Abstract
Simple SummaryCalcium phosphate is an important component in natural bone. Bone defects caused by trauma or resection of tumors demand bone substitutes to close the defect. One viable option is to use a scaffold material seeded with stem cells. Stem cells can differentiate towards bone cells (osteoblasts), among other cell types. To trigger stem cells towards the osteoblast type of cell, particular supplementation of the culture medium is needed. This study tests whether calcium phosphate nanoparticles can induce a commitment towards the osteoblast type of cell. In addition, we test for other commitments, such as endothelial cell, chondrocyte, or adipocyte commitment. After 1 or 2 weeks, with either 5 or 50 µg/mL nanoparticles in the culture medium, gene expression is analyzed. We find a significant increase of two specific bone marker genes after two weeks in 50 µg/mL nanoparticles compared to 5 µg/mL for two out of three tested human donors of adipose-derived stem cells. Moreover, endothelial cell commitment is also induced. Hence, such nanoparticles have the potential to trigger osteogenic and endothelial cell commitment.Amorphous calcium phosphate (aCaP) nanoparticles may trigger the osteogenic commitment of adipose-derived stem cells (ASCs) in vitro. The ASCs of three human donors are investigated using basal culture medium DMEM to either 5 or 50 µg/mL aCaP nanoparticles suspension (control: no nanoparticles). After 7 or 14 days, stem cell marker genes, as well as endothelial, osteogenic, chondrogenic, and adipogenic genes, are analyzed by qPCR. Free calcium and phosphate ion concentrations are assessed in the cell culture supernatant. After one week and 5 µg/mL aCaP, downregulation of osteogenic markers ALP and Runx2 is found, and averaged across the three donors. Our results show that after two weeks, ALP is further downregulated, but Runx2 is upregulated. Endothelial cell marker genes, such as CD31 and CD34, are upregulated with 50 µg/mL aCaP and a 2-week exposure. Inter-donor variability is high: Two out of three donors show a significant upregulation of ALP and Runx2 at day 14 with 50 µg/mL aCaP compared to 5 µg/mL aCaP. Notably, all changes in stem cell commitment are obtained in the absence of an osteogenic medium. While the chemical composition of the culture medium and the saturation status towards calcium phosphate phases remain approximately the same for all conditions, gene expression of ASCs changes considerably. Hence, aCaP nanoparticles show the potential to trigger osteogenic and endothelial commitment in ASCs.
Highlights
IntroductionCalcium phosphate (Ca3 (PO4 )2 ) is a biomaterial that is often used in orthopedic surgery, either in the form of cement [1,2], ceramic [3], or coating of titanium implants [4]
Calcium phosphate (Ca3 (PO4 )2 ) is a biomaterial that is often used in orthopedic surgery, either in the form of cement [1,2], ceramic [3], or coating of titanium implants [4].Nanoparticles of calcium phosphate may originate from mechanical abrasion of implant materials, as well as through degradation of larger entities
Regarding the two endothelial cell markers, CD31 and CD34, respectively, gene expression averaged over the three donors showed a significant increase if the value at day 14 with 50 μg/mL amorphous tricalcium phosphate (aCaP) was compared to the control
Summary
Calcium phosphate (Ca3 (PO4 )2 ) is a biomaterial that is often used in orthopedic surgery, either in the form of cement [1,2], ceramic [3], or coating of titanium implants [4]. Nanoparticles of calcium phosphate may originate from mechanical abrasion of implant materials, as well as through degradation of larger entities. Calcium phosphate phases differ in terms of solubility, where crystalline forms like alpha-tricalcium phosphate (α-TCP) and beta-tricalcium phosphate (β-TCP) are less soluble in aqueous solution than amorphous tricalcium phosphate (aCaP) [7], resulting in higher free calcium (Ca2+ ) and phosphate ion (Pi) concentrations in equilibrium with solid aCaP. Amorphous calcium phosphate has gained a lot of attention, due to its bioactivity; it transforms to hydroxyapatite (HAp) [8], the main inorganic component of natural bone [9]. ACaP is an interesting material for bone tissue engineering purposes, if combined with an organic phase to give nanocomposites, such as PLGA/aCaP [10,11,12]
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