Evaluation of decellularization process for developing osteogenic bovine cancellous bone scaffolds in-vitro
Current immunological issues in bone grafting regarding the transfer of xenogeneic donor bone cells into the recipient are challenging the industry to produce safer acellular natural matrices for bone regeneration. The aim of this study was to investigate the efficacy of a novel decellularization te...
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| Veröffentlicht in: | PloS one 2023-04, Vol.18 (4), p.e0283922 |
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| creator | Al Qabbani, Ali Rani, K G Aghila Syarif, Junaidi AlKawas, Sausan Sheikh Abdul Hamid, Suzina Samsudin, A R Azlina, Ahmad |
| description | Current immunological issues in bone grafting regarding the transfer of xenogeneic donor bone cells into the recipient are challenging the industry to produce safer acellular natural matrices for bone regeneration. The aim of this study was to investigate the efficacy of a novel decellularization technique for producing bovine cancellous bone scaffold and compare its physicochemical, mechanical, and biological characteristics with demineralized cancellous bone scaffold in an in-vitro study. Cancellous bone blocks were harvested from a bovine femoral head (18-24 months old) subjected to physical cleansing and chemical defatting, and further processed in two ways. Group I was subjected to demineralization, while Group II underwent decellularization through physical, chemical, and enzymatic treatments. Both were then freeze-dried, and gamma radiated, finally producing a demineralized bovine cancellous bone (DMB) scaffold and decellularized bovine cancellous bone (DCC) scaffold. Both DMB and DCC scaffolds were subjected to histological evaluation, scanning electron microscopy/energy-dispersive X-ray spectroscopy (SEM/EDS), fourier-transform infrared spectroscopy (FTIR), quantification of lipid, collagen, and residual nucleic acid content, and mechanical testing. The osteogenic potential was investigated through the recellularization of scaffolds with human osteoblast cell seeding and examined for cell attachment, proliferation, and mineralization by Alizarin staining and gene expression. DCC produced a complete acellular extracellular matrix (ECM) with the absence of nucleic acid content, wider pores with extensive interconnectivity and partially retaining collagen fibrils. DCC demonstrated a higher cell proliferation rate, upregulation of osteogenic differentiation markers, and substantial mineralized nodules production. Our findings suggest that the decellularization technique produced an acellular DCC scaffold with minimal damage to ECM and possesses osteogenic potential through the mechanisms of osteoconduction, osteoinduction, and osteogenesis in-vitro. |
| doi_str_mv | 10.1371/journal.pone.0283922 |
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The aim of this study was to investigate the efficacy of a novel decellularization technique for producing bovine cancellous bone scaffold and compare its physicochemical, mechanical, and biological characteristics with demineralized cancellous bone scaffold in an in-vitro study. Cancellous bone blocks were harvested from a bovine femoral head (18-24 months old) subjected to physical cleansing and chemical defatting, and further processed in two ways. Group I was subjected to demineralization, while Group II underwent decellularization through physical, chemical, and enzymatic treatments. Both were then freeze-dried, and gamma radiated, finally producing a demineralized bovine cancellous bone (DMB) scaffold and decellularized bovine cancellous bone (DCC) scaffold. Both DMB and DCC scaffolds were subjected to histological evaluation, scanning electron microscopy/energy-dispersive X-ray spectroscopy (SEM/EDS), fourier-transform infrared spectroscopy (FTIR), quantification of lipid, collagen, and residual nucleic acid content, and mechanical testing. The osteogenic potential was investigated through the recellularization of scaffolds with human osteoblast cell seeding and examined for cell attachment, proliferation, and mineralization by Alizarin staining and gene expression. DCC produced a complete acellular extracellular matrix (ECM) with the absence of nucleic acid content, wider pores with extensive interconnectivity and partially retaining collagen fibrils. DCC demonstrated a higher cell proliferation rate, upregulation of osteogenic differentiation markers, and substantial mineralized nodules production. Our findings suggest that the decellularization technique produced an acellular DCC scaffold with minimal damage to ECM and possesses osteogenic potential through the mechanisms of osteoconduction, osteoinduction, and osteogenesis in-vitro.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0283922</identifier><identifier>PMID: 37018321</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Acids ; Alcohol ; Alizarin ; Animals ; Biology and Life Sciences ; Biomedical engineering ; Bone (cancellous) ; Bone demineralization ; Bone grafts ; Bone growth ; Cancellous Bone ; Cattle ; Cell adhesion ; Cell Differentiation ; Cell proliferation ; Child, Preschool ; Collagen ; Collagens ; Demineralization ; Differentiation (biology) ; Electron microscopy ; Evaluation ; Extracellular matrix ; Fibrils ; Fourier transforms ; Gene expression ; Humans ; Immunology ; Infant ; Infrared spectroscopy ; Lipids ; Mechanical properties ; Mechanical tests ; Medicine and Health Sciences ; Methods ; Mineralization ; Nodules ; Nucleic Acids ; Osteoconduction ; Osteogenesis ; Osteogenesis - physiology ; Physiological aspects ; Protein seeding ; Regeneration ; Regeneration (physiology) ; Research and Analysis Methods ; Scaffolds ; Scanning electron microscopy ; Skin & tissue grafts ; Substitute bone ; Tissue Engineering - methods ; Tissue Scaffolds - chemistry ; Topography ; Trabecular bone ; X-ray spectroscopy ; Xenografts</subject><ispartof>PloS one, 2023-04, Vol.18 (4), p.e0283922</ispartof><rights>Copyright: © 2023 Al Qabbani et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</rights><rights>COPYRIGHT 2023 Public Library of Science</rights><rights>2023 Al Qabbani et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2023 Al Qabbani et al 2023 Al Qabbani et al</rights><rights>2023 Al Qabbani et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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The aim of this study was to investigate the efficacy of a novel decellularization technique for producing bovine cancellous bone scaffold and compare its physicochemical, mechanical, and biological characteristics with demineralized cancellous bone scaffold in an in-vitro study. Cancellous bone blocks were harvested from a bovine femoral head (18-24 months old) subjected to physical cleansing and chemical defatting, and further processed in two ways. Group I was subjected to demineralization, while Group II underwent decellularization through physical, chemical, and enzymatic treatments. Both were then freeze-dried, and gamma radiated, finally producing a demineralized bovine cancellous bone (DMB) scaffold and decellularized bovine cancellous bone (DCC) scaffold. Both DMB and DCC scaffolds were subjected to histological evaluation, scanning electron microscopy/energy-dispersive X-ray spectroscopy (SEM/EDS), fourier-transform infrared spectroscopy (FTIR), quantification of lipid, collagen, and residual nucleic acid content, and mechanical testing. The osteogenic potential was investigated through the recellularization of scaffolds with human osteoblast cell seeding and examined for cell attachment, proliferation, and mineralization by Alizarin staining and gene expression. DCC produced a complete acellular extracellular matrix (ECM) with the absence of nucleic acid content, wider pores with extensive interconnectivity and partially retaining collagen fibrils. DCC demonstrated a higher cell proliferation rate, upregulation of osteogenic differentiation markers, and substantial mineralized nodules production. Our findings suggest that the decellularization technique produced an acellular DCC scaffold with minimal damage to ECM and possesses osteogenic potential through the mechanisms of osteoconduction, osteoinduction, and osteogenesis in-vitro.</description><subject>Acids</subject><subject>Alcohol</subject><subject>Alizarin</subject><subject>Animals</subject><subject>Biology and Life Sciences</subject><subject>Biomedical engineering</subject><subject>Bone (cancellous)</subject><subject>Bone demineralization</subject><subject>Bone grafts</subject><subject>Bone growth</subject><subject>Cancellous Bone</subject><subject>Cattle</subject><subject>Cell adhesion</subject><subject>Cell Differentiation</subject><subject>Cell proliferation</subject><subject>Child, Preschool</subject><subject>Collagen</subject><subject>Collagens</subject><subject>Demineralization</subject><subject>Differentiation (biology)</subject><subject>Electron 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of decellularization process for developing osteogenic bovine cancellous bone scaffolds in-vitro</title><author>Al Qabbani, Ali ; Rani, K G Aghila ; Syarif, Junaidi ; AlKawas, Sausan ; Sheikh Abdul Hamid, Suzina ; Samsudin, A R ; Azlina, Ahmad</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c693t-b9046a07d58af232908586238c87fd1f0a6ffa5265aad2e11b207c89f2147c793</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Acids</topic><topic>Alcohol</topic><topic>Alizarin</topic><topic>Animals</topic><topic>Biology and Life Sciences</topic><topic>Biomedical engineering</topic><topic>Bone (cancellous)</topic><topic>Bone demineralization</topic><topic>Bone grafts</topic><topic>Bone growth</topic><topic>Cancellous Bone</topic><topic>Cattle</topic><topic>Cell adhesion</topic><topic>Cell Differentiation</topic><topic>Cell proliferation</topic><topic>Child, 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One</addtitle><date>2023-04-05</date><risdate>2023</risdate><volume>18</volume><issue>4</issue><spage>e0283922</spage><pages>e0283922-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Current immunological issues in bone grafting regarding the transfer of xenogeneic donor bone cells into the recipient are challenging the industry to produce safer acellular natural matrices for bone regeneration. The aim of this study was to investigate the efficacy of a novel decellularization technique for producing bovine cancellous bone scaffold and compare its physicochemical, mechanical, and biological characteristics with demineralized cancellous bone scaffold in an in-vitro study. Cancellous bone blocks were harvested from a bovine femoral head (18-24 months old) subjected to physical cleansing and chemical defatting, and further processed in two ways. Group I was subjected to demineralization, while Group II underwent decellularization through physical, chemical, and enzymatic treatments. Both were then freeze-dried, and gamma radiated, finally producing a demineralized bovine cancellous bone (DMB) scaffold and decellularized bovine cancellous bone (DCC) scaffold. Both DMB and DCC scaffolds were subjected to histological evaluation, scanning electron microscopy/energy-dispersive X-ray spectroscopy (SEM/EDS), fourier-transform infrared spectroscopy (FTIR), quantification of lipid, collagen, and residual nucleic acid content, and mechanical testing. The osteogenic potential was investigated through the recellularization of scaffolds with human osteoblast cell seeding and examined for cell attachment, proliferation, and mineralization by Alizarin staining and gene expression. DCC produced a complete acellular extracellular matrix (ECM) with the absence of nucleic acid content, wider pores with extensive interconnectivity and partially retaining collagen fibrils. DCC demonstrated a higher cell proliferation rate, upregulation of osteogenic differentiation markers, and substantial mineralized nodules production. Our findings suggest that the decellularization technique produced an acellular DCC scaffold with minimal damage to ECM and possesses osteogenic potential through the mechanisms of osteoconduction, osteoinduction, and osteogenesis in-vitro.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>37018321</pmid><doi>10.1371/journal.pone.0283922</doi><tpages>e0283922</tpages><orcidid>https://orcid.org/0000-0002-7226-3403</orcidid><orcidid>https://orcid.org/0000-0002-2766-6596</orcidid><orcidid>https://orcid.org/0000-0002-1393-5775</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2023-04, Vol.18 (4), p.e0283922 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2796097223 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Acids Alcohol Alizarin Animals Biology and Life Sciences Biomedical engineering Bone (cancellous) Bone demineralization Bone grafts Bone growth Cancellous Bone Cattle Cell adhesion Cell Differentiation Cell proliferation Child, Preschool Collagen Collagens Demineralization Differentiation (biology) Electron microscopy Evaluation Extracellular matrix Fibrils Fourier transforms Gene expression Humans Immunology Infant Infrared spectroscopy Lipids Mechanical properties Mechanical tests Medicine and Health Sciences Methods Mineralization Nodules Nucleic Acids Osteoconduction Osteogenesis Osteogenesis - physiology Physiological aspects Protein seeding Regeneration Regeneration (physiology) Research and Analysis Methods Scaffolds Scanning electron microscopy Skin & tissue grafts Substitute bone Tissue Engineering - methods Tissue Scaffolds - chemistry Topography Trabecular bone X-ray spectroscopy Xenografts |
| title | Evaluation of decellularization process for developing osteogenic bovine cancellous bone scaffolds in-vitro |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-02T22%3A37%3A12IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Evaluation%20of%20decellularization%20process%20for%20developing%20osteogenic%20bovine%20cancellous%20bone%20scaffolds%20in-vitro&rft.jtitle=PloS%20one&rft.au=Al%20Qabbani,%20Ali&rft.date=2023-04-05&rft.volume=18&rft.issue=4&rft.spage=e0283922&rft.pages=e0283922-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0283922&rft_dat=%3Cgale_plos_%3EA744390277%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2796097223&rft_id=info:pmid/37018321&rft_galeid=A744390277&rft_doaj_id=oai_doaj_org_article_375bf17339eb4c3cbaf6b758008a02ac&rfr_iscdi=true |