Document Type : Original Article

Authors

1 Postgarduate Student, Dept. of Periodontics, Faculty of Dentistry, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran.

2 Dept. of Periodontics, Faculty of Dentistry, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran.

Abstract

Statement of the Problem: Predictable bone regeneration is an objective in implant and periodontal treatments and barrier membranes may play a significant role in osteogenic reconstruction and differentiation.
Purpose: We compared the osteoblastic differentiation level of bone marrow stem cells in the vicinity of different barrier membranes.
Materials and Method: In this experimental in vitro study, human collagen membrane (HCM; Regen), xenogeneic collagen membrane (XCM; Jason), human acellular dermal matrix (HADM; Regen), and xenogeneic acellular dermal matrix (XADM) were used in 4 groups. No membranes were used in the control group (5th group). Bone marrow stem cells with 150,000 cells/well density were added to the culture medium. Cellular differentiation was assessed through real-time quantitative reverse transcription polymerase chain reaction (QRT-PCR) for alkaline phosphatase (ALP) and osteopontin (OPN) gene expression, and Alizarin Red staining after 21 days. Data were analyzed using Kruskal- Wallis and Mann–Whitney statistical tests on SPSS 20 software (p Value< 0.05).
Results: ALP gene expression was significantly higher in HCM group compared to other four groups (p< 0.009) followed by XADM, control, HADM and XCM groups, respectively (p< 0.001). OPN gene expression was significantly more prominent in HCM group compared to other groups (p< 0.01) followed by XADM group in which OPN gene was expressed significantly more than XCM group. OPN gene expression was not significantly different in HADM and control groups (p= 0.52). Light absorption rate was higher in HCM group compared with other groups (p< 0.012). Light absorption rate was not significantly different among HADM, XADM, and control groups (p> 0.05), though it was higher in XCM group (p= 0.009).
Conclusion: Bone marrow stem cells show different levels of differentiation in the vicinity of different membranes. Generally, cell differentiation was more prominent in the vicinity of human collagen membrane.

Keywords

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  • Wang HL, Miyauchi M, Takata T. Initial attachment of osteoblasts to various guided bone regeneration membranes: an in vitro study. J Periodontal Res. 2002; 37: 340-344.
  • Jacob SA, Amudha D. Guided Tissue Regeneration: A Review. J Dent Health Oral Disord Ther. 2017; 6: 00197.
  • Soheilifar S, Soheilifar S, Bidgoli M, Torkzaban P. Barrier Membrane, a Device for Regeneration: Properties and Applications. Avicenna J Dent Res. 2014; 6: e21343.
  • Burridge K, Molony L, Kelly T. Adhesion plaques: sites of transmembrane interaction between the extracellular matrix and the actin cytoskeleton. J Cell Sci Suppl. 1987; 8: 211-229.
  • Shanmugam M, Sivakumar V, Anitha V, Sivakumar B. Clinical evaluation of alloderm for root coverage and colour match. J Indian Soc Periodontol. 2012; 16: 218-223.
  • Hammerle CHF. Membranes and bone substitutes in guided bone regeneration. Proceedings of the 3rd European Workshop on Periodontology. 1999; 3: 468-499.
  • Rakhmatia YD, Ayukawa Y, Furuhashi A, Koyano K. Current barrier membranes: Titanium mesh and other membranes for guided bone regeneration in dental applications. J. Prosthodont Res. 2013; 57: 3–14.
  • Monteiro AS, Macedo LG, Macedo NL, Balducci I. Polyurethane and PTFE membranes for guided bone regeneration: histopathological and ultrastructural evaluation. Med Oral Patol Oral Cir Bucal. 2010; 15: 401-406.
  • Erdag G, Sheridan RL. Fibroblasts improve performance of cultured composite skin substitutes on athymic mice. Burns. 2004; 30: 322-328.
  • Allen EP. AlloDerm: an effective alternative to palatal donor tissue for treatment of gingival recession. Dent Today. 2006; 25: 48, 50-52; quiz 52.
  • Shirakata Y, Sculean A,Shinohara Y, Sena K, Takeuchi N, Bosshardt Healing of localized gingival recession treated with a coronally advanced flap alone or combined with an enamel matrix derivative and a porcine acellular dermal matrix: a preclinical study. Clin Oral Invest. 2015; 10: 1680-1684.
  • Pabst AM, Lehmann KM, Walter C, Krüger M, Stratul SI, Kasaj A. Influence of porcine-derived collagen matrix on endothelial progenitor cells: an in vitro study. Odontology. 2016; 104: 19-26.
  • Rothamel D, Benner M, Fienitz T, Happe A, Kreppel M, Nickenig HJ, et al. Biodegradation pattern and tissue integration of native and cross-linked porcine collagen soft tissue augmentation matrices- an experimental study in the rat. Head Face Med. 2014; 10: 10.
  • Chang T, Liu Q, Marino V, Bartold PM. Attachment of periodontal fibroblasts to barrier membranes coated with platelet-rich plasma. Aust Dent J. 2007; 52: 227-233.
  • Bartold PM, McCulloch CA, Narayanan AS, Pitaru S. Tissue engineering: a new paradigm for periodontal regeneration based on molecular and cell biology. Periodontol 2000. 2000; 24: 253-269.
  • Pejcic A, Kojovic D, Mirkovic D, Minic Stem Cells for Periodontal Regeneration. Balkan J Med Genet. 2013; 16: 7–12.
  • Hanna H, Mir LM, Andre F. In vitro osteoblastic differentiation of mesenchymal stem cells generates cell layers with distinct properties. Stem Cell Research & Therapy. 2018; 9: 203.
  • Basudan A, Babay N, Ramalingam S, Nooh N, Al-Kindi M, Al-Rasheed A, et al. Efficacy of mucograft vs conventional resorbable collagen membranes in guided bone regeneration around standardized calvarial defects in rats: an in vivo microcomputed tomographic analysis. Int J Periodontics Restorative Dent. 2016; 36 Suppl: 109-21.
  • An YZ, Kim YK, Lim SM, Heo YK, Kwon MK, Cha JK, et al. Physiochemical properties and resorption progress of porcine skin-derived collagen membranes: In vitro and in vivo analysis. Dent Mater J. 2018; 37: 332-340.
  • Pappalardo S, Guarnieri R. Proliferation Assessment of Mesenchymal Stem Cells on an Acellular Dermal Matrix (AlloDerm®GBR) Used for Guided Bone Regeneration. J Biomate Tissue Eng. 2013; 3: 589-596.
  • Miron RJ, Bosshardt DD, Gemperli AC, Dard M, Buser D, Gruber R, et al. In vitro characterization of a synthetic calcium phosphate bone graft on periodontal ligament cell and osteoblast behavior and its combination with an enamel matrix derivative. Clin Oral Invest. 2014; 18: 443-451.
  • Pabst AM, Happe A, Callaway A, Ziebart T, Stratul SI, Ackermann M, et al. In vitro and in vivo characterization of porcine acellular dermal matrix for gingival augmentation procedures. J Periodontal Res. 2014; 49: 371-381.
  • Rothamel D, Schwarz F, Fienitz T, Smeets R, Dreiseidler T, Ritter L, et al. Biocompatibility and biodegradation of a native porcine pericardium membrane: results of in vitro and in vivo examinations. Int J Oral Maxillofac Implants. 2012; 27: 146-154. 
  • Liu M, Wang SA, Mo A, Meng Y, Hu J, Li XY. Study on the microstructure of acellular dermal matrix and its biocompatibility with MG63 osteoblast-like cells. Hua Xi Kou Qiang Yi Xue Za Zhi. 2008; 26: 129-132.
  • Rothamel D, Schwarz F, Sculean A, Herten M, Scherbaum W, Becker J. Biocompatibility of various collagen membranes in cultures of human PDL fibroblasts and human osteoblast-like cells. Clin Oral Implants Res. 2004; 15: 443-449.
  • Papaioannou KA, Markopoulou CE, Gioni V, Mamalis AA, Vayouraki HN, Kletsas D. Attachment and proliferation of human osteoblast like cells on guided bone regeneration (GBR) membranes in the absence or presence of nicotine: an in vitro study. Int J Oral Maxillofac Implants. 2011; 26: 509-519.
  • Miron RJ, Fujioka-Kobayashi M, Buser D, Zhang Y, Bosshardt DD, Sculean A. Combination of Collagen Barrier Membrane with Enamel Matrix Derivative-Liquid Improves Osteoblast Adhesion and Differentiation. Int J Oral Maxillofac Implants. 2017; 32: 196-203.
  • Miron RJ, Saulacic N, Buser D, Iizuka T, Sculean A. Osteoblast proliferation and differentiation on a barrier membrane in combination with BMP2 and TGFβ1. Clin Oral Investig. 2013; 17: 981-988. 
  • Kobayashi M, Serrano J, Bosshardt D, Gruber R, Buser D, Miron R. Bone conditioned media (BCM) improves osteoblast adhesion and differentiation on collagen barrier membranes. BMC Oral Health. 2017; 17: 7.
  • Ziccardi VB, Buchbinder D. Guided tissue regeneration in dentistry. N Y State Dent J. 1996; 62: 48-51.
  • Wang HL, Miyauchi M, Takata T. Initial attachment of osteoblasts to various guided bone regeneration membranes: an in vitro study. J Periodontal Res. 2002; 37: 340-344.
  • Jacob SA, Amudha D. Guided Tissue Regeneration: A Review. J Dent Health Oral Disord Ther. 2017; 6: 00197.
  • Soheilifar S, Soheilifar S, Bidgoli M, Torkzaban P. Barrier Membrane, a Device for Regeneration: Properties and Applications. Avicenna J Dent Res. 2014; 6: e21343.
  • Burridge K, Molony L, Kelly T. Adhesion plaques: sites of transmembrane interaction between the extracellular matrix and the actin cytoskeleton. J Cell Sci Suppl. 1987; 8: 211-229.
  • Shanmugam M, Sivakumar V, Anitha V, Sivakumar B. Clinical evaluation of alloderm for root coverage and colour match. J Indian Soc Periodontol. 2012; 16: 218-223.
  • Hammerle CHF. Membranes and bone substitutes in guided bone regeneration. Proceedings of the 3rd European Workshop on Periodontology. 1999; 3: 468-499.
  • Rakhmatia YD, Ayukawa Y, Furuhashi A, Koyano K. Current barrier membranes: Titanium mesh and other membranes for guided bone regeneration in dental applications. J. Prosthodont Res. 2013; 57: 3–14.
  • Monteiro AS, Macedo LG, Macedo NL, Balducci I. Polyurethane and PTFE membranes for guided bone regeneration: histopathological and ultrastructural evaluation. Med Oral Patol Oral Cir Bucal. 2010; 15: 401-406.
  • Erdag G, Sheridan RL. Fibroblasts improve performance of cultured composite skin substitutes on athymic mice. Burns. 2004; 30: 322-328.
  • Allen EP. AlloDerm: an effective alternative to palatal donor tissue for treatment of gingival recession. Dent Today. 2006; 25: 48, 50-52; quiz 52.
  • Shirakata Y, Sculean A,Shinohara Y, Sena K, Takeuchi N, Bosshardt Healing of localized gingival recession treated with a coronally advanced flap alone or combined with an enamel matrix derivative and a porcine acellular dermal matrix: a preclinical study. Clin Oral Invest. 2015; 10: 1680-1684.
  • Pabst AM, Lehmann KM, Walter C, Krüger M, Stratul SI, Kasaj A. Influence of porcine-derived collagen matrix on endothelial progenitor cells: an in vitro study. Odontology. 2016; 104: 19-26.
  • Rothamel D, Benner M, Fienitz T, Happe A, Kreppel M, Nickenig HJ, et al. Biodegradation pattern and tissue integration of native and cross-linked porcine collagen soft tissue augmentation matrices- an experimental study in the rat. Head Face Med. 2014; 10: 10.
  • Chang T, Liu Q, Marino V, Bartold PM. Attachment of periodontal fibroblasts to barrier membranes coated with platelet-rich plasma. Aust Dent J. 2007; 52: 227-233.
  • Bartold PM, McCulloch CA, Narayanan AS, Pitaru S. Tissue engineering: a new paradigm for periodontal regeneration based on molecular and cell biology. Periodontol 2000. 2000; 24: 253-269.
  • Pejcic A, Kojovic D, Mirkovic D, Minic Stem Cells for Periodontal Regeneration. Balkan J Med Genet. 2013; 16: 7–12.
  • Hanna H, Mir LM, Andre F. In vitro osteoblastic differentiation of mesenchymal stem cells generates cell layers with distinct properties. Stem Cell Research & Therapy. 2018; 9: 203.
  • Basudan A, Babay N, Ramalingam S, Nooh N, Al-Kindi M, Al-Rasheed A, et al. Efficacy of mucograft vs conventional resorbable collagen membranes in guided bone regeneration around standardized calvarial defects in rats: an in vivo microcomputed tomographic analysis. Int J Periodontics Restorative Dent. 2016; 36 Suppl: 109-21.
  • An YZ, Kim YK, Lim SM, Heo YK, Kwon MK, Cha JK, et al. Physiochemical properties and resorption progress of porcine skin-derived collagen membranes: In vitro and in vivo analysis. Dent Mater J. 2018; 37: 332-340.
  • Pappalardo S, Guarnieri R. Proliferation Assessment of Mesenchymal Stem Cells on an Acellular Dermal Matrix (AlloDerm®GBR) Used for Guided Bone Regeneration. J Biomate Tissue Eng. 2013; 3: 589-596.
  • Miron RJ, Bosshardt DD, Gemperli AC, Dard M, Buser D, Gruber R, et al. In vitro characterization of a synthetic calcium phosphate bone graft on periodontal ligament cell and osteoblast behavior and its combination with an enamel matrix derivative. Clin Oral Invest. 2014; 18: 443-451.
  • Pabst AM, Happe A, Callaway A, Ziebart T, Stratul SI, Ackermann M, et al. In vitro and in vivo characterization of porcine acellular dermal matrix for gingival augmentation procedures. J Periodontal Res. 2014; 49: 371-381.
  • Rothamel D, Schwarz F, Fienitz T, Smeets R, Dreiseidler T, Ritter L, et al. Biocompatibility and biodegradation of a native porcine pericardium membrane: results of in vitro and in vivo examinations. Int J Oral Maxillofac Implants. 2012; 27: 146-154. 
  • Liu M, Wang SA, Mo A, Meng Y, Hu J, Li XY. Study on the microstructure of acellular dermal matrix and its biocompatibility with MG63 osteoblast-like cells. Hua Xi Kou Qiang Yi Xue Za Zhi. 2008; 26: 129-132.
  • Rothamel D, Schwarz F, Sculean A, Herten M, Scherbaum W, Becker J. Biocompatibility of various collagen membranes in cultures of human PDL fibroblasts and human osteoblast-like cells. Clin Oral Implants Res. 2004; 15: 443-449.
  • Papaioannou KA, Markopoulou CE, Gioni V, Mamalis AA, Vayouraki HN, Kletsas D. Attachment and proliferation of human osteoblast like cells on guided bone regeneration (GBR) membranes in the absence or presence of nicotine: an in vitro study. Int J Oral Maxillofac Implants. 2011; 26: 509-519.
  • Miron RJ, Fujioka-Kobayashi M, Buser D, Zhang Y, Bosshardt DD, Sculean A. Combination of Collagen Barrier Membrane with Enamel Matrix Derivative-Liquid Improves Osteoblast Adhesion and Differentiation. Int J Oral Maxillofac Implants. 2017; 32: 196-203.
  • Miron RJ, Saulacic N, Buser D, Iizuka T, Sculean A. Osteoblast proliferation and differentiation on a barrier membrane in combination with BMP2 and TGFβ1. Clin Oral Investig. 2013; 17: 981-988. 
  • Kobayashi M, Serrano J, Bosshardt D, Gruber R, Buser D, Miron R. Bone conditioned media (BCM) improves osteoblast adhesion and differentiation on collagen barrier membranes. BMC Oral Health. 2017; 17: 7.