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Fig 1.

Flow diagram of systematic literature search and screening.

Screening step 1: Hits from 3 online bibliographic literature sources were combined, primary studies were selected, and duplicates were removed. Title and abstracts were screened for the presence of OB-OC co-cultures. Screening step 2: OB-OC co-cultures were screened in full text for relevant outcome measures. All studies in which at least one relevant outcome measure was studied were included into Database 1. Screening step 3: Papers in which both cell types were studied with relevant outcome measures were included into Database 2. Screening step 4: Papers included into Database 2 and reviews were screened for potentially missing relevant studies and identified studies were screened in the same manner as above. Each screening step is marked with a separate background color. Each selection step within the screening steps is marked with a colored header. Blue header: used as input for the review. Grey header: selection step. Red header: excluded studies. Yellow header: Database as presented in this systematic map. Abbreviations: outcome measures (OM), Database 2 (DB2), osteoblast (OB), osteoclast (OC).

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Fig 2.

Schematic overview of Databases 1 and 2.

All identified studies were searched for OB-OC co-cultures, where co-culture was defined as OB and OC being present simultaneously and able to exchange biochemical signals. In addition to direct-contact cultures, cultures such as transwell cultures, 3D or scaffold cultures and bioreactor cultures were allowed as well. OB-OC co-culture studies which used relevant outcome measures were included into Database 1. Of these, only the relevant outcome measures were analyzed. All studies where relevant outcome measures were used for both OB and OC were included into Database 2 as well. Of these, cells and culture conditions were analyzed. The figure was modified from Servier Medical Art, licensed under a Creative Common Attribution 3.0 Generic License (http://smart.servier.com, accessed on 2 July 2021).

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Fig 3.

Relevant publications per year.

A) All 255 publications that contain relevant outcome measures counted by year ranging from 1983 to 2019 (Database 1). B) The 39 selected publications of Database 2 counted by year ranging from 1998 to 2019 (Database 2).

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Table 1.

Combinations and frequencies of primary and secondary outcome measures.

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Table 2.

a. Occurrences of resorption on different types of substrates and subsequent analyses. b. Supernatant resorption techniques.

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Table 3.

Occurrences of different methods of formation detection and subsequent analyses.

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Table 4.

TRAP measurement techniques and analyses.

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Table 5.

ALP measurement techniques and analysis.

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Table 6.

Osteoblast origins and occurrences.

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Fig 4.

Seeding densities and seeding ratios.

Violin plots of 2D and 3D seeding ratios of OB (A+D), OC (B+E) and respective seeding ratios in co-cultures (C+F). Values are calculated based on reported seeding numbers of the cells or precursors thereof per surface are or volume. No distinction was made between different (precursor) cell types in these figures, resulting in a considerable spread in data that could be attributed to proliferation and cell fusion after seeding The ranges along the Y-axis are not the same for each figure. Each seeding density of each study is represented by a blue dot.

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Table 7.

Osteoclast origins and occurrences.

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Fig 5.

Medium components used by studies in Database 2.

A) The occurrence of all identified base and complete media used during the co-culture phase of each study. B) Serum concentrations during the co-culture phase of each study. C) OC supplements administered during the co-culture phase of each study. Please note that the x-axis has a linear distribution. D) Osteogenic supplements during the co-culture phase of each study. Please note that the x-axis has a logarithmic scale. Individual concentrations or molarities are shown as blue dots.

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