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Three-dimensional osteogenic differentiation of bone marrow mesenchymal stem cells promotes matrix metallopeptidase 13 (Mmp13) expression in type i collagen hydrogels
Umeå universitet, Medicinska fakulteten, Institutionen för integrativ medicinsk biologi (IMB).
Umeå universitet, Medicinska fakulteten, Institutionen för integrativ medicinsk biologi (IMB).ORCID-id: 0000-0003-2596-5936
Umeå universitet, Medicinska fakulteten, Institutionen för integrativ medicinsk biologi (IMB).ORCID-id: 0000-0002-6181-9904
Umeå universitet, Medicinska fakulteten, Institutionen för integrativ medicinsk biologi (IMB). Umeå universitet, Medicinska fakulteten, Institutionen för kirurgisk och perioperativ vetenskap, Handkirurgi.
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2021 (Engelska)Ingår i: International Journal of Molecular Sciences, ISSN 1661-6596, E-ISSN 1422-0067, Vol. 22, nr 24, artikel-id 13594Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Autologous bone transplantation is the principal method for reconstruction of large bone defects. This technique has limitations, such as donor site availability, amount of bone needed and morbidity. An alternative to this technique is tissue engineering with bone marrow-derived mesenchymal stem cells (BMSCs). In this study, our aim was to elucidate the benefits of culturing BMSCs in 3D compared with the traditional 2D culture. In an initial screening, we combined BMSCs with four different biogels: unmodified type I collagen (Col I), type I collagen methacrylate (ColMa), an alginate and cellulose-based bioink (CELLINK) and a gelatin-based bioink containing xanthan gum (GelXA-bone). Col I was the best for structural integrity and maintenance of cell morphology. Osteogenic, adipogenic, and chondrogenic differentiations of the BMSCs in 2D versus 3D type I collagen gels were investigated. While the traditional pellet culture for chondrogenesis was superior to our tested 3D culture, Col I hydrogels (i.e., 3D) favored adipogenic and osteogenic differentiation. Further focus of this study on osteogenesis were conducted by comparing 2D and 3D differentiated BMSCs with Osteoimage® (stains hydroxyapatite), von Kossa (stains anionic portion of phosphates, carbonates, and other salts) and Alizarin Red (stains Ca2+ deposits). Multivariate gene analysis with various covariates showed low variability among donors, successful osteogenic differentiation, and the identification of one gene (matrix metallopeptidase 13, MMP13) significantly differentially expressed in 2D vs. 3D cultures. MMP13 protein expression was confirmed with immunohistochemistry. In conclusion, this study shows evidence for the suitability of type I collagen gels for 3D osteogenic differentiation of BMSCs, which might improve the production of tissue-engineered constructs for treatment of bone defects.

Ort, förlag, år, upplaga, sidor
MDPI, 2021. Vol. 22, nr 24, artikel-id 13594
Nyckelord [en]
3D culture, Biogel, Cell differentiation, Mesenchymal stem cells, MMP13, MSCs, Osteogenesis, Type I collagen
Nationell ämneskategori
Medicinska material och protesteknik
Identifikatorer
URN: urn:nbn:se:umu:diva-190846DOI: 10.3390/ijms222413594ISI: 000737916100001Scopus ID: 2-s2.0-85121319833OAI: oai:DiVA.org:umu-190846DiVA, id: diva2:1623460
Tillgänglig från: 2021-12-29 Skapad: 2021-12-29 Senast uppdaterad: 2024-07-02Bibliografiskt granskad
Ingår i avhandling
1. Development and optimization of a 3D in vitro model for osteogenic biomaterial evaluation
Öppna denna publikation i ny flik eller fönster >>Development and optimization of a 3D in vitro model for osteogenic biomaterial evaluation
2023 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Despite the innate regenerative capabilities of bone tissue, self-repair is impaired when an injury exceeds the critical size threshold because of trauma, congenital, or pathological conditions. Autologous transplantation is the gold standard to reconstruct large bone defects. However, this method has drawbacks such as limited amount of graftable material, limited accessibility, and donor site morbidity. For these reasons, alternative regenerative medicine and tissue engineering approaches are being developed, including implantable scaffolds.The use of in vitro-made scaffolds containing biomaterials that mimic the functional characteristics and composition of extracellular bone matrix has been favored in 3D vs 2D in vitro culture systems. Adult stem cells such as mesenchymal stem cells (MSCs), that give rise to bone building cells, have been used in combination with various biomaterials. The development of an implantable scaffold with or without cells requires extensive in vitro validation and optimization prior to its testing in vivo. Thus, the primary aim of this thesis was to develop a 3D model for the optimization of MSC differentiation. Further aims were to utilize this 3D model to evaluate the MSC response to a novel osteogenic biomaterial. To achieve these objectives human bone marrow MSCs (BMSCs) were utilized in various hydrogels in combination with chemical differentiation factors or biomaterials. Moreover, the osteogenic capability of the tested biomaterials and their induced inflammatory/angiogenic responses were investigated, and the culture conditions were optimized for clinical application. In this thesis, a comparison between 2D and 3D (hydrogels) in vitro culture models was developed with the purpose of studying osteogenic differentiation in BSMCs. Testing various hydrogels revealedt he superiority of type I collagen hydrogels for the osteogenic 3D in vitro culture system. Further, cell culture conditions were improved for the expansion and differentiation of BMSCs to fulfill clinically approved standards according to Good Manufacturing Practice (GMP) conditions. Comparisons between fetal bovine serum (FBS) and human platelet lysate (PLT) showed superior cellular differentiation in FBS, while PLT enhanced cell proliferation. Based on the developed 3D model, the osteogenic properties of a novel nanoporous silica calcium phosphate (nSCP) material were investigated. The results indicated that nSCP was osteoinductive, involving different pathways compared with the traditional chemical differentiation protocols or other tested osteogenic biomaterials. Finally, the inflammatory and angiogenic responses from human BMSCs and an immortalized monocyte cell line (THP-1) exposed to nSCP in the established 3D model were assessed.The results indicated limited inflammatory effect of nSCP, while inducing the secretion of pro-angiogenic cytokines. The bioactivity of these released factors was confirmed in an assay using human endothelial cells.Taken together, this thesis presents a 3D in vitro model for studying osteogenic differentiation in MSCs, which can be utilized to evaluate, validate, and optimize biomaterial candidates for bone regeneration applications.

Ort, förlag, år, upplaga, sidor
Umeå: Umeå University, 2023. s. 68
Serie
Umeå University medical dissertations, ISSN 0346-6612 ; 2265
Nyckelord
angiogenesis, biomaterial, bone marrow mesenchymal stem cells, bone regeneration, hydrogel, osteoinduction, osteogenesis, scaffold, type I collagen, 3D culture
Nationell ämneskategori
Cellbiologi
Forskningsämne
biologi; biomedicinsk laboratorievetenskap; medicinsk cellbiologi
Identifikatorer
urn:nbn:se:umu:diva-216944 (URN)978-91-8070-189-1 (ISBN)978-91-8070-188-4 (ISBN)
Disputation
2023-12-15, NAT.D.360, Naturvetarhuset, Umeå, 09:00 (Engelska)
Opponent
Handledare
Forskningsfinansiär
Umeå universitetRegion Västerbotten, 7003459, and 7003589
Tillgänglig från: 2023-11-24 Skapad: 2023-11-20 Senast uppdaterad: 2023-11-23Bibliografiskt granskad

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Anerillas, Luis OliverosKingham, Paul J.Lammi, MikkoWiberg, MikaelKelk, Peyman

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