백서 두개골 결손모델에서 골형성단백질이 함유된 헤파린 결합 박테리아 셀룰로오스 지지체가 골재생에 미치는 영향

정성린; 안성준; 김세은; 이소현; 박종석; 권희정; 강성수; 허중보; 임윤묵; Sung In Jeong; Sung-Jun An; Se Eun Kim; So-Hyoun Lee; Jong-Seok Park; Hui-Jeong Gwon; Seong Soo Kang; Jung-Bo Huh; Youn-Mook Lim

Polymer(Korea), Vol.41, No.2, 339-345, March, 2017

DOI10.7317/pk.2017.41.2.339 Export Citation

백서 두개골 결손모델에서 골형성단백질이 함유된 헤파린 결합 박테리아 셀룰로오스 지지체가 골재생에 미치는 영향

Efficacy of rhBMP-2 on Heparin Immobilized Bacterial Cellulose Scaffolds for Bone Tissue Engineering in Rat Calvarial Defect Model

정성린, 안성준, 김세은¹, 이소현², 박종석, 권희정, 강성수¹, 허중보², 임윤묵^†

한국원자력연구원 첨단방사선연구소 공업환경연구부
¹전남대학교 수의대학
²부산대학교 치과대학

Sung In Jeong, Sung-Jun An, Se Eun Kim¹, So-Hyoun Lee², Jong-Seok Park, Hui-Jeong Gwon, Seong Soo Kang¹, Jung-Bo Huh², and Youn-Mook Lim^†

Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, 1266 Sinjeong-dong Jeongeup-si Jeollabuk-do 56212, Korea
¹Department of Veterinary Surgery, College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Korea
²Department of Prosthodontics, School of Dentistry, Dental Research Institute, Pusan National University, Beomeo-ri, Mulgeum-eup, Yangsan, Gyeongnam 50612, Korea

E-mail:

초록

조직공학용 나노섬유 지지체의 3차원 고공극 구조는 세포의 부착과 성장에 적합하고, 지지체 내에 영양분과 불순물이 통과되어야 한다. 최근에 우리 연구실에서는 감마선과 EDC/NHS 화학반응을 이용하여 조직공학용 bacterial cellulose(BC) 지지체에 heparin과 2-aminoethyl methacrylate(AEMA)을 표면 개질하였다. 본 연구에서는 백서 두개골 결손 모델을 이용하여 recombinant human bone morphogenic protein-2(rhBMP-2) 함유 유무에 따른 헤파린 결합 박테리아 셀룰로오스 지지체가 신생골 형성에 미치는 영향을 평가하려고 한다. 지지체의 골형성 효과를 평가하기 위해서 지지체를 백서 두개골에 식립 8주 후에 방사선학적 및 조직학적 평가를 통해 지지체에 대한 임상학적인 측면에서 골결손부 크기, 신생골 재생속도, 염증반응 등을 평가하였다. 골형성 단백질이 함유된 헤파린이 결합 박테리아 셀룰로오스는 두개골 결손 모델에서 골형성을 촉진하였다. 이는 조직공학적으로나 임상학적으로 우수한 특성을 가지고 있다고 사료된다.

Nanofibrous scaffolds for tissue regeneration need to be three-dimensional and highly porous to support uniform cell attachment and proliferation, and need to have an interconnected and permeable pore network to promote nutrient and waste exchange. Our laboratory recently developed heparin and 2-aminoethyl methacrylate (AEMA) immobilized bacterial cellulose (BC) scaffolds for tissue engineering using gamma-irradiation and EDC/NHS chemistry. Here, we examined in vivo efficiency of a heparin immobilized AEMA-BC nanofibrous scaffolds under the condition of with or without recombinant human bone morphogenic protein-2 (rhBMP-2) for a therapeutic substrate in bone healing. To evaluate the bone healing effect of the scaffolds, they were evaluated at 8 weeks post-operatively according to the following criteria: size of residual defect area, speed of new bone formation, inflammation, and clinical applicability of the material. This heparin immobilized AEMA-BC scaffolds and rhBMP-2 independently facilitated a bone healing of the calvarial defect for its good biocompatibility. It holds a great potential for a clinical application and tissue engineering.

Keywords:bacterial cellulose;gamma-irradiation;2-aminoethyl methacrylate;heparin;bone tissue engineering

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Jeon O, Powell C, Solorio LD, Krebs MD, Alsberg E, J. Control. Release, 154, 258 (2011)
Jeon O, Song SJ, Kang SW, Putnam AJ, Kim BS, Biomaterials, 28, 2763 (2007)
Jeong SI, Jeon O, Krebs MD, Hill MC, Alsberg E, Eur. Cell. Mater., 24, 331 (2012)
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Shin YM, Lim JY, Park JS, Gwon HJ, Jeong SI, Ahn SJ, Shin H, Lim YM, RSC Advances, 5, 55948 (2015)
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Kim SE, Song SH, Yun YP, Choi BJ, Kwon IK, Bae MS, Moon HJ, Kwon YD, Biomaterials, 32, 366 (2011)
Darwis D, Khusniya T, Hardiningsih L, Nurlidar F, Winarno H, Atom Indonesia, 38, 78 (2012)

[1] Mckay WF, Peckham SM, Badura JM, Int. Orthop., 31, 729 (2007)

[2] Nohe A, Hassel S, Ehrlich M, Neubauer F, Sebald W, Henis YI, Knaus P, J. Biol. Chem., 277, 5330 (2002)

[3] Ryoo HM, Lee MH, Kim YJ, Gene, 366, 51 (2006)

[4] Kim SE, Song SH, Yun YP, Choi BJ, Kwon IK, Bae MS, Moon HJ, Kwon YD, Biomaterials, 32, 366 (2011)

[5] Jeon O, Song SJ, Kang SW, Putnam AJ, Kim BS, Biomaterials, 28, 2763 (2007)

[6] Li C, Vepari C, Jin HJ, Kim HJ, Kaplan DL, Biomaterials, 27, 3115 (2006)

[7] Jeon O, Powell C, Solorio LD, Krebs MD, Alsberg E, J. Control. Release, 154, 258 (2011)

[8] Jeon O, Song SJ, Kang SW, Putnam AJ, Kim BS, Biomaterials, 28, 2763 (2007)

[9] Jeong SI, Jeon O, Krebs MD, Hill MC, Alsberg E, Eur. Cell. Mater., 24, 331 (2012)

[10] Gao T, Cui W, Wang Z, Wang Y, Liu Y, Malliappan PS, Ito Y, Zhang P, RSC Advances, 6, 20202 (2016)

[11] Shin YM, Kim KS, Lim YM, Nho YC, Shin H, Biomacromolecules, 9(7), 1772 (2008)

[12] Bolbasov EN, Rybachuk M, Golovkin AS, Antonova LV, Shesterikov EV, Malchikhina AI, Novikov VA, Anissimov YG, Tverdokhlebov SI, Mater. Lett., 132, 281 (2014)

[13] Shin YM, La WG, Lee MS, Yang HS, Lim YM, J. Mater. Chem. B, 3, 8375 (2015)

[14] Shin YM, Lim JY, Park JS, Gwon HJ, Jeong SI, Ahn SJ, Shin H, Lim YM, RSC Advances, 5, 55948 (2015)

[15] Embuscado ME, Marks JS, BeMiller JN, Food Hydrocolloids, 8, 419 (1994)

[16] Jeong SI, Jeong JO, Choi JB, Shin YM, Park JS, Gwon HJ, Nho YC, Ahn SJ, Park MY, Lim YM, Tissue Eng. Regen. Med., 11, 56 (2014)

[17] Lee SH, Lim YM, Jeong SI, Ahn SJ, Kang SS, Jeong CM, Huh JB, J. Adv. Prosthodont, 7, 484 (2015)

[18] Czaja WK, Young DJ, Kawecki M, Brown RM, Biomacromolecules, 8(1), 1 (2007)

[19] Muller FA, Muller L, Hofmann I, Greil P, Wenzel MM, Staudenmaier R, Biomaterials, 27, 3955 (2006)

[20] Zhou C, Shi Q, Guo W, Terrell L, Qureshi AT, Hayes DJ, Wu Q, ACS Appl. Mater. Inter,, 5, 3847 (2013)

[21] Kadokawa J, Saitou S, Shoda S, Carbohydr. Polym., 60, 253 (2005)

[22] Figueiredo ARP, Figueiredo AGPR, Silva NHCS, Barros-Timmons A, Almeida A, Silvestre AJD, Freire CSR, Carbohydr. Polym., 123, 443 (2015)

[23] Topfer O, Schmidt-Naake G, Macromol. Symp., 248, 239 (2007)

[24] Kim SE, Song SH, Yun YP, Choi BJ, Kwon IK, Bae MS, Moon HJ, Kwon YD, Biomaterials, 32, 366 (2011)

[25] Darwis D, Khusniya T, Hardiningsih L, Nurlidar F, Winarno H, Atom Indonesia, 38, 78 (2012)