pribenszky C, Molnar M, Zhang X, Yang H, Kuwayama M, Pedersen AM, Villemoes K, Bolund L, Vajta G. High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification. Reproduction. 2008;135(1):13-17.PubMed
24. Fowler A. Cryo-injury and biopreservation. Ann N Y Acad Sci. 2005 Dec;1066: 119-135.Article PubMed
25. Gao D, Critser JK. Mechanisms of cryoinjury in living cells. ILAR J. 2000;41(4):187-196.Article PubMed
26. Mazur P, Leibo SP, Chu EH. A two-factor hypothesis of freezing injury. Evidence from Chinese hamster tissue-culture cells. Exp Cell Res. 1972;71(2):345-355.PubMed
27. Hyon SH. A Non-frozen living tissue bank for allotransplantation using green tea polyphenols. Yonsei Med J. 2004;45(6):1025-1034.PubMed
28. Bridgman P.W. Water in the liquid and five solid forms under pressure. Proc Amer Acad Arts and Sci. 1911;47: 441-558.Article
29. Pribenszky C, Molnár M, Cseh S, Solti L. Survival of mouse blastocysts after low-temperature preservation under high pressure. Acta Vet Hung. 2004;52(4):479-487.Article PubMed
30. Baek DY, Lee SJ, Jung HS, Kim ES. Comparison of viability of oral epithelial cells stored by different freezing methods. J Korean Acad Conserv Dent. 2009;34: 491-499.Article
31. Andreasen JO, Schwartz O. Atlas of replantation and transplantation of teeth. 1992;Fribourg, Switzerland: Mediglobe SA.
32. Kawata T. Tooth transplantation by teeth bank-approach to human-Hiroshima. 2005;Department of Orthodontics, Hiroshima University School of Dentistry.
33. Kaku M, Kamata H, Kawata T. Cryopreservation of PDL cells by use of program freezer with magnetic field for tooth banking. Dentistry in Japan. 2007;43: 82-86.
34. Ashwood-smith MJ. Low temperature preservation of cells, tissues and organs. Pitman Medicals. 1980;19-45.
35. Farrant J. Water transport and cell survival in cryobiological procedures. Philos Trans R Soc Lond B Biol Sci. 1977;278(959):191-205.Article PubMed PDF
36. Kim JW, Kim ES, Kim J, Lee SJ. Evaluation of periodontal ligament cell viability in rat teeth after frozen preservation using in-vivo MTT assay. J Korean Acad Conserv Dent. 2006;3(3):192-202.Article
37. Abe F, Kato C, Horikoshi K. Pressure regulated metabolism in microorganisms. Trends Microbiol. 1999;7: 447-453.PubMed
38. Aldridge BE, Bruner LJ. Pressure effects on mechanisms of charge transport across bilayer membranes. Biochem Biophys Acta. 1985;817: 343-354.Article PubMed
39. Huang SY, Pribenszky C, Kuo YH, et al. The effect of hydrostatic pressure treatment on the protein profile of boar spermatozoa before and after freezing. Proceedings of the 6th International Conference on Boar Semen Preservation. 2007;Alliston, Ontario, Cananda: I.-34.Article
40. Kaarniranta K, Elo M, Sironen R, Lammi MJ, Goldring MB, Eriksson JE, Sistonen L, Helminen HJ. Hsp 70 accumulation in chondrocytic cells exposed to high continuous hydrostatic pressure coincides with mRNA stabilization rather than transcriptional activation. Proc Natl Acad Sci USA. 1998;95: 2319-2324.Article PubMed PMC
41. Elo MA, Sironen RK, Karjalainen HM, Kaarniranta KK, Helminen HJ, Lammi MJ. Specific induction of heat shock protein 90 beta by high hydrostatic pressure. Biorheology. 2003;40: 141-146.PubMed
42. Alahiotis SN. Heat shock proteins. A new view on the temperature compensation. Comp Biochem Physiol B. 1983;75: 379-387.Article PubMed
43. Parsell DA, Lindquist S. The function of heat shock proteins in stress tolerance: degradation and reactivation of damaged proteins. Ann Rev Genet. 1993;27: 437-496.PubMed
44. Kwak HJ, Jun CD, Pae HO, Yoo JC, Park YC, Choi BM, Na YG, Park RK, Chung HT, Park WJ, Seo JS. The role of inducible 70-kDa heat shock protein in cell cycle control, differentiation and apoptotic cell death of the human myeloid leukemic HL-60 cells. Cell Immunol. 1998;187: 1-12.PubMed
45. Carlson MA. Technical note: assay of cell quantity in the fibroblast populated collagen matrix with a tetrazolium reagent. Eur Cell Mater. 2006;12: 44-48.PubMed
46. Kim ES, Jeon IS, Kim JW, Kim J, Juhn HS, Lee SJ. An MTT-based method for quantification of periodontal ligament cell viability. Oral Diseases. 2007;13(5):495-499.Article PubMed
47. Ahn HJ, Kim ES, Kim J, Kim DW, Kim KY, Lee CY, Lee SJ. Evaluation of viability of periodontal ligament cell in rat teeth using slow cryoperservation method with magnetic field. J Korean Acad Conserv Dent. 2008;33(4):332-340.
48. Alotto D, Ariotti S, Graziano S, Verrua R, Stella M, Magliacani G, Castagnoli C. The role of quality control in a skin bank: tissue viability determination. Cell and Tissue banking. 2002;3: 3-10.PubMed

Tables & Figures

Figure 1

Schematic diagram of program freezer with pressure vessel.

a. Oxygen container: 2 or 3 MPa of pressure, b. Program freezer, c. Pressure bottle, d. 2 ml Cryotube: 1 ml F medium + 10% DMSO, e. Tooth treated 10% DMSO, f. Pressure valve, g. Thermometer.

Table 1

Experimental Groups

Table 2

The averages and standard deviations of optical density of MTT

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

Table 3

The averages and standard deviations of optical density of WST-1

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

REFERENCES

1. Schwartz O, Andreasen JO. Cryopreservation of mature teeth before replantation in monkeys(I). Effect of different cryopreservation agents and freezing devices. Int J Oral Surg. 1983;12: 425-436.PubMed
2. Schwartz O, Andreasen JO, Greve T. Cryopreservation before replantation of mature teeth in monkeys(II). Effect of preincubation, different freezing and equilibration rates and endodontic treatment upon periodontal healing. Int J Oral Surg. 1985;14: 350-361.PubMed
3. Kristerson L. Autotransplantation of human premolars. A clinical and radiographic study of 100 teeth. Int J Oral Surg. 1985;14: 200-213.PubMed
4. Lindskog S, Blomlof L, Hammarstrom L. Repair of periodontal tissues in vivo and in vitro. J Clin Periodontol. 1983;10: 188-205.PubMed
5. Melcher AH. Repair of wounds in the periodontium of the rat. Influence of periodontal ligament on osteogenesis. Arch Oral Biol. 1970;15: 1183-1204.Article PubMed
6. Andreasen JO. Interrelation between alveolar bone and periodontal ligament repair after replantation of mature permanent incisors in monkeys. J Periodont Res. 1981;16: 228-235.Article PubMed
7. Andreasen J.O0, Kristerson L. "The effect of limited drying or removal of the periodontal ligament. Periodontal healing after replantation of mature permanent incisors in monkeys". Acta Odontol Scand. 1981;39(1):1-13.PubMed
8. Schwartz O, Rank C.P. "Autotransplantation of cryopreserved tooth in connection with orthodontic treatment". Am J Orthod Dentofacial Orthop. 1986;90(1):67-72.Article PubMed
9. Schwartz O. Cryopreservation of teeth before replantation or transplantation. Atlas of replantation and transplantation of teeth. 1992;Medglobe; 241-256.
10. Rubinsky B. Principles of low temperature cell preservation. Heart Fail Rev. 2003;8(3):277-284.PubMed
11. Pegg DE. The history and principles of cryopreservation. Seminars in reproductive medicine. 2002;20(1):5-13.PubMed
12. Chesne C, Guillouzo A. Crypreservation of isolated rat hepatocytes: a clinical evaluation of freezing and thawing conditions. Cryobiology. 1988;25(4):323-330.Article PubMed
13. Kawasaki N, Hamamoto Y, Nakajima T, Irie K, Ozawa H. Periodontal regeneration of transplanted rat molars after cryopreservation. Archives of Oral Biology. 2004;49: 59-69.Article PubMed
14. Lee YE, Kim ES, Kim J, Han SH, Lee SJ. Efficacy of programmed cryo-preservation under pressure in rat periodontal ligament cells. J Korean Acad Conserv Dent. 2009;34: 356-363.Article
15. Schluter O, Urrutia Benet G, Heinz V, Knorr D. Metastable states of water and ice during pressure-supported freezing of potato tissue. Biotechnol Prog. 2004;20: 799-810.PubMed
16. Arav A, Natan Y. Directional freezing: a solution to the methodological challenges to preserve large organs. Semin Reprod Med. 2009 11;27(6):438-442 Epub Oct 5, 2009.Article PubMed
17. Inuzuka K, Unno N, Yamamoto N, Sagara D, Suzuki M, Nishiyama M, Konno H. Effect of hyperbarically oxygenated-perfluorochemical with university of Wisconsin solution on preservation of rat small intestine using an original pressure-resistant portable apparatus. Surgery. 2007;142(1):57-66.Article PubMed
18. Takahashi T, Kakita A, Takahashi Y, Sakamoto I, Yokoyama K, Fujiu T, Yamashina S, Tamaki T, Takazawa Y, Muratsubaki R. Funtional integrity of the rat liver after subzero preservation under high pressure. Transplantation Proceedings. 2000;32: 1634-1636.Article PubMed
19. Takahashi T, Kakita A, Takahashi Y, Yokoyama K, Sakamoto I, Yamashina S. Preservation of rat livers by supercooling under high pressure. Transplantation Proceedings. 2001;33: 916-919.PubMed
20. Pribenszky C, Molnar M, Cseh S, Solti L. Improving post-thaw survival of cryopreserved mouse blastocysts by hydrostatic pressure challenge. Anim Reprod Sci. 2005;87(1-2):143-150.Article PubMed
21. Pribenszky C, Du Y, Molnar M, Harnos A, Vajta G. Increased stress tolerance of matured pig oocytes after high hydrostatic pressure treatment. Anim Reprod Sci. 2008;106(1-2):200-207.Article PubMed
22. Du Y, Lin L, Schmidt M, Bogh IB, Kragh PM, Sorensen CB, Li J, Purup S, Pribenszky C, Molnar M, Kuwayama M, Zhang X, Yang H, Bolund L, Vajta G. High hydrostatic pressure treatment of porcine oocytes before handmade cloning improves developmental competence and cryosurvival. Cloning and stem cells. 2008;10(3):325-330.Article PubMed
23. Du Y, Pribenszky C, Molnar M, Zhang X, Yang H, Kuwayama M, Pedersen AM, Villemoes K, Bolund L, Vajta G. High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification. Reproduction. 2008;135(1):13-17.PubMed
24. Fowler A. Cryo-injury and biopreservation. Ann N Y Acad Sci. 2005 Dec;1066: 119-135.Article PubMed
25. Gao D, Critser JK. Mechanisms of cryoinjury in living cells. ILAR J. 2000;41(4):187-196.Article PubMed
26. Mazur P, Leibo SP, Chu EH. A two-factor hypothesis of freezing injury. Evidence from Chinese hamster tissue-culture cells. Exp Cell Res. 1972;71(2):345-355.PubMed
27. Hyon SH. A Non-frozen living tissue bank for allotransplantation using green tea polyphenols. Yonsei Med J. 2004;45(6):1025-1034.PubMed
28. Bridgman P.W. Water in the liquid and five solid forms under pressure. Proc Amer Acad Arts and Sci. 1911;47: 441-558.Article
29. Pribenszky C, Molnár M, Cseh S, Solti L. Survival of mouse blastocysts after low-temperature preservation under high pressure. Acta Vet Hung. 2004;52(4):479-487.Article PubMed
30. Baek DY, Lee SJ, Jung HS, Kim ES. Comparison of viability of oral epithelial cells stored by different freezing methods. J Korean Acad Conserv Dent. 2009;34: 491-499.Article
31. Andreasen JO, Schwartz O. Atlas of replantation and transplantation of teeth. 1992;Fribourg, Switzerland: Mediglobe SA.
32. Kawata T. Tooth transplantation by teeth bank-approach to human-Hiroshima. 2005;Department of Orthodontics, Hiroshima University School of Dentistry.
33. Kaku M, Kamata H, Kawata T. Cryopreservation of PDL cells by use of program freezer with magnetic field for tooth banking. Dentistry in Japan. 2007;43: 82-86.
34. Ashwood-smith MJ. Low temperature preservation of cells, tissues and organs. Pitman Medicals. 1980;19-45.
35. Farrant J. Water transport and cell survival in cryobiological procedures. Philos Trans R Soc Lond B Biol Sci. 1977;278(959):191-205.Article PubMed PDF
36. Kim JW, Kim ES, Kim J, Lee SJ. Evaluation of periodontal ligament cell viability in rat teeth after frozen preservation using in-vivo MTT assay. J Korean Acad Conserv Dent. 2006;3(3):192-202.Article
37. Abe F, Kato C, Horikoshi K. Pressure regulated metabolism in microorganisms. Trends Microbiol. 1999;7: 447-453.PubMed
38. Aldridge BE, Bruner LJ. Pressure effects on mechanisms of charge transport across bilayer membranes. Biochem Biophys Acta. 1985;817: 343-354.Article PubMed
39. Huang SY, Pribenszky C, Kuo YH, et al. The effect of hydrostatic pressure treatment on the protein profile of boar spermatozoa before and after freezing. Proceedings of the 6th International Conference on Boar Semen Preservation. 2007;Alliston, Ontario, Cananda: I.-34.Article
40. Kaarniranta K, Elo M, Sironen R, Lammi MJ, Goldring MB, Eriksson JE, Sistonen L, Helminen HJ. Hsp 70 accumulation in chondrocytic cells exposed to high continuous hydrostatic pressure coincides with mRNA stabilization rather than transcriptional activation. Proc Natl Acad Sci USA. 1998;95: 2319-2324.Article PubMed PMC
41. Elo MA, Sironen RK, Karjalainen HM, Kaarniranta KK, Helminen HJ, Lammi MJ. Specific induction of heat shock protein 90 beta by high hydrostatic pressure. Biorheology. 2003;40: 141-146.PubMed
42. Alahiotis SN. Heat shock proteins. A new view on the temperature compensation. Comp Biochem Physiol B. 1983;75: 379-387.Article PubMed
43. Parsell DA, Lindquist S. The function of heat shock proteins in stress tolerance: degradation and reactivation of damaged proteins. Ann Rev Genet. 1993;27: 437-496.PubMed
44. Kwak HJ, Jun CD, Pae HO, Yoo JC, Park YC, Choi BM, Na YG, Park RK, Chung HT, Park WJ, Seo JS. The role of inducible 70-kDa heat shock protein in cell cycle control, differentiation and apoptotic cell death of the human myeloid leukemic HL-60 cells. Cell Immunol. 1998;187: 1-12.PubMed
45. Carlson MA. Technical note: assay of cell quantity in the fibroblast populated collagen matrix with a tetrazolium reagent. Eur Cell Mater. 2006;12: 44-48.PubMed
46. Kim ES, Jeon IS, Kim JW, Kim J, Juhn HS, Lee SJ. An MTT-based method for quantification of periodontal ligament cell viability. Oral Diseases. 2007;13(5):495-499.Article PubMed
47. Ahn HJ, Kim ES, Kim J, Kim DW, Kim KY, Lee CY, Lee SJ. Evaluation of viability of periodontal ligament cell in rat teeth using slow cryoperservation method with magnetic field. J Korean Acad Conserv Dent. 2008;33(4):332-340.
48. Alotto D, Ariotti S, Graziano S, Verrua R, Stella M, Magliacani G, Castagnoli C. The role of quality control in a skin bank: tissue viability determination. Cell and Tissue banking. 2002;3: 3-10.PubMed

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Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method
Sun Mi Jang, Sin-Yeon Cho, Eui-Seong Kim, Il-Young Jung, Seung Jong Lee
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The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure

J Korean Acad Conserv Dent. 2010;35(4):285-294. Published online July 31, 2010

DOI: https://doi.org/10.5395/JKACD.2010.35.4.285

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Figure

The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure

Figure 1 Schematic diagram of program freezer with pressure vessel. a. Oxygen container: 2 or 3 MPa of pressure, b. Program freezer, c. Pressure bottle, d. 2 ml Cryotube: 1 ml F medium + 10% DMSO, e. Tooth treated 10% DMSO, f. Pressure valve, g. Thermometer.

Figure 1

The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure

Table 1

Experimental Groups

Table 2

The averages and standard deviations of optical density of MTT

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

Table 3

The averages and standard deviations of optical density of WST-1

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

Table 1 Experimental Groups

Table 2 The averages and standard deviations of optical density of MTT

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

Table 3 The averages and standard deviations of optical density of WST-1

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).