Email Alert | RSS

Journal of Peking University (Health Sciences) ›› 2021, Vol. 53 ›› Issue (4): 776-784. doi: 10.19723/j.issn.1671-167X.2021.04.026

Previous Articles     Next Articles

Biocompatibility and effect on bone formation of a native acellular porcine pericardium: Results of in vitro and in vivo

YOU Peng-yue,LIU Yu-hua(),WANG Xin-zhi,WANG Si-wen,TANG Lin   

  1. Department of Prosthodontics, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing 100081, China
  • Received:2021-06-07 Online:2021-08-18 Published:2021-08-25
  • Contact: Yu-hua LIU E-mail:lyhdentist@163.com

RICH HTML

 15   

PDF (PC)

328

Abstract:

Objective: To examine the morphology and biocompatibility of a native acellular porcine pericardium (APP) in vitro and to evaluate its barrier function and effects on osteogenesis when used in guided bone regeneration (GBR) in vivo. Methods: First, the morphology of APP (BonanGen®) was detected using a scanning electron microscope (SEM). Next, for biocompatibility test, proliferation of human bone marrow mesenchymal stem cells (hBMSCs) were determined using cell counting kit-8 (CCK-8) after being seeded 1, 3 and 7 days. Meanwhile, the cells stained with phalloidine and 4,6-diamidino-2-phenylindole (DAPI) were observed using a confocal laser scanning microscopy (CLSM) to view the morphology of cell adhesion and pattern of cell proliferation on day 5. A 3-Beagle dog model with 18 teeth extraction sockets was used for the further research in vivo. These sites were randomly treated by 3 patterns below: filled with Bio-Oss® and coverd by APP membrane (APP group), filled with Bio-Oss® and covered by Bio-Gide® membrane (BG group) and natural healing (blank group). Micro-CT and hematoxylin-eosin (HE) were performed after 4 and 12 weeks. Results: A bilayer and three-dimensional porous ultrastructure was identified for APP through SEM. In vitro, APP facilitated proliferation and adhesion of hBMSCs, especially after 7 days (P<0.05). In vivo, for the analysis of the whole socket healing, no distinct difference of new bone ratio was found between all the three groups after 4 weeks (P>0.05), however significantly more new bone regeneration was detected in APP group and BG group in comparison to blank group after 12 weeks (P<0.05). The radio of bone formation below the membrane was significantly higher in APP group and BG group than blank group after 4 and 12 weeks (P<0.05), however, the difference between APP group and BG group was merely significant in 12 weeks (P<0.05). Besides, less resorption of buccal crest after 4 weeks and 12 weeks was observed in APP group of a significant difference compared in blank group (P<0.05). The resorption in BG group was slightly lower than blank group (P>0.05). Conclusion: APP showed considerable biocompatibility and three-dimentional structure. Performing well as a barrier membrane in the dog alveolar ridge preservation model,APP significantly promoted bone regeneration below it and reduced buccal crest resorption. On the basis of this study, APP is a potential osteoconductive and osteoinductive biomaterial.

Key words: Pericardium, Resorbable membrane, Human bone marrow mesenchymal stem cell, Guided bone regeneration

CLC Number: 

  • R783.1

Figure 1

Choosing method of ROI and measurement of the vertical distance between the buccal and lingual(palatal) crest A, coronal view of region of interest (ROI); B, saggital view of ROI; C, central vertical line (CVL). BC, buccal crest; LC, lingual crest; VD, vertical distance."

Figure 2

Microstructure of APP (×300) A, smooth side of acellular porcine pericardium (APP); B, rough side of APP; C, cross section of APP."

Figure 3

CLSM view of the morphology of hBMSCs cultured on BG or APP membranes The left three rows were viewed at 250 magnification and the right three were at 5 000 magnification. Actins of cells were shown in green and nuclei were blue. APP, acellular porcine pericardium; BG, Bio-Gide."

Table 1

Percentage of different areas in ROI-1 at 4 weeks and 12 weeks /%"

Areas Time APP group BG group Blank group
Bone area 4 weeks 24.05±1.05 26.48±2.42 29.81±5.46
12 weeks 44.05±5.74c 41.50±6.22c 21.55±1.56ab
Bone marrow area 4 weeks 48.52±3.16 47.78±0.55 40.46±4.07ab
12 weeks 32.97±2.22c 33.06±4.63c 42.56±2.88ab
Soft tissue area 4 weeks 14.15±2.57 13.53±1.51 31.12±4.75ab
12 weeks 8.93±1.56c 12.56±3.45 35.88±3.03ab
Material area 4 weeks 13.24±3.07 13.58±2.42 -
12 weeks 14.46±3.91 13.49±1.93 -

Table 2

Percentage of different areas in ROI-2 at 4 and 12 weeks /%"

Areas Time APP group BG group Blank group
Bone area 4 weeks 31.26±2.76 30.36±1.72 11.26±0.91ab
12 weeks 42.56±2.88bc 37.62±1.87ac 15.09±1.81abc
Bone marrow area 4 weeks 30.05±1.70 30.59±1.20 10.67±0.71ab
12 weeks 25.69±1.96c 28.17±2.87 10.57±1.88ab
Soft tissue area 4 weeks 27.36±2.57 27.98±3.63 78.07±8.54ab
12 weeks 20.35±4.29 22.37±2.69 74.33±1.74ab
Material area 4 weeks 11.30±1.50 10.74±2.84 -
12 weeks 11.39±2.21 11.83±2.72 -

Table 3

Vertical distance of buccal and lingual crest /mm"

Time APP group BG group Blank group
4 weeks 0.89±0.09 1.32±0.33 1.53±0.29a
12 weeks 1.14±0.20 1.51±0.33 1.78±0.39a

Figure 4

Histological observations of extraction sockets healing after 4 weeks or 12 weeks APP group after 4 weeks’ healing (A, ×40; D, ×200), after 12 weeks’ healing (G, ×40; J, ×200); BG group after 4 weeks’ healing (B, ×40; E, ×200), after 12 weeks’ healing (H, ×40; K, ×200); BLANK group after 4 weeks’ healing (C, ×40; F, ×200); after 12 weeks’ healing (I, ×40; L, ×200). Asterisk, graft particles; NB, new bone; OB, old bone; blue arrow, osteoblast grew into the graft particles; black arrow, osteoclast; green arrow, osteoid."

Figure 5

Histological observations of coronal part of extraction sockets after healing of 4 weeks or 12 weeks APP group after 4 weeks’ healing (A, ×40; D, ×200), after 12 weeks’ healing (G, ×40; J, ×200); BG group after 4 weeks’ healing (B, ×40; E, ×200), after 12 weeks’ healing (H, ×40; K, ×200); blank group after 4 weeks’ healing (C, ×40; F, ×200), after 12 weeks’ healing (I, ×40; L, ×200). Asterisk, graft particles; NB, new bone; OB, old bone; blue arrow, osteoblast grew into the graft particles; green arrow, osteoid; M, membrane."

Figure 6

Schematic diagram of ROI area A, alveolar socket after tooth extraction; B, blank group after healing, soft tissue grew in and vertical bone lost; C, APP group or BG group after healing, less soft tissue grew in and less vertical bone lost."

[1] 赵丽萍, 胡文杰, 徐涛, 等. 罹患重度牙周病变磨牙拔牙后两种牙槽嵴保存方法的比较 [J]. 北京大学学报(医学版), 2019, 51(3):579-585.
[2] Karring T, Nyman S, Gottlow J, et al. Development of the biological concept of guided tissue regeneration-animal and human studies [J]. Periodontol 2000, 1993, 1(1):26-35.
pmid: 8401858
[3] Retzepi M, Donos N. Guided Bone Regeneration: biological principle and therapeutic applications [J]. Clin Oral Implants Res, 2010, 21(6):567-576.
doi: 10.1111/(ISSN)1600-0501
[4] Gruber R, Stadlinger B, Terheyden H. Cell-to-cell communication in guided bone regeneration: molecular and cellular mechanisms [J]. Clin Oral Implants Res, 2017, 28(9):1139-1146.
doi: 10.1111/clr.2017.28.issue-9
[5] Caridade SG, Mano JF. Engineering membranes for bone rege-neration [J]. Tissue Eng Part A, 2017, 23(23/24):1502-1533.
doi: 10.1089/ten.tea.2017.0094
[6] Sheikh Z, Hamdan N, Ikeda Y, et al. Natural graft tissues and synthetic biomaterials for periodontal and alveolar bone reconstructive applications: a review [J]. Biomater Res, 2017, 9(21):1-20.
[7] Badylak SF, Freytes DO, Gilbert TW. Extracellular matrix as a biological scaffold material: structure and function [J]. Acta Biomater, 2009, 5(1):1-13.
doi: 10.1016/j.actbio.2008.09.013
[8] Rothamel D, Benner M, Fienitz T, et al. Biodegradation pattern and tissue integration of native and cross-linked porcine collagen soft tissue augmentation matrices: an experimental study in the rat [J]. Head Face Med, 2014, 10(1):1-10.
doi: 10.1186/1746-160X-10-1
[9] Wang J, Wang L, Zhou Z, et al. Biodegradable polymer membranes applied in guided bone/tissue regeneration: a review [J]. Polymers (Basel), 2016, 8(4):115-135.
doi: 10.3390/polym8040115
[10] An YZ, Kim YK, Lim SM, et al. Physiochemical properties and resorption progress of porcine skin-derived collagen membranes: in vitro and in vivo analysis [J]. Dent Mater J, 2018, 37(2):332-340.
doi: 10.4012/dmj.2017-065
[11] Brown BN, Badylak SF. Extracellular matrix as an inductive scaffold for functional tissue reconstruction [J]. Transl Res, 2014, 163(4):268-285.
doi: 10.1016/j.trsl.2013.11.003
[12] Badylak SF. The extracellular matrix as a biologic scaffold material [J]. Biomaterials, 2007, 28(25):3587-3593.
doi: 10.1016/j.biomaterials.2007.04.043
[13] Shahabipour F, Banach M, Johnston TP, et al. Novel approaches toward the generation of bioscaffolds as a potential therapy in cardiovascular tissue engineering [J]. Int J Cardiol, 2017, 228:319-326.
doi: S0167-5273(16)33685-3 pmid: 27866022
[14] Zhang J, Wang GY, Xiao YP, et al. The biomechanical behavior and host response to porcine-derived small intestine submucosa, pericardium and dermal matrix acellular grafts in a rat abdominal defect model [J]. Biomaterials, 2011, 32(29):7086-7095.
doi: 10.1016/j.biomaterials.2011.06.016 pmid: 21741703
[15] 闫建伟. 牙种植引导骨再生心包胶原膜的制备及理化性能研究[D]. 山东大学, 2017.
[16] Gauvin R, Marinov G, Mehri Y, et al. A comparative study of bovine and porcine pericardium to highlight their potential advantages to manufacture percutaneous cardiovascular implants [J]. J Biomater Appl, 2013, 28(4):552-565.
doi: 10.1177/0885328212465482
[17] Khorramirouz R, Go JL, Noble C, et al. In vivo response of acellular porcine pericardial for tissue engineered transcatheter aortic valves [J]. Sci Rep, 2019, 9(1):1094-1105.
doi: 10.1038/s41598-018-37550-2 pmid: 30705386
[18] Rothamel D, Schwarz F, Fienitz T, et al. Biocompatibility and biodegradation of a native porcine pericardium membrane results of in vitro and in vivo examinations [J]. Int J Oral Maxillofac Implants, 2012, 27(1):146-154.
pmid: 22299091
[19] Meyer M. Processing of collagen based biomaterials and the resulting materials properties [J]. Biomed Eng Online, 2019, 18(1):24-98.
doi: 10.1186/s12938-019-0647-0
[20] Song C, Li S, Zhang J, et al. Controllable fabrication of porous PLGA/PCL bilayer membrane for GTR using supercritical carbon dioxide foaming [J]. Appl Surf Sci, 2019, 472:82-92.
doi: 10.1016/j.apsusc.2018.04.059
[21] Talebi Ardakani MR, Hajizadeh F, Yadegari Z. Comparison of attachment and proliferation of human gingival fibroblasts on different collagen membranes [J]. Ann Maxillofac Surg, 2018, 8(2):218-223.
doi: 10.4103/ams.ams_150_17
[22] Mendoza-Novelo B, Castellano LE, Padilla-Miranda RG, et al. The component leaching from decellularized pericardial bioscaffolds and its implication in the macrophage response [J]. J Biomed Mater Res A, 2016, 104(11):2810-2822.
doi: 10.1002/jbm.a.35825 pmid: 27387409
[23] Rajabi-Zeleti S, Jalili-Firoozinezhad S, Azarnia M, et al. The behavior of cardiac progenitor cells on macroporous pericardium-derived scaffolds [J]. Biomaterials, 2014, 35(3):970-982.
doi: 10.1016/j.biomaterials.2013.10.045 pmid: 24183165
[24] Megerle K, Woon C, Kraus A, et al. Flexor tendon sheath engineering using decellularized porcine pericardium [J]. Plast Reconstr Surg, 2016, 138(4):630e-641e.
doi: 10.1097/PRS.0000000000002459
[25] Pizzicannella J, Pierdomenico SD, Piattelli A, et al. 3D human periodontal stem cells and endothelial cells promote bone development in bovine pericardium-based tissue biomaterial [J]. Materials (Basel), 2019, 12(13):2157-2172.
doi: 10.3390/ma12132157
[26] Saulacic N, Schaller B, Munoz F, et al. Recombinant human BMP9 (RhBMP9) in comparison with rhBMP2 for ridge augmentation following tooth extraction: an experimental study in the Beagle dog [J]. Clin Oral Implants Res, 2018, 29(10):1050-1059.
doi: 10.1111/clr.2018.29.issue-10
[27] Kim JJ, Schwarz F, Song HY, et al. Ridge preservation of extraction sockets with chronic pathology using Bio-Oss® collagen with or without collagen membrane: an experimental study in dogs [J]. Clin Oral Implants Res, 2017, 28(6):727-733.
doi: 10.1111/clr.2017.28.issue-6
[28] Sun Y, Wang CY, Wang ZY, et al. Test in canine extraction site preservations by using mineralized collagen plug with or without membrane [J]. J Biomater Appl, 2016, 30(9):1285-1299.
doi: 10.1177/0885328215625429 pmid: 26721867
[29] Lozano-Carrascal N, Delgado-Ruiz RA, Gargallo-Albiol J, et al. Xenografts supplemented with pamindronate placed in postextraction sockets to avoid crestal bone resorption. Experimental study in Fox hound dogs [J]. Clin Oral Implants Res, 2016, 27(2):149-155.
[30] Araujo MG, Lindhe J. Dimensional ridge alterations following tooth extraction. An experimental study in the dog [J]. J Clin Periodontol, 2005, 32(2):212-218.
doi: 10.1111/cpe.2005.32.issue-2
[31] Macbeth N, Trullenque-Eriksson A, Donos N, et al. Hard and soft tissue changes following alveolar ridge preservation: a sys-tematic review [J]. Clin Oral Implants Res, 2017, 28(8):982-1004.
doi: 10.1111/clr.2017.28.issue-8
[1] WANG Si-wen,YOU Peng-yue,LIU Yu-hua,WANG Xin-zhi,TANG Lin,WANG Mei. Efficacy of two barrier membranes and deproteinized bovine bone mineral on bone regeneration in extraction sockets: A microcomputed tomographic study in dogs [J]. Journal of Peking University (Health Sciences), 2021, 53(2): 364-370.
[2] Wei-yi WU,Bo-wen LI,Yu-hua LIU,Xin-zhi WANG. Biodegradation properties of multi-laminated small intestinal submucosa [J]. Journal of Peking University (Health Sciences), 2020, 52(3): 564-569.
[3] Dong SHI,Jie CAO,Shi-ai DAI,Huan-xin MENG. Short-term outcome of regenerative surgery treating peri-implantitis [J]. Journal of Peking University(Health Sciences), 2020, 52(1): 58-63.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Author. English Title Test[J]. Journal of Peking University(Health Sciences), 2010, 42(1): 1 -10 .
[2] . [J]. Journal of Peking University(Health Sciences), 2009, 41(2): 188 -191 .
[3] . [J]. Journal of Peking University(Health Sciences), 2009, 41(3): 376 -379 .
[4] . [J]. Journal of Peking University(Health Sciences), 2009, 41(4): 459 -462 .
[5] . [J]. Journal of Peking University(Health Sciences), 2009, 41(6): 682 -686 .
[6] . [J]. Journal of Peking University(Health Sciences), 2010, 42(1): 82 -84 .
[7] . [J]. Journal of Peking University(Health Sciences), 2007, 39(3): 319 -322 .
[8] . [J]. Journal of Peking University(Health Sciences), 2007, 39(3): 333 -336 .
[9] . [J]. Journal of Peking University(Health Sciences), 2007, 39(3): 337 -340 .
[10] . [J]. Journal of Peking University(Health Sciences), 2007, 39(3): 225 -328 .
Copyright © Journal of Peking University (Health Sciences), All Rights Reserved.
Powered by Beijing Magtech Co. Ltd