Transplantation of Autologous Bone Marrow Mesenchymal Stem Cells with Platelet-Rich Plasma Accelerate Distraction Osteogenesis in a Canine Model

Original Article

Transplantation of Autologous Bone Marrow Mesenchymal Stem Cells with Platelet-Rich Plasma Accelerate Distraction Osteogenesis in A Canine Model

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*Corresponding Address: P.O.Box: 16635-148, Department of Stem Cells and Developmental Biology at Cell Sciences Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran Email: [email protected] 5HFHLYHG'HF$FFHSWHG$SU Abstract 2EMHFWLYHDistraction osteogenesis (DO) is a surgical procedure used to generate large volumes of new bone for limb lengthening. 0DWHULDOVDQG0HWKRGV In this animal experimental study, a 30% lengthening of the left tibia (mean distraction distance: 60.8 mm) was performed in ten adult male dogs by callus GLVWUDFWLRQDIWHURVWHRWRP\DQGDSSOLFDWLRQRIDQ,OL]DURY¿[DWRU'LVWUDFWLRQZDVVWDUWHGRQ postoperative day seven with a distraction rate of 0.5 mm twice per day and carried out at a rate of 1.5 mm per day until the end of the study. Autologous bone marrow mesenchymal stem cells (BM-MSCs) and platelet-rich plasma (PRP) as the treatment group (n=5) or PRP alone (control group, n=5) were injected into the distracted callus at the middle and end of the distraction period. At the end of the consolidation period, the dogs were VDFUL¿FHGDIWHUZKLFKFRPSXWHUL]HGWRPRJUDSK\ &7 DQGKLVWRPRUSKRPHWULFHYDOXDWLRQV were performed. 5HVXOWV Radiographic evaluationsrevealed that the amount and quality of callus for- mations were significantly higher in the treatment group (P<0.05). As measured by CT scan, the healing parametersin dogs of the treatment group were significantly greater (P<0.05). New bone formation in the treatment group was significantly higher (P<0.05). &RQFOXVLRQThe present study showed that the transplantation of BM-MSCs positively affects early bony consolidation in DO. The use of MSCs might allow a shortened period of consolidationArchive and therefore permit earlier device removal.of SID Keywords: Distraction Osteogenesis, Bone Lengthening, Mesenchymal Stem Cells, Autologous Transplantation, Platelet-Rich Plasma

Cell Journal(Yakhteh), Vol 17, No 2, Summer 2015, Pages: 243- 252

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ZHUHLQVHUWHGDWWKHFHQWHURIWKHGLVWUDFWHGFDOOXV Table 2:/ŶĚĞǆŽĨĞůŝŵŝŶĂƟŽŶŽĨƚŚĞŐĂƉ IDFHWRIDFHZLWKHDFKWLS:HVLPXOWDQHRXVO\LQ Description Score MHFWHGRQHPORIDFDOFLXPWKURPELQPL[WXUHDQG PORI06&VDQGWKH353VXVSHQVLRQLQWRWKH &OHDUO\REVHUYDWLRQRIWKHJDS FDOOXV VR WKDW WKH 353 JHO PLJKW GHYHORS ZLWKLQ ¿OOHGJDS WKHLQMHFWHGVLWH 6LPLODUO\LQFRQWUROJURXS GRJV353DORQHZDVORFDOO\DSSOLHGWRWKHSULPDU\ ¿OOHGJDS FDOOXV)ROORZLQJWKH¿UVWWUDQVSODQWDWLRQWKHVHF ¿OOHGJDS ond session of distraction was carried out at rate of &RPSOHWH¿OOHGJDS PPWZLFHGDLO\ PPGD\ XQWLOOHQJWKHQ LQJZDVDFKLHYHGWRRIWKHWRWDOWLELDOHQJWK The second transplantation was performed at the HQGRIWKHGLVWUDFWLRQSHULRGWKHGLVWUDFWRUZDVUH PRYHGDQGDQLQWHUORFNLQJQDLO PP ZDVSXW Table 3:/ŶĚĞǆŽĨĨŽƌŵĂƟŽŶŽĨƚŚĞĞǆƚĞƌŶĂůĐĂůůƵƐ in its place. Description Score

Radiography and computerized tomography No callus formation &7 VFDQ Beginning of callus formation

,PPHGLDWHO\ DIWHU VXUJHU\ DQG WZLFH ZHHNO\ %ULGJHIRUPDWLRQRIWKHH[WHUQDOFDOOXVZLWKDQREYLRXVJDS there after we obtained radiographs of the left %ULGJHIRUPDWLRQRIWKHH[WHUQDOFDOOXVZLWKDPXIÀHJDS WLELD7KHDPRXQWRIPLQHUDOL]DWLRQRISULPDU\ callus (immature bone) and mature bone tis &RPSOHWHGFDOOXVDQGHOLPLQDWLRQRIWKHJDS VXH ZHUH VHPLTXDQWLWDWLYHO\ DVVHVVHG E\ UD GLRJUDSKV%HIRUHERQHVHJPHQWVZHUH¿[HGLQ IRUPDOLQ&7VFDQLPDJHVZHUHREWDLQHGLQWKH ORQJLWXGLQDODQGWUDQVYHUVHDVSHFWVRIWKHOHIWLQ Table 4:/ŶĚĞǆŽĨĨŽƌŵĂƟŽŶŽĨŝŶƚĞƌĐŽƌƟĐĂůĐĂůůƵƐ tact tibia. The critical outcomes (healing param HWHUV VXFK DV WKH IRUPDWLRQ RI H[WHUQDO SHUL Description Score RVWHDO FDOOXVLQWHUFRUWLFDOFDOOXVWKHGHQVLW\RI No callus formation WKHFDOOXVDQGWKHHOLPLQDWLRQRIWKHJDSZHUH VHPLTXDQWLWDWLYHO\GH¿QHGE\DUDGLRORJLVWZKR Beginning of the callus formation was blinded to the group assignment of the ani Intercortical formation of callus and its recognition from PDOV7KHVFRUHVIRUHDFKLQGH[DUHWDEXODWHGLQ WKHFRUWH[ WDEOHV&7VFDQHYDOXDWLRQZDVSHUIRUPHGLQ Intercortical formation of callus and possibility of the same manner. FDOOXVUHFRJQLWLRQIURPWKHFRUWH[ &RPSOHWHGFDOOXVZLWKRXWSRVVLELOLW\RIFDOOXV UHFRJQLWLRQIURPWKHFRUWH[

Table 1:/ŶĚĞǆŽĨĐĂůůƵƐĚĞŶƐŝƚǇ Description ArchiveScore +LVWRSDWKRORJLFDQDO\VLV of SID No callus density 6L[PRQWKVDIWHUWKHLQLWLDOVXUJHU\WKHGRJVZHUH HXWKDQDWL]HGZLWKDEDUELWXUDWHRYHUGRVHDQGWKHLQ WHUORFNLQJ QDLO ZDV UHPRYHG 7KH OHIW WLELDV ZHUH 0LOGFDOOXVGHQVLW\ KDUYHVWHGIROORZLQJLQWUDDUWHULDOYLWDOSHUIXVLRQRID QHXWUDOEXIIHUHGIRUPDOGHK\GHVROXWLRQ 6LJPD 0RGHUDWHFDOOXVGHQVLW\ 86$ 7KHWLELDVZHUHURXWLQHO\¿[HGDQGGHFDOFL¿HG The distraction area was sectioned from the central 6HYHUHRUKLJKFDOOXVGHQVLW\ ]RQHWRWKHSHULSKHUDO]RQH,QWKHSUHVHQWVWXG\ZH LQYHVWLJDWHG WKH DPRXQW RI ERQH LQ WKH GLVWUDFWLRQ &DOOXVGHQVLW\VLPLODUWRLQWDFWSHULSKHUDOERQH gaps in four areas that were equally cut from the cent HU RI WKH JDS WRZDUG WKH PDUJLQV ZLWK KHPDWR[\OLQ

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$GGLWLRQDOO\WKHVHJURZWKIDFWRUVLQKLELWRVWHRFODVW tion and soft-tissue preservation. Clin Orthop Relat Res. 1989; (238): 249-281. formation and bone resorption and possibly increase 2. Feng X, Tuo X, Chen F, Wu W, Ding Y, Duan Y, et al. Ultra- WKHQXPEHURIRVWHREODVWVWKHUHE\DFFHOHUDWLQJERQH structural cell response to tension stress during mandibu- IRUPDWLRQ 2QWKHZKROHWKHDGGLWLRQRI06& lar distraction osteogenesis. Br J Oral Maxillofac Surg. 2008; 46(7): 527-532 LQ353VFDIIROGKDVLQFUHDVHGQHZERQHIRUPDWLRQ 3. Codivilla A. On the means of lengthening, in the lower PLQHUDOL]DWLRQ DQG PHFKDQLFDO SURSHUW\ FRPSDUHG limbs, the muscles and tissues which are shortened WRWKH353RQO\JURXS ,QWKHSUHVHQWVWXG\ZH through deformity. Am J Orthop Surg. 1905; 2: 353–363. 4. Ilizarov GA. Clinical application of the tension-stress effect XVHGDXWRORJRXVIUHVK3537KHUDWHRIGLVWUDFWLRQLQ for limb lengthening. Clin Orthop Relat Res. 1990; (250): WKH'2SURFHVVKDGDQLPSRUWDQWUROHLQDFKLHYLQJ 8-26. WKHEHVWUHVXOWV)URPDPHFKDQRELRORJLFDOSRLQWRI 5. Kinoshita K, Hibi H, Yamada Y, Ueda M. Promoted new bone formation in Maxillary distraction osteogenesis us- YLHZDSRRURVWHRWRP\IUDPHLQVWDELOLW\DQGDKLJK ing a tissue-engineered osteogenic material. J Craniofac GLVWUDFWLRQUDWHPLJKWDOOGLVWXUEYDVFXODUL]DWLRQDQG Surg. 2008; 19(1): 80-87. ORFDOEORRGVXSSO\WRWKHUHJHQHUDWLQJWLVVXHWKHUHE\ 6. Marino JT, Bruce BH. Use of solid and cancellous autologous bone graft for fractures and nonunions. Orthop Clin FDXVLQJGHOD\HGERQHKHDGLQJ ,WKDVEHHQVKRZQ North Am. 2010; 41(1): 15-26. that a rapid distraction rate causes the disruption of 7. Hagiwara T, Bell WH. Effect of electrical stimulation on EORRGYHVVHOVLQWKHGLVWUDFWLRQ]RQH ,OL]DURYGH mandibular distraction osteogenesis. J Craniomaxillofac Surg. 2000; 28(1): 12-19. WHUPLQHGWKDWRQHPPSHUGD\ PPOHQJWKHQLQJ 8. Kitakoji T, Takashi S, Ono Y, Hattori T, Takahashi H, Iwata four times a day) resulted in acceptable new bone for H. Effect of hyperbaric oxygenation treatment on length- PDWLRQ ,QGRJVDGLVWUDFWLRQUDWHRIPPSHU ened callus. Undersea Hyperb Med.1999; 26(3): 165-168. 9. Eberson CP, Hogan KA, Moore DC, Ehrlich MG. Effect day has been shown to result in premature consoli of low-intensity ultrasound stimulation on consolidation of GDWLRQDIWHUDSSUR[LPDWHO\GD\V ,WVKRXOGEH the regenerate zone in a rat model of distraction osteo- mentioned that this rate could be changed depending genesis. J Pediatr Orthop. 2003; 23(1): 46-51. 10. 0R¿G00,QRXH1$WDEH\$0DUWL*&KDR(<0DQVRQ on the status of the callus and tension within the soft PN, et al. Callus stimulation in distraction osteogenesis. WLVVXHV 7KHVHVWDQGDUGVKDYHEHHQXVHGLQWKH Plast Reconstr Surg. 2002; 109(5): 1621-1629. SUHVHQWVWXG\2QWKHZKROHZHFDQFRQFOXGHWKDW 11. Kim IS, Park JW, Kwon IC, Baik BS, Cho BC. Role of BMP, betaig-h3, and chitosan in early bony consolidation autologous cell therapy for bone regeneration by a in distraction osteogenesis in a dog model. Plast Reconstr FRPELQDWLRQRI06&VDQG353KDVPDQ\DGYDQWDJHV Surg. 2002; 109(6): 1966-1977. IRUFOLQLFDOIHDVLELOLW\7KHVH¿QGLQJVPD\EHDSSOL 12. Nolan PC, Nicholas RM, Mulholland BJ, Mollan RA, Wil- son DJ.Culture of human osteoblasts on demineralised cable in the repair of bone defects and can be a use human bone. Possible means of graft enhancement. J IXODOWHUQDWLYHWRDOORJHQLFRUDXWRORJRXVERQHJUDIWV Bone Joint Surg Br. 1992; 74(2): 284-286. EHFDXVH WKH SURSRVHG WUHDWPHQW LV VDIH PLQLPDOO\ 13. Qi MC, Hu J, Zou SJ, Chen HQ, Zhou HX, Han LC. Me- chanical strain induces osteogenic differentiation: Cbfa1 LQYDVLYHHDV\WRSHUIRUPDQGKDVJUHDWSRWHQWLDOIRU and Ets-1 expression in stretched rat mesenchymal stem clinical applications. cells. Int J Oral Maxillofac Surg. 2008; 37(5): 453-458. 14. Jones E, Yang X. Mesenchymal stem cells and bone regeneration: current status. Injury. 2011; 42(6): 562-568. Conclusion 15. Zomorodian E, Baghaban Eslaminejad M. Mesenchymal stem cells as a potent cell source for bone regeneration. 7DNHQWRJHWKHUWKHWUDQVSODQWDWLRQRI%006&V Stem Cells Int. 2012; 2012: 980353. SRVLWLYHO\DIIHFWVHDUO\ERQ\FRQVROLGDWLRQLQ'2 16. Granero-Molto F, Weis JA, Miga MI, Landis B, Myers 7KH XVH RI 06&V DORQJ ZLWK 353 PD\ DOORZ D TJ, ORear L, et al. Regenerative effects of transplanted mesenchymal stem cells in fracture healing. Stem Cells. shortened period of consolidation and therefore 2009; 27(8): 1887-1898. SHUPLWHDUOLHUGHYLFHUHPRYDOArchive17. Wroblewskiof AP, MejiaSID HA, Wright VJ. Application of platelet-rich plasma to enhance tissue repair. Oper Tech Or- thop. 2010; 20(2): 98-105. Acknowledgments 18. Jiang ZQ, Liu HY, Zhang LP, Wu ZQ, Shang DZ. Repair of calvarial defects in rabbits with platelet-rich plasma as the This research was financially supported by an scaffold for carrying bone marrow stromal cells. Oral Surg ,QGXVWULDO'HYHORSPHQWDQG5HQRYDWLRQ2UJDQ Oral Med Oral Pathol Oral Radiol. 2012; 113(3): 327-333. 19. Hernandez-Fernandez A, Velez R, Soldado F, Carlos L]DWLRQRI,UDQ ,'52 /,'&2JUDQW7KHUHLV Saenz-Ríos J, Barber I, Aguirre-Canyadell M. Effect of ad- no conflict of interest in this article. ministration of platelet-rich plasma in early phases of distraction osteogenesis: an experimental study in an ovine References femur model. Injury. 2013; 44(7): 901-907. 20. Latalski M, Elbatrawy YA, Thabet AM, Gregosiewicz A, 1. Ilizarov GA. The tension–stress effect on the genesis and Raganowicz T, Fatyga M. Enhancing bone healing during JURZWKRIWLVVXHV3DUW,7KHLQÀXHQFHRIVWDELOLW\RI¿[D- distraction osteogenesis with platelet-rich plasma. Injury.

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