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Time-sequential modulation in expression of growth factors from platelet-rich plasma (PRP) on the chondrocyte cultures

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Abstract

Platelets are involved in hemostasis, wound healing, and tumor growth. Autologous blood products are commonly used to facilitate healing in a variety of clinical surgery applications. Recently, it was shown that platelet-rich plasma (PRP) has more specific growth factors that participate in the healing process. This study investigated the expression of PRP growth factors and evaluated their potential role in the cartilage regeneration using primary isolated chondrocytes. PRP obtained from New Zealand White rabbit by low speed centrifugation. Extracted PRPs contained 6–10 × 106 platelet/μl and concentration of platelets was slightly variable. Primary isolated chondrocytes from the same rabbits were cultured and treated with 0.1–20% PRP. The cells were collected and examined by reverse transcription-polymerase chain reaction and cytochemical staining. The expression of sex determining region Y-box 9, transforming growth factor-beta, vascular endothelial growth factor, and chondromdulin-I was increased in chondrocyte cultures with 10% PRP by time-dependent manner. To maintain the integrity of the cartilage, the proteoglycan contents were also up-regulated from the mRNA of aggrecan and positive Safranin-O staining in PRP concentration- and time-dependent manner. PRP provides crucial growth factors related to chondrocyte proliferation and differentiation through time-sequential modulation. Controlled in vivo trials for cartilage regeneration are needed.

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References

  1. Woolf AD, Pfleger B (2003) Burden of major musculoskeletal conditions. Bull World Health Organ 81:646–656

    PubMed  Google Scholar 

  2. Anitua M, Sanchez E, Nurden A, Nurden P, Orive G, Andia I (2006) New insights into and novel applications for platelet-rich fibrin therapies. Trends Biotechnol 24:227–234

    Article  PubMed  CAS  Google Scholar 

  3. Folkman J, Browder T, Palmblad J (2001) Angiogenesis research: guidelines for translation to clinical application. Thromb Haemost 86:23–33

    PubMed  CAS  Google Scholar 

  4. Pietramaggiori G, Scherer SS, Mathews JC, Gennaoui T, Lancerotto L, Ragno G, Valeri CR, Orgill DP (2010) Quiescent platelets stimulate angiogenesis and diabetic wound repair. J Surg Res 160:169–177

    Article  PubMed  CAS  Google Scholar 

  5. Eppley BL, Pietrzak WS, Blanton M (2006) Platelet-rich plasma: a review of biology and applications in plastic surgery. Plast Reconstr Surg 118:147e–159e

    Article  PubMed  CAS  Google Scholar 

  6. Knighton DR, Ciresi KF, Fiegel VD, Austin LL, Butler EL (1986) Classification and treatment of chronic nonhealing wounds, successful treatment with autologous platelet-derived wound healing factors (PDWHF). Ann Surg 204:322–330

    Article  PubMed  CAS  Google Scholar 

  7. Ma L, Perini R, McKnight W, Dicay M, Klein A, Hollenberg MD, Wallace JL (2005) Proteinase-activated receptors 1 and 4 counter-regulate endostatin and VEGF release from human platelets. Proc Natl Acad Sci USA 102:216–220

    Article  PubMed  CAS  Google Scholar 

  8. Andreas K, Lübke C, Häupl T, Dehne T, Morawietz L, Ringe J, Kaps C, Sittinger M (2008) Key regulatory molecules of cartilage destruction in rheumatoid arthritis: an in vitro study. Arthritis Res Ther 10:R9

    Article  PubMed  Google Scholar 

  9. Ringe J, Sittinger M (2009) Tissue engineering in the rheumatic diseases. Arthritis Res Ther 11:211

    Article  PubMed  Google Scholar 

  10. Marx RE (2004) Patelet-rich plasma: evidence to support its use. J Oral Maxillofac Surg 62:489–496

    Article  PubMed  Google Scholar 

  11. Kim DJ, Moon SH, Kim H, Kwon UH, Park MS, Han KJ, Hahn SB, Lee HM (2003) Bone morphogenetic protein-2 facilitates expression of chondrogenic, not osteogenic, phenotype of human intervertebral disc cells. Spine 28:2679–2684

    Article  PubMed  Google Scholar 

  12. Thompson JP, Oegema TR Jr, Bradford DS (1991) Stimulation of mature canine intervertebral disc by growth factors. Spine 16:253–260

    Article  PubMed  CAS  Google Scholar 

  13. Versijil N, DeGroot J, Thorpe SR, Bank RA, Shaw JN, Lyons TJ, Bijlsma JW, Lafeber FP, Baynes JW, TeKoppele JM (2000) Effect of collagen turnover on the accumulation of advanced glycation end products. J Biol Chem 275:39027–39031

    Article  Google Scholar 

  14. Paul R, Haydon RC, Cheng H, Ishikawa A, Nenadovich N, Jiang W, Zhou L, Breyer B, Feng T, Gupta P, He TC, Phillips FM (2003) Potential use of Sox9 gene therapy for intervertebral degenerative disc disease. Spine 28:755–763

    PubMed  Google Scholar 

  15. Kuhne M, John T, El-Sayed K, Marzahn U, Aue A, Kohl B, Stoelzel K, Ertel W, Blottner D, Haisch A, Schulze-Tanzil G (2010) Characterization of auricular chondrocytes and auricular/articular chondrocyte co-cultures in terms of an application in articular cartilage repair. Int J Mol Med 25:701–708

    PubMed  CAS  Google Scholar 

  16. Lefebvre V, Behringer RR, de Crombrugghe B (2001) L-Sox5, Sox6 and Sox9 control essential steps of the chondrocyte differentiation pathway. Osteoarthritis Cartil 9(Suppl A):S69–S75

    Article  Google Scholar 

  17. Aigner T, Gebhard PM, Schmid E, Bau B, Harley V, Pöschl E (2003) SOX9 expression does not correlate with type II collagen expression in adult articular chondrocytes. Matrix Biol 22:363–372

    Article  PubMed  CAS  Google Scholar 

  18. Zhao Q, Eberspaecher H, Lefebvre V, De Crombrugghe B (1997) Parallel expression of Sox9 and Col2a1 in cells undergoing chondrogenesis. Dev Dyn 209:377–386

    Article  PubMed  CAS  Google Scholar 

  19. Rabie AB, Leung FY, Chayanupatkul A, Hagg U (2002) The correlation between neovascularization and bone vormation in the condyle during forward mandibular positioning. Angle Orthod 72:431–438

    PubMed  CAS  Google Scholar 

  20. Rabie AB, Hagg U (2002) Factors regulating mandibular condylar growth. Am J Orthod Dentrofacial Orthop 122:401–409

    Article  CAS  Google Scholar 

  21. Hashimoto S, Creighton-Achermann L, Takahashi K, Amiel D, Coutts RD, Lotz M (2002) Development and regulation of osteophyte formation during experimental osteoarthritis. Osteoarthritis Cartil 10:180–187

    Article  CAS  Google Scholar 

  22. Bluteau G, Julien M, Magne D, Mallein-Gerin F, Weiss P, Daculsi G, Guicheux J (2007) VEGF and VEGF receptors are differentially expressed in chondrocytes. Bone 40:568–576

    Article  PubMed  CAS  Google Scholar 

  23. Brew CJ, Clegg PD, Boot-Handford RP, Andrew JG, Hardingham T (2010) Gene expression in human chondrocytes in late osteoarthritis is changed in both fibrillated and intact cartilage without evidence of generalised chondrocyte hypertrophy. Ann Rheum Dis 69:234–240

    Article  PubMed  CAS  Google Scholar 

  24. Yee G, Yu U, Walsh WR, Linderman R, Poole MD (2003) The immunolocalisation of VEGF in the articular cartilage of sheep mandibular condyles. J Craniomaxillofac Surg 31:244–251

    Article  PubMed  Google Scholar 

  25. Emons J, Chagin AS, Malmlöf T, Lekman M, Tivesten A, Ohlsson C, Wit JM, Karperien M, Sävendahl L (2010) Expression of vascular endothelial growth factor in the growth plate is stimulated by estradiol and increases during pubertal development. J Endocrinol 205:61–68

    Article  PubMed  CAS  Google Scholar 

  26. Ashraf S, Walsh DA (2008) Angiogenesis in osteoarthritis. Curr Opin Rheumatol 20:573–580

    Article  PubMed  Google Scholar 

  27. Kitahara H, Hayami T, Tounga K, Endo N, Funaki H, Yoshida Y, Yaoita E, Yamamoto T (2003) Chondromodulin-1 expression in rat articular cartilage. Arch Histol Cytol 66:221–228

    Article  PubMed  CAS  Google Scholar 

  28. Hiraki Y, Tanaka H, Inoue H, Kondo J, Kamizono A, Suzuki F (1991) Molecular cloning of a new class of cartilage-specific matrix, chondromodulin-1, which stimulates growth of cultured chondrocytes. Biochem Biophys Res Commun 175:971–977

    Article  PubMed  CAS  Google Scholar 

  29. Wtahiki J, Yamaguchi T, Enomoto A, Irie T, Yoshie K, Tachikawa T, Maki K (2008) Identification of differentially expressed genes in mandibular condylar and tibial growth cartilages using laser microdissection and fluorescent differential display: chondromodulin-I (ChM-1) and tenomodulin (TeM) are differentially expressed in mandibular condylar and other growth cartilages. Bone 42:1053–1060

    Article  Google Scholar 

  30. Hiraki Y, Inoue H, Iyama K, Kamizono A, Ochiai M, Shukunami C, Iijima S, Suzuki F, Kondo J (1997) Identification of chondromodulin I as a novel endothelial cell growth inhibitor, purification and its localization in the avascular zone of epiphyseal cartilage. J Biol Chem 272:32419–32426

    Article  PubMed  CAS  Google Scholar 

  31. Shukunami C, Hiraki Y (1998) Expression of cartilage-specific functional matrix chondromodulin-I mRNA in rabbit growth plate chondrocytes and its responsiveness to growth stimuli in vitro. Biochem Biophys Res Commun 249:885–890

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

This study was supported by grants of the Nano-Biotechnology Project (Regenomics), Ministry of Science and Technology, Republic of Korea (B020214) and the National Research Foundation of Korea Grant, Ministry of Education, Science and Technology, Korean Government (NRF-2009-351-E00060).

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Authors and Affiliations

  1. Department of Orthopedic Surgery, College of Medicine, Yeungnam University, Daegu, 705-717, Korea

    Se-Il Park & Myun-Whan Ahn

  2. Department of Clinical Pathology, College of Veterinary Medicine, Konkuk University, Seoul, 143-701, Korea

    Hye-Rim Lee & Sun Hee Do

  3. School of Materials Science and Engineering, Yeungnam University, Gyeongsan, 712-749, Korea

    Sukyoung Kim

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  1. Se-Il Park
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  2. Hye-Rim Lee
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  3. Sukyoung Kim
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  4. Myun-Whan Ahn
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  5. Sun Hee Do
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Correspondence to Sun Hee Do.

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Se-Il Park and Hye-Rim Lee contributed equally to this study.

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Park, SI., Lee, HR., Kim, S. et al. Time-sequential modulation in expression of growth factors from platelet-rich plasma (PRP) on the chondrocyte cultures. Mol Cell Biochem 361, 9–17 (2012). https://doi.org/10.1007/s11010-011-1081-1

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  • DOI: https://doi.org/10.1007/s11010-011-1081-1

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