Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs

Abstract Background Three-dimensional (3D) spheroid culture can promote the osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSC). 3D printing offers the possibility to produce customized scaffolds for complex bone defects. The aim of this study was to compare the potential of h...

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Autores principales:	Siddharth Shanbhag, Salwa Suliman, Samih Mohamed-Ahmed, Carina Kampleitner, Mohamed Nageeb Hassan, Patrick Heimel, Toni Dobsak, Stefan Tangl, Anne Isine Bolstad, Kamal Mustafa
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Xeno-free Platelet lysate MSC Spheroid culture Bone tissue engineering Medicine (General) R5-920 Biochemistry QD415-436
Acceso en línea:	https://doaj.org/article/a6354fed2ab0472b96c17a790423244f
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spelling	oai:doaj.org-article:a6354fed2ab0472b96c17a790423244f2021-11-21T12:04:43ZBone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs10.1186/s13287-021-02642-w1757-6512https://doaj.org/article/a6354fed2ab0472b96c17a790423244f2021-11-01T00:00:00Zhttps://doi.org/10.1186/s13287-021-02642-whttps://doaj.org/toc/1757-6512Abstract Background Three-dimensional (3D) spheroid culture can promote the osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSC). 3D printing offers the possibility to produce customized scaffolds for complex bone defects. The aim of this study was to compare the potential of human BMSC cultured as 2D monolayers or 3D spheroids encapsulated in constructs of 3D-printed poly-L-lactide-co-trimethylene carbonate scaffolds and modified human platelet lysate hydrogels (PLATMC-HPLG) for bone regeneration. Methods PLATMC-HPLG constructs with 2D or 3D BMSC were assessed for osteogenic differentiation based on gene expression and in vitro mineralization. Subsequently, PLATMC-HPLG constructs with 2D or 3D BMSC were implanted in rat calvarial defects for 12 weeks; cell-free constructs served as controls. Bone regeneration was assessed via in vivo computed tomography (CT), ex vivo micro-CT and histology. Results Osteogenic gene expression was significantly enhanced in 3D versus 2D BMSC prior to, but not after, encapsulation in PLATMC-HPLG constructs. A trend for greater in vitro mineralization was observed in constructs with 3D versus 2D BMSC (p > 0.05). In vivo CT revealed comparable bone formation after 4, 8 and 12 weeks in all groups. After 12 weeks, micro-CT revealed substantial regeneration in 2D BMSC (62.47 ± 19.46%), 3D BMSC (51.01 ± 24.43%) and cell-free PLATMC-HPLG constructs (43.20 ± 30.09%) (p > 0.05). A similar trend was observed in the histological analysis. Conclusion Despite a trend for superior in vitro mineralization, constructs with 3D and 2D BMSC performed similarly in vivo. Regardless of monolayer or spheroid cell culture, PLATMC-HPLG constructs represent promising scaffolds for bone tissue engineering applications.Siddharth ShanbhagSalwa SulimanSamih Mohamed-AhmedCarina KampleitnerMohamed Nageeb HassanPatrick HeimelToni DobsakStefan TanglAnne Isine BolstadKamal MustafaBMCarticleXeno-freePlatelet lysateMSCSpheroid cultureBone tissue engineeringMedicine (General)R5-920BiochemistryQD415-436ENStem Cell Research & Therapy, Vol 12, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Xeno-free Platelet lysate MSC Spheroid culture Bone tissue engineering Medicine (General) R5-920 Biochemistry QD415-436
spellingShingle	Xeno-free Platelet lysate MSC Spheroid culture Bone tissue engineering Medicine (General) R5-920 Biochemistry QD415-436 Siddharth Shanbhag Salwa Suliman Samih Mohamed-Ahmed Carina Kampleitner Mohamed Nageeb Hassan Patrick Heimel Toni Dobsak Stefan Tangl Anne Isine Bolstad Kamal Mustafa Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs
description	Abstract Background Three-dimensional (3D) spheroid culture can promote the osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSC). 3D printing offers the possibility to produce customized scaffolds for complex bone defects. The aim of this study was to compare the potential of human BMSC cultured as 2D monolayers or 3D spheroids encapsulated in constructs of 3D-printed poly-L-lactide-co-trimethylene carbonate scaffolds and modified human platelet lysate hydrogels (PLATMC-HPLG) for bone regeneration. Methods PLATMC-HPLG constructs with 2D or 3D BMSC were assessed for osteogenic differentiation based on gene expression and in vitro mineralization. Subsequently, PLATMC-HPLG constructs with 2D or 3D BMSC were implanted in rat calvarial defects for 12 weeks; cell-free constructs served as controls. Bone regeneration was assessed via in vivo computed tomography (CT), ex vivo micro-CT and histology. Results Osteogenic gene expression was significantly enhanced in 3D versus 2D BMSC prior to, but not after, encapsulation in PLATMC-HPLG constructs. A trend for greater in vitro mineralization was observed in constructs with 3D versus 2D BMSC (p > 0.05). In vivo CT revealed comparable bone formation after 4, 8 and 12 weeks in all groups. After 12 weeks, micro-CT revealed substantial regeneration in 2D BMSC (62.47 ± 19.46%), 3D BMSC (51.01 ± 24.43%) and cell-free PLATMC-HPLG constructs (43.20 ± 30.09%) (p > 0.05). A similar trend was observed in the histological analysis. Conclusion Despite a trend for superior in vitro mineralization, constructs with 3D and 2D BMSC performed similarly in vivo. Regardless of monolayer or spheroid cell culture, PLATMC-HPLG constructs represent promising scaffolds for bone tissue engineering applications.
format	article
author	Siddharth Shanbhag Salwa Suliman Samih Mohamed-Ahmed Carina Kampleitner Mohamed Nageeb Hassan Patrick Heimel Toni Dobsak Stefan Tangl Anne Isine Bolstad Kamal Mustafa
author_facet	Siddharth Shanbhag Salwa Suliman Samih Mohamed-Ahmed Carina Kampleitner Mohamed Nageeb Hassan Patrick Heimel Toni Dobsak Stefan Tangl Anne Isine Bolstad Kamal Mustafa
author_sort	Siddharth Shanbhag
title	Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs
title_short	Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs
title_full	Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs
title_fullStr	Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs
title_full_unstemmed	Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs
title_sort	bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs
publisher	BMC
publishDate	2021
url	https://doaj.org/article/a6354fed2ab0472b96c17a790423244f
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Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs

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