Human Serum Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells In Vitro and In Vivo
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{"title"=>"Human Serum Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells In Vitro and In Vivo", "type"=>"journal", "authors"=>[{"first_name"=>"Alessandra", "last_name"=>"Pisciotta", "scopus_author_id"=>"37057907200"}, {"first_name"=>"Massimo", "last_name"=>"Riccio", "scopus_author_id"=>"7005221944"}, {"first_name"=>"Gianluca", "last_name"=>"Carnevale", "scopus_author_id"=>"35315390500"}, {"first_name"=>"Francesca", "last_name"=>"Beretti", "scopus_author_id"=>"6602234694"}, {"first_name"=>"Lara", "last_name"=>"Gibellini", "scopus_author_id"=>"17343252100"}, {"first_name"=>"Tullia", "last_name"=>"Maraldi", "scopus_author_id"=>"6507881128"}, {"first_name"=>"Gian Maria", "last_name"=>"Cavallini", "scopus_author_id"=>"7102295836"}, {"first_name"=>"Adriano", "last_name"=>"Ferrari", "scopus_author_id"=>"57106025400"}, {"first_name"=>"Giacomo", "last_name"=>"Bruzzesi", "scopus_author_id"=>"6504050203"}, {"first_name"=>"Anto", "last_name"=>"de Pol", "scopus_author_id"=>"6701780342"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84870562178", "pui"=>"366195672", "pmid"=>"23209773", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0050542", "sgr"=>"84870562178"}, "id"=>"f9a5927d-2550-38b8-aeab-2f37f5f256e7", "abstract"=>"Human dental pulp is a promising alternative source of stem cells for cell-based tissue engineering in regenerative medicine, for the easily recruitment with low invasivity for the patient and for the self-renewal and differentiation potential of cells. So far, in vitro culture of mesenchymal stem cells is usually based on supplementing culture and differentiation media with foetal calf serum (FCS). FCS is known to contain a great quantity of growth factors, and thus to promote cell attachment on plastic surface as well as expansion and differentiation. Nevertheless, FCS as an animal origin supplement may represent a potential means for disease transmission besides leading to a xenogenic immune response. Therefore, a significant interest is focused on investigating alternative supplements, in order to obtain a sufficient cell number for clinical application, avoiding the inconvenients of FCS use. In our study we have demonstrated that human serum (HS) is a suitable alternative to FCS, indeed its addition to culture medium induces a high hDPSCs proliferation rate and improves the in vitro osteogenic differentiation. Furthermore, hDPSCs-collagen constructs, pre-differentiated with HS-medium in vitro for 10 days, when implanted in immunocompromised rats, are able to restore critical size parietal bone defects. Therefore these data indicate that HS is a valid substitute for FCS to culture and differentiate in vitro hDPSCs in order to obtain a successful bone regeneration in vivo.", "link"=>"http://www.mendeley.com/research/human-serum-promotes-osteogenic-differentiation-human-dental-pulp-stem-cells-vitro-vivo", "reader_count"=>33, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>2, "Researcher"=>2, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>1, "Student > Bachelor"=>6, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>2, "Researcher"=>2, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>1, "Student > Bachelor"=>6, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Biochemistry, Genetics and Molecular Biology"=>4, "Medicine and Dentistry"=>14, "Agricultural and Biological Sciences"=>10, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>14}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>10}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>4}}, "group_count"=>3}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/535491"], "description"=>"<p>Images show transversal sections carried out through the central area of the implants. A–C: cranial defect filled with hDPSCs/collagen pre-differentiated with FCS containing medium; D–F: cranial defect closed with hDPSCs/collagen pre-differentiated with HS containing medium; (dotted line delimitates the areas of bone resection; arrowheads indicate vasa; * indicate areas of active bone deposition). Bar: 100 µm. G: morphometric analysis of new-formed bone areas in controls (C), FCS and HS implants. Values are mean ± SD of the percentage of regenerated bone respect to the whole resected bone area. HS and FCS n = 6; C n = 4; white * inside the column indicate values of ANOVA test of HS and FCS vs. C (**<i>p<</i>0.01, ***<i>p<</i>0.001); black * indicate values of ANOVA test of HS vs. FCS (***<i>p<</i>0.001). H: number of vasa in the scaffold not yet reabsorbed and in new-formed bone areas. Data, normalized to areas of the scaffold not yet reabsorbed and of new-formed bone areas respectively, were presented as mean ± SD (vasa number respect to the total implant area) of each experimental group (controls n = 4; treated n = 6). White * inside the column indicate values of ANOVA test of HS and FCS vs. C (***<i>p<</i>0.001); black * indicate values of ANOVA test of HS vs. FCS (*<i>p<</i>0.05).</p>", "links"=>[], "tags"=>["histological", "cranial", "defect", "reconstruction", "constructs", "40", "days"], "article_id"=>205971, "categories"=>["Physiology", "Cell Biology", "Developmental Biology"], "users"=>["Alessandra Pisciotta", "Massimo Riccio", "Gianluca Carnevale", "Francesca Beretti", "Lara Gibellini", "Tullia Maraldi", "Gian Maria Cavallini", "Adriano Ferrari", "Giacomo Bruzzesi", "Anto De Pol"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050542.g004", "stats"=>{"downloads"=>5, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparative_histological_analysis_haematoxylin_eosin_staining_of_the_critical_size_cranial_defect_reconstruction_by_hDPSCs_collagen_constructs_40_days_post_surgery_/205971", "title"=>"Comparative histological analysis (haematoxylin/eosin staining) of the critical size cranial defect reconstruction by hDPSCs/collagen constructs 40 days post-surgery.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-29 01:39:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/287753"], "description"=>"<div><p>Human dental pulp is a promising alternative source of stem cells for cell-based tissue engineering in regenerative medicine, for the easily recruitment with low invasivity for the patient and for the self-renewal and differentiation potential of cells. So far, <em>in vitro</em> culture of mesenchymal stem cells is usually based on supplementing culture and differentiation media with foetal calf serum (FCS). FCS is known to contain a great quantity of growth factors, and thus to promote cell attachment on plastic surface as well as expansion and differentiation. Nevertheless, FCS as an animal origin supplement may represent a potential means for disease transmission besides leading to a xenogenic immune response. Therefore, a significant interest is focused on investigating alternative supplements, in order to obtain a sufficient cell number for clinical application, avoiding the inconvenients of FCS use. In our study we have demonstrated that human serum (HS) is a suitable alternative to FCS, indeed its addition to culture medium induces a high hDPSCs proliferation rate and improves the <em>in vitro</em> osteogenic differentiation. Furthermore, hDPSCs-collagen constructs, pre-differentiated with HS-medium <em>in vitro</em> for 10 days, when implanted in immunocompromised rats, are able to restore critical size parietal bone defects. Therefore these data indicate that HS is a valid substitute for FCS to culture and differentiate <em>in vitro</em> hDPSCs in order to obtain a successful bone regeneration <em>in vivo</em>.</p> </div>", "links"=>[], "tags"=>["serum", "promotes", "osteogenic", "differentiation", "dental", "pulp", "cells"], "article_id"=>116681, "categories"=>["Physiology", "Cell Biology", "Developmental Biology"], "users"=>["Alessandra Pisciotta", "Massimo Riccio", "Gianluca Carnevale", "Francesca Beretti", "Lara Gibellini", "Tullia Maraldi", "Gian Maria Cavallini", "Adriano Ferrari", "Giacomo Bruzzesi", "Anto De Pol"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050542", "stats"=>{"downloads"=>2, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Human_Serum_Promotes_Osteogenic_Differentiation_of_Human_Dental_Pulp_Stem_Cells_In_Vitro_and_In_Vivo__/116681", "title"=>"Human Serum Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells <em>In Vitro</em> and <em>In Vivo</em>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-29 01:51:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/534960"], "description"=>"<p>A: CFDA vital staining of hDPSCs cultured for 1, 4 and 7 days. Green signal indicates viable cells. Bar: 100 µm. B: Proliferation rate of hDPSCs cultured for a week; Values are mean reported in a Log scale, n = 3; * indicates values of paired <i>t</i>-test HS vs. FCS (**<i>p<</i>0.01, ***<i>p<</i>0.001). C: Cumulative population doubling (CPD) of hDPSCs cultured for a total of 5 passages. At each passage cells cultured in HS-medium show a CPD significantly higher than FCS-medium cultured cells (n = 3; ***<i>p<</i>0.001). D shows β-galactosidase activity staining in confluent culture of hDPSCs grown for 5 passages in FCS-medium or in HS-medium as indicated. Arrowheads indicate cells positive to β-galactosidase activity staining. Bar 10 µm. E: western blot analysis of PARP in hDPSCs cultured in FCS-medium and in HS-medium at passages 1, 3 and 5. HL60, treated with etoposide, were loaded as positive control of the presence of cleaved PARP (cPARP). Actin bands were presented as control of the protein loading. Densitometry of cPARP bands was shown on the bottom of western blot images.</p>", "links"=>[], "tags"=>["hs", "serum"], "article_id"=>205448, "categories"=>["Physiology", "Cell Biology", "Developmental Biology"], "users"=>["Alessandra Pisciotta", "Massimo Riccio", "Gianluca Carnevale", "Francesca Beretti", "Lara Gibellini", "Tullia Maraldi", "Gian Maria Cavallini", "Adriano Ferrari", "Giacomo Bruzzesi", "Anto De Pol"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050542.g001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_hDPSCs_growing_in_FCS_HS_and_serum_free_SF_media_/205448", "title"=>"hDPSCs growing in FCS, HS and serum free (SF) media.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-29 01:30:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/535299"], "description"=>"<p>A: Alkaline phosphatase assay (AP) and Alizarin Red staining; B: densitometric analysis of AP and Alizarin Red staining. Values are mean ± SD of gray levels (0–255 scale). HS n = 3; FCS n = 3; ** indicate values of paired t-test HS vs. FCS (<i>p<</i>0.01). C: Confocal analysis of osteogenic differentiation of hDPSCs. Double immunofluorescence confocal images showing signals from anti-OPN (green) and anti-Runx2 (red); DAPI (blue) and anti-OCN (green); DAPI (blue) and anti-Osx (red). Bar: 50 µm. D: Western blot (WB) analysis of Coll-I and OCN expression in whole cell lysates of differentiated hDPSCs. Whole cell lysates were collected from three plates of human dental pulp stem cells for each differentiation protocol. Actin bands demonstrate that an equal amount of protein was loaded in each line. Undiff samples show hDPSCs cultured in FCS-medium alone. The same results were obtained in HS-medium culture (data not shown).</p>", "links"=>[], "tags"=>["osteogenic", "differentiation", "hdpscs", "24", "days", "supplemented", "fcs"], "article_id"=>205785, "categories"=>["Physiology", "Cell Biology", "Developmental Biology"], "users"=>["Alessandra Pisciotta", "Massimo Riccio", "Gianluca Carnevale", "Francesca Beretti", "Lara Gibellini", "Tullia Maraldi", "Gian Maria Cavallini", "Adriano Ferrari", "Giacomo Bruzzesi", "Anto De Pol"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050542.g003", "stats"=>{"downloads"=>2, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_osteogenic_differentiation_of_hDPSCs_after_24_days_of_culture_in_osteogenic_medium_supplemented_with_FCS_or_HS_/205785", "title"=>"<i>In vitro</i> osteogenic differentiation of hDPSCs after 24 days of culture in osteogenic medium, supplemented with FCS or HS.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-29 01:36:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/535683"], "description"=>"<p>A–D: Double fluorescence signals from DAPI (blue) and anti-hMit Ab (green) superimposed to pseudo-phase contrast images (B, D). Arrowheads indicate cells entrapped in calcified bone matrix clearly stained by anti-human mitochondria antibody. E–H: triple fluorescence signals from DAPI (blue) and anti-hMit (green) and anti-OCN (red) Abs superimposed to pseudo-phase contrast images (F, H). Yellow arrowheads indicate osteocytes labelled by the two Abs; cyan arrowheads indicate OCN deposits in extracelluar bone matrix. I–L: triple fluorescence signals from DAPI (blue) and anti-hMit (green) and anti-von Willebrand factor (red) Abs superimposed to pseudo-phase contrast images (J, L). Arrowheads indicate vasa double stained by the two Abs. Bar: 10 µm.</p>", "links"=>[], "tags"=>["images", "implants", "pre-differentiated"], "article_id"=>206168, "categories"=>["Physiology", "Cell Biology", "Developmental Biology"], "users"=>["Alessandra Pisciotta", "Massimo Riccio", "Gianluca Carnevale", "Francesca Beretti", "Lara Gibellini", "Tullia Maraldi", "Gian Maria Cavallini", "Adriano Ferrari", "Giacomo Bruzzesi", "Anto De Pol"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050542.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Confocal_images_of_implants_pre_differentiated_in_both_the_conditions_/206168", "title"=>"Confocal images of implants pre-differentiated in both the conditions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-29 01:42:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/535100"], "description"=>"<p>A: Cytofluorimetric analysis of Stro-1, c-Kit and CD34 expression in hDPSCs cultured in FCS-medium and in HS-medium. Dot plots reporting SSC vs fluorescence are shown. In the histograms, the net fluorescence value was calculated by linearizing the fluorescence value from the logarithmic scale and subtracting the linearized value of the unstained sample to the linearized value of the stained one. Data represent the mean±SD of three different experiments. * indicates values of paired <i>t</i>-test HS vs. FCS (*<i>p<</i>0.05). B: In the first line are shown double immunofluorescence images of hDPSCs/C<sub>2</sub>C<sub>12</sub> co-culture stained by anti-hMit (green) and anti-myosin (red) Abs. DAPI staining is shown in blue. The second line shows oil red staining of hDPSCs differentiated for two weeks towards adipogenic lineage with HS or FCS supplemented medium. Cells were counterstained with Harris haematoxylin. Images, in third line represent anti-β3-Tubulin immunofluorescence labelling on hDPSCs differentiated in culture neurogenic media supplemented with FCS or HS. Bar 50 µm.</p>", "links"=>[], "tags"=>["antigens", "multilineage", "differentiation"], "article_id"=>205587, "categories"=>["Physiology", "Cell Biology", "Developmental Biology"], "users"=>["Alessandra Pisciotta", "Massimo Riccio", "Gianluca Carnevale", "Francesca Beretti", "Lara Gibellini", "Tullia Maraldi", "Gian Maria Cavallini", "Adriano Ferrari", "Giacomo Bruzzesi", "Anto De Pol"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050542.g002", "stats"=>{"downloads"=>2, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_hDPSCs_surface_antigens_expression_and_in_vitro_multilineage_differentiation_ability_/205587", "title"=>"hDPSCs surface antigens expression and <i>in vitro</i> multilineage differentiation ability.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-29 01:33:07"}

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Relative Metric

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