From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluation

This paper describes a promising candidate molecule, investigates the pattern of scaffold composition which arises and assesses the effect of the agent on its mechanical properties. Methods: Scaffold samples were fabricated using a commercial extrusion bioprinter equipped with a pneumatic printhead...

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Main Authors: Shahad Ahmed Daood, Martha Then Xin Yi, Nicole Wen Ce Mun, Sharjeel Ilyas, Lee Yin Shien, Oh Jia En, Syed Saad Bin Qasim, Yichen Dai, Galvinderjeet Kaur Grewal, Ng Mei Liit, Gopu Sriram, Malikarjuna Rao Pichika, Kit-Kay Mak, Ranjeet Ajit Bapat, Zeeshan Sheikh, Umer Daood
Format: Article
Language:English
Published: Elsevier 2025-12-01
Series:Regenerative Therapy
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352320425001695
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author Shahad Ahmed Daood
Martha Then Xin Yi
Nicole Wen Ce Mun
Sharjeel Ilyas
Lee Yin Shien
Oh Jia En
Syed Saad Bin Qasim
Yichen Dai
Galvinderjeet Kaur Grewal
Ng Mei Liit
Gopu Sriram
Malikarjuna Rao Pichika
Kit-Kay Mak
Ranjeet Ajit Bapat
Zeeshan Sheikh
Umer Daood
author_facet Shahad Ahmed Daood
Martha Then Xin Yi
Nicole Wen Ce Mun
Sharjeel Ilyas
Lee Yin Shien
Oh Jia En
Syed Saad Bin Qasim
Yichen Dai
Galvinderjeet Kaur Grewal
Ng Mei Liit
Gopu Sriram
Malikarjuna Rao Pichika
Kit-Kay Mak
Ranjeet Ajit Bapat
Zeeshan Sheikh
Umer Daood
author_sort Shahad Ahmed Daood
collection DOAJ
description This paper describes a promising candidate molecule, investigates the pattern of scaffold composition which arises and assesses the effect of the agent on its mechanical properties. Methods: Scaffold samples were fabricated using a commercial extrusion bioprinter equipped with a pneumatic printhead fitted with a 21G conical nozzle. The Pore Printability Index, and the area and perimeter of the pore within the grid patterns were quantified using ImageJ software (NIH, USA). Mechanical properties of scaffolds were assessed using atomic force microscopy. The phase composition and crystal structures were analyzed using X-ray diffraction and Raman mapping. Morphologies of human gingival fibroblastic cells were examined using scanning electron microscopy. Lactobacillus biofilms were generated for cytolysin peptide cleavage. A rabbit bone defect model with scaffold implantations was used to provide histologic specimens for measuring percentages of bone trabeculae, collagen fibers and inflammatory cells along with granulation tissue. The Primeway Total RNA Extraction Kit was used for RNA extraction. Results: All bioink formulations demonstrated successful printing of 3D grid and solid square patterned scaffolds achieving Pr values exceeding 0.9. 0.1 %K21 group showed the highest elastic modulus. XRD revealed a pattern producing around 90 % β-tricalcium phosphate displaying two peaks at 2θ angles. 0.1 % K21 and 0.1 %CHX did not alter scaffold's pore size and porosity. 0.1 %K21 group exhibited highest ratio (62.5 ± 6.1 θ), significantly surpassing control. Surface morphologies of cells were also well retained. TEM image shows a sequence of structural changes in fibroblastic cell structure when exposed to K21. 0.1 % K21 proved to be critical in completely eradicating the biofilm. 0.K21 group closed the openings of wound areas completely. Correlation coefficient of gene expression levels demonstrates sample variations and recurring instances among groupings. Conclusion: 3D-printing technologies with 0.1 %K21 represent a significant advancement over conventional regenerative medicine techniques for bone-related treatments.
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spelling doaj-art-3e7ea73257bc45d6bb64f721aae4729c2025-08-20T03:04:57ZengElsevierRegenerative Therapy2352-32042025-12-013055857410.1016/j.reth.2025.08.001From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluationShahad Ahmed Daood0Martha Then Xin Yi1Nicole Wen Ce Mun2Sharjeel Ilyas3Lee Yin Shien4Oh Jia En5Syed Saad Bin Qasim6Yichen Dai7Galvinderjeet Kaur Grewal8Ng Mei Liit9Gopu Sriram10Malikarjuna Rao Pichika11Kit-Kay Mak12Ranjeet Ajit Bapat13Zeeshan Sheikh14Umer Daood15Restorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaRestorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaRestorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaOral Biology, Akhtar Saeed Medical and Dental College, Lahore, PakistanRestorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaRestorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaDepartment of Bioclinical Sciences, College of Dentistry, Kuwait University, Kuwait City, KuwaitDiscipline of Oral Sciences, Faculty of Dentistry, National University of Singapore, SingaporeRestorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaRestorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaDiscipline of Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore; NUS Centre for Additive Manufacturing (AM.NUS), National University of Singapore, SingaporeSchool of Pharmacy, IMU University KualaLumpur, KualaLumpur, MalaysiaSchool of Pharmacy, IMU University KualaLumpur, KualaLumpur, MalaysiaRestorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaDepartments of Biomaterials & Applied Oral Sciences and Dental Clinical Science, Faculty of Dentistry, Dalhousie University, 5981 University Ave, Halifax, NS, B3H 1W2, Canada; School of Biomedical Engineering (SBME), Faculty of Medicine, Dalhousie University, 5981 University Ave, Halifax, NS, B3H 1W2, Canada; Faculty of Dental Medicine and Oral Health Sciences-McGill University, 2001 Av. McGill College, Montreal, QC, H3A 1G1, Canada; Faculty of Dentistry, University of Toronto, 101 Elm St, Toronto, ON, M5G 2L3, Canada; Corresponding author. Applied Oral Sciences & Dental Clinical Sciences, Faculty of Dentistry, Dalhousie University, 5981 University Ave, Halifax, Nova Scotia, B3H 1W2, Canada.Restorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, Malaysia; Dental Materials Science, Applied Oral Sciences & Community Dental Care, Faculty of Dentistry, The University of Hong Kong, 34 Hospital Road, Sai Ying Pun, Hong Kong Special Administrative Region; Corresponding author. Restorative Dentistry Division, School of Dentistry, International Medical University Kuala Lumpur, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000, Wilayah Persekutuan Kuala Lumpur, MalaysiaThis paper describes a promising candidate molecule, investigates the pattern of scaffold composition which arises and assesses the effect of the agent on its mechanical properties. Methods: Scaffold samples were fabricated using a commercial extrusion bioprinter equipped with a pneumatic printhead fitted with a 21G conical nozzle. The Pore Printability Index, and the area and perimeter of the pore within the grid patterns were quantified using ImageJ software (NIH, USA). Mechanical properties of scaffolds were assessed using atomic force microscopy. The phase composition and crystal structures were analyzed using X-ray diffraction and Raman mapping. Morphologies of human gingival fibroblastic cells were examined using scanning electron microscopy. Lactobacillus biofilms were generated for cytolysin peptide cleavage. A rabbit bone defect model with scaffold implantations was used to provide histologic specimens for measuring percentages of bone trabeculae, collagen fibers and inflammatory cells along with granulation tissue. The Primeway Total RNA Extraction Kit was used for RNA extraction. Results: All bioink formulations demonstrated successful printing of 3D grid and solid square patterned scaffolds achieving Pr values exceeding 0.9. 0.1 %K21 group showed the highest elastic modulus. XRD revealed a pattern producing around 90 % β-tricalcium phosphate displaying two peaks at 2θ angles. 0.1 % K21 and 0.1 %CHX did not alter scaffold's pore size and porosity. 0.1 %K21 group exhibited highest ratio (62.5 ± 6.1 θ), significantly surpassing control. Surface morphologies of cells were also well retained. TEM image shows a sequence of structural changes in fibroblastic cell structure when exposed to K21. 0.1 % K21 proved to be critical in completely eradicating the biofilm. 0.K21 group closed the openings of wound areas completely. Correlation coefficient of gene expression levels demonstrates sample variations and recurring instances among groupings. Conclusion: 3D-printing technologies with 0.1 %K21 represent a significant advancement over conventional regenerative medicine techniques for bone-related treatments.http://www.sciencedirect.com/science/article/pii/S2352320425001695BoneCollagenAntimicrobialTri-calcium phosphateRamanCytotoxicity
spellingShingle Shahad Ahmed Daood
Martha Then Xin Yi
Nicole Wen Ce Mun
Sharjeel Ilyas
Lee Yin Shien
Oh Jia En
Syed Saad Bin Qasim
Yichen Dai
Galvinderjeet Kaur Grewal
Ng Mei Liit
Gopu Sriram
Malikarjuna Rao Pichika
Kit-Kay Mak
Ranjeet Ajit Bapat
Zeeshan Sheikh
Umer Daood
From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluation
Regenerative Therapy
Bone
Collagen
Antimicrobial
Tri-calcium phosphate
Raman
Cytotoxicity
title From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluation
title_full From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluation
title_fullStr From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluation
title_full_unstemmed From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluation
title_short From an Antimicrobial Agent to a constituent of 3D Printed Heterogenous Scaffolds Stimulating Bone Characteristics: An In-vitro and Animal model evaluation
title_sort from an antimicrobial agent to a constituent of 3d printed heterogenous scaffolds stimulating bone characteristics an in vitro and animal model evaluation
topic Bone
Collagen
Antimicrobial
Tri-calcium phosphate
Raman
Cytotoxicity
url http://www.sciencedirect.com/science/article/pii/S2352320425001695
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